• Journal of Microbiology and Biotechnology
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• v.9 no.3
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• pp.340-345
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• 1999

A novel simultaneous saccharification and fermentation (SSF) process from the microcrystalline cellulose to ethanol was developed by using $\delta$-integrated recombinant cellulolytic Saccharomyces cerevisiae L2612$L2612\deltaGC$, which can utilize cellulose as carbon and energy sources. The optimum amount of enzymes needed for the efficient conversion of cellulose to ethanol at $30^{\circ}C$ was determined with commercial cellulolytic enzymes. By fed-batch cultivation, the heterologous cellulolytic enzymes were accumulated up to 42.67% of the total cellulase and 29% of the $\beta$-glucosidase needed for the efficient SSF process. When this $\delta$-integrated recombinant yeast was applied to the successive SSF step for ethanol production, 20.35 g/l of ethanol was produced after 12 h from 50 g/l of microcrystalline cellulose. By using this novel SSF process, a considerable amount of commercial enzymes was reduced.

• KSBB Journal
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• v.11 no.3
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• pp.347-352
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• 1996

The enzyme deactivation in a membrane recycling system for the simultaneous saccharification and fermentation(SSF) was studied under various temperature and pressure. The optimum molecular weight cut off(MWCO) of the ultrafiltration membrane for recycling cellulase and ${\beta}$-glucosidase was 50,000. When the cellulase was recycled continuously through the membrane system, it was not deactivated. But the activity of ${\beta}$-glucosidase was decreased with an increase in operating temperature and transmembrane pressure. After 720 minutes at $42^{\circ}C$ and 24.8 psig , the activity of ${\beta}$-glucosidase was reduced by 35% of the initial activity. Such tendencies could be well explained by the results of highly induced shear at the fiber surface of membrane when temperature and transmembrane pressure became higher.

• Applied Biological Chemistry
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• v.28 no.2
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• pp.98-105
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• 1985

In the sheep and cattle's rumen, facultative anaerobic cellulolytic bacteria were isolated by using Hungate's roll tube technique. In the 21 isolated species, one was screened by its strong cellulolytic activity and identified as Cellulomonas fimi C-14 by investigate morphological, cultural, physiological characteristics and electron microgram. Optimum conditions of the cell growth and enzyme production were pH 6.5 an $30^{\circ}C$, Thiamine and biotin support a good growth of C. fimi C-14. In the enzyme activities, Crystalline cellulose hydrolyzing activity, CMCase activity and ${\beta}-glucosidase$ activity were 20.6, 226.6 and 0.56$(unit{\times}10^3/ml)$ at pH 6.0, $40^{\circ}C$. By addition of fungal cellulase, enzyme activity was increased. Simultaneous Saccharification Fermentation is better than two step fermentation in ethanol yield with Saccharomyces cerevisiae DY2.

• Journal of Microbiology and Biotechnology
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• v.20 no.1
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• pp.101-109
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• 2010

Efficient L-lactic acid production from Jerusalem artichoke tubers, by Lactobacillus casei G-02, using simultaneous saccharification and fermentation (SSF) in a fed-batch culture, is demonstrated. A kinetic analysis of the SSF revealed that the inulinase activity was subjected to product inhibition, whereas the fermentation activity of G-02 was subjected to substrate inhibition. It was also found that the intracellular NADH oxidase (NOX) activity was enhanced by the citrate metabolism, which dramatically increased the carbon flux of the Embden-Meyerhof-Parnas (EMP) pathway, along with the production of ATP. As a result, when the SSF was carried out at $40^{\circ}C$ after an initial hydrolysis of 1 h and included a sodium citrate supplement of 10 g/l, an L-lactic acid concentration of 141.5 g/l was obtained after 30 h, with a volumetric productivity of 4.7 g/l/h. The conversion efficiency and product yield were 93.6% of the theoretical lactic acid yield and 52.4 g lactic acid/l00 g Jerusalem artichoke flour, respectively. Such a high concentration of lactic acid with a high productivity from Jerusalem artichokes has not been reported previously, making G-02 a potential candidate for the economic production of L-lactic acid from Jerusalem artichokes on a commercial scale.

• Journal of Industrial Technology
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• v.39 no.1
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• pp.15-19
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• 2019

This work focused on characterization of the starch degradation activity from extremophile strain Deinococcus geothermalis. Glucoamylase gene from D. geothermalis was cloned and overexpressed by pET-21a vector using E. coli BL21 (DE3). In order to characterize starch degrading activity of recombinant glucoamylase, enzyme was purified using HisPur Ni-NTA column. The recombinant glucoamylase from D. geothermalis exhibited the optimum temperature as $45^{\circ}C$ for starch degradation activity. And highly acido-stable starch degrading activity was shown at pH 2. For further optimization of starch degrading activity with metal ion, various metal ions ($AgCl_2$, $HgCl_2$, $MnSO_4{\cdot}4H_2O$, $CoCl_2{\cdot}6H_2O$, $MgSO_4$, $ZnSO_4{\cdot}7H_2O$, $K_2SO_4$, $FeCl_2{\cdot}4H_2O$, NaCl, or $CuSO_4$) were added for enzyme reaction. As results, it was found that $FeCl_2{\cdot}4H_2O$ or $MnSO_4{\cdot}4H_2O$ addition resulted in 17% and 9% improved starch degrading activity, respectively. The recombinant glucoamylase from D. geothermalis might be used for simultaneous saccharification and fermentation (SSF) process at high acidic conditions.

• Korean Chemical Engineering Research
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• v.52 no.4
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• pp.430-435
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• 2014

Physical and chemical barriers, caused by the close association of the main components of cellulosic biomass, hinder the hydrolysis of cellulose to fermentable sugars. Since the main goal of pretreatment is to increase the enzyme accessibility improving digestibility of cellulose, development of an effective pretreatment process has been considered to be important. In this study, SAA (Soaking in Aqueous Ammonia) was chosen as pretreatment because this is the simple and low-cost method. Rice straw of which the production is outstandingly high in domestic agriculture residues in Korea was chosen as raw material. SSA pretreatment with various reaction time of 3 h to 72 h was tested. The enzymatic hydrolysis and SSF (Simultaneous Saccharification and Fermentation) were performed at three different temperature (30, 40 and $50^{\circ}C$) to investigate performance of SSF upon various pretreatment conditions. As a result, this SAA treated-rice straw was found to have great potential for effective enzymatic hydrolysis and SSF with lower enzyme dosage at lower temperature ($30^{\circ}C$) than its conventional SSF. In SAA addition, SAA reduced fermentation time to 24 h owing to increase the initial hydrolysis rate substantially.

• Korean Journal of Weed Science
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• v.32 no.2
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• pp.85-97
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• 2012

To investigate the usefulness of freshwater alga Water-net (Hydrodictyon reticulatum, HR) as resources for production of fermentable sugars, the easiness of enzymatic saccharification was evaluated at first. When 6 plant materials (HR, Spirulina, Chlorella, Scenedesmus, Cladophora, Corn stover) were enzymatically hydrolyzed with 2% solid loading at the same condition, HR showed the highest ratio of saccharification based on glucose production. No milled HR was also completely saccharified at the amounts of optimal enzyme mixture. Glucose yield was not changed though the citrate buffer strength for saccharification was decreased from 0.1 M to 0.1 mM. Only about 10% yield reduction was observed compared to that of $120^{\circ}C$ treatment when HR was enzymatically hydrolyzed at room temperature. The saccharification was normally occurred at $37^{\circ}C$ and pH 6.5 which is general growth condition of fermentable microrganisms, suggesting that HR have a biomass characteristics applicable for the simultaneous saccharification and fermentation. The saccharification was occurred by more than 70~80% of one of the best condition although the supplied enzyme amounts was reduced to 1/10 volume. And the glucose yield by enzymatic hydrolysis was not decreased by 10% HR solid loading and began to decrease at more than 15% solid contents. Above these results show that HR is an interesting algal biomass which is relatively easy to be saccharified by hydrolyzing enzymes. In addition, HR is a flilamentous alga and very easy to be collected. Therefore, HR seems to be an useful and valuable resources in the economical production of fermentable sugars for manufacture of bio-chemical products.

• Applied Chemistry for Engineering
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• v.23 no.3
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• pp.326-332
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• 2012

Among the samples prepared by various pre-treatment methods, the one pretreated by dilute sulfuric acid showed the highest glucose yield in the enzymatic hydrolysis. Statistical analysis of enzymatic hydrolysis revealed that the glucose yield was in proportion to the enzyme dosage, the ratio of the pre-treated sample to the buffer solution, and the reaction time and that the effect of enzyme dosage was predominant in the experiment range. In addition, the glucose yield was estimated to be 76.1% at an optimal enzymatic hydrolysis condition. In a separate hydrolysis and fermentation (SHF), Saccharomyces cerevisiae converted over 80% of glucose from the enzymatic hydrolysis of pre-treated wasted corn stalk by dilute sulfuric acid to bioethanol with 37% of ethanol yield and 0.42 $g/L{\cdot}hr$ of productivity. In the simultaneous saccharification and fermentation (SSF), 59.5% of conversion from glucan to ethanol and 0.20 $g/L{\cdot}hr$ of productivity were achieved. In both SHF and SSF, approximately 88 g of bioethanol could be obtained from 1 kg of wasted corn stalk. The possible amount of bioethanol in Gangwon province were estimated to be 1.9 kiloton with the assumption of the 50% of collection ratio.

• Journal of Life Science
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• v.26 no.8
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• pp.976-982
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• 2016

Seaweed has high growth rate, low land usage, high CO₂ absorption and no competition for food resources. Therefore, the use of lignin-free seaweed as a raw material is arising as a third generation biomass for bioethanol production. Various pretreatment techniques have been introduced to enhance the overall hydrolysis yield, and can be categorized into physical, chemical, biological, enzymatic or a combination. Thermal acid hydrolysis pretreatment is one of the most popular methods to attain high sugar yields from seaweed biomass for economic reasons. At thermal acid hydrolysis conditions, the 3,6-anhydro-galactose (AHG) from biomass could be converted to 5-hydroxymethylfurfural (HMF), which might inhibit the cell growth and decrease ethanol production. AHG is prone to decomposition into HMF, due to its acid-labile character, and subsequently into weak acids such as levulinic acid and formic acid. These inhibitors can retard yeast growth and reduce ethanol productivity during fermentation. Thus, the carbohydrates in seaweed require effective treatment methods to obtain a high concentration of monosaccharides and a low concentration of inhibitor HMF for ethanol fermentation. The efficiency of bioethanol production from the seaweed biomass hydrolysate is assessed by separate hydrolysis and fermentation (SHF). To improve the efficiency of the ethanol fermentation of mixed monosaccharides, the adaptation of yeast to high concentration of sugar could make simultaneous utilization of mixed monosaccharides for the production of ethanol from seaweed.

• The Korean Journal of Food And Nutrition
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• v.26 no.1
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• pp.29-34
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• 2013

The objective of this study was to manufacture three kinds of domestic sweet potato Makgeolli using a mixture design and an optimization technique. The effects of four different manufacture methods, such as simultaneous saccharification and fermentation (SSF) with or without malt and separate hydrolysis and fermentation (SHF) with or without malt were determined. The SSF methods of Makgeolli produced higher alcohol content than that of SHF methods. The sensory score was not influenced by different making methods. Fourteen experimental points were selected, and rice (10~50%), sweet potato (10~50%) and water (40~60%) were chosen as independent variables. The measured responses were sensory preference, total polyphenol content, and DPPH radical scavenging activities. The ratio of the optimum sweet potato Makgeolli mixture formulation was developed as 15.11 (rice): 44.89 (sweet potato): 40 (water) using the optimization technique. The desirability of the optimum mixture formulation was 0.839. Yellow sweet potato Makgeolli using the optimum mixture formulation produced higher soluble sugar content compared to others. Regular sweet potato Makgeolli produced higher pH. The purple sweet potato Makgeolli's total polyphenol content and DPPH radical scavenging activity were measured to be the highest at $771.91{\pm}1.42mg\;GAE/{\ell}$, $131.55{\pm}4.03%$.