Tmp252e TMP

Applied Energy 155 (2015) 585598
Contents lists available at ScienceDirect
Applied Energy
journal homepage: www.elsevier.com/locate/apenergy
Review
Microalgal biofuel revisited: An informatics-based analysis

of developments to date and future prospects
Hui Chen a, Tian Qiu b, Junfeng Rong c, Chenliu He a, Qiang Wang a,
a
Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, Hubei, China
Wuhan Library, Chinese Academy of Sciences, Wuhan 430071, Hubei, China
c
SINOPEC Research Institute of Petroleum Processing, Beijing 100083, China
b
h i g h l i g h t s
Microalgal biofuel studies between 1900 and mid-2015 were analyzed informatically.
Burst interest since 20062012 stimulated mass culture and biotechnology studies.
Unremitting study and investment is expected for better understanding of microalgae.
Integrated application of energy microalgae could be a possible solution.
Recent advances of approaches to bypass the production bottleneck were reviewed.
a r t i c l e
i n f o
Article history:
Received 11 February 2015
Received in revised form 15 June 2015
Accepted 17 June 2015
Keywords:
Microalgal biofuel
Microalgal biotechnology
Microalgal mass culture
Informatics
a b s t r a c t
Microalgae have reported to be one of the most promising feedstock for biofuel production. To obtain a
comprehensive and systematic overview of the current state of microalgal research, particularly microalgal biofuel research, we retrieved and analyzed manuscripts and patents related to this topic and published between 1900 and mid-2015. We found that there was a burst in microalgal biofuel research
from 2006 to 2011 that signicantly stimulated the development of microalgal biotechnology for the production of high value-added commodities and for environmental applications and microalgal mass culturing, in an attempt to make the entire process of biofuel production economically viable for
industrialization. However, a lag in basic microalgal research has kept production costs high, resulting
in a decline in investments, funding, and research efforts in the elds of microalgal biofuel production,
microalgal biotechnology, and mass culturing since 2012. Based on a review of the challenges/problems
of microalgae biofuel production and recent advances of their solution, the perspective view of the future
R&D needs and trends were proposed. To bypass the price bottleneck of microalgae-based biofuel production, it has been proposed that energy-producing microalgal biotechnological applications be synergistically combined with microalgal biofuel production. Future investments and funding will most likely
be directed toward basic studies that aim to elucidate the microorganisms characteristics and toward the
development of microalgal biotechnology and its environmental applications, which have potential economic and social benets. This review represents a theoretical reference for both algal researchers and
decision makers regarding the future directions of microalgal research, particularly that involving
microalgal-based biofuel production.
2015 Elsevier Ltd. All rights reserved.
Contents
1.
2.
3.
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Literature retrieval and analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Trends in microalgal biofuel research and its biotechnological applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3.1.
The intensity of research in the field of microalgal biofuel has decreased since 2012 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Corresponding author. Tel.: +86 27 68780790.

E-mail address: wangqiang@ihb.ac.cn (Q. Wang).
http://dx.doi.org/10.1016/j.apenergy.2015.06.055
0306-2619/ 2015 Elsevier Ltd. All rights reserved.
586
586
586
586
586
H. Chen et al. / Applied Energy 155 (2015) 585598
3.2.
High capital and operating costs hamper commercial-scale algal biofuel production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 588
3.3.
Research into microalgal biotechnology for the production of high value-added products and for environmental applications was stimulated
by the economic demand for microalgal biofuel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 590
3.4.
The demands for the industrialization of microalgal biofuel production have driven studies on mass microalgal culturing . . . . . . . . . . 591
3.5.
Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 592
4.
The challenges of microalgae biofuel production and recent advances of their solution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 593
4.1.
Species . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 593
4.2.
Cultivation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 594
4.3.
Harvesting and extraction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 594
4.4.
Co-production by biorefinery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 595
5.
Perspective view of the future R&D needs and trends. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 595
6.
Conclusions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 596
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 596
Appendix A.
Supplementary material . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 596
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 596
1. Introduction
The extensive use of fossil fuels has led to global climate
change, environmental pollution, health problems, and an energy
crisis associated with the irreversible depletion of traditional
sources of fossil fuels [1]. Many countries are thus turning their
attention to the development of new, clean, and sustainable energy
sources [2,3]. Among the various potential sources of renewable
energy, biofuel, the fuels obtained from biomass (i.e., organic
matter derived from plants, animals, and microorganisms), are of
great interest and are expected to play a crucial role in the global
energy infrastructure in the future. In contrast to other forms of
renewable energy (e.g., wind, tidal, and solar energy), energy is
chemically stored in biofuels and biofuels can be used in existing
engines after blending to various degrees with petroleum diesel,
resulting in lower carbon monoxide (CO) and sulfur oxide (SOx)
emission levels [4,5].
Microalgae are the fastest growing photosynthesizing
organisms and, in addition to consuming carbon dioxide (CO2)
and nitrogen (N)-based compounds such as ammonium, microalgae are one of the most important producers of oxygen on earth
[6,7]. Microalgal biomass contains approximately 50% carbon by
dry weight [8]. During the production of 100 tons of microalgal
biomass using natural or articial light, approximately 180 tons
of CO2 can be xed [9]. Although all CO2 absorbed by an alga to
produce fuel oil is released back into the environment as soon as
the oil is burnt and the residual biomass degrades [10], no more
extra CO2 is produced in the production and consumption of
microalgal biofuel, which maintains carbon balance in environment and may relax global warming associated with the consumption of fossil fuels and emission of CO2. Given their rapid growth
and ability to convert solar energy into chemical energy via CO2
xation, microalgae have been considered one of the most promising sources of oil for the production of biodiesel [11,12]. In
addition, microalgae are used as feedstock for a wide variety of
practical and potential metabolic products, such as food supplements, pharmacological substances, lipids, enzymes, biomass,
polymers, toxins, and pigments [13,14]. Microalgal cultivation is
a costly process due to the large amounts of water, inorganic nutrients (mainly N and phosphate (P)), and CO2 needed [15,16].
Compared to the microalgal cultivation by using freshwater, N
and P fertilizer, and purchased CO2, biodiesel production from
microalgae may be more environmentally sustainable,
cost-effective, and protable if combined with processes such as
wastewater and ue gas treatments, in which freshwater is
replaced with wastewater and ue gases are used as a source of
carbon and inorganic nutrients for microalgal culture [17,18].
However, the availability of sufcient concentrated CO2 from
burning coal or other fossil fuel is still a major impediment to
production of algal fuel oils at a meaningful scale [10], and other

CO2 source (e.g., oil renery and biomass power plant, etc.) should
be further taken into account.
Although the entire chain of microalgal biofuel production,
including culture selection [1,1925], cultivation and pest control
[12,17,2633], harvest [18,31,34,35], and lipid production and processing [18,21,3640], has been extensively studied and reviewed
during the past few years, the production cost is too high for industrialization, and the price of algae-based biodiesel ($2.76/kg)
remains strikingly higher than that of normal diesel ($0.95/kg)
[41]. To obtain a comprehensive and systematic overview of trends
in microalgal research, particularly in microalgal biofuel research,
we retrieved and extensively analyzed manuscripts and patents
related to this topic and published between 1900 and May 2015.
This informatics study provides useful guidance for future research
in the eld of microalgae, particularly microalgal biofuel. Moreover,
the challenges/problems of microalgae biofuel production and
recent advances of their solution were reviewed, and the perspective view of the future R&D needs and trends are proposed.
2. Literature retrieval and analysis
Manuscripts and patents were retrieved from the Wuhan
Documentation and Information Center of the Chinese Academy
of Sciences on May 28, 2015 (see Retrieval method of manuscripts
and patents in le of Supplemental material). We analyzed the data
collected from the retrieved manuscripts and patents using
Thomson Data Analyzer software (TDA, Thomson Reuters, New
York, NY, USA). For the manuscripts, we derived, edited, and analyzed manuscript titles, keywords listed in the manuscript, year
in which the study was published, and country in which the study
was conducted. The keyword analysis and keyword-correlation
analysis were performed using cluster analysis. For the retrieved
patents, we derived, edited, and analyzed the patent titles, the year
and country in which the study was published and conducted, and
technical components. The technical directions of the patents were
classied according to the International Patent Classication (IPC)
criteria.
3. Trends in microalgal biofuel research and its
biotechnological applications
3.1. The intensity of research in the eld of microalgal biofuel has
decreased since 2012
We retrieved a total of 6194 published manuscripts and 1230
patents related to microalgal biofuel for the period 1900 to May
2015, and the results are presented in Table 1.
Fig. 1. Statistical analysis of trends in the number of published manuscripts related

to microalgal biofuel. (A) Annual number of published manuscripts; (B) distribution
in the number of manuscripts published in the 10 countries/regions in which the
study was conducted with the highest number of publications on the topic;
(C) annual number of manuscripts published per top 10 countries during the past
decade (20062015). The data for 2015 in the current and following gures and
tables shows only manuscripts/patents published between January 1 and May 28.
The rst manuscript on microalgal biofuel appeared in 1970,

and the number of manuscripts on this topic subsequently showed
an overall annual increase (Fig. 1A). Notably, a sharp increase in the
number of such manuscripts occurred between 2007 and 2013,
with 1018 manuscripts being published on this topic in 2013. In
20142015, the increased trend in number of manuscripts on
microalgal biofuel was relatively low, with 1220 and 519 manuscripts being published by 2014 and May 28 of 2015. Moreover,
we found that the number of manuscripts on microalgal biofuel
published between 2007 and 2013 was greater than that published
587
Fig. 2. Trends in the number of published patents pertaining to microalgal biofuel.

(A) Annual number of published patents; (B) number of published patents in the top
10 countries/regions; (C) annual number of published patents in the top 10
countries/regions in the past decade (20062015). Europe with the exception of
France, Spain, and Germany.
on the topic in all previous years combined, suggesting that

microalgal biofuel emerged as a hot topic in the eld of microalgal
research during this period. We found that published studies
related to microalgal biofuel were derived from more than 80
countries/regions throughout the world (Table 1), indicating that
scientists in more than 80 countries/regions have participated in
microalgal biofuel research. Furthermore, the USA, China, and
Spain were the top three countries/regions in terms of the number
of published manuscripts pertaining to microalgal biofuel
(Fig. 1B and C).
588
Supplemental material). The major advances in microalgal biofuel

production made since 2006 are highlighted below.
Table 1
Number of manuscripts and patents retrieved.
Research direction
Microalgal biofuel
Microalgal
biotechnologya
Microalgal mass
culturing
Published manuscripts
Published patents
Number
Country/region
Number
Country/region
6194
4820
85
85
1230
859
48
46
2426
65
944
50
a
Microalgal biotechnology for the production of high value-added products and
environmental applications (e.g., ue gas and wastewater treatments).
To elucidate the major foci of research on microalgal biofuel, we

analyzed the published manuscripts for various keywords and
obtained a list of high-frequency keywords. Based on the keywords
with the highest frequency, we divided the research foci into the
following four categories: breeding algal species, mass culturing,
processing technologies, and types of energy (Table S1 in le of
Supplemental material). We then conducted a cluster analysis of
these high-frequency keywords, and identied key focus and
bottlenecks in microalgal biofuel research, with a focus on mass
culturing, biodiesel production, and environmental applications
(Fig. S1 in le of Supplemental material). In particular, large-scale
microalgal culturing was a major emphasis of these studies, and
related topics included photobioreactor and culturing conditions
and focused on the algal species, light intensity, temperature, level
of CO2, photosynthetic efciency, and biomass, among other issues.
As a more practical assessment of the success of these studies,
we analyzed the research trends by retrieving and analyzing published patents related to microalgal biofuel. Similar to the results
obtained from the analysis of published manuscripts pertaining
to microalgal biofuel, the number of patents published on the topic
showed an increasing trend (Fig. 2A). The number of patents significantly increased in 2006, indicating that research on microalgal
biofuel had moved into high gear. However, the number of annual
patents peaked in 2011, and declined sharply after 2012. Thus,
research on microalgal biofuel was intensive between 2006 and
2011, but started to decline in 2012. Nearly 50 countries/regions
were involved in published patents pertaining microalgal biofuel
(Table 1), and the top three countries/regions were China, the
USA, and Korea (Fig. 2B and C). In accordance with the general
trend in the number of published patents (Fig. 2A), the number
of relevant published patents in most of these countries has
recently declined (Fig. 2C), indicating that the downturn in the
intensity of research on microalgal biofuel was a global trend
and occurred in most countries/regions in which research on
microalgal biofuel had been conducted.
We used the IPC criteria to elucidate the focus of the research on
microalgal biofuel (Table S2 in le of Supplemental material), and
identied the top 15 technical classications of patents on this
topic. We found that more than 80% of the patents included in
the top 15 categories were published between 2006 and 2012
and that microalgal mass culturing and microalgal metabolite production and their environmental applications dominated the
research on microalgal biofuel (Fig. S2 in le of Supplemental
material), which was consistent with the conclusions drawn from
the analysis of high-frequency keywords in the published manuscripts (Fig. S1 in le of Supplemental material). As the burst in
microalgal biofuel research started in 20062007 (Figs. 1 and 2),
we further analyzed the research published on microalgal biofuel
in 2006 by conducting a temporal distribution analysis and cluster
analysis of the high-frequency keywords (Fig. S3 in le of
Supplemental material). In accordance with the data shown in
Fig. S1 in le of Supplemental material, mass culture was the main
focus of microalgal biofuel research since 2006 (Fig. S3 in le of
3.2. High capital and operating costs hamper commercial-scale algal

biofuel production
An in-depth knowledge of the recent advances in microalgal
biofuel, especially the progress related to mass culture, is essential
for microalgae-based biofuel industrialization.
The rst and one of the most important steps of the algae-based
biofuel industry is microalgal strain selection [21], as this affects
the potential biofuel yield and determines which nutrients and cultivation units must be used for their growth. Since 2006, much
effort has focused on collecting and optimizing strains for biofuel
production, and green algae, including Chlorella, Chlamydomonas,
Scenedesmus, Nannochloropsis, and Botryococcus, some diatoms,
and cyanobacteria, are the most frequently used strains in microalgal biofuel production (Figs. S1 and S3 in le of Supplemental
material). The ideal microalga for biodiesel feedstock would have
high biomass productivities and would exhibit efcient lipid
biosynthesis. Thus, not just a single parameter (i.e., growth rate
or lipid content), but the volumetric lipid productivity of the
microalga should be the main selective criterion [12,18,25,42]. In
particular, knowledge of the biochemical composition of microalgae, which can provide information about their potential use as
an alternative fuel feedstock, is important for developing efcient
processing technology at an industrial scale [43].
Photoautotrophic cultivation, typically in outdoor environments
where sunlight is abundant and free, could provide a technically
and economically feasible method for culturing microalgae at the
commercial scale [30]. However, slow growth rate and low cell density are the main limiting factors of photoautotrophic cultivation
[7]. Relatively high lipid yields and biomass productivity can be
attained through heterotrophic or mixotrophic cultivation methods, providing good opportunities for large-scale production.
However, the limited number of available heterotrophic or
mixotrophic algal species, potential contamination by bacteria,
inhibition of growth by soluble organic substrates at low concentrations, and high cost of nutrient medium are the main factors limiting large-scale production [44]. Microalgae are primarily cultured
on open ponds or in closed photobioreactors (PBRs) for
large-scale production. However, low culture density, low biomass
productivity, contamination, susceptibility to weather events such
as rainfall, and high evaporative losses in open ponds are major
drawbacks hindering commercialization of this approach [45,46].
PBRs are closed to the atmosphere and have several advantages
over ponds: (1) the risk of contamination is reduced; (2) the growth
parameters (e.g., temperature) can be better controlled; (3) they
have higher volumetric productivities and cell concentrations due
to a higher surface-to-volume (S/V) ratio; and (4) these closed systems eliminate or strongly reduce evaporation and thus conserve
water [47,48]. Although few PBR designs have been explored at
the pilot level, the high capital and operating costs of PBRs are
the major hurdles for their large-scale implementation for biofuel
production and they still await evaluation at real scale [18].
After cultivation, microalgal biomass has to be separated from
its growth medium and recovered for downstream processing.
The traditional harvesting method usually involves two steps, bulk
harvesting (also known as primary harvesting) to separate microalgae from their growth medium, by methods such as sedimentation,
occulation, and oatation, and thickening (known as secondary
dewatering) to concentrate the microalgal slurry after bulk harvesting, by methods such as centrifugation and ltration [44,49].
Among these bulk harvesting methods, occulation is frequently
used to increase the efciency of gravity sedimentation [34].
However, conventional occulants are toxic and expensive, and
589
Fig. 3. Trends in published manuscripts related to microalgal biotechnology for high value-added product production and environmental applications. (A) Annual statistics
for the published manuscripts; (B) statistics for manuscripts published in the top 10 countries/regions; (C) annual statistics for the manuscripts published in the top 10
countries/regions during the past decade (20062015); (D) annual statistics for published manuscripts pertaining to both microalgal biofuel and microalgal biotechnology for
the production of high-value-added products and environmental applications; 1, published manuscripts pertaining to microalgal biofuel; 2, published manuscripts pertaining
to microalgal biotechnology for the production of high value-added products and environmental applications.
nontoxic occulants (e.g., organic polymers) have been intensively

investigated but are too expensive for large-scale applications [50].
Autoocculation by increasing the pH of the medium and electrolytic occulation may be used to separate algae without the
addition of chemicals and would be more economical than other
harvesting techniques for marine microalgae [50,51]. Moreover,
unlike other harvesting technologies, the cost contribution to algal
lipid from electrolytic occulation was unaffected by the concentration of algae in the algal water [52]. Thickening by centrifugation and ltration are too costly and energy intensive for
large-scale biofuel production. Coons et al. [52] reported that the
cost contribution to algal lipid from membrane ltration could be
signicantly higher than electroocculation processes operating
with inert electrodes, and the cost of centrifugation was generally
higher than the cost of membrane ltration. In particular, ultrasonic and electrolytic harvesting technologies show the most
potential for energy and cost reduction. Notably, Coons et al. [52]
also showed that ultrasonic harvesting, which functioned from
the principle of a standing wave created by forward and reverse
propagating pressure waves in a body of water that contains algae,
could well be lower than other harvesting methods in energy cost.
However, ultrasonic technologies also require further development
and scale-up before they can achieve low-cost performance at
industrially relevant scales. Thus, the development of bulk harvesting technologies may play an important role in reducing energy
consumption during microalgal thickening [53].
For lipid production, extensive drying of microalgal biomass is

essential, as the presence of water interferes with the extraction
and/or conversion of algal lipids to biodiesel [54]. Spray drying,
drum drying, freeze drying, and solar drying are common methods
for drying microalgae. The rst three drying methods are not economical and lead to a negative energy balance in large-scale
microalgal biodiesel production because of their heavy dependency on fossil fuels. Solar drying is considered to be an economical method, but it is not feasible in temperate countries where
sunlight is limited at certain times of the year, and it requires signicant land areas for large-scale operations [44].
Lipids and fatty acids are commonly extracted from the
microalgal biomass before biodiesel production. Two general
extraction processes, the Hexane Soxhlet method and the Bligh
Dyer method [55,56], are effective means of extracting microalgal
lipids, with hexane-based oil extraction being more energy efcient and therefore preferred for scaling up efforts [57]. However,
the disadvantages of using chemical solvents are mostly related
to their high toxicity to humans and the environment. Also, drying
and then solvent extraction are high energy cost steps in any
extraction process. For example, even the high energy is required
in the dimethyl ether (DME) system, a solvent extraction method
on wet algae, its energy requirements is the lowest at about half
that needed to dry the algae [52]. Several supercritical uids, but
particularly supercritical-CO2, have recently been used in microalgal lipid extraction for biodiesel production. Although supercritical
590
ultrasonic processes require further development and scale-up,

which will lead to lower capital costs.
After lipid extraction, transesterication, the most accessible
technology reported for biodiesel production to date, is generally
used to produce biodiesel [64]. By contrast, in-situ transesterication, in which lipid extraction and transesterication are carried
out simultaneously, promises to both simplify the process and
reduce the cost of producing biodiesel from microalgal biomass
[30]. The algal biomass needs to be dried prior to the in-situ transesterication procedure, because when the moisture content of
algal biomass exceeds 20%, biodiesel recovery using in-situ transesterication is greatly reduced [65]. An improved in-situ transesterication process that directly converts wet oil-bearing
microalgal biomass into biodiesel was recently explored [66].
Besides microalgal biodiesel (the most studied microalgal biofuel
type), microalgae can provide other several types of renewable biofuels, including biomethane (also known as biogas) [67], biohydrogen [68], and bioethanol [62].
In summary, among the impediments to commercialization of
algal fuels (impossibly high demands on certain key resources;
the high cost of production; and the need to achieve an energy
ratio of well above unity) [10], the high capital and operating costs
(i.e., the cost of water, CO2, and nutrient supply, and of biomass
harvesting and processing) render algae-based biodiesel much
more expensive than regular diesel, which may be one key impediment to limit commercial applications of algae-based biodiesel.
Currently, algal biodiesel is not industrially produced anywhere
in the world, and even a widespread availability of algal fuels is
certainly not likely in the near term [10].
3.3. Research into microalgal biotechnology for the production of high
value-added products and for environmental applications was
stimulated by the economic demand for microalgal biofuel
Fig. 4. Trends in published patents related to microalgal biotechnology for the

production of high value-added products and environmental applications.
(A) Annual statistics for the published patents; (B) statistics for the patents
published in the top 10 countries/regions; (C) annual statistics for the patents
published in the top 10 countries/regions during the past decade (20062015).
Europe with the exception of France, Spain, and Germany.
extraction is non-toxic and provides a non-oxidizing environment

that avoids degradation of the extracts [58], it is prohibitively
expensive. In the lipid extraction process, effective cell disruption
allows access to intracellular lipids. Among a variety of mechanical
technologies, including high pressure homogenization (HPH) [59],
bead mills [60], ultrasonic disruption or sonication [61,62], and
electroporation [63], ultrasonic disruption is reported to be the
disruption technology with lowest energy requirement [52]. Be
similar to harvesting technologies, extraction technologies with
We retrieved 4820 manuscripts and 859 patents in the area of

microalgal biotechnology for the production of high value-added
products and for environmental applications that were published
from 1900 to May 2015 (Table 1).
The rst manuscript related to microalgal biotechnology for the
production of high value-added products and for environmental
applications was published in 1982, and the number of published
manuscripts on this topic has since increased exponentially
(Fig. 3A). Similar to our ndings for microalgal biofuel research,
the analysis revealed that research on microalgal biotechnology
for the production of high value-added products and for environmental applications was performed in more than 80 countries/regions (Table 1), and the USA, China, and Spain were
consistently the top three countries in terms of the number of published manuscripts on this topic (Fig. 3B and C). Although published manuscripts related to microalgal biotechnology for the
production of high value-added products and environmental applications showed a similar increasing trend (Fig. 3A), this trend
exhibited a hysteretic quality compared with that found for published manuscripts pertaining to microalgal biofuel (Fig. 1A).
Whereas the annual number of published manuscripts pertaining
to microalgal biofuel rose sharply at the beginning of 2007, that
pertaining to microalgal biotechnology for the production of high
value-added products and environmental applications increased
signicantly in 2008. These results suggest that research on
products and environmental applications was stimulated by
research on microalgal biofuel. Notably, the concept of using
oil-producing microalgae for the production of high value-added
commodities and environmental protective applications became
popular in 1994 and increased after 2008, as revealed by our analysis of 2504 manuscripts involving both microalgal biofuel and
591
Fig. 5. Trends in published manuscripts related to microalgal mass culture. (A) Annual statistics of published manuscripts; and (B) annual statistics of published manuscripts
in the top 10 countries/regions from 1990 to 2015.

products and environmental applications (Fig. 3D). Because the
high price of algae-based biofuel hampers its large-scale manufacturing [41,69], studies that focus on using oil-producing microalgal
biotechnology to make microalgal biofuel production economically
viable have been encouraged.
We then analyzed keywords in published manuscripts pertaining to microalgal biotechnology for the production of high
value-added products and environmental applications to elucidate
the research foci. Consistent with the corresponding analysis of
keywords used in publications pertaining to microalgal biofuel,
our analysis of the keywords with the highest frequency revealed
that the research foci of this eld could also be divided into four
categories, namely breeding algal species, mass culturing, processing technologies, and types of energy, of which mass culturing and
processing technologies were the most prominent (Table S3 in le
of Supplemental material). A cluster analysis of the high-frequency
keywords (Fig. S4 in le of Supplemental material) indicated that
this research focused on three directions: the rst direction was
mass culturing, including the culture conditions, photosynthesis,
photobioreactors, and biodiesel production; the second direction
involved the accumulation and extraction of high added-value
products (e.g., b-carotenoid and astaxanthin); and the third direction focused on the application of microalgae for environmental
protection, such as for the biosorption of toxic heavy metals.
Similar to the results obtained from the analysis of published
patents pertaining to microalgal biofuel (Fig. 2A), the number of
published patents related to microalgal biotechnology for the
production of high value-added products and environmental
applications was relatively stable and then increased signicantly
during the period from 2006 to 2011, but started to decrease in
2012 (Fig. 4A). Nearly 50 countries/regions were involved in
publishing patents pertaining to microalgal biotechnology for the
production of high value-added products and environmental applications (Table 1), and the top three countries/regions were China,
the USA, and Korea (Figs. S5B and S5C in le of Supplemental material). However, the total output of research pertaining to microalgal
biotechnology for the production of high value-added products and
environmental applications also began to decrease recently, in
accordance with the marked downturn in research on microalgal
biofuel (Fig. 2A).
Using the IPC criteria (Table S2 in le of Supplemental material),
we identied and analyzed the top 15 technical classications of
patents related to microalgal biotechnology for the production of
high value-added products and environmental applications, and
found that more than 80% of the patents included in this group
were published between 2006 and 2012 (Fig. S5 in le of
Supplemental material). Similar to the conclusions derived from
the analysis of the high-frequency keywords of the published
manuscripts (Fig. S4 in le of Supplemental material), microalgal
mass culturing and environmental protection were found to be
the primary foci of this research (Fig. S5 in le of Supplemental
material).
3.4. The demands for the industrialization of microalgal biofuel
production have driven studies on mass microalgal culturing
According to the analysis of research foci in the elds of both
microalgal biofuel and microalgal biotechnology for the production
of high value-added products and environmental applications
(Tables S1 and S3, and Figs. S1S5 in le of Supplemental material),
microalgal mass culturing was always an important research focus,
particularly in publications pertaining to microalgal biofuel. To further understand the developmental trend found for studies on
microalgal mass culturing, we analyzed the 2426 published manuscripts and 944 published patents retrieved (Table 1).
Similar to the results found for published manuscripts related
to microalgal biofuel, the published manuscripts concerning
microalgal mass culturing exhibited a sustained increasing trend,
and increased sharply between 2008 and 2013, but the increase
in annual number of published manuscripts was relatively low in
20142015 (Fig. 5). The published patents related to microalgal
mass culturing (Fig. 6) showed a trend similar to that found in
the analysis of patents pertaining to microalgal biofuel (Fig. 2).
Moreover, the main countries in which these manuscripts and
patents were produced and the relevant trends in most of these
countries were consistent with the results obtained from our analysis of manuscripts and patents pertaining to microalgal biofuel.
In addition, a hysteretic quality during the burst stage was
found for publications related to microalgal mass culturing
(Figs. 5A and 6A), as was found for publications related to microalgal biofuel (Figs. 1A and 2A) and microalgal biotechnology for the
production of high value-added products and environmental applications (Figs. 3A and 4A), indicating that research on microalgal
mass culturing was also stimulated by research on microalgal biofuel. The practical application of microalgal biofuel production
requires industrialization; therefore, advances in the technology
and equipment needed for microalgal mass culturing, which is
the key challenge, have received much attention and were consequently promoted, indicating that advances in microalgal mass
592
keywords associated with this topic (Table S4 in le of

Supplemental material), we conclude that recent studies on
microalgal mass culturing focused on the breeding of algal species,
culture technologies, and metabolite production, as was also
revealed using the IPC criteria (Fig. S6 in le of Supplemental
material).
3.5. Discussion
Fig. 6. Trends in published patents pertaining to microalgal mass culturing. (A)

Annual statistics for the published patents; (B) statistics for the patents published in
the top 10 countries/regions; (C) annual statistics for the patents published in the
top 10 countries/regions during the past decade (20062015). Europe with the
exception of France, Spain, and Germany.
culturing may play an important role in reducing the cost of

algae-based biofuel. However, with the decrease in the number
of publications related to microalgal biofuel (Fig. 2A), the total
number of studies on microalgal mass culturing also began to
decline (Fig. 6A).
Our comprehensive analysis of the research foci of publications
pertaining to microalgal mass culturing is helpful for understanding and further promoting relevant research developments. Based
on the results of our comprehensive analysis of the countries in
which the authors of published studies resided, manuscripts, and
Based on the analysis of the retrieved manuscripts and patents,

the three research directions in the eld of microalgae, including
microalgal biofuel, microalgal biotechnology for the production of
high value-added products and environmental applications, and
microalgal mass culturing, presented similar trends (Figs. 16).
The published patents can be used to better indicate research
trends (Figs. 2, 4 and 6). These analyses revealed that research in
microalgal biofuel occurred more rapidly and entered the burst
stage relatively earlier than research in the other two areas, which
gradually entered the period of rapid development. Because the
greatest challenge for the manufacturing and application of
algae-based biodiesel is the high cost of cultivation, harvesting,
and biofuel extraction and production [41,69], research into
microalgal mass culturing and microalgal biotechnology for the
production of high value-added products and environmental applications has been greatly promoted in an attempt to make the entire
process economically feasible for industrialization. However, the
unacceptably high cost of microalgal biofuel production will continue to be a bottleneck for the development of further methods
[70], and has resulted in a recent sharp decrease in research on
microalgal biofuel and a further reduction in investment in the
other two research directions and even in overall microalgal
research. Researchers are thus attempting to study and develop
economically viable microalgal biofuel production technologies.
For example, Zhou et al. [71] introduced and analyzed a novel system for the production of algal biofuel that synergistically integrated algal wastewater treatment and the production of biofuel
co-products to provide a viable and advantageous pathway to sustainable, carbonneutral energy independence. In a previous study,
we achieved a 60% efciency of nitrite removal using Chlorella sp.
C2 cultured using ue gas-xed salts (FGFSs) in a 3-L reactor column with the simultaneous production of 33% algal lipids, suggesting that the synergistic combination of microalgal biological DeNOx
(bio-DeNOx) of industrial ue gas and the production of a biofuel,
which further suggested an utilization potential for the cost reduction of both ue-gas treatment and biodiesel production [18]. As
shown in Section 3, compared with the results regarding the
recently reduced rate in output of manuscripts concerning microalgal biofuel (Fig. 1), the output of manuscripts concerning microalgal
biotechnological applications for the production of high valueadded products and environmental-protection applications
(Fig. 3), particularly their synergistic application for the production
of microalgal biofuel (Fig. 3D), also exhibited an exponential rate of
growth, suggesting that the development of microalgal biotechnology continued to receive the attention of many researchers.
Although the decline in the rate of published patents demonstrated
a reduction in relevant technological advances (Figs. 2, 4 and 6), the
comprehensive utilization of energy-producing microalgal
resources, particularly the use of energy-producing microalgae for
the production of high value-added products and environmental
protection applications as a possibly economic and feasible mode
of biofuel production, merit further investigation.
In the face of the global energy crisis, many countries are
turning their attention to the development of new, clean, and sustainable energy sources [2], among which microalgal biofuel has
been recognized as a possible primary energy source [11,12]. As
presented in Section 3.1, microalgal biofuel research (Table 1)
has been conducted in more than 80 countries/regions, and a similar level of progress in this eld of research has occurred in most of
these countries (Fig. 1C), indicating that the production of microalgal biofuel is a global issue that has received the attention of many
countries/regions. However, research on microalgal biofuel has
been relatively concentrated on the national/regional level, and
most of the research ndings arose from a few countries. For example, more than 70% and 90% of the total manuscripts and patents
were, respectively, derived from the 10 countries/regions with
the
highest
number
of
publications
on
the
topic
(Figs. 1B and 2B). Because these 10 countries/regions are major
economically developed or developing countries/regions in the
world, investments in new energy sources, such as microalgal biofuel, are driven by their huge energy demands. By contrast, we
found that the rate of published patents in the eld of microalgal
biofuel was signicantly lower than that of the corresponding
number of published manuscripts in terms of both the quantity
and the geographic distribution (Table 1), suggesting a lower level
of transformation into achievements. Thus, the current level of
understanding of microalgal biofuel production is not sufciently
deep, and this results in the high cost of research and production
processes and is a key factor causing the bottleneck regarding
microalgal biofuel production. A similar trend was found for
research directions related to microalgal biotechnology for the production of high value-added products and environmental applications and microalgal mass culturing (Table 1), indicating that our
current understanding of these two directions as well as of the
entire eld of microalgal research is not sufciently deep and that
the solutions to various problems in the area of microalgae
research may depend on the development of a more in-depth
understanding.
As the most populous and largest developing country in the
world, China has invested a large amount of money and research
effort in studies of these three directions of microalgal research.
Consequently, China has the most and second most published
patents and published manuscripts related to microalgal research,
respectively, in the world (Figs. 16). Interestingly, recent research
conducted in China appeared to follow a unique development
trend compared with research efforts conducted in other countries.
The downturn in the number of studies on microalgal biofuel
occurred in most countries in 2012 or 2013, when the number of
published patents in this eld declined precipitously. By contrast,
the number of such patents in China appeared to greatly increase
in 2012 and remained at a high level until 2013 (Fig. 2C).
Regarding the research directions of microalgal mass culturing
and microalgal biotechnology for the production of high
value-added products and environmental applications, China led
the world in the output of related patents from 2009 to 2010,
and most of the total output of global related patents was obtained
from China during 20122013 (Figs. 4 and 6). The explanation for
this phenomenon is mainly that Chinas economic and scientic
plans are both pre-arranged as Five-Year Plans (FYPs), resulting
in most of the microalgal biofuel research projects being launched
in its 12th FYP (20102014). For example, the ENN Group Co. Ltd.
initiated microalgal biofuel projects from 2008 to 2010 [72].
However, it was not until 2011 and 2012 that the National
Program on Key Basic Research Project of China (973 Program)
and the National High Technology Research and Development
Program of China (863 Program) ofcially launched their respective national research projects on microalgal biofuel [72,73].
Although a decline in microalgal biofuel research throughout the
world occurred in 2012 or 2013, the projects in China were being
executed and most of the related research was still ongoing, resulting in a relatively high output of publications, particularly in patent
publication. In addition, researchers in China appeared to pay earlier attention than other countries to microalgal biotechnology for
593
the production of high value-added products and environmental

applications and microalgal mass culturing and to look forward
to economically viable microalgal biofuel industrialization based
on the comprehensive utilization of energy-producing microalgae.
4. The challenges of microalgae biofuel production and recent
advances of their solution
The microalgal biofuels witness an obvious and serious
dilemma, where it is found that there is no commercial production
on a large scale for bulk application due to the overwhelming
investments in capital and operation [74]. Most of all, our limited
understanding of microalgae, including microalgal biofuel, will
remain the most fundamental cause of the unacceptably high cost
of research and production (as well as the discussion in
Section 3.2), which is a bottleneck for the further development of
microalgal biofuel for a long time to come. The high costs as well
as microalgal biology, specic cultivation, harvesting, dewatering,
and extraction requirements have limited the development of this
technology for many years [75]. With the aim of commercialization, the potential solutions to some impediments, including
impossibly high demands on certain key resources, the high cost
of production, and the need to achieve an energy ratio of well
above unity, are being investigated [10].
As energy efciency and cost-effective processing are two major
challenges in the commercialization of biofuels from algae,
research addressing these challenges ought to be intensied [76].
Economics are currently the main barrier to produce microalgae
on a large scale for the application of lower values such as biofuels,
since industries are still in R&D phase [77]. Large scale microalgal
cultivation for the production of biofuels requires a combination of
technical breakthroughs and innovative pathways. Based on the
analysis and discussion in Section 3, researchers have increasingly
focused on increasing microalgal biomass while reducing costs and
on using energy-producing microalgae to produce biofuel and high
value-added products (Figs. S1S6, and Tables S1, S3 and S4 in le
of Supplemental material). One option to reduce costs in the
microalgae-based biofuel industry is to develop the process of
microalgal cultivation. Efforts, such as optimization of cultivation
conditions (e.g., nutrient deprivation, light control, mixing, etc.),
harvest method development and utilization of residues, have been
made. In addition, if the alga can be selected, engineered, or otherwise coaxed into increasing lipid productivity or other high
value-added products, the production economics might be substantially improved.
4.1. Species
The physical properties of microalgae, e.g., the detailed descriptions of the physical properties of microalgae affected by growth
and environmental conditions and the key physical properties that
are useful in designing methods to separate cells from water, are
important when understanding of microalgae and considering processing options [78]. In addition, the pace of the available understanding of microalgae is obviously increasing with sequencing
and genomes technologies, post-genomics technologies, genetic
tools, proteomic technologies and metabonomics technologies,
which have already made major contributions to fundamental
research on photosynthetic microalgae (e.g., Chlamydomonas reinhardtii) in the elds of functional biology and the exploration of
metabolic pathways and biological processes, and will accelerate
the commercialisation of alga-derived compounds by providing a
framework for hypothesis-based strain improvement programs
built on an improved fundamental understanding of the specic
pathways and regulation networks [79].
594
Among the wild species available in formal collections, high

oil-producing microalgal species often have a slow growth rate.
In such cases, mutagenic or genetic methods can be used to
develop organisms that grow rapidly and assimilate large amounts
of energy and are suitable for commercial biodiesel production
[80,81]. However, several shortcomings have hindered the broader
use of genetically engineered microalgae. For instance, few
microalgal strains have been genetically modied due to a lack of
specic molecular biology tools and the mechanisms underlying
the regulation of both gene expression and the metabolic network
are unclear. Furthermore, concerns of biological contamination
have materialized in restrictive legislation [1]. Monocultures,
which are used in most microalgal studies, are susceptible to contamination, especially in a complex environment (e.g., wastewater), and strains with high lipid productivity are more likely to be
outperformed by other faster growing species [82,83]. Mixed cultures of algae can sustain themselves in complex environments,
and diverse microalgal communities are considered to be more
stable and have a lower invasion risk than monocultures [83].
Some of the main species of algae used in large-scale commercial
production are restricted to geographic locations with warm climate and would be unable to grow at acceptable rates during the
hot or cold season of certain geographical region [84]. One solution
to this problem is to identify indigenous algae that are adapted to
the local environment [84]. In addition, native microalgal species
also performed better than most other species in commercial scale
cultivation with native environment [85].
4.2. Cultivation
Microalgae grown at a large scale, especially in open pond systems, are susceptible to contamination by undesirable algae, bacteria, protozoa, and zooplankton, which could be very costly to
control. Bacterial contamination may be limited by increasing the
pH or by treatment with detergent and phenol [86,87].
Contamination by protozoa, zooplankton, and other algae may be
reduced by using highly selective culture conditions, such as high
salinity [88] and high alkalinity [87]. In addition, closed systems
are more likely to be run as continuous systems due to their higher
efciency and signicantly lower amounts of contamination of
undesirable algae and other organisms [86].
The large consumption of water resources, inorganic nutrients
(mainly N and phosphate) and CO2 in microalgal cultivation is
costly and hinders commercialization of algal biofuel production
[15]. Biofuel production from microalgae may be more environmentally sustainable, cost-effective, and protable if combined
with processes such as ue gas and wastewater treatments [71].
CO2 bio-xation from ue gas by microalgae has drawn much
attention as an environmentally friendly CO2 mitigation and biomass production strategy. However, due to the low solubility of
CO2 in water, a limited amount of CO2 is absorbed in the liquid,
resulting in a low carbon utilization efciency [89]. Compared with
open culture systems, closed PBRs could reduce CO2 losses by prolonging CO2 retention time and improving mass transfer efciency
[90]. To improve the efciency of CO2 uptake by algae, CO2 can be
absorbed by alkaline solutions, which stabilize the CO2 as carbonate, and then fed to algae [91].
Nitrogen oxides (NOx), another signicant component of ue
gas, could serve as a N source for microalgae and could be metabolized by microalgae [18]. NO, which is the main component of
NOx, is sparingly soluble in water, and the dissolution of NO into
the microbial culture is the rate-limiting step for NO removal
[92]. Some attempts have been made to enhance the solubility of
NO gas using an efcient complexing agent such as
metal-chelated EDTA [93]. However, these complexing agents are
harmful to the algae and expensive when used in
commercial-scale cultures. In addition to enhancing the dissolution of NO in water, the initial xation of massive amounts of NO
or NOx into alkaline solution and then the cultivation of algal cells
using xed nutrients is possibly an effective way of improving NO
or NOx removal efciency [18]. In our previous study, the actual
NOx xed salts, which xed by using alkaline solution, were
applied to Chlorella sp. C2 cultivation, and a 60% nitrite removal
efciency was obtained together with the production of 33% algae
lipids [18].
The large volumes of fresh water required for large-scale
microalgal culture represent a serious problem. As large amounts
of N and P can be recovered from wastewater, microalgal biomass
could be produced using wastewater as the nutrient source to conserve fresh water, while offering added environmental advantages
[94]. A recent study suggested that coupling biofuel generation to
wastewater treatment is an attractive option for reducing the
energy, nutrient, and freshwater costs involved in biofuel production [95]. Due to the complex nature of wastewater, issues such as
contamination, inconsistent wastewater components, and unstable
biomass production hinder efforts to use wastewater for
large-scale algal cultivation [96]. The industrial production of
microalgal biofuels in wastewater will depend on the development
of microalgal cultivation methods that address issues such as
nutrient removal from wastewater under outdoor and cold climate
conditions [97].
In eukaryotic green microalgae, manipulation of metabolic
pathways by altering the culture medium and/or culture condition
is a powerful tool for physiological control and may be also generally cost-efcient, environmentally friendly, applicable on a large
scale and exible for various industrially attractive microalgae species [98]. Several cultivation strategies, of which N shortage is the
most effective, can be used for increasing TAG synthesis rates [22].
N limitation (generally a continuous cultivation mode) and N starvation (generally a batch process) are two conditions with respect
to N shortage [99]. Maximum biomass productivity and target
compound content usually cannot be obtained simultaneously
under the same cultivation conditions. Although starvation processes yield the highest average TAG production rates and nal
TAG contents, continuous cultivation could offer important advantages over batch operation are those the operation exibility under
varying outdoor conditions and the increased opportunities to
make value of all cell components by adjusting cultivation conditions [100]. Thus, further exploration on continuous cultivation
for the production of microalgal oil is deserved on a commercial
scale.
4.3. Harvesting and extraction
Conventional harvesting and extraction processes, which were
thought to be costly and energy intensive, still in general remain
small-scale, traditional and primitive, and are in need of
innovation.
High cost of microalgal biomass harvesting is one of the bottlenecks for commercialization of microalgae-based industrial processes. Wan et al. [101] thought that occulation was a
promising method to harvest microalgae with low cost from mass
culture when considering harvesting efciency, operation economics and technological feasibility. However, there are still a lot
of challenges in microalgae biomass harvesting using efcient
and cost effective occulating technologies. Various novel occulation technologies have been developed. The occulation process
induced by microorganism, extracellular polymer substances, or
bioocculants (or called occulating agents) produced by microbes
is termed as bioocculation, which distinguishes itself by being
environmentally friendly among the occulation methods used
for microalgae harvesting [101]. Fungal/bacterial-assisted
occulation is currently receiving increased attention because of

its high harvesting efciency. A study by Wang et al. [102] optimized the co-culture conditions of Chlorella and bioocculantproducing bacteria in synthetic wastewater using response surface
methodology (RSM) for enhancing Chlorella harvesting and lipid
content, and the results showed that the increase of Chlorella harvesting efciency reached 450500% and the increase of lipid content was over 210%. Cell self-occulation, which widely occurs in
microorganisms including several self-occulating microalgae,
refers to cell aggregation and adhesion of cells to each other in liquid culture due to special cell surface properties [103]. The greatest
advantages of cell self-occulation for cell harvest are that it does
not need chemical addition, and thus is cost effective, free of chemical contaminations and environmentally friendly [101].
Among the various cell-disruption methods for extraction,
ultrasonic or sonication is one of the most widely employed, which
also could be combined efciently with other production processes. Coons et al. [52] showed that ultrasonic harvesters and
extractors were uniquely capable of meeting the requirement of
low energy cost. In a study by Park et al. [104], the feasibility of
a sonication-assisted homogenization system for lipid extraction
from Chlorella vulgaris was evaluated, which showed that, by combination of sonication and homogenization, the lipid-recovery
yields were increased compared with that by solo use of either
homogenization or sonication. In addition, most mechanical
energy is dissipated to produce heat rather than disruption of cell
walls, and biological means are needed for future development
[76]. A study proposed a bacterial (enzymatic) destruction pretreatment of the microalgae cell wall to render the microalgal oil
extraction more efcient, which has been found that a nearly
100% increase in lipid extraction efciency was obtained [40].
4.4. Co-production by biorenery
Biorenery is an industrial process, where biomass is converted
into arrange of biochemicals, materials and energy products, which
aims to produce multiple products and maximizing the value
derived from different microalgal components [74]. Besides biofuels, various high-value chemical compounds, such as pigments,
antioxidants, b-carotenes, polysaccharides, vitamins, and biomass,
can be extracted from microalgae, depending on the microalgal
compositions, and are largely used as bulk commodities in different industrial sectors [12]. Co-producing high value-added products with biofuels can provide a promising opportunity to
commercialize microalgal biofuels. Williams and Laurens [70]
noted that 30% and 50% of the algal primary product mass was lost
during protein and lipid production, respectively, and that the
increased lipid content reduces other valuable compounds in the
biomass. Thus, biofuel production with the biofuel only option
is unlikely to be economically viable [70]. The large-scale production of algae-based biofuel depends not only on efforts to scale-up
algal growth, harvesting and oil extraction, but also on the
availability of revenue streams from the oil and by-products that
are likely to be of higher value than the fuel itself. Even the revenue
from lower yield by-products, such as biofertilizers, may tip the
balance between viable and non-viable commercialization [105].
With the aim at maximizing the value derived from different
microalgal components, Zhu [74] proposed that the innovative
microalgal biorenery concept includes four pathways:
high-value productsbiodieselbioethanolbiogas; high-value pro
ductsbioethanolbiogas; high-value productsbiodieselbiogas;
and high-value productsbiogas, which are expected to promote
the economics of microalgal biofuels. Bohutskyi et al. [106] also
presented to be maximizing the utilization of microalgal biomass
that converted into fuel, and generation of waste products could
be minimized through the potential conversion of lipid extracted
595
algal into additional renewable energy products (e.g., methane)

by integration with other technologies. For example, anaerobic
digestion represents a useful approach for conversion of algal residues into complementary biopower and provides the capacity to
recycle essential nutrients from lipid extracted algal back into
the cultivation stage, reducing the requirement for fertilizers to
lower the overall costs of algal biofuels [107]. By coupling lipid
extraction and anaerobic digestion, an innovative, mixed trophic
state process with high productivity was implemented to generate
microalgae with high lipid content for generating biodiesel and
biogas, which increased the overall process energy output up to
40% [106]. Similarly, the biodiesel-derived co-production (e.g.,
glycerol or xylose) could also be recycled for increased growth
and lipid production. Leite et al. [108] reported that some microalgal strains were capable of mixotrophic and heterotrophic growth
on biodiesel-derived glycerol and xylose with the signicant
increase in growth rate and lipid productivity.
5. Perspective view of the future R&D needs and trends
After a period of intensive development, the level of interest in
research on microalgal biofuel recently declined, as did that on
microalgal biotechnology and mass culturing. The perspective of
algal biodiesel is currently poor due to several biological and technical restrictions. Due to our limited understanding of microalgae,
including microalgal biofuel, the unacceptably high cost of
research and production will remain a bottleneck for the further
development of microalgal biofuel for a long time to come. It is
urgent to bring evident breakthroughs to produce economically
viable biofuels for future development. For algal biofuel to become
a commercial reality, unremitting research and sustained capital
investment are required. Much work remains to be done on the
basic biology of microalgae, such as species selection, genetic
manipulation and molecular characterization of the metabolic
switch for carbon sequestering and storage. The future research
should not concentrate on single aspect, and the effective and economic processes for microalgal biofuels will require the integration
of microalgal biology said above, as well as the reactor design and
integration into environmental mass and energy uxes, and the
processes will be identied and further promoted by integrating
life cycle analysis [109]. Based on the knowledge of basic biological
functions and technical methods for controlling microalgal metabolism, the aim of increasing the content of target compounds or
enhancing their productivity will be achievable. Furthermore, culturing systems accompanying water, nutrient and CO2 requirements, the method of cell harvesting, cell disruption and
subsequent lipid extraction, and the biodiesel production from
the crude lipid fraction require further investigation. All economic
assessments of above technique processes in algal biofuels at
large-scale or pilot-scale are necessarily and relevant advances
must be proven in large-scale or pilot-scale production systems.
To date, advances on the selection and modication of optimized algal species for biofuel production have been slow.
Genetic manipulation of alga species, particular the algae with
specic characters, should be drew more attention in future R&D
activities. For example, harvesting microalgae using cell
self-occulation has distinguished itself as a promising method
for low-cost harvesting technology, but only a few natural
self-occulating microalgae have been documented, which apparently cannot meet the demand for commercial application of
microalgae harvesting, and enhancement of self-occulation in
microalgae via genetic engineering is necessary. [101]. Taken into
consideration, optimization of genetic elements and transformation methods during further investigation in genetic modication
is great signicance to obtain optimized algal species microalgae
with biomass growth and/or lipid production, and biosafety of
596
transgenic microalgae should be considered as well. For genetic

manipulation of alga species, the regulation of TAG accumulation
and the relation between TAG metabolism and photosynthesis
are also important knowledge gaps in future R&D activities. In
addition, the high-throughput microalgal screening methods for
obtaining optimized algal species are indispensable in future
researches.
The cost of microalgal biofuel production may be reduced by the
following strategies: (1) coupling microalgae-based biofuel production with processes such as wastewater and ue gas treatments;
and (2) bio-rening microalgae for the production of high
value-added products. Aimed to economically commercial microalgal biofuel on the basis these two strategies, some breakthroughs
and technical requirements should be concerned in the future
R&D activities. As the ability of CO2 uptake by microalgae varies
dramatically among algal species, it is necessary to select or reform
suitable algal candidates to x CO2 from actual ue gases under a
large collection of algal cultures. Besides CO2 and NOx, ue gases
contain many other compounds such as H2O, O2, N2, sulfur oxides
(SOx), unburned carbohydrates (CxHy), CO, heavy metals, halogen
acids and particulate matter (PM). Several chemical compounds
(SOx, heavy metals, etc.) have shown to be toxic to some microalgae
[110]. To better engineer ue gas-fed microalgal cultures, the
effects of all ue gas compounds on microalgae, tolerance of various
microalgae to ue gas compounds and the interaction of ue gas
compounds and microalgae need to be assessed [111,112]. In addition, the tests of growing algae in wastewater are mostly at laboratory scale, which require conrmation in long term and at
large-scale production. Improve the stable performance of microalgal cultivation and nutrient removal under outdoor and cold climate conditions is an urgent issue needed to be solved to achieve
sustainable development for the algal industry. For the prosperity
of microalgal biorenery in future, R&D activities should be conducted on the basis of the desired end product, the suitable design
of culture conditions induced and activated the accumulation of
some targeted constituents, and the genetic and metabolic engineering to improve the targeted ingredients [74]. Notably, researchers in many countries, including China, have continued research
into effective ways to reduce the production costs of microalgal biofuel, and the biotechnological applications of energy-producing
microalgae for the production of high value-added products and
environmental protection appear to be an effective means of
achieving economically viable biofuel production.
In view of the above, current and future research for potential
solutions to the existing barriers in commercially microalgal biofuel production should focus on the following: (1) screening and
breeding energy-producing microalgal species for the production
of high value-added products and environmental applications;
(2) exploring the metabolic mechanisms underlying the production of high value-added metabolites in energy-producing microalgae, as well as the mechanisms through which energy-producing
microalgae adapt, x, and remove pollutants, including those in
ue gas, waste water, and waste residues; (3) examining mass culturing conditions for energy-producing microalgae, focusing on the
optimization of conditions, such as nutrition, light, temperature,
pH, and ventilation, to reduce the production costs of microalgal
biomass; and, most importantly, (4) basic research on the physiology and molecular biology of microalgae to improve our understanding of these microorganisms. Biotechnology advances,
well-funded R&D researches on biotechnology and policy support
can help realize this scenario.
6. Conclusions
Through an informatics analysis of papers and patents published
between 1900 and May 2015 in microalgal research, particularly in
microalgal biofuel research, a burst in microalgal biofuel research

from 2006 to 2011 that signicantly stimulated the development
of microalgal biotechnology for the production of high
value-added commodities and for environmental applications and
microalgal mass culturing. However, a lag in basic microalgal
research has kept production costs high, which was also shown
from an in-depth knowledge of the recent advances, resulting in a
decline in investments, funding, and research efforts in the elds
of microalgal biofuel production, microalgal biotechnology, and
mass culturing since 2012. Faced to the challenges of commercial
microalgae-based biofuel production, much work remains to be
done on the basic biology of microalgae, and energy-producing
microalgal biotechnological applications should be synergistically
combined with microalgal biofuel production. In future R&D activities, the effective and economic processes for microalgal biofuels
will require the integration of microalgal biology, such as species
selection, genetic manipulation and molecular characterization of
the metabolic switch for carbon sequestering and storage, as well
as the reactor design and integration into environmental mass
and energy uxes. Future investments and funding will most likely
be directed toward basic studies that aim to elucidate the microorganisms characteristics and toward the development of microalgal
biotechnology and its environmental applications. Once our understanding of basic microalgae biology is equivalent to that of crops
such as corn, we will be able to farm microalgae for biofuel production and other purposes in an economically feasible manner.
Acknowledgements
This work was supported jointly by the National Program on
Key Basic Research Project (2012CB224803), the National Natural
Science Foundation of China (31300030), Sinopec (S213049), the
Natural Science Foundation of Hubei Province of China
(2013CFA109), and the Knowledge Innovation Program of the
Chinese Academy of Sciences (Y35E05).
Appendix A. Supplementary material
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.apenergy.2015.
06.055.
References
[1] Amaro HM, Guedes AC, Malcata FX. Advances and perspectives in using
microalgae to produce biodiesel. Appl Energy 2011;88:340210.
[2] Farrell AE, Plevin RJ, Turner BT, Jones AD, OHare M, Kammen DM. Ethanol can
contribute to energy and environmental goals. Science 2006;311:5068.
[3] Liang MH, Jiang JG. Advancing oleaginous microorganisms to produce lipid
via metabolic engineering technology. Prog Lipid Res 2013;52:395408.
[4] Miao XL, Wu QY. Biodiesel production from heterotrophic microalgal oil.
Bioresour Technol 2006;97:8416.
[5] Singh J, Cu S. Commercialization potential of microalgae for biofuels
production. Renew Sust Energy Rev 2010;14:2596610.
[6] Loureno SO. Cultivo de microalgas marinhas: princpios e aplicaes. So
Carlos Brazil: Editora Rima; 2006.
[7] Perez-Garcia O, Escalante FME, de-Bashan LE, Bashan Y. Heterotrophic
cultures of microalgae: metabolism and potential products. Water Res
2011;45:1136.
[8] Miron AS, Garciia MCC, Gomez AC, Camacho FG, Grima EM, Chisti Y. Shear
stress tolerance and biochemical characterization of Phaeodactylum
tricornutum in quasi steady-state continuous culture in outdoor
photobioreactors. Biochem Eng J 2003;16:28797.
[9] Converti A, Casazza AA, Ortiz EY, Perego P, Del Borghi M. Effect of temperature
and nitrogen concentration on the growth and lipid content of
Nannochloropsis oculata and Chlorella vulgaris for biodiesel production.
Chem Eng Process 2009;48:114651.
[10] Chisti Y. Constraints to commercialization of algal fuels. J Biotechnol
2013;167:20114.
[11] Ahmad AL, Yasin NHM, Derek CJC, Lim JK. Microalgae as a sustainable energy
source for biodiesel production: a review. Renew Sust Energy Rev
2011;15:58493.

[12] Mata TM, Martins AA, Caetano NS. Microalgae for biodiesel production and
other applications: a review. Renew Sust Energy Rev 2010;14:21732.
[13] Araujo GS, Matos LJBL, Fernandes JO, Cartaxo SJM, Goncalves LRB, Fernandes
FAN, et al. Extraction of lipids from microalgae by ultrasound application:
prospection of the optimal extraction method. Ultrason Sonochem
2013;20:958.
[14] Gong Y, Wan X, Jiang M, Hu C, Hu H, Huang F. Metabolic engineering of
microorganisms to produce omega-3 very long-chain polyunsaturated fatty
acids. Prog Lipid Res 2014;56:1935.
[15] Markou G, Georgakakis D. Cultivation of lamentous cyanobacteria (blue
green algae) in agro-industrial wastes and wastewaters: a review. Appl
Energy 2011;88:3389401.
[16] Ketheesan B, Nirmalakhandan N. Development of a new airlift-driven
raceway reactor for algal cultivation. Appl Energy 2011;88:33706.
[17] Jiang LL, Luo SJ, Fan XL, Yang ZM, Guo RB. Biomass and lipid production of
marine microalgae using municipal wastewater and high concentration of
CO2. Appl Energy 2011;88:333641.
[18] Zhang X, Chen H, Chen W, Qiao Y, He C, Wang Q. Evaluation of an OilProducing Green Alga Chlorella sp. C2 for biological DeNOx of industrial ue
gases. Environ Sci Technol 2014;48:10497504.
[19] Chisti Y. Biodiesel from microalgae. Biotechnol Adv 2007;25:294306.
[20] Li YQ, Horsman M, Wang B, Wu N, Lan CQ. Effects of nitrogen sources on cell
growth and lipid accumulation of green alga Neochloris oleoabundans. Appl
Microbiol Biotechnol 2008;81:62936.
[21] Scott SA, Davey MP, Dennis JS, Horst I, Howe CJ, Lea-Smith DJ, et al. Biodiesel
from algae: challenges and prospects. Curr Opin Biotech 2010;21:27786.
[22] Hu Q, Sommerfeld M, Jarvis E, Ghirardi M, Posewitz M, Seibert M, et al.
Microalgal triacylglycerols as feedstocks for biofuel production: perspectives
and advances. Plant J 2008;54:62139.
[23] Abou-Shanab RAI, Hwang JH, Cho YC, Min B, Jeon BH. Characterization of
microalgal species isolated from fresh water bodies as a potential source for
biodiesel production. Appl Energy 2011;88:33006.
[24] Sydney EB, da Silva TE, Tokarski A, Novak AC, de Carvalho JC, Woiciecohwski
AL, et al. Screening of microalgae with potential for biodiesel production and
nutrient removal from treated domestic sewage. Appl Energy
2011;88:32914.
[25] Nascimento IA, Marques SSI, Cabanelas ITD, Pereira SA, Druzian JI, de Souza
CO, et al. Screening microalgae strains for biodiesel production: lipid
productivity and estimation of fuel quality based on fatty acids proles as
selective criteria. Bioenergy Res 2013;6:113.
[26] Huang GH, Chen F, Wei D, Zhang XW, Chen G. Biodiesel production by
microalgal biotechnology. Appl Energy 2010;87:3846.
[27] Mandal S, Mallick N. Microalga Scenedesmus obliquus as a potential source for
biodiesel production. Appl Microbiol Biotechnol 2009;84:28191.
[28] Li XF, Xu H, Wu QY. Large-scale biodiesel production from microalga Chlorella
protothecoides through heterotrophic cultivation in bioreactors. Biotechnol
Bioeng 2007;98:76471.
[29] Gao CF, Zhai Y, Ding Y, Wu QY. Application of sweet sorghum for biodiesel
production by heterotrophic microalga Chlorella protothecoides. Appl Energy
2010;87:75661.
[30] Lam MK, Lee KT. Microalgae biofuels: a critical review of issues, problems and
the way forward. Biotechnol Adv 2012;30:67390.
[31] Pires JCM, Alvim-Ferraz MCM, Martins FG, Simoes M. Carbon dioxide capture
from ue gases using microalgae: engineering aspects and biorenery
concept. Renew Sust Energy Rev 2012;16:304353.
[32] Moreno-Garrido I, Canavate JP. Assessing chemical compounds for controlling
predator ciliates in outdoor mass cultures of the green algae Dunaliella salina.
Aquacult Eng 2001;24:10714.
[33] Guschina IA, Harwood JL. Lipids and lipid metabolism in eukaryotic algae.
Prog Lipid Res 2006;45:16086.
[34] Brennan L, Owende P. Biofuels from microalgae-A review of technologies for
production, processing, and extractions of biofuels and co-products. Renew
Sust Energy Rev 2010;14:55777.
[35] Grima EM, Belarbi EH, Fernandez FGA, Medina AR, Chisti Y. Recovery of
microalgal biomass and metabolites: process options and economics.
Biotechnol Adv 2003;20:491515.
[36] Mendes RL, Reis AD, Palavra AF. Supercritical CO2 extraction of gammalinolenic acid and other lipids from Arthrospira (Spirulina) maxima:
Comparison with organic solvent extraction. Food Chem 2006;99:5763.
[37] Vijayaraghavan K, Hemanathan K. Biodiesel production from freshwater
algae. Energy Fuel 2009;23:544853.
[38] Ehimen EA, Sun ZF, Carrington CG. Variables affecting the in situ
transesterication of microalgae lipids. Fuel 2010;89:67784.
[39] Chen H, Zhang Y, He C, Wang Q. Ca2+ signal transduction related to neutral
lipid synthesis in an oil-producing green alga Chlorella sp. C2. Plant Cell
Physiol 2014;55:63444.
[40] Zhang Y-M, Chen H, He C-L, Wang Q. Nitrogen starvation induced oxidative
stress in an oil-producing green alga Chlorella sorokiniana C3. PLoS ONE
2013;8:e69225.
[41] Carioca JOB, Hiluy JJ, Leal MRLV, Macambira FS. The hard choice for
alternative biofuels to diesel in Brazil. Biotechnol Adv 2009;27:104350.
[42] Bogen C, Klassen V, Wichmann J, La Russa M, Doebbe A, Grundmann M, et al.
Identication of Monoraphidium contortum as a promising species for liquid
biofuel production. Bioresour Technol 2013;133:6226.
[43] Lpez-Gonzlez D, Fernandez-Lopez M, Valverde JL, Sanchez-Silva L. Kinetic
analysis and thermal characterization of the microalgae combustion process
[44]
[45]
[46]
[47]
[48]
[49]
[50]
[51]
[52]
[53]
[54]
[55]
[56]
[57]
[58]
[59]
[60]
[61]
[62]
[63]
[64]
[65]
[66]
[67]
[68]
[69]
[70]
[71]
[72]
597
by thermal analysis coupled to mass spectrometry. Appl Energy

2014;114:22737.
Zhang X, Rong J, Chen H, He C, Wang Q. Current status and outlook in the
application of microalgae in biodiesel production and environmental
protection. Front Energy Res 2014;2.
Chen Y, Wang JF, Zhang W, Chen L, Gao LL, Liu TZ. Forced light/dark
circulation operation of open pond for microalgae cultivation. Biomass
Bioenergy 2013;56:46470.
Richardson JW, Johnson MD, Outlaw JL. Economic comparison of open pond
raceways to photo bio-reactors for protable production of algae for
transportation fuels in the Southwest. Algal Res 2012;1:93100.
Arbib Z, Ruiz J, Alvarez-Diaz P, Garrido-Perez C, Barragan J, Perales JA. Long
term outdoor operation of a tubular airlift pilot photobioreactor and a high
rate algal pond as tertiary treatment of urban wastewater. Ecol Eng
2013;52:14353.
Moazami N, Ashori A, Ranjbar R, Tangestani M, Eghtesadi R, Nejad AS. Largescale biodiesel production using microalgae biomass of Nannochloropsis.
Biomass Bioenergy 2012;39:44953.
Lam MK, Lee KT. Potential of using organic fertilizer to cultivate Chlorella
vulgaris for biodiesel production. Appl Energy 2012;94:3038.
Lee YC, Kim B, Farooq W, Chung J, Han JI, Shin HJ, et al. Harvesting of
oleaginous Chlorella sp by organoclays. Bioresour Technol 2013;132:4405.
Beuckels A, Depraetere O, Vandamme D, Foubert I, Smolders E, Muylaert K.
Inuence of organic matter on occulation of Chlorella vulgaris by calcium
phosphate precipitation. Biomass Bioenergy 2013;54:10714.
Coons JE, Kalb DM, Dale T, Marrone BL. Getting to low-cost algal biofuels: a
monograph on conventional and cutting-edge harvesting and extraction
technologies. Algal Res 2014;6:25070.
Uduman N, Qi Y, Danquah MK, Forde GM, Hoadley A. Dewatering of
microalgal cultures: a major bottleneck to algae-based fuels. J Renew Sust
Energy 2010;2.
Kumar A, Ergas S, Yuan X, Sahu A, Zhang Q, Dewulf J, et al. Enhanced CO2
xation and biofuel production via microalgae: recent developments and
future directions. Trends Biotechnol 2010;28:37180.
Demirbas A. Production of biodiesel from algae oils. Energy Sour, Part A:
Recov, Util, Environ Effects 2008;31:1638.
Kanda H, Li P, Yoshimura T, Okada S. Wet extraction of hydrocarbons from
Botryococcus braunii by dimethyl ether as compared with dry extraction by
hexane. Fuel 2013;105:5359.
Peralta-Ruiz Y, Gonzlez-Delgado AD, Kafarov V. Evaluation of alternatives for
microalgae oil extraction based on exergy analysis. Appl Energy
2013;101:22636.
Mouahid A, Crampon C, Toudji SAA, Badens E. Supercritical CO2 extraction of
neutral lipids from microalgae: experiments and modelling. J Supercrit Fluids
2013;77:716.
Samarasinghe N, Fernando S, Lacey R, Faulkner WB. Algal cell rupture using
high pressure homogenization as a prelude to oil extraction. Renewable
Energy 2012;48:3008.
Doucha J, Lvansky K. Inuence of processing parameters on disintegration of
Chlorella cells in various types of homogenizers. Appl Microbiol Biotechnol
2008;81:43140.
Adam F, Abert-Vian M, Peltier G, Chemat F. Solvent-free ultrasoundassisted extraction of lipids from fresh microalgae cells: a green, clean and
scalable process. Bioresour Technol 2012;114:45765.
Bigelow TA, Xu J, Stessman DJ, Yao L, Spalding MH, Wang T. Lysis of
Chlamydomonas reinhardtii by high-intensity focused ultrasound as a
function of exposure time. Ultrason Sonochem 2014;21:125864.
Sheng J, Vannela R, Rittmann BE. Evaluation of cell-disruption effects of
pulsed-electric-eld treatment of Synechocystis PCC 6803. Environ Sci
Technol 2011;45:3795802.
Lam MK, Lee KT, Mohamed AR. Homogeneous, heterogeneous and enzymatic
catalysis for transesterication of high free fatty acid oil (waste cooking oil) to
biodiesel: a review. Biotechnol Adv 2010;28:50018.
Sathish A, Smith BR, Sims RC. Effect of moisture on in situ transesterication
of microalgae for biodiesel production. J Chem Technol Biotechnol
2014;89:13742.
Dang-Thuan T, Chen CL, Chang JS. Effect of solvents and oil content on direct
transesterication of wet oil-bearing microalgal biomass of Chlorella vulgaris
ESP-31 for biodiesel synthesis using immobilized lipase as the biocatalyst.
Frigon J-C, Matteau-Lebrun F, Hamani Abdou R, McGinn PJ, OLeary SJB, Guiot
SR. Screening microalgae strains for their productivity in methane following
anaerobic digestion. Appl Energy 2013;108:1007.
Zhang Y, Fan X, Yang Z, Wang H, Yang D, Guo R. Characterization of H2
photoproduction by a new marine green alga, Platymonas helgolandica var.
tsingtaoensis. Appl Energy 2012;92:3843.
Bai MD, Cheng CH, Wan HM, Lin YH. Microalgal pigments potential as
byproducts in lipid production. J Taiwan Inst Chem E 2011;42:7836.
Williams PJIB, Laurens LML. Microalgae as biodiesel & biomass feedstocks:
review & analysis of the biochemistry energetics & economics. Energy
Environ Sci 2010;3:55490.
Zhou Y, Schideman L, Yu G, Zhang Y. A synergistic combination of algal
wastewater treatment and hydrothermal biofuel production maximized by
nutrient and carbon recycling. Energy Environ Sci 2013;6:376579.
Wang L, Zhu Z, Xu C, Liu M. Analysis of CO2 consolidation and bioenergy
producrion via microalage. J China Agric Univ (Chinese) 2012;17:24752.
598
[73] Wang H, Gao LL, Chen L, Guo FJ, Liu TZ. Integration process of biodiesel
production from lamentous oleaginous microalgae Tribonema minus.
[74] Zhu LD. Biorenery as a promising approach to promote microalgae industry:
an innovative framework. Renew Sust Energy Rev 2015;41:137684.
[75] Ziolkowska JR, Simon L. Recent developments and prospects for algae-based
fuels in the US. Renew Sust Energy Rev 2014;29:84753.
[76] Show KY, Lee DJ, Tay JH, Lee TM, Chang JS. Microalgal drying and cell
disruption recent advances. Bioresour Technol 2015;184:25866.
[77] Carriquiry MA, Du XD, Timilsina GR. Second generation biofuels: economics
and policies. Energy Policy 2011;39:422234.
[78] Greenwell HC, Laurens LML, Shields RJ, Lovitt RW, Flynn KJ. Placing
microalgae on the biofuels priority list: a review of the technological
challenges. J R Soc Interface 2010;7:70326.
[79] Cadoret JP, Garnier M, Saint-Jean B. Microalgae, functional genomics and
biotechnology. Adv Bot Res 2012;64:285341.
[80] Anandarajah K, Mahendraperumal G, Sommerfeld M, Hu Q. Characterization
of microalga Nannochloropsis sp. mutants for improved production of
biofuels. Appl Energy 2012;96:3717.
[81] Iwai M, Ikeda K, Shimojima M, Ohta H. Enhancement of extraplastidic oil
synthesis in Chlamydomonas reinhardtii using a type-2 diacylglycerol
acyltransferase with a phosphorus starvation-inducible promoter. Plant
Biotechnol J 2014;12:80819.
[82] Vasudevan PT, Briggs M. Biodiesel production-current state of the art and
challenges. J Ind Microbiol Biotech 2008;35:42130.
[83] Chen G, Zhao L, Qi Y. Enhancing the productivity of microalgae cultivated in
wastewater toward biofuel production: a critical review. Appl Energy
2015;137:28291.
[84] Holbrook GP, Davidson Z, Tatara RA, Ziemer NL, Rosentrater KA, Scott
Grayburn W. Use of the microalga Monoraphidium sp. grown in wastewater
as a feedstock for biodiesel: cultivation and fuel characteristics. Appl Energy
2014;131:38693.
[85] Zhou WG, Li YC, Min M, Hu B, Chen P, Ruan R. Local bioprospecting for highlipid producing microalgal strains to be grown on concentrated municipal
wastewater for biofuel production. Bioresour Technol 2011;102:690919.
[86] Rawat I, Ranjith Kumar R, Mutanda T, Bux F. Biodiesel from microalgae: a
critical evaluation from laboratory to large scale production. Appl Energy
2013;103:44467.
[87] Abu GO, Ogbonda KH, Aminigo RE. Optimization studies of biomass
production and protein biosynthesis in a Spirulina sp isolated from an oilpolluted ame pit in the Niger Delta. Afr J Biotechnol 2007;6:25504.
[88] Chen H, Lao YM, Jiang JG. Effects of salinities on the gene expression of a
(NAD(+))-dependent glycerol-3-phosphate dehydrogenase in Dunaliella
salina. Sci Total Environ 2011;409:12917.
[89] Soletto D, Binaghi L, Ferrari L, Lodi A, Carvalho J, Zilli M, et al. Effects of carbon
dioxide feeding rate and light intensity on the fed-batch pulse-feeding
cultivation of Spirulina platensis in helical photobioreactor. Biochem Eng J
2008;39:36975.
[90] Li SW, Luo SJ, Guo RB. Efciency of CO2 xation by microalgae in a closed
raceway pond. Bioresour Technol 2013;136:26772.
[91] Iancu P, Plesu V, Velea S. Flue Gas CO2 Capture by Microalgae in
Photobioreactor: a Sustainable Technology. In: Varbanov PS, Lam HL,
Klemes JJ, Pierucci S, Klemes JJ, editors. Pres 2012: 15th international
conference on process integration, modelling and optimisation for energy
saving and pollution reduction. Milano: Aidic Servizi Srl; 2012. p. 799-804.
[92] Santiago DEO, Jin HF, Lee K. The inuence of ferrous-complexed EDTA as a
solubilization agent and its auto-regeneration on the removal of nitric oxide
gas through the culture of green alga Scenedesmus sp. Process Biochem
2010;45:194953.
[93] Jin H-F, Santiago DEO, Park J, Lee K. Enhancement of nitric oxide solubility
using Fe(II)EDTA and its removal by green algae Scenedesmus sp. Biotechnol
Bioprocess Eng 2008;13:4852.
[94] Gomez C, Escudero R, Morales MM, Figueroa FL, Fernandez-Sevilla JM, Acien
FG. Use of secondary-treated wastewater for the production of Muriellopsis
sp. Appl Microbiol Biotechnol 2013;97:223949.
[95] Sun X, Wang C, Li Z, Wang W, Tong Y, Wei J. Microalgal cultivation in
wastewater from the fermentation efuent in Riboavin (B2) manufacturing
for biodiesel production. Bioresour Technol 2013;143:499504.
[96] Cai T, Park SY, Li Y. Nutrient recovery from wastewater streams by
microalgae: status and prospects. Renew Sustain Energy Rev 2013;19:3609.
[97] Zhu L, Hiltunen E, Shu Q, Zhou W, Li Z, Wang Z. Biodiesel production from
algae cultivated in winter with articial wastewater through pH regulation
by acetic acid. Appl Energy 2014;128:10310.
[98] Prochazkova G, Branyikova I, Zachleder V, Branyik T. Effect of nutrient supply
status on biomass composition of eukaryotic green microalgae. J Appl Phycol
2014;26:135977.
[99] Rodol L, Zittelli GC, Bassi N, Padovani G, Biondi N, Bonini G, et al. Microalgae
for oil: strain selection, induction of lipid synthesis and outdoor mass
cultivation
in
a
low-cost
photobioreactor.
Biotechnol
Bioeng
2009;102:10012.
[100] Klok AJ, Lamers PP, Martens DE, Draaisma RB, Wijffels RH. Edible oils from
microalgae: insights in TAG accumulation. Trends Biotechnol 2014;32:5218.
[101] Wan C, Alam MA, Zhao XQ, Zhang XY, Guo SL, Ho SH, et al. Current progress
and future prospect of microalgal biomass harvest using various occulation
technologies. Bioresour Technol 2015;184:2517.
[102] Wang Y, Yang Y, Ma F, Xuan L, Xu Y, Huo H, et al. Optimization of Chlorella
vulgaris and bioocculant-producing bacteria co-culture: enhancing
microalgae harvesting and lipid content. Lett Appl Microbiol
2015;60:497503.
[103] Alam MA, Wan C, Guo SL, Zhao XQ, Huang ZY, Yang YL, et al. Characterization
of the occulating agent from the spontaneously occulating microalga
Chlorella vulgaris JSC-7. J Biosci Bioeng 2014;118:2933.
[104] Park JY, Lee K, Choi SA, Jeong MJ, Kim B, Lee JS, et al. Sonication-assisted
homogenization system for improved lipid extraction from Chlorella vulgaris.
Renewable Energy 2015;79:38.
[105] McMillan JR, Watson IA, Ali M, Jaafar W. Evaluation and comparison of algal
cell disruption methods: microwave, waterbath, blender, ultrasonic and laser
treatment. Appl Energy 2013;103:12834.
[106] Bohutskyi P, Kula T, Kessler B, Hong Y, Bouwer EJ, Betenbaugh MJ, et al. Mixed
trophic state production process for microalgal biomass with high lipid
content for generating biodiesel and biogas. Bioenergy Res 2014;7:117485.
[107] Sialve B, Bernet N, Bernard O. Anaerobic digestion of microalgae as a
necessary step to make microalgal biodiesel sustainable. Biotechnol Adv
2009;27:40916.
[108] Leite GB, Paranjape K, Abdelaziz AEM, Hallenbeck PC. Utilization of biodieselderived glycerol or xylose for increased growth and lipid production by
indigenous microalgae. Bioresour Technol 2015;184:12330.
[109] Collet P, Lardon L, Helias A, Bricout S, Lombaert-Valot I, Perrier B, et al.
Biodiesel from microalgae life cycle assessment and recommendations for
potential improvements. Renew Energy 2014;71:52533.
[110] Lee JH, Lee JS, Shin CS, Park SC, Kim SW. Effects of NO and SO2 on growth of
highly-CO2,-tolerant microalgae. J Microbiol Biotechnol 2000;10:33843.
[111] Van den Hende S, Vervaeren H, Boon N. Flue gas compounds and microalgae:
(Bio-)chemical interactions leading to biotechnological opportunities.
Biotechnol Adv 2012;30:140524.
[112] de-Bashan LE, Bashan Y. Immobilized microalgae for removing pollutants:
review of practical aspects. Bioresour Technol 2010;101:161127.

Tmp252e TMP

Enviado por

Dados do documento

Descrição original:

Título original

Direitos autorais

Formatos disponíveis

Compartilhar este documento

Compartilhar ou incorporar documento

Opções de compartilhamento

Você considera este documento útil?

Este conteúdo é inapropriado?

Direitos autorais:

Formatos disponíveis

Tmp252e TMP

Enviado por

Direitos autorais:

Formatos disponíveis

Applied Energy 155 (2015) 585598

Contents lists available at ScienceDirect

Microalgal biofuel revisited: An informatics-based analysis

Corresponding author. Tel.: +86 27 68780790.

H. Chen et al. / Applied Energy 155 (2015) 585598

production of algal fuel oils at a meaningful scale [10], and other

H. Chen et al. / Applied Energy 155 (2015) 585598

Fig. 1. Statistical analysis of trends in the number of published manuscripts related

The rst manuscript on microalgal biofuel appeared in 1970,

Fig. 2. Trends in the number of published patents pertaining to microalgal biofuel.

on the topic in all previous years combined, suggesting that

H. Chen et al. / Applied Energy 155 (2015) 585598

Supplemental material). The major advances in microalgal biofuel

To elucidate the major foci of research on microalgal biofuel, we

3.2. High capital and operating costs hamper commercial-scale algal

H. Chen et al. / Applied Energy 155 (2015) 585598

nontoxic occulants (e.g., organic polymers) have been intensively

For lipid production, extensive drying of microalgal biomass is

H. Chen et al. / Applied Energy 155 (2015) 585598

ultrasonic processes require further development and scale-up,

Fig. 4. Trends in published patents related to microalgal biotechnology for the

Europe with the exception of France, Spain, and Germany.

extraction is non-toxic and provides a non-oxidizing environment

We retrieved 4820 manuscripts and 859 patents in the area of

H. Chen et al. / Applied Energy 155 (2015) 585598

microalgal biotechnology for the production of high value-added

H. Chen et al. / Applied Energy 155 (2015) 585598

keywords associated with this topic (Table S4 in le of

Fig. 6. Trends in published patents pertaining to microalgal mass culturing. (A)

culturing may play an important role in reducing the cost of

Based on the analysis of the retrieved manuscripts and patents,

H. Chen et al. / Applied Energy 155 (2015) 585598

the production of high value-added products and environmental

H. Chen et al. / Applied Energy 155 (2015) 585598

Among the wild species available in formal collections, high

H. Chen et al. / Applied Energy 155 (2015) 585598

occulation is currently receiving increased attention because of

algal into additional renewable energy products (e.g., methane)

H. Chen et al. / Applied Energy 155 (2015) 585598

transgenic microalgae should be considered as well. For genetic

microalgal biofuel research, a burst in microalgal biofuel research

H. Chen et al. / Applied Energy 155 (2015) 585598

by thermal analysis coupled to mass spectrometry. Appl Energy

H. Chen et al. / Applied Energy 155 (2015) 585598

Você também pode gostar