• Journal of Microbiology and Biotechnology
• /
• v.4 no.3
• /
• pp.215-221
• /
• 1994

Two strains of yeast (RA-74-2 and RA-912) showing superior fermenting ability at a high temperature were isolated from soils and wastewaters by an enrichment culture method. Based on the morphological and physiological charateristics, the two strains were identified as Saccharomyces cerevisiae and Kluyveromyces marxianus, respectively. RA-74-2 was able to grow upto $43^{\circ}C$ and sustain similar fermenting ability in the temperatures range from 30 to $40^{\circ}C$. In addition, the sugar- and ethanol-tolerance of RA-74-2 were 30% (w/v) glucose and 10% (v/v) ethanol, which appeared to be higher than those of nine other industrial yeast strains currently being used in the alcohol factories. The thermotolerant ethanol fermenting yeast RA-912 showed identical growth in the temperatures range from 35 to $45^{\circ}C$ and was resistant to various heavy metals. The quality and quantity of byproducts of the isolated yeast strains in fermentation broth after fermentation at $40^{\circ}C$ and $45^{\circ}C$ were similiar with those obtained at $30^{\circ}C$. These results show that RA-74-2 can be adopted for the ethanol fermentation process where the expenses for cooling system is significant, and suggest that RA-912 may be applied in either SSF(simultaneous saccharification and fermentation) or Flash-fermentation process and RA-912 may be used as a gene donor for the development of thermotolerant ethanol-fermenting yeasts.

• Microbiology and Biotechnology Letters
• /
• v.17 no.2
• /
• pp.88-93
• /
• 1989

To improve a new yeast strain capable of converting xylan to ethanol directly, we tried protoplast fusion between xylose fermenting yeast (Candida sp. X-6-41) and xylan assimilating yeast (Crypto-coccus sp. XB-33), finally selected the most promising two fusants (XFU-1 and XFU-2). As the optimum conditions for protoplast formation, the yeast cells were cultured to exponential phase in YPD and YPX containing 0.6M KCI, respectively, and then treated with zymolyase (0.25mg/$m\ell$), cellulase(4mg/$m\ell$) and 100mM 2-mercaptoethanol at pH 8 and 3$0^{\circ}C$. The protoplasts of parental auxotrophs were fused in the presence of 20mM CaCl$_2$and 40% polyethylene glycol(M.W.4000). The physiological and morphological characteristics of the fusants, such as assimilation of carbon sources, cell size, growth rate, xylanase activity and xylan fermentation ability were investigated. Xylanase activity of fusants that cultured in chemically minimal medium was higher than that of fusants that cultured in completed medium, because xylanase producing activity of xylose fermenting yeast(X-6-41) was inhibited by isoleucine.

• Journal of Microbiology and Biotechnology
• /
• v.11 no.4
• /
• pp.598-606
• /
• 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 Microbiology and Biotechnology
• /
• v.27 no.9
• /
• pp.1649-1656
• /
• 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.

• The Korean Journal of Food And Nutrition
• /
• v.12 no.3
• /
• pp.219-225
• /
• 1999

In order to improve the quality of nuruk a Korean traditional fermenting agent for brewing nuruk was modified by a new method inoculating Rhizopus japonicus T2 Aspergillus oryzae L2 and hansenula sp. BC26 which had been isolated from traditional Nuruk. And the characteristics of modified Nuruk were investimgated as compared with current fermenting agents such as commercial Nuruk and rice koji of As-pergillus kawachii. The odor of modified Nuruk was savory but those of commercial Nuruk and rice koji were fetid andinodorous repectively. The extracted waters of modified Nuruk commercial Nuruk and rice koji were yellow pale yellow and colorless and showed 6.15, 6.01 and 3.30 of pH respectively. Mod-ified Nuruk had 7.6${\times}$106CFU/g of yeast but commercial uruk and rice koji had no yeast. Commercial Nuruk had 1.0${\times}$102CFU/g of lactic acid bacteria but modified Nuruk and rice koji had no lactic acid bacteria. The amylolytic and proteolytic activities of modified Nuruk were much higher than those of commercial Nuruk or rice koji. Seed mash of modified Nuruk had ester aroma but that of commercial Nuruk rather offensive odor and that of rice koji neither ester aroma or offensive odor. It seemed that if the modified Nuruk is used in seed mashing the supplement of acidulant is need to lower pH. The quality of modified Nuruk was thought to be much better than that of commercial Nuruk or rice koji.

• Journal of Microbiology and Biotechnology
• /
• v.32 no.1
• /
• pp.117-125
• /
• 2022

Until recently, four types of cellobiose-fermenting Saccharomyces cerevisiae strains have been developed by introduction of a cellobiose metabolic pathway based on either intracellular β-glucosidase (GH1-1) or cellobiose phosphorylase (CBP), along with either an energy-consuming active cellodextrin transporter (CDT-1) or a non-energy-consuming passive cellodextrin facilitator (CDT-2). In this study, the ethanol production performance of two cellobiose-fermenting S. cerevisiae strains expressing mutant CDT-2 (N306I) with GH1-1 or CBP were compared with two cellobiose-fermenting S. cerevisiae strains expressing mutant CDT-1 (F213L) with GH1-1 or CBP in the simultaneous saccharification and fermentation (SSF) of cellulose under various conditions. It was found that, regardless of the SSF conditions, the phosphorolytic cellobiose-fermenting S. cerevisiae expressing mutant CDT-2 with CBP showed the best ethanol production among the four strains. In addition, during SSF contaminated by lactic acid bacteria, the phosphorolytic cellobiose-fermenting S. cerevisiae expressing mutant CDT-2 with CBP showed the highest ethanol production and the lowest lactate formation compared with those of other strains, such as the hydrolytic cellobiose-fermenting S. cerevisiae expressing mutant CDT-1 with GH1-1, and the glucose-fermenting S. cerevisiae with extracellular β-glucosidase. These results suggest that the cellobiose-fermenting yeast strain exhibiting low energy consumption can enhance the efficiency of the SSF of cellulosic biomass.

• Microbiology and Biotechnology Letters
• /
• v.16 no.6
• /
• pp.505-509
• /
• 1988

Ethanol productivity of a xylose fermenting yeast (Candida sp. X-6-4l) isolated from soil was investigated in laboratory scale using Erlenmeyer flask and mini-jar tormentor. The optimal conditions of xylose fermentation in flask experiment were pH 4, asparagine as nitrogen source, xylose 20g/$\ell$, and in these condition, ethanol yield was about 80% to theoretical yield. Using mini-jar fermentor containing 5% total sugar with 2.5% xylose and 2.5% glucose, we obtained 2.3%(v/ v) ethanol and the corresponding efficiency was 72.3% of total sugar. In this case, the consumming speed of sugar under aerobic condition was faster than that of anaerobic condition, and glucose was used previously to xylose. The optimum concentration of xylose for ethanol fermentation in mini-jar fer-mentor scale was 5%, and the efficiency was 69% of total sugar(Alc.2.2% v/v).

• Microbiology and Biotechnology Letters
• /
• v.36 no.2
• /
• pp.158-164
• /
• 2008

The fusants which contain starch utilizing ability and alcohol fermenting ability were developed by protoplast fusion of Saccharomyces cerevisiae KOY-1 and Saccharomyces diastaticus KCTC 1804. Sacharomyces cerevisiae KH-12 was obtained by haploid induction from Saccharomyces cerevisiae KOY-1. The auxotropic mutants of yeast were obtained by using an ethylmethane sulfonate (EMS). The frequency of protoplast formation in Saccharomyces cerevisiae KOY-1 $(Met^-)$ and Saccharomyces diastaticus KCTC 1804 $(Trp^-)$ were 90.5% and 97.7%, respectively. The frequency of fusant formation was $1.79{\times}10^{-4 }$ for the regenerated protoplast and the 1,000 fusants were obtained. Fusant FA 776 was selected as a potential yeast which contain an alcohol fermenting ability in the starch medium. The genetic stability was 4.64% for 10 passages of generation. Fusant FA 776 produced 13mg/ml of alcohol in 24% starch medium and showed 1.86-fold higher alcohol fermenting ability than Saccharomyces diastaticus KCTC 1804.

• Journal of Korean Society of Forest Science
• /
• v.98 no.3
• /
• pp.305-310
• /
• 2009

Yellow poplar was selected a promising biomass resources for bio-ethanol production through alkali prehydrolysis, enzymatic saccharification and fermentation using commercial cellulase mixtures (Celluclast 1.5L and Novozym 342 mixtures) and fermenting yeast. In alkali prehydrolysis, 51.1% of Yellow poplar biomass remained as residues, which chemical compositions were 82.2% of cellulose, 17.6% of xylan and 2.0% of lignin. In alkali prehydrolysis process, 96.9% of cellulose, 38.0% of xylan and 5.7% of lignin were remained. Enzymatic saccharification by commercial cellulases led to 87.0% of cellulose to glucose and 87.2% of xylan to xylose conversion. Produced glucose and xylose were fermented with fermenting yeast (Saccharomycess cerevisiae), which resulted in selective fermentation of glucose only to bio-ethanol. Residual monosaccharides after fermentation were consisted to 0.4-1.4% of glucose and 92.1-99.5% of xylose. Ethanol concentration was highest for 24 h fermentation as 57.2 g/L, but gradually decreased to 56.2 g/L for 48 h fermentation and 54.3 g/L for 72 h fermentation, due to the ethanol consumption by fermenting yeast.

• Applied Biological Chemistry
• /
• v.31 no.3
• /
• pp.267-273
• /
• 1988

Transformed, hybrid strains of the yeast Saccharomyces capable of simultaneous secretion of both glucoamylase and ${\alpha}-amylase$ have been produced. These strains can carry out direct, one-step assimilation of starch with conversion efficiency greater than 93% during a 5 day growth period. One of the transformants converts 92.8% of available starch into reducing sugars in only 2 days. Glucoamylase secretion by these strains results from expression of one or more chromosomal STA genes derived from Saccharomyces diastaticus. The strains were transformed by a plasmid(pMS12) containing mouse salivary ${\alpha}-amylase$ cDNA in an expression vector containing yeast alcohol dehydrogenase promoter and a segment of yeast $2{\mu}$ plasmid. The major starch hydrolysis product produced by crude amylases found in culture broths is glucose, indicating that ${\alpha}-amylase$ and glucoamylase act cooperatively.