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Our work represents the only study of the pilot-scale production of pectinases in
which samples have been removed from several different locations within the bed in
order to evaluate the spatial uniformity of activity levels. Activity levels varied
significantly when compaction occurred (from 11 to 28103 U kg-1 in the third pilot
fermentation and from 15 to 26 in the fourth pilot fermentation). In the second pilot
fermentation, for which compaction of the bed did not occur, activity levels were
more uniform throughout the bed (17 to 20103 U kg-1). Interestingly, the moisture
contents measured in the bed for this fermentation confirmed the prediction of the
two-phase model of packed-bed operation developed by von Meien and Mitchell
[19] that the inner regions of the bed would dry more quickly than the upper regions
and the regions very near to the base.
Pekerjaan kami merupakan satu-satunya studi tentang produksi skala pilot
pectinases di mana sampel telah dihapus dari beberapa lokasi yang berbeda dalam
tbed untuk mengevaluasi keseragaman spasial tingkat aktivitas. Tingkat aktivitas
bervariasi secara signifikan ketika pemadatan terjadi (11-28 103 U kg-1 di
fermentasi percontohan ketiga dan 15-26 dalam fermentasi percontohan keempat).
Dalam pilot fermentasi kedua, yang pemadatan bed tidak terjadi, tingkat aktivitas
yang lebih seragam di seluruh bed (17-20 103 U kg-1). Menariknya, isi
kelembaban diukur dalam bed untuk fermentasi ini menegaskan prediksi dari model
dua-tahap operasi packed bed yang dikembangkan oleh von Meien dan Mitchell [19]
bahwa daerah bagian dari bed akan kering lebih cepat dari pada daerah atas dan
daerah sangat dekat dengan pangkalan.
On the basis of our results, we propose a strategy for scaling up of pectinase
production by SSF to commercial scale, using a mixture of 90% wheat bran and 10%
sugarcane bagasse: a bed height of 40 cm should be maintained, with the scale of
production being increased by increasing the diameter of the bioreactor and
maintaining the superficial air velocity at 0.1 m s-1. As with our fourth pilot-scale
fermentation, the temperature of the inlet air should be controlled, with cooler air
being supplied when the bed temperature rises above 35C. The process would be
similar to koji production in the soy sauce industry, where the bioreactors have
diameters of up to 12 m and bed heights of around 20 cm [20]. With this strategy, if
the fermentation is harvested at 10 h, then the results of the fourth pilot-scale
fermentation suggest that it is possible to avoid the problems with bed compaction
that occur later in the fermentation and that a pectinase productivity of 1840 U kg-1
h-1 can be obtained.
Atas dasar hasil kami, kami mengusulkan strategi untuk scaling up produksi
pektinase oleh SSF untuk skala komersial, menggunakan campuran 90% dedak
gandum dan 10% ampas tebu: ketinggian bed 40 cm harus dipertahankan, dengan
skala produksi yang meningkat dengan meningkatkan diameter bioreaktor dan
mempertahankan kecepatan udara superfisial pada 0,1 m s-1. Seperti fermentasi
skala pilot keempat kami, suhu udara masuk harus dikontrol, dengan udara dingin
yang disediakan ketika suhu naik di atas bed 35 C. Proses ini akan sama dengan
produksi koji dalam industri kecap, di mana bioreaktor memiliki diameter hingga 12
m dan ketinggian bed sekitar 20 cm [20]. Dengan strategi ini, jika fermentasi
dipanen pada 10 h, maka hasil fermentasi skala pilot keempat menunjukkan bahwa
adalah mungkin untuk menghindari masalah dengan pemadatan bed yang terjadi
kemudian di fermentasi dan produktivitas pektinase dari 1840 U kg -1 h-1 dapat
diperoleh.
Previous pilot-scale studies of the production of pectinases by SSF have not
provided the basis for clear scale-up strategies (Table 2). For example, although
Huerta et al. [11] claimed that the use of sugarcane bagasse avoided the problems
of heat transfer that are usually encountered in packed-bed bioreactors, they did
not provide any experimental temperature measurements to support this claim.
Additionally, they did not specify the bed height that was used. In the case of the
work of He and Chen et al. [12], the question of scale up was not addressed.
However, since their bioreactor is a tray-type system, albeit with pressure
pulsations inside the tray chamber, the implicit scale-up rule would be to build a
larger reactor with more trays. However, their bioreactor has a low volumetric
loading capacity, with 15 kg of dry solids per 800 L of bioreactor volume. In our
case, at 40 cm bed height, our bioreactor uses 27 kg of dry wheat bran (and 3 kg of
dry sugarcane bagasse) for a total volume of 400 L, namely a 200 L working volume
plus 200 L headspace.
Studi skala pilot sebelumnya produksi pectinases oleh SSF belum memberikan dasar
bagi strategi skala-up yang jelas (Tabel 2). Sebagai contoh, meskipun Huerta dkk.
[11] menyatakan bahwa penggunaan ampas tebu menghindari masalah
perpindahan panas yang biasanya dihadapi dalam packed-bed bioreaktor, mereka
tidak memberikan pengukuran suhu eksperimental untuk mendukung klaim ini.
Selain itu, mereka tidak menentukan tinggi yang digunakan. Dalam kasus
pekerjaan Dia dan Chen et al. [12], pertanyaan skala up tidak dibahas. Namun,
karena bioreaktor mereka adalah sistemjenis tray, meskipun dengan tekanan
denyutan di dalam ruang nampan, aturan skala-up implisit akan membangun
reaktor yang lebih besar dengan lebih nampan. Namun, bioreaktor mereka memiliki
kapasitas memuat volumetrik rendah, dengan 15 kg padatan kering per 800 L
volume bioreaktor. Dalam kasus kami, pada 40 cm tinggi bed, bioreaktor kami
menggunakan 27 kg dedak gandum kering (dan 3 kg ampas tebu kering) untuk total
volume 400 L, yaitu 200 L volume kerja ditambah 200 L headspace.
Only Rodrguez-Fernndez et al. [21] have suggested a strategy for the scale up of
pectinase production by SSF. On the basis of experiments undertaken with 0.8 kg of
dry substrate at four different aeration rates in a horizontal cylinder, they suggested
that it would be sufficient to maintain the air flow (L min-1) per kg of solid constant
during scale up. It should be noted that no mention was made of the bed being
agitated during the fermentation, so presumably the bed was static, and therefore it
is questionable as to whether their scale-up strategy would work. Due to the circular
cross section of the bioreactor, the bed height, and therefore the resistance to air
flow, varies across the bed. Consequently, the air would tend to flow preferentially
through the regions of lesser height. This would lead to serious overheating
problems in the thicker regions of the bed if such a bioreactor were operated with
several tonnes of substrate.
Although workable scale-up strategies have not been proposed previously for the
production of pectinases, various modeling and optimization studies have been
done with the intention of guiding the scale up of SSF processes in packed-bed
bioreactors [19,22,23]. However, although these models give insights into the
performance of packed-bed bioreactors, they are of limited usefulness for guiding
the scale up of processes involving specific microorganism/substrate combinations.
This is due to a lack of knowledge with respect to heat and mass transfer in packedbed bioreactors and growth kinetics of microorganisms in SSF, as discussed in the
following paragraphs.
Meskipun strategi skala-up yang bisa diterapkan belum diusulkan sebelumnya untuk
produksi pectinases, berbagai model dan optimasi penelitian telah dilakukan
dengan maksud membimbing skala up proses SSF di packed-bed bioreaktor
[19,22,23]. Namun, meskipun model ini memberikan wawasan ke dalam kinerja
packed=bed bioreaktor, kegunaan terbatas untuk membimbing skala up dari
proses yang melibatkan kombinasi mikroorganisme / substrat tertentu. Hal ini
disebabkan kurangnya pengetahuan terhadap perpindahan panas dan perpindahan
massa di packed-bed bioreaktor dan kinetika pertumbuhan mikroorganisme di SSF,
seperti yang dibahas dalam paragraf berikut.
Firstly, with respect to heat and mass transfer in packed beds, although the general
principles are well established, in SSF processes the properties of the bed of solids
in the bioreactor change significantly during growth of the microorganism. For
example, the growth of aerial hyphae of fungi into the interparticle spaces and the
reduction of particle size affect the bed porosity in a complex manner, while the
presence of biomass in the solid affects the isotherm of the solids [24,25]. Some
efforts have been made to characterize these phenomena, but they are quite
specific for organism and substrate. For example, the void fraction did not change
significantly during the growth of Myceliophthora thermophila on a 3:7 mixture of
wheat bran and sugarcane bagasse, but it increased from 0.52 to 0.76 during the
growth of A. niger on wheat bran [26,27].
Pertama, sehubungan dengan panas dan perpindahan massa di bed penuh,
meskipun prinsip-prinsip umum yang terpancang, di SSF memproses properti dari
bed padatan dalam perubahan bioreaktor secara signifikan selama pertumbuhan
mikroorganisme. Misalnya, pertumbuhan hifa aerial jamur ke dalam ruang
interparticle dan pengurangan ukuran partikel mempengaruh porositas bed secara
kompleks, sementara kehadiran biomassa dalam padat mempengaruhi isoterm dari
padatan [24,25]. Beberapa upaya telah dilakukan untuk mengkarakterisasi
fenomena ini, tetapi mereka cukup spesifik untuk organisme dan substrat. Misalnya,
fraksi kekosongan tidak berubah secara signifikan selama pertumbuhan
Myceliophthora thermophila pada 3: 7 campuran dedak gandum dan ampas tebu,
Secondly, with respect to the growth kinetics of microorganisms in SSF, the key
variables that affect growth change in a complex manner during the process. There
can be significant variations in the temperature and the water activity of the solid
[28]. Additionally, there are complex patterns of induction of enzymes for
hydrolyzing the range of polymeric substrates present within the solid particle and
the concentrations of soluble hydrolysis products and O2 vary significantly [29,30].
It would be too complicated to try to describe all these phenomena mechanistically
within bioreactor models and, as a result, these models use simple empirical
equations to describe the growth kinetics [31]. These empirical equations can be
fitted to specific results, but often fail to make accurate predictions for new
situations.
Kedua, sehubungan dengan kinetika pertumbuhan mikroorganisme di SSF, variabel
kunci yang mempengaruhi perubahan pertumbuhan secara kompleks selama proses
tersebut. Ada variasi yang signifikan dalam suhu dan aktivitas air padat [28]. Selain
itu, ada pola yang kompleks induksi enzim untuk hidrolisis berbagai substrat
polimer hadir dalam partikel padat dan konsentrasi produk hidrolisis larut dan O2
bervariasi secara signifikan [29,30]. Ini akan menjadi terlalu rumit untuk mencoba
menjelaskan semua fenomena ini secara mekanis dalam model bioreaktor dan,
sebagai hasilnya, model ini menggunakan persamaan empiris sederhana untuk
menggambarkan kinetika pertumbuhan [31]. Persamaan-persamaan empiris dapat
dipasang untuk hasil yang spesifik, tetapi sering gagal untuk membuat prediksi
yang akurat untuk situasi baru.