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1.
The major constraint in the enzymatic saccharification of biomass for ethanol production is the cost of cellulase enzymes.
Production cost of cellulases may be brought down by multifaceted approaches which includes the use of cheap lignocellulosic
substrates for fermentation production of the enzyme, and the use of cost efficient fermentation strategies like solid state
fermentation (SSF). The current study investigated the production of cellulase by Trichoderma reesei RUT C30 on wheat bran under SSF. Process parameters important in cellulase production were identified by a Plackett and Burman
design and the parameters with significant effects on enzyme production were optimized for maximal yield using a central composite
rotary design (CCD). Higher initial moisture content of the medium had a negative effect on production whereas incubation
temperature influenced cellulase production positively in the tested range. Optimization of the levels of incubation temperature
and initial moisture content of the medium resulted in a 6.2 fold increase in production from 0.605 to 3.8 U/gds of cellulase.
The optimal combination of moisture and temperature was found to be 37.56% and 30 °C, respectively, for maximal cellulase
production by the fungus on wheat bran. 相似文献
2.
Enzymatic saccharification and fermentation of xylose-optimized dilute acid-treated lignocellulosics
Yun-Chin Chung Alan Bakalinsky Michael H. Penner 《Applied biochemistry and biotechnology》2005,124(1-3):947-961
The cellulose reactivity of two lignocellulosic feedstocks, switchgrass and poplar, was evaluated under straight saccharification
(SS) and simultaneous saccharification and fermentation (SSF) conditions following dilute sulfuric acid pretreatments designed
for optimum xylose yields. The optimum pretreatment conditions, within the constraints of the experimental system (Parr batch
reactor), were 1.2% acid, 180°C, and 0.5 min for switchgrass and 1% acid, 180°C, and 0.56 min for poplar. The cellulase enzyme
preparation was from Trichoderma reesei and fermentations were done with Saccharomyces cerevisiae. Time courses for SS were monitored as the sum of glucose and cellobiose; those for SSF as the sum of glucose, cellobiose,
and ethanol. Percentage conversions under SS conditions were 79.1% and 91.4% for the pretreated poplar and switchgrass feedstocks,
respectively. Analogous values under SSF conditions were 73.0% and 90.3% for pretreated poplar and switchgrass, respectively. 相似文献
3.
Ethanol production from Jerusalem artichoke was studied using inulinase and Z.mobilis by simultaneous saccharification and fermentation (SSF) process. The SSF process showed higher ethanol yield and productivity
than the acid or enzymatic prehydrolyzed two-step process. The optimum temperature and inulinase concentration for SSF were
35°C and 0.25% (v/w, 4.4 units/g of sugar), respectively. In order to operate the SSF process in a continuous mode, inulinase
and Z.mobilis cells were coimmobilized in alginate beads, using chitin as a matrix for enzyme immobilization. The maximum ethanol productivity
of the continuous SSF process was 55.1 g/L/h, with 55% conversion yield. At the conversion yield of 90%, the productivity
was 32.7 g/L/h. The continuous SSF system could be operated stably over 2 wk with an ethanol concentration of 48.6 g/L (95%
of theoretical yield). 相似文献
4.
Paper mill sludge is a solid waste material composed of pulp residues and ash generated from pulping and paper making processes. The carbohydrate portion of the sludge has chemical and physical characteristics similar to pulp. Because of its high carbohydrate content and well-dispersed structure, the sludges can be biologically converted to value-added products without pretreatment. In this study, two different types of paper mill sludges, primary sludge and recycle sludge, were evaluated as a feedstock for bioconversion to ethanol. The sludges were first subjected to enzymatic conversion to sugars by commercial cellulase enzymes. The enzymatic conversion was inefficient because of interference by ash in the sludges with the enzymatic reaction. The main cause was that the pH level is dictated by CaCO3 in ash, which is two units higher than the pH optimum of cellulase. To alleviate this problem, simultaneous saccharification and cofermentation (SSCF) using cellulase (Spezyme CP) and recombinant Escherichia coli (ATCC-55124), and simultaneous saccharification and fermentation (SSF) using cellulase and Saccharomyces cerevisiae (ATCC-200062) were applied to the sludges without any pretreatment. Ethanol yields of 75–81% of the theoretical maximum were obtained from the SSCF on the basis of total carbohydrates. The yield from the SSF was also found to be in the range of 74–80% on the basis of glucan. The SSCF and SSF proceeded under stable condition with the pH staying near 5.0, close to the optimum for cellulase. Decrease of pH occurred due to carbonic acid and other organic acids formed during fermentation. The ash was partially neutralized by the acids produced from the SSCF and SSF and acted as a buffer to stabilize the pH during fermentation. When the SSF and SSCF were operated in fed-batch mode, the ethanol concentration in the broth increased from 25.5 and 32.6 g/L (single feed) to 45 and 42 g/L, respectively. The ethanol concentration was limited by the tolerance of the microorganism in the case of SSCF. The ethanol yield in fed-batch operation decreased to 68% for SSCF and 70% for SSF. The high-solids condition in the bioreactor appears to create adverse effects on the cellulase reaction. 相似文献
5.
Kwang Il Kim Woo Kyung Kim Deok Ki Seo In Sang Yoo Eun Ki Kim Hyon Hee Yoon 《Applied biochemistry and biotechnology》2003,107(1-3):637-647
Conversion of food wastes into lactic acid by simultaneous saccharification and fermentation (SSF) was investigated. The process
involves saccharification of the starch component in food wastes by a commercial amylolytic enzyme preparation (a mixture
of amyloglucosidase, α-amylase, and protease) and fermentation by Lactobacillus delbrueckii. The highest observed overall yield of lactic acid in the SSF was 91% of theoretical. Lactic acid concentration as high as
80 g/L was attainable in 48 h of the SSF. The optimum operating conditions for the maximum productivity were found to be 42°C
and pH 6.0. Without supplementation of nitrogen-containing nutrients, the lactic acid yield in the SSF decreased to 60%: 27
g/L of lactic acid from 60 g/L of food waste. The overall performance of the SSF, however, was not significantly affected
by the elimination of mineral supplements. 相似文献
6.
Ethanol production was studied in simultaneous saccharification and fermentation (SSF) of steam-pretreated spruce at 42°C,
using a thermotolerant yeast. Three yeast strains of Kluyveromyces marxianus were compared in test fermentations. SSF experiments were performed with the best of these on 5% (w/w) of substrate at a
cellulase loading of 37 filter paper units/g of cellulose, and a β-glucosidase loading of 38 IU/gof cellulose. The detoxification
of the substrate and the lack of pH control in the experiments increased the final ethanol concentration. The final ethanol
yield was 15% lower compared to SSF with Saccharomyces cerevisiae at 37°C, owing to the cessation of ethanol fermentation after the first 10 h. 相似文献
7.
Isci A Himmelsbach JN Strohl J Pometto AL Raman DR Anex RP 《Applied biochemistry and biotechnology》2009,157(3):453-462
Aqueous-ammonia-steeped switchgrass was subject to simultaneous saccharification and fermentation (SSF) in two pilot-scale
bioreactors (50- and 350-L working volume). Switchgrass was pretreated by soaking in ammonium hydroxide (30%) with solid to
liquid ratio of 5 L ammonium hydroxide per kilogram dry switchgrass for 5 days in 75-L steeping vessels without agitation
at ambient temperatures (15 to 33 °C). SSF of the pretreated biomass was carried out using Saccharomyces cerevisiae (D5A) at approximately 2% glucan and 77 filter paper units per gram cellulose enzyme loading (Spezyme CP). The 50-L fermentation
was carried out aseptically, whereas the 350-L fermentation was semiaseptic. The percentage of maximum theoretical ethanol
yields achieved was 73% in the 50-L reactor and 52–74% in the 350-L reactor due to the difference in asepsis. The 350-L fermentation
was contaminated by acid-producing bacteria (lactic and acetic acid concentrations approaching 10 g/L), and this resulted
in lower ethanol production. Despite this problem, the pilot-scale SSF of aqueous-ammonia-pretreated switchgrass has shown
promising results similar to laboratory-scale experiments. This work demonstrates challenges in pilot-scale fermentations
with material handling, aseptic conditions, and bacterial contamination for cellulosic fermentations to biofuels. 相似文献
8.
van Walsum G. Peter Allen Stephen G. Spencer Mark J. Laser Mark S. Antal Michael J. Lynd Lee R. 《Applied biochemistry and biotechnology》1996,(1):157-170
Lignocellulosic materials pretreated using liquid hot water (LHW) (220°C, 5 MPa, 120 s) were fermented to ethanol by batch
simultaneous saccharification and fermentation (SSF) usingSaccharomyces cerevisiae in the presence ofTrichoderma reesei cellulase. SSF of sugarcane bagasse (as received), aspen chips (smallest dimension 3 mm), and mixed hardwood flour (−60 +70
mesh) resulted in 90% conversion to ethanol in 2–5 d at enzyme loadings of 15–30 FPU/g. In most cases, 90% of the final conversion
was achieved within 75 h of inoculation. Comminution of the pretreated substrates did not affect the conversion to ethanol.
The hydrolysate produced from the LHW pretreatment showed slight inhibition of batch growth ofS. cerevisiae. Solids pretreated at a concentration of 100 g/L were as reactive as those pretreated at a lower concentration, provided
that the temperature was maintained at 220°C. 相似文献
9.
Pretreatment of corn stover by soaking in aqueous ammonia 总被引:1,自引:0,他引:1
Soaking in aqueous ammonia (SAA) was investigated as a pretreatment method for corn stover. In this method, the feedstock
was soaked in aqueous ammonia over an extended period (10–60 d) at room temperature. It was done without agitation at atmospheric
pressure. SAA treatment removed 55–74% of the lignin, but retained nearly 100% of the glucan and 85% of the xylan. The xylan
remaining in the corn stover after SAA treatment was hydrolyzed along with the glucan by xylanase present in the Spezyme CP
enzyme. In the simultaneous saccharification and fermentation (SSF) test of SAA-treated corn stover, using S. cerevisiae (D5A), an ethanol yield of 73% of theoretical maximum was obtained on the basis of the glucan content in the treated corn stover.
The accumulation of xylose in the SSF appears to inhibit the cellulase activity on glucan hydrolysis, which limits the yield
of ethanol. In the simultaneous saccharification and co-fermentation (SSCF) test, using recombinant E. coli (KO11), both the glucan and xylose were effectively utilized, resulting in on overall ethanol yield of 77% based on the glucan
and xylan content of the substrate. When the SSCF process is used, the fact that the xylan fraction is retained during pretreatment
is a desirable feature since the overall bioconversion can be carried out in a single step without separate recovery of xylose
from the pretreatment liquid. 相似文献
10.
Cao Ningjun Xia Youkun Gong Cheng S. Tsao George T. 《Applied biochemistry and biotechnology》1997,(1):129-139
A simple and effective method of treatment of lignocellulosic material was used for the preparation of corn cob for the production
of 2,3-butanediol byKlebsiella oxytoca ATCC 8724 in a simultaneous saccharification and fermentation process. During the treatment, lignin, and alkaline extractives
were solubilized and separated from cellulose and hemicellulose fractions by dilute ammonia (10%) steeping. Hemicellulose
was then hydrolyzed by dilute hydrochloric acid (1%, wJv) hydrolysis at 100°C at atmospheric pressure and separated from cellulose
fraction. The remaining solid, with 90% of cellulose, was then used as the substrate. A butanediol concentration of 25 g/L
and an ethanol concentration of 7 g/L were produced byK. oxytoca from 80 g/L of corn cob cellulose with a cellulase dosage of 8.5 IFPU/g corn cob cellulose after 72 h of SSF. With only dilute
acid hydrolysis, a butanediol production rate of 0.21 g/L/h was obtained that is much lower than the case in which corn cob
was treated with ammonia steeping prior to acid hydrolysis. The butanediol production rate for the latter was 0.36 g/L/h. 相似文献
11.
Statistical Optimization of Recycled-Paper Enzymatic Hydrolysis for Simultaneous Saccharification and Fermentation Via Central Composite Design 总被引:1,自引:0,他引:1
Qing Liu Ke-ke Cheng Jian-an Zhang Jin-ping Li Ge-hua Wang 《Applied biochemistry and biotechnology》2010,160(2):604-612
A central composite design of the response surface methodology (RSM) was employed to study the effects of temperature, enzyme
concentration, and stirring rate on recycled-paper enzymatic hydrolysis. Among the three variables, temperature and enzyme
concentration significantly affected the conversion efficiency of substrate, whereas stirring rate was not effective. A quadratic
polynomial equation was obtained for enzymatic hydrolysis by multiple regression analysis using RSM. The results of validation
experiments were coincident with the predicted model. The optimum conditions for enzymatic hydrolysis were temperature, enzyme
concentration, and stirring rate of 43.1 °C, 20 FPU g−1 substrate, and 145 rpm, respectively. In the subsequent simultaneous saccharification and fermentation (SSF) experiment under
the optimum conditions, the highest 28.7 g ethanol l−1 was reached in the fed-batch SSF when 5% (w/v) substrate concentration was used initially, and another 5% added after 12 h fermentation. This ethanol output corresponded
to 77.7% of the theoretical yield based on the glucose content in the raw material. 相似文献
12.
Bradley A. Saville Chunbei Huang Vince Yacyshyn Andrew Desbarats 《Applied biochemistry and biotechnology》2006,129(1-3):180-194
Studies were conducted on maltodextrin saccharification and on simultaneous saccharification and fermentation (SSF) with various
commercial glucoamylases. In kinetics studies, none of the glucoamylases were able to completely convert maltodextrin into
glucose. Typically, about 85% conversion was obtained, and glucose yields were about 75%. Typically, the kinetics were biphasic,
with 1 h of rapid conversion, then a significant reduction in rate. Data were consistent with strong product inhibition and/or
enzyme inactivation. Some glucoamylases followed first-order kinetics, initially slower at dextrin conversion, but eventually
achieving comparable conversion and glucose concentrations. Most of the glucoamylases were more active at 55°C than at 35°C,
but pH had little effect on activity. Screening studies in an SSF system demonstrated little difference between the glucoamylases,
with a few exceptions. Subsequent targeted studies showed clear differences in performance, depending on the fermentation
temperature and yeast used, suggesting that these are key parameters that would guide the selection of a glucoamylase. 相似文献
13.
Mark S. Ou Nazimuddin Mohammed L. O. Ingram K. T. Shanmugam 《Applied biochemistry and biotechnology》2009,155(1-3):76-82
Ethanol production from lignocellulosic biomass depends on simultaneous saccharification of cellulose to glucose by fungal cellulases and fermentation of glucose to ethanol by microbial biocatalysts (SSF). The cost of cellulase enzymes represents a significant challenge for the commercial conversion of lignocellulosic biomass into renewable chemicals such as ethanol and monomers for plastics. The cellulase concentration for optimum SSF of crystalline cellulose with fungal enzymes and a moderate thermophile, Bacillus coagulans, was determined to be about 7.5 FPU g?1 cellulose. This is about three times lower than the amount of cellulase required for SSF with Saccharomyces cerevisiae, Zymomonas mobilis, or Lactococcus lactis subsp. lactis whose growth and fermentation temperature optimum is significantly lower than that of the fungal cellulase activity. In addition, B. coagulans also converted about 80% of the theoretical yield of products from 40 g/L of crystalline cellulose in about 48 h of SSF with 10 FPU g?1 cellulose while yeast, during the same period, only produced about 50% of the highest yield produced at end of 7 days of SSF. These results show that a match in the temperature optima for cellulase activity and fermentation is essential for decreasing the cost of cellulase in cellulosic ethanol production. 相似文献
14.
Ballesteros Ignacio Oliva José Miguel Negro Maria José Manzanares Paloma Ballesteros Mercedes 《Applied biochemistry and biotechnology》2002,98(1-9):717-732
The olive pulp fraction contained in the residue generated in olive oil extraction by a two-step centrifugation process can
be upgraded by using the cellulose fraction to produce ethanol and recovering high value phenols (tyrosol and hydroxytyrosol).
Olive pulp was pretreated in a laboratory scale stirred autoclave at different temperatures (150–250°C). Pretreatment was
evaluated regarding cellulose recovery, enzymatic hydrolysis effectiveness ethanol production by a simultaneous saccharification
and fermentation process (SSF), and phenols recovery in the filtrate. The pretreatment of olive pulp using water at temperatures
between 200°C and 250°C enhanced enzymatic hydrolysis. Maximum ethanol production (11.9 g/L) was obtained after pretreating
pulp at 210°C in a SSF fed-batch procedure. Maximum hydroxytyrosol recovery was obtained in the liquid fraction when pretreated
at 230°C. 相似文献
15.
Continuous Production of Ethanol from Starch Using Glucoamylase and Yeast Co-Immobilized in Pectin Gel 总被引:1,自引:0,他引:1
This work presents a continuous simultaneous saccharification and fermentation (SSF) process to produce ethanol from starch
using glucoamylase and Saccharomyces cerevisiae co-immobilized in pectin gel. The enzyme was immobilized on macroporous silica, after silanization and activation of the
support with glutaraldehyde. The silica–enzyme derivative was co-immobilized with yeast in pectin gel. This biocatalyst was
used to produce ethanol from liquefied manioc root flour syrup, in three fixed bed reactors. The initial reactor yeast load
was 0.05 g wet yeast/ml of reactor (0.1 g wet yeast/g gel), used in all SSF experiments. The enzyme concentration in the reactor
was defined by running SSF batch assays, using different amount of silica–enzyme derivative, co-immobilized with yeast in
pectin gel. The chosen reactor enzyme concentration, 3.77 U/ml, allowed fermentation to be the rate-limiting step in the batch
experiment. In this condition, using initial substrate concentration of 166.0 g/l of total reducing sugars (TRS), 1 ml gel/1 ml
of medium, ethanol productivity of 8.3 g/l/h was achieved, for total conversion of starch to ethanol and 91% of the theoretical
yield. In the continuous runs, feeding 163.0 g/l of TRS and using the same enzyme and yeast concentrations used in the batch
run, ethanol productivity was 5.9 g ethanol/l/h, with 97% of substrate conversion and 81% of the ethanol theoretical yield.
Diffusion effects in the extra-biocatalyst film seemed to be reduced when operating at superficial velocities above 3.7 × 10−4 cm/s. 相似文献
16.
Danielle da Silveira dos Santos Anna Carolina Camelo Kelly Cristina Pedro Rodrigues Luís Cláudio Carlos Nei Pereira Jr. 《Applied biochemistry and biotechnology》2010,161(1-8):93-105
Considerable efforts have been made to utilize agricultural and forest residues as biomass feedstock for the production of second-generation bioethanol as an alternative fuel. Fermentation utilizing strains of Zymomonas mobilis and the use of simultaneous saccharification and fermentation (SSF) process has been proposed. Statistical experimental design was used to optimize the conditions of SSF, evaluating solid content, enzymatic load, and cell concentration. The optimum conditions were found to be solid content (30%), enzymatic load (25 filter paper units/g), and cell concentration (4 g/L), resulting in a maximum ethanol concentration of 60 g/L and a volumetric productivity of 1.5 g L?1?h?1. 相似文献
17.
Experimental results are presented for continuous conversion of pretreated hardwood flour to ethanol. A simultaneous saccharification
and fermentation (SSF) system comprised ofTrichoderma reesei cellulase supplemented with additional β-glucosidase and fermentation bySaccharomyces cerevisiae was used for most experiments, with data also presented for a direct microbial conversion (DMC) system comprised ofClostridium thermocellum. Using a batch SSF system, dilute acid pretreatment of mixed hardwood at short residence time(10 s, 220°C, 1% H2SO4) was compared to poplar wood pretreated at longer residence time (20 min, 160°C, 0.45% H2SO4). The short residence time pretreatment resulted in a somewhat (10–20%) more reactive substrate, with the reactivity difference
particularly notable at low enzyme loadings and/or low agitation. Based on a preliminary screening, inhibition of SSF by byproducts
of short residence time pretreatment was measurable, but minor. Both SSF and DMC were carried out successfully in well-mixed
continuous systems, with steady-state data obtained at residence times of 0.58–3 d for SSF as well as 0.5 and 0.75 d for DMC.
The SSF system achieved substrate conversions varying from 31% at a 0.58-d residence time to 86% at a 2-d residence time.
At comparable substrate concentrations (4–5 g/l) and residence times (0.5–0.58 d), substrate conversion in the DMC system
(77%) was significantly higher than that in the SSF system (31%). Our results suggest that the substrate conversion in SSF
carried out in CSTR is relatively insensitive to enzyme loading in the range 7–25 U/g cellulose and to substrate concentration
in the range of 5–60 g/L cellulose in the feed. 相似文献
18.
Borden Jacob R. Lee Youn Y. Yoon Hyon-Hee 《Applied biochemistry and biotechnology》2000,84(1-9):963-970
Astrain of Clostridium thermoaceticum (ATCC 49707) was evaluated for its homoacetate potential. This thermophilic anaerobe best produces acetate from glucose at
pH 6.0 and 59°C with a yield of 83% of theoretical. Enzyme hydrolysis of two substrates, a-cellulose and a pulp mill sludge,
yielded 68% and 70% digestion, respectively. The optimum conditions for the simultaneous saccharification and fermentation
(SSF) were substrate dependent: 55°C, pH 6.0 for α-cellulose, and 55°C, pH 5.5 for the pulp mill sludge. In the SSF with α-cellulose,
the overall yield of acetate was strongly influenced by the enzyme loading. In a fed-batch operation of SSF with α-cellulose,
an overall acetic acid yield of 60 wt% was obtained. Among the factors limiting the yields were incomplete digestion by the
enzyme and the end-product inhibition. In the SSF of pulp mill sludge, inhibitors present in the sludge severely limited bacterial
action. A large accumulation of glucose developed over the entire process, changing the intended SSF operation into a separate
hydrolysis and fermentation operation. Despite a long lag phase of microbial growth, a terminal yield of 85% was obtained
with this substrate. 相似文献
19.
A pretreatment method using aqueous ammonia was investigated with the intent of minimizing the liquid throughput. This process
uses a flow-through packed column reactor (or percolation reactor). In comparison to the ammonia recycle percolation (ARP)
process developed previously in our laboratory, this process significantly reduces the liquid throughput to one reactor void
volume in packed bed (2.0–4.7 mL of liquid/g of corn stover) and, thus, is termed low-liquid ARP (LLARP). In addition to attaining
short residence time and reduced energy input, this process achieves 59–70% of lignin removal and 48–57% of xylan retention.
With optimum operation of the LLARP to corn stover, enzymatic digestibilities of 95, 90 and 86% were achieved with 60, 15,
and 7.5 filter paper units/g of glucan, respectively. In the simultaneous saccharification and fermentation test of the LLARP
samples using Saccharomyces cerevisiae (NREL-D5A), an ethanol yield of 84% of the theoretical maximum was achieved with 6% (w/v) glucan loading. In the simultaneous saccharification
and cofermentation (SSCF) test using recombinant Escherichia coli (KO11), both the glucan and xylan in the solid were effectively utilized, giving an overall ethanol yield of 109% of the
theoretical maximum based on glucan, a clear indication that the xylan content was converted into ethanol. The xylooligomers
existing in the LLARP effluent were not effectively hydrolyzed by cellulase enzyme, achieving only 60% of digestibility. SSCF
of the treated corn stover was severely hampered when the substrate was supplemented with the LLARP effluent, giving only
56% the overall yield of ethanol. The effluent appears to significantly inhibit cellulase and microbial activities. 相似文献
20.
Ballesteros I. Ballesteros M. CabaÑas A. Carrasco J. MartÍn C. Negro M. J. Saez F. Saez R. 《Applied biochemistry and biotechnology》1991,(1):307-315
A total of 27 yeast strains belonging to the groupsCandida, Saccharomyces, andKluyveromyces were screened for their ability to grow and ferment glucose at temperatures ranging 32-45°C.
K. marxianus andK. fragilis were found to be the best ethanol producing organisms at the higher temperature tested and, so, were selected for subsequent
simultaneous saccharification and fermentation (SSF) studies. 相似文献