OIL & GAS FEED FOR FISH?

BIOPROTEIN, A NEW HIGH QUALITY SINGLE CELL PROTEIN BASED ON NATURAL GAS

BY

Einar Moen, Norferm, Stavanger Norway.

SUMMARY

A production process has been developed for the commercial production of a high quality
single cell protein based on natural gas. The process is based on the use of natural
methanotropic bacteria, which is able to utilize methane as the energy and carbon source in a
fermentation process. A unique loop reactor has been developed which gives a very high mass
transport of the gaseous raw materials into the water phase. The final product BioProtein
contains approximately 70% protein of high quality in terms of its amino acid profile.
Through a very extensive documentation of the nutritional properties as a feedstuff,
BioProtein obtained an EU approval for use in feed for Atlantic salmon and domestic
animals in 1995. Feeding trials with salmon has shown that BioProtein can be added up to a
inclusion level of 20% of the total salmon feed replacing more than 50% of the fish meal,
resulting in improved or equal growth rate for the salmon compared with standard diets. The
presentation will also include some results from feeding trials with swine and broiler chicks.
Commercial production of BioProtein started in 2001.

1. INTRODUCTION.
The title of my presentation is OIL & GAS FEED FOR FISH?. It is to be a
presentation of a new protein raw material based on a novel biotechnological approach. The
topics I am going to deal with in my presentation are:
-

Background
Production of BioProtein
Product description
Nutritional properties
BioProtein in fish feed
Product development
Concluding remarks

Bacteria, which utilize methane for growth has been known for a long time. They are referred
to as methanotropic bacteria and are found in nature often associated with methane producing
bacteria (methanogenic). These microorganisms plays a very important part in the global turn
over of carbon and are normally found in soil, waste waters, lakes and marine sediments. The
best-known ecosystem for the methanotropic bacteria is the borderline between oxic and
anoxic layers in stagnant waters. Due to the fact that the methane utilizing bacteria is
recognized as being part of the food chain in espesially fresh water systems, these bacteria
have for decades had commercial interest, mainly because of its high content of proteins
which might be used for feed and food purposes.
Several attempts to start commercial production of single cell proteins based on paraffins,
methanol and natural gas have so far failed due to technical or economical reasons.
Norferms single cell protein process was originally developed by the Danish company Dansk
BioProtein AS, which is now a subsidiary of Norferm. The process is based on a new
approach where the bioreactor is designed for gaseous raw materials such as methane,

ammonia and oxygen. The methanotropic bacteria Methylococcus capsulatus is the driving
force in the process due to its ability to use methane as its carbon and energy source.
Statoil as the 100% owner of Norferm was attracted to the technology since Statoil operates
large gas fields on the Norwegian continental shelf and is constantly looking for opportunities
to establish land based industry based on natural gas.

The goal behind the commercial process development was to produce, preferentially in
Norway, a natural gas based biomass with a high protein content that could be applied in feed
as well as for human consumption.

2. PROCESS DEVELOPMENT
2.1 Production organism.

The Norferm process in based on the use of the natural methanotropic bacteria Methylococcus
capsulatus, as the organism able to convert natural gas into biomass in a very delicate process
involving many enzymes and metabolites. Nature has designed this bacterium with internal
membranes and associated enzyme systems that can bind and use methane, oxygen and
ammonia for its growth. These are actually the basic elements needed for creating life. It is
the bacteria itself which constitute the final product; i.e. the BioProtein. Consequently the
BioProtein product can be classified as a true single cell protein product. The main chemical
reaction catalysed by the bacteria is the conversion of methane to methanol with subsequent
production of biomass.
The specific bacteria used in the Norferm process was originally found in Scotland and was
selected due to its high content of protein as well as its ability to grow at elevated
temperatures.

2.2. Process design.

The major task in developing a commercial production process based on a gas-utilizing

bacterium is to design a reactor that results in a high productivity with an efficient use of the
gasses involved. The idea is to create a surrounding atmosphere for the bacteria where all the
growth requirements are present in optimal concentrations. For Norferm, the strategy was
from the start to develop a reactor, which operates on a continuous manner with a mixing

system allowing a very high mass transport. Mass transport is the diffusion of gases into the
water phase.
After a rather extensive development period the final reactor design is that of a water filled,
continuous loop reactor where the gas phase is mixed with the water phase using static mixers
distributed along the tubular reactor. The liquid is driven through the fermentor by an internal
pump. This ensures high mass transfer and subsequent high productivity. When the bacteria is
introduced into the water phase, it will start to grow and divide due to the fact that water
phase will supply it with all the nutrients it requires.
When the bacteria reach a steady state growth, the biomass is continuously harvested through
a side stream, at a rate, which reflects the growth rate of the bacteria. The liquid phase
separated from the biomass is returned to the fermentor thus reducing the effluent from the
process to a very limited volume. From there the biomass is concentrated, heat inactivated
and, as a final step, dried in a spray drier into a nicely granulated powder (and subsequently
pellets).

3. THE PRODUCT.
As mentioned before, the BioProtein product is the biomass from the gas based fermentation
process. The major component of the product is protein constituting close to 70% of the total
dry weight. However, like all types of biomass, BioProtein also contains other natural
components like lipids, carbohydrates, vitamins and minerals.
Compared with other commercial protein sources, the protein content of BioProtein is rather
high making the product of great interest in feed as well as in food. When looking at the
amino acid profile, BioProtein for most nutritional purposes contains all the essential amino
acids in adequate amounts. Of great interest, especially for some animal feed is the relatively
high content of tryptophan, a high cost amino acid, which can be used in special diets
espesially for swine.
The spray dried BioProtein will be supplied as a granulated product (350 kg/m3) or as a highdensity pellet (900 kg/m3) form thereof. Initial tests have proved that the basic BioProtein
product has a rather neutral taste and functions well, even at high inclusion levels, in pellets
for domestic animals as well as salmons.
Comparative studies have shown that the BioProtein product is very similar to the single cell
protein Pruteen which was a methanol based commercial feed protein from ICI up to 1985.

4. BIOPROTEIN IN FEED.
4.1

Strategy.

Once the process for making commercial quantities of BioProtein was developed, a rather
extensive program for qualifying the use of BioProtein in feed was initiated. This process had
two major goals:
a) To qualify the protein as a feedstuff for different animals including salmon and
b) To obtain an EU approval for the BioProtein as a protein ingredient in feed.
These qualifying activities were performed in parallel in a rather costly and time-consuming
process over a period of several years with activities mainly in Denmark and in Norway. The
toxicological tests required for approval were performed in Denmark while most of the
feeding trials on target animals were performed in Norway. It should be said that Norferm
quite early in this project believed that a major market for the BioProtein was the rapidly
growing fish farming industry where the demand for high quality protein was expected to
grow into large numbers.
4.2 BioProtein for Atlantic salmon.
It was natural for Norferm to perform all the necessary tests and trials on Atlantic salmon in
Norway. The local commercial feed industry was very enthusiastic about the project. Norferm
got very valuable support from this industry in technical as well as the more nutritional
aspects of our product testing. The major feeding experiments on Atlantic salmon were
performed at the Norwegian research institute Akvaforsk AS. The main purpose of the
experiment was to evaluate the nutritional value of BioProtein in the diet of on growing
salmons with respect to growth, feed conversion, digestibility and body composition. The test
was performed over a 12 month period involving approximately 1000 fish fed on four
different diets. As can be seen from the growth curve, the general trend is an enhanced growth
of the fish fed on the diets containing 10 and 20% BioProtein. It was also observed an
increased feed uptake for fish fed on the intermediary BioProtein containing diets especially
in the early part of the feeding period. The feed/gain results from the feeding trials gave a
positive tendency for the intermediate diets containing 10 and 20% BioProtein.
In terms of product quality there was a tendency towards lower fat content in the fish fed
BioProtein, while the carotenoid concentration was similar to the one found in the control
group fed on LT fishmeal.
In a second major test on Atlantic salmon with an average initial weight on 1,4 kg fed on to
the slaughter weight, the general observation on growth rate for the fish fed on BioProtein
containing diets, was better or equal to the control group.

The conclusion from the tests on Atlantic salmon is that BioProtein performs very well in
salmon feed. The recommended inclusion level can be up to 25% of the total dietary amino
acids. The BioProtein seems to stimulate feed intake in the early salt-water period and an
improved Feed/gain ratio is observed compared to standard LT fish meal diets.

4.3. BioProtein for domestic animals.

Encouraged by the positive results from the salmon feeding tests, it became evident that
Norferms strategy had to be extended also to qualify the BioProtein for different domestic
animals. In a collaboration program with the Department of Animal Science at the
Agricultural University of Norway, different test programs were performed mainly with pigs
and broiler chicks using mink as a reference animal for studying digestibility of the
BioProtein
The results from a test on growing finishing pigs with initial weight 25 kg. The results shows
an increased growth rate in terms of daily gain of pigs fed on BioProtein compared to the
control diet. This was also observed in tests performed on weanling pigs from 7 to 25 kg. A
clear tendency towards increased feed intake was observed for the pigs fed on the diets
containing BioProtein.
Feeding tests on Broiler chicks: Following a difficult design process for the diets for the
chicks, it was apparent that inclusion levels of up to 9 % BioProtein in the diet resulted in
equal growth rate as the control. However, the feed/gain ratio of the chicks was improved
relatively to the content of BioProtein in the diet. A very interesting observation was a
significant, concentration dependent, reduction of the content of abdominal fat found in the
chicks fed BioProtein.

4.4. Feed quality and EU approval of BioProtein.

In parallel with the feeding trials on salmon and domestic animals, a more basic study on the
digestibility of the BioProtein was performed. In general, a total digestibility of approximately
85% is observed for BioProtein with some variations between animals as well as among the
essential amino acids.
Based on the obtained nutritional as well as toxicological data for BioProtein, an application
for EU approval was filed through Danish authorities in 1993. Following a rather time
consuming and complicated dialogue with Brussels, approval for BioProtein was obtained in

1995. The approval includes the use in feed for salmon (one generation), swine between 25
and 60 kg and calves from 80 kg. The inclusion levels for the different feeds are indicated.
Norferm has filed an EU application in which we hope to get approval for cat and dogs,
broiler chicks and swine from 7 kg and up to slaughter early year 2003.

5. FUTURE ASPECTS.
The first commercial production plant has a yearly capacity of 10.000 tonnes of BioProtein.
The commercial production started year 2001. Due to the limited capacity of this first plant,
we see it mainly as a large-scale pilot plant where the major tasks are to prove the technology
as well as introducing the product to different markets. Initially most of the produced
quantities of BioProtein will be sold for feed purposes in Norway through an agreement with
EWOS. Based on the European market volumes for BioProtein in feed it is evident that we
have to plan for the future. Ideal production plants with capacity of 40.000 tonnes will have to
be built in order to produce commercial quantities of BioProtein for feed. We think that there
is a market for several hundred thousand tonnes of BioProtein in Europe and espesially the pet
market seems to be growing very fast.

It should also to be mentioned that we believe that BioProtein also has a future in food
products as well as a substrate for the fermentation industry.

6. CONCLUDING REMARKS.

Norferm has developed a commercial process for the production of BioProtein.

BioProtein represents a high quality protein approved within EU for use in feed.

BioProtein has acceptable digestibility and palatability for use in salmon feed at inclusion
levels up to 50% of total protein.

BioProtein is a very stable product with no seasonal variations.

Norferm has a collaboration agreement with EWOS for commercial use of BioProtein in
salmon feed in Norway.

BioProtein can be produced in unlimited quantities.