• KSBB Journal
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• v.27 no.2
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• pp.86-90
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• 2012

Ethanol fermentations were carried out using simultaneous saccharification and fermentation (SSF) and separated hydrolysis and fermentation (SHF) processes with monosaccharides from seaweed, Saccharina japonica (sea tangle, Dasima) as the biomass. The pretreatment was carried out by thermal acid hydrolysis with $H_2SO_4$ or HCl. Optimal pretreatment condition was determined at 10% (w/v) seaweed slurry with 37.5 mM $H_2SO_4$ at $121^{\circ}C$ for 60 min. To increase the yield of saccharfication, isolated marine bacteria Bacillus sp. JS-1 was used and 48 g/L of reducing sugar were produced. Ethanol fermentation was performed using SSF and SHF process with Pachysolen tannophilus KCTC 7937. The ethanol concentration was 6.5 g/L by SSF and 6.0 g/L by SHF.

• Journal of Microbiology and Biotechnology
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• v.27 no.9
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• pp.1649-1656
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• 2017

In simultaneous saccharification and fermentation (SSF) for production of cellulosic biofuels, engineered Saccharomyces cerevisiae capable of fermenting cellobiose has provided several benefits, such as lower enzyme costs and faster fermentation rate compared with wild-type S. cerevisiae fermenting glucose. In this study, the effects of an alternative intracellular cellobiose utilization pathway-a phosphorolytic pathway based on a mutant cellodextrin transporter (CDT-1 (F213L)) and cellobiose phosphorylase (SdCBP)-was investigated by comparing with a hydrolytic pathway based on the same transporter and an intracellular ${\beta}$-glucosidase (GH1-1) for their SSF performances under various conditions. Whereas the phosphorolytic and hydrolytic cellobiose-fermenting S. cerevisiae strains performed similarly under the anoxic SSF conditions, the hydrolytic S. cerevisiae performed slightly better than the phosphorolytic S. cerevisiae under the microaerobic SSF conditions. Nonetheless, the phosphorolytic S. cerevisiae expressing the mutant CDT-1 showed better ethanol production than the glucose-fermenting S. cerevisiae with an extracellular ${\beta}$-glucosidase, regardless of SSF conditions. These results clearly prove that introduction of the intracellular cellobiose metabolic pathway into yeast can be effective on cellulosic ethanol production in SSF. They also demonstrate that enhancement of cellobiose transport activity in engineered yeast is the most important factor affecting the efficiency of SSF of cellulose.

• KSBB Journal
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• v.28 no.6
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• pp.366-371
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• 2013

Ethanol productions were performed by separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) processes using seaweed, Enteromorpha intestinalis (sea lettuce). Pretreatment conditions were optimized by the performing thermal acid hydrolysis and enzymatic hydrolysis for the increase of ethanol yield. The pretreatment by thermal acid hydrolysis was carried out with different sulfuric acid concentrations in the range of 25 mM to 75 mM $H_2SO_4$, pretreatment time from 30 to 90 minutes and solid contents of seaweed powder in the range of 10~16% (w/v). Optimal pretreatment conditions were determined as 75 mM $H_2SO_4$ and 13% (w/v) slurry at $121^{\circ}C$ for 60 min. For the further saccharification, enzymatic hydrolysis was performed by the addition of commercial enzymes, Celluclast 1.5 L and Viscozyme L, after the neutralization. A maximum reducing sugar concentration of 40.4 g/L was obtained with 73% of theoretical yield from total carbohydrate. The ethanol concentration of 8.6 g/L of SHF process and 7.6 g/L of SSF process were obtained by the yeast, Saccharomyces cerevisiae KCTC 1126, with the inoculation cell density of 0.2 g dcw/L.

• KSBB Journal
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• v.14 no.2
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• pp.212-219
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• 1999

lactic acid production from cellulose by simultaneous saccharification and fermentation(SSF) was studied. The SSF using cellulase enzyme Cytolase CL and Lactobacillus delbrueckii was strongly inhibited by the end product(lactic acid). An ion-exchange resin(RA-400) was used for in-situ product removal during SSF. The sorption capacity of the resin was 200mg/g-resin. The simple SSF and the extractive SSF resulted in lactic acid concentrations of 30.4g/L and 32.0g/L, respectively, at the initial substrate concentration of 50g/L. A model was developed to simulate the extractive SSF. The lactic acid conversion for the initial substrate of 100g/L was estimated to be improved from 60% to 09% by in-situ product removal. The experimentally determined kinectic parameters were pH dependent, and fitted as empirical expressions to establish their values at different pH's. Lactic acid productivity was predicted to be maximum at pH 4.5-5.0.

• KSBB Journal
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• v.15 no.5
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• pp.458-463
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• 2000

Production of acetic acid from cellulosic biomass by Simultaneous Saccharification and Extractive Fermentation (SSEF) was investigated. The homoacetate organism used in this study was a strain of Clostridium thermoaceticum, ATCC # 49707. A batch operation of Simultaneous Saccharification and Fermentation(SSF) using ${\alpha}$-cellulose at pH 5.5 and 55$^{\circ}C$ yielded 40% conversion of cellulose to acetic acid, while a fed-batch SSF operation produced a maximum acetic acid concentration of 25 g/L, with 50% overall yield. In-situ extractive fermentation to reduce the end-product inhibition on both bacteria and enzyme was carried out. in a batch SSEF using 200 g/L IRA-400 resin, acetic acid concentration reached to 23.9 g/L and acetic acid yield and productivity were observed to be 48% and 0.20 g/L-hr, respectively.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1161-1168
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• 2009

Ethanol production by the simultaneous saccharification and fermentation (SSF) of low-value rice wine cake (RWC) without cooking was investigated. RWC is the filtered solid waste of fermented rice wine mash and contains 53% raw starch. For the SSF, the RWC slurry was mixed with the raw-starch-digesting enzyme of Rhizopus sp. and yeast, where the yeast strain was selected from 300 strains and identified as Saccharomyces cerevisiae KV25. The highest efficiency (94%) of ethanol production was achieved when the uncooked RWC slurry contained 23.03% starch. The optimal SSF conditions were determined as 1.125 units of the raw-starch-digesting enzyme per gram of RWC, a fermentation temperature of $30^{\circ}C$, slurry pH of 4.5, 36-h-old seeding culture, initial yeast cell number of $2{\times}10^7$ per ml of slurry, 17 mM of urea as the nitrogen additive, 0.25 mM of $Cu^{2+}$ as the metal ion additive, and a fermentation time of 90 h. Under these optimal conditions, the ethanol production resulting from the SSF of the uncooked RWC slurry was improved to 16.8% (v/v) from 15.1% (v/v) of pre-optimization.

• Biotechnology and Bioprocess Engineering:BBE
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• v.2 no.2
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• pp.101-104
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• 1997

Lactic acid production from ${\alpha}$-cellulose by simultaneous saccharification and fermentation (SSF) was studied. The cellulose was converted in a batch SSF using cellulase enzyme Cytolase CL to produce glucose sugar and Lactobacillus delbrueckii to ferment the glucose to lactic acid. The effects of temperature, PH, yeast extract loading, and lactic acid inhibition were studied to determine the optimum conditions for the batch processing. Cellulose was converted efficiently to lactic acid, and enzymatic hydrolysis was the rate controlling step in the SSF. The highest conversion rate was obtained at 46$^{\circ}C$ and pH 5.0. The observed yield of lactic acid from ${\alpha}$-cellulose was 0.90 at 72 hours. The optimum pH of the SSF was coincident with that of enzymatic hydrolysis. The optimum temperature of the SSF was chosen as the highest temperature the microoraganism could withstand. The optimum yeast extract loading was found to be 2.5g/L. Lactic acid was observed to be inhibitory to the microorganisms' activity.

• Journal of Microbiology and Biotechnology
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• v.11 no.4
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• pp.598-606
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• 2001

A mathematical model is proposed that can depict the kinetics of simultaneous saccharification and fermentation (SSF) using steam-exploded wood(SEW) with a glucose- and cellobiose-fermenting yeast strain. Brettanomyces custersii. An expression to describe the reduction of the relative digestibility during the hydrolysis of the SEW is introduced in the hydrolysis model. The fermentation model also takes two new factors into account, that is, the effects of the inhibitory compounds present in the SEW hydrolysates on the microorganism and the fermenting ability of Brettanomyces custersii, which can use both glucose and cellobiose as carbon sources. The model equations were used to simulate the hydrolysis of the SEW, the fermentation of the SEW hydrolysates, and a batch SSF, and the results were compared with the experimental data. The model was found to be capable of representing ethanol production over a range of substrate concentrations. Accordingly, the limiting factors in ethanol production by SSF under the high concentration of the SEW were identified as the effect of inhibitory compounds present in the SEW, the enzyme deactivation, and a limitation in the digestibility based on the physical condition of the substrate.

• Journal of the Korean Wood Science and Technology
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• v.39 no.6
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• pp.490-497
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• 2011

In this study, we determined optimal conditions for simultaneous saccharification and fermentation (SSF) using corncob biomass pretreated with oxalic acid. The effect of SSF temperature ($25.8{\sim}34.2^{\circ}C$) and agitation speed (80~220 rpm) were significant at a 99% confidence level in its effect on ethanol production. The highest ethanol production was expected when SSF was performed at $30^{\circ}C$, 170 rpm (22.5 g/L). The ethanol production was improved by mixture of yeast extract (1.25 g/L) and urea (1.25 g/L) as nitrogen source. However, addition of trace metal components and vitamin for SSF was not affected in the ethanol production. Optimal concentration of $KH_2PO_4$, $MgSO_4{\cdot}7H_2O$ for SSF was 1 g/L, 0.25 g/L respectively.

• Microbiology and Biotechnology Letters
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• v.44 no.2
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• pp.145-149
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• 2016

Thermal acid hydrolysis pretreatment of Kappaphycus alvarezii was carried out with 12% (w/v) seaweed slurry and 180 mM H₂SO₄ at 140°C for 5 min. Utility of the thermotolerant yeast Kluyveromyces marxianus KCTC7150 was evaluated with respect to cell growth and ethanol fermentation at 40°C was close to optimal for enzymatic hydrolysis. This could lead to the integration of both the saccharification and fermentation processes. The levels of ethanol production by simultaneous saccharification and fermentation (SSF) with non-adapted and adapted K. marxianus KCTC7150 were 9.1 g/l with an ethanol yield (Y_EtOH) of 0.24 and 10.2 g/l with an ethanol yield (Y_EtOH) of 0.27 at 156 h, respectively. The two-phase SSF process was employed in this study to improve the efficiency of ethanol fermentation. Adapted K. marxianus KCTC7150 using the two-phase SSF process produced 13.5 g/l with an ethanol yield (Y_EtOH) of 0.35 at 96 h. Development of the two-phase SSF process could enhance the overall ethanol fermentation yields of the seaweed K. alvarezii.