• Title/Summary/Keyword: fermenting conditions

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Effects of Engineered Saccharomyces cerevisiae Fermenting Cellobiose through Low-Energy-Consuming Phosphorolytic Pathway in Simultaneous Saccharification and Fermentation

  • Choi, Hyo-Jin;Jin, Yong-Su;Lee, Won-Heong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.1
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    • pp.117-125
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    • 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.

Evaluation of Ethanol Production Activity by Engineered Saccharomyces cerevisiae Fermenting Cellobiose through the Phosphorolytic Pathway in Simultaneous Saccharification and Fermentation of Cellulose

  • Lee, Won-Heong;Jin, Yong-Su
    • 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.

Optimum Alcohol Fermenting Conditions for Kiwi (Actinidia chinensis) Wine

  • Jang, Se-Young;Woo, Seung-Mi;Kim, Ok-Mi;Choi, In-Wook;Jeong, Yong-Jin
    • Food Science and Biotechnology
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    • v.16 no.4
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    • pp.526-530
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    • 2007
  • The objective of this study was to establish the optimum alcohol fermenting conditions for the processing of kiwi wine and vinegar products. Six yeast strains were examined for their alcohol production from kiwi at $30^{\circ}C$ for 72 hr with continuous shaking at 100 rpm. Under these conditions, Saccharomyces kluyveri DJ97 produced the highest alcohol content of 10.2%. As the fermentation time extended to 96 hr, the alcohol content reached a maximum of 12.75%. The optimum alcohol fermenting conditions for kiwi fruit were accomplished when kiwi was added to an equal amount of water, inoculated with S. kluyveri DJ97 and fermented at $30^{\circ}C$ for 96 hr with continuous shaking. The content of soluble solids decreased as the alcohol concentration increased, whereas little change was observed in the pH and titratable acidity during the low temperature aging process. Other alcoholic compounds, such as methanol, isopropanol, n-propanol, isobutanol, and isoamylalcohol, tended to increase as fermentation progressed.

Fungi-rice bran based Fermentation of Coptis Chinensis and Curcuma Longa Root and its Influence of Silk Dyeing

  • Park, Young Mi;Choi, Jae Hong
    • Fashion & Textile Research Journal
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    • v.15 no.4
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    • pp.635-641
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    • 2013
  • This study examined the dye-properties of natural fabrics dyed with Coptis chinensis and Curcuma longa root fermented with fungi. The optimum culture conditions for the fermentation of microorganisms, the relationship between natural dye color and fermentation conditions were investigated. Two different medical herbs (ground to 80-100 mesh in size) were used as a natural dyeing source. Phellinus linteus (P. linteus), which can grow in different media, such as Agarmedium (only agar containing medium), maltose extract agar (MA) and potato dextrose extract agar (PDA) culture media, were isolated from the medium. P. linteus was confirmed to be the optimum microorganism for the fermentation of Coptis chinensis and Curcuma longa, and the MA medium was confirmed to be the best for culturing. When using the microorganism as the fermenting agent, $32^{\circ}C$ was found to be the optimum fermenting temperature for both natural colorants. Regarding the dyeing property of the fermented natural dye, silk was dyed quite darkly in an appearance by naked eye estimation and the K/S value in the color strength of silk reached a high level of 16 after the fermenting process. The washing fastness of dyed silk after treatment washing was reduced from 4 to under4 and indicates that dyed silk with fermented plant was not unsubstantial. The light fastness was 1 to 2, showing intended to maintain due to the fermentation process.

Studies on the Selection of Microorganism for Food Wastes and Optimization of Fermentation Process (음식물찌꺼기 소멸효율 재고를 위한 발효균 및 발효 공정 최적화 연구)

  • Kim, Young-Kwon;Hong, Myung-Pyo;Kim, Myung-Jin;Hong, Suk-Il;Park, Myung-Suk;Kim, Jong-Suk;Chang, Ho-Geun
    • Journal of the Korea Organic Resources Recycling Association
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    • v.6 no.2
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    • pp.95-112
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    • 1998
  • For the effective disposal of organic food wastes, we seleted 4 strains of microorganism from 186 microbial candidate via enzyme activity test, salt tolerance, food decomposition rate, stability and safety of strains. The identity of these 4 strains are as follows : Fungi is Rhizopus sp., yeasts are Galactomyces sp., Pichia sp. and Hyphopichia sp., In the 50L fermenter scale, we tested various fermenting factor for the optimization of conditions of food waste decomposition using 4 selected strains. The optimum fomenting conditions were as follows : BIO-CHIP Volume 25-30 L, BIO CHIP size 2.0-6.0mm, air flow 200-280L/min, mixing intensity 2-4rpm, temperature $30-45^{\circ}C$. In these fermenting conditions, the efficiency of decomposition(rate of weight loss of food wastes) were 93%. Also the quality of fermenting output were assayed at the basis of fertilizer, and the results were as good as general compost.

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Studies on the Optimum Fermenting Conditions of Dongchimi for Production of Ion Beverage (이온음료 제조를 위한 동치미의 최적 담금 조건에 관한 연구)

  • Ko, Eun-Jung;Hur, Sang-Sun;Park, Man;Choi, Yong-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.1
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    • pp.141-146
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    • 1995
  • The study was conducted for optimum fermenting conditions of Dongchimi(pony tailed chinese radish kimchi) in production of ion beverage. The changes of pH and total acidity were increased as the temperature increased. Non-volatile organic acids, such as lactic acid, citric acid, malic acid and succinic acid were produced in Dongchimi fermentation. The amount of lactic acid was increased higher, followed by citric acid and malic acid. However succininc acid was produced a little of amount at $0^{\circ}C$. Lactic acid producing bacteria number increased in initial period and then decreased in last period of fermentation. During lactic acid producing bacteria was increased, the amouont of lactic acid was increased. The flavor components were tentatively identified as methyl pentane, ethyl thioethene 2, 3-diazaindolizine, dimethyl disulfide. The optimum fermenting conditions of Dongchimi for production of ion beverage were 24~29 days at $0^{\circ}C$, 9~12dyas at $5^{\circ}C$ and 16~22days at $10^{\circ}C$, respectively.

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Observation of Cellodextrin Accumulation Resulted from Non-Conventional Secretion of Intracellular β-Glucosidase by Engineered Saccharomyces cerevisiae Fermenting Cellobiose

  • Lee, Won-Heong;Jin, Yong-Su
    • Journal of Microbiology and Biotechnology
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    • v.31 no.7
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    • pp.1035-1043
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    • 2021
  • Although engineered Saccharomyces cerevisiae fermenting cellobiose is useful for the production of biofuels from cellulosic biomass, cellodextrin accumulation is one of the main problems reducing ethanol yield and productivity in cellobiose fermentation with S. cerevisiae expressing cellodextrin transporter (CDT) and intracellular β-glucosidase (GH1-1). In this study, we investigated the reason for the cellodextrin accumulation and how to alleviate its formation during cellobiose fermentation using engineered S. cerevisiae fermenting cellobiose. From the series of cellobiose fermentation using S. cerevisiae expressing only GH1-1 under several culture conditions, it was discovered that small amounts of GH1-1 were secreted and cellodextrin was generated through trans-glycosylation activity of the secreted GH1-1. As GH1-1 does not have a secretion signal peptide, non-conventional protein secretion might facilitate the secretion of GH1-1. In cellobiose fermentations with S. cerevisiae expressing only GH1-1, knockout of TLG2 gene involved in non-conventional protein secretion pathway significantly delayed cellodextrin formation by reducing the secretion of GH1-1 by more than 50%. However, in cellobiose fermentations with S. cerevisiae expressing both GH1-1 and CDT-1, TLG2 knockout did not show a significant effect on cellodextrin formation, although secretion of GH1-1 was reduced by more than 40%. These results suggest that the development of new intracellular β-glucosidase, not influenced by non-conventional protein secretion, is required for better cellobiose fermentation performances of engineered S. cerevisiae fermenting cellobiose.

Taste Components of Soy Sauce Manufactured by Bacillus Species SSA3-2M1 and Fused ST723-F31

  • Kim, Haeng Ja;Eun Ju Lee;Ok Sun Shin;Myeong Rak Choi;Jong Kyu Kim
    • Journal of Microbiology and Biotechnology
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    • v.6 no.3
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    • pp.202-208
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    • 1996
  • In order to investigate fermenting conditions and the microorganisms necessary for factory production of traditional Korean soy sauce, we manufactured soy sauce made by Bacillus species SSA3-2M1 and fused ST723-F31 with aeration (1/30 vvm, 113 vvm and 2/3 vvm) at $30^{\circ}C$ for 40 days. This method was chosen to investigate the changes of dissolved oxygen, pH, cell number, flavor and the taste components during fermentation. When air was supplied (2/3 vvm) to the fermentor during fermentation, the flavor of the soy sauce and the composition of taste components (free amino acids, free sugars and organic acids) were similar to that of traditional Korean soy sauce after 22 days. The results of our experiments indicates that the mass production of traditional Korean soy sauce is possible using Bacillus species SSA3-2M1 and fused ST723-F31 given sufficient aeration.

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Screening and Characterization of Potential Bacillus Starter Cultures for Fermenting Low-Salt Soybean Paste (Doenjang)

  • Jeon, Hye Hee;Jung, Ji Young;Chun, Byung-Hee;Kim, Myoung-Dong;Baek, Seong Yeol;Moon, Ji Young;Yeo, Soo-Hwan;Jeon, Che Ok
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.666-674
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    • 2016
  • The bacterial strains were screened as potential starters for fermenting low-salt doenjang (a Korean traditional fermented soybean paste) using Korean doenjang based on proteolytic and antipathogenic activities under 6.5-7.5% NaCl conditions. Phylogenetic analysis based on 16S rRNA gene sequences showed that they all belonged to the genus Bacillus. Proteolytic and antipathogenic activities against Escherichia coli, Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, and Aspergillus flavus, as well as fibrinolytic, amylase, and cellulase activities of the 10 strains were quantitatively evaluated. Of these, strains D2-2, JJ-D34, and D12-5 were selected, based on their activities. The functional, phenotypic, and safety-related characteristics of these three strains were additionally investigated and strains D2-2 and D12-5, which lacked antibiotic resistance, were finally selected. Strains D2-2 and D12-5 produced poly-γ-glutamic acid and showed various enzyme activities, including α-glucosidase and β-glucosidase. Growth properties of strains D2-2 and D12-5 included wide temperature and pH ranges, growth in up to 16% NaCl, and weak anaerobic growth, suggesting that they facilitate low-salt doenjang fermentation. Strains D2-2 and D12-5 were not hemolytic, carried no toxin genes, and did not produce biogenic amines. These results suggest that strains D2-2 and D12-5 can serve as appropriate starter cultures for fermenting low-salt doenjang with high quality and safety.

Cell Fusion Between Xylose Fermenting Yeast and Xylanase Secreting Yeast (Xylanase 분비효모와 Xylose 발효효모의 Protoplast Fusion)

  • 김남순;배명애;서정훈
    • Microbiology and Biotechnology Letters
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    • v.17 no.2
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    • pp.88-93
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    • 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.

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