• Asian-Australasian Journal of Animal Sciences
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• 제30권2호
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• pp.171-180
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• 2017

Objective: This study investigated the association of enzyme-producing microbes and their enzymes with starch and hemicellulose degradation during fermentation of total mixed ration (TMR) silage. Methods: The TMRs were prepared with soybean curd residue, alfalfa hay (ATMR) or Leymus chinensis hay (LTMR), corn meal, soybean meal, vitamin-mineral supplements, and salt at a ratio of 25:40:30:4:0.5:0.5 on a dry matter basis. Laboratory-scale bag silos were randomly opened after 1, 3, 7, 14, 28, and 56 days of ensiling and subjected to analyses of fermentation quality, carbohydrates loss, microbial amylase and hemicellulase activities, succession of dominant amylolytic or hemicellulolytic microbes, and their microbial and enzymatic properties. Results: Both ATMR and LTMR silages were well preserved, with low pH and high lactic acid concentrations. In addition to the substantial loss of water soluble carbohydrates, loss of starch and hemicellulose was also observed in both TMR silages with prolonged ensiling. The microbial amylase activity remained detectable throughout the ensiling in both TMR silages, whereas the microbial hemicellulase activity progressively decreased until it was inactive at day 14 post-ensiling in both TMR silages. During the early stage of fermentation, the main amylase-producing microbes were Bacillus amyloliquefaciens (B. amyloliquefaciens), B. cereus, B. licheniformis, and B. subtilis in ATMR silage and B. flexus, B. licheniformis, and Paenibacillus xylanexedens (P. xylanexedens) in LTMR silage, whereas Enterococcus faecium was closely associated with starch hydrolysis at the later stage of fermentation in both TMR silages. B. amyloliquefaciens, B. licheniformis, and B. subtilis and B. licheniformis, B. pumilus, and P. xylanexedens were the main source of microbial hemicellulase during the early stage of fermentation in ATMR and LTMR silages, respectively. Conclusion: The microbial amylase contributes to starch hydrolysis during the ensiling process in both TMR silages, whereas the microbial hemicellulase participates in the hemicellulose degradation only at the early stage of ensiling.

• Korean Chemical Engineering Research
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• 제46권5호
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• pp.858-862
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• 2008

분리 당화발효(Separate Hydrolysis and Fermentation, SHF)는 당화와 발효공정을 따로 수행하는 방법으로 최종 생성물인 글루코오스에 의해 억제 영향을 받기 때문에 반응에 진행됨에 따라 축적된 글루코오스의 농도가 높아지면 반응이 종결되는 단점이 있다. 이를 극복하기 위해 효소의 양을 늘리는 방법이 있지만, 효소의 생산비용이 비싸기 때문에 경제적인 방법이 될 수 없다. 이러한 분리 당화발효 공정의 단점을 극복하기 위해서 동시당화발효 공정(Simultaneous Saccharification and Fermentation, SSF)은 하나의 반응기에서 당화와 발효를 동시에 수행한다. 동시당화발효 공정에서는 당화과정에서 글루코오스가 생성되자마자 효모가 발효과정을 통해 글루코오스를 바로 제거하기 때문에 반응기내에서 당의 축적을 최소화할 수 있다. 따라서 동시당화발효 공정은 최종 생성물의 억제 작용을 방지할 수 있고, 효소의 가수분해 반응을 향상시킬 수 있다. 본 연구에서는 동시당화발효에서 에탄올의 수율에 관여하는 조건들(pH, 반응온도, 효소 투입량, 반응시간)의 최적 조건을 찾는 연구를 수행하였다. 기질로는 감자전분을 사용하였고, 효소는 glucoamylase, 균주는 Saccharomyces cerevisiae가 각각 사용되었다. 동시당화발효의 최적의 조건은 pH 4, 온도 38로 나타났다. 최적의 조건으로 감자전분을 동시당화발효하였을 때 반응 18시간 후에 에탄올은 최대 수율 86%에 도달하였다.

• 한국미생물·생명공학회지
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• 제47권3호
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• pp.412-422
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• 2019

The present study reports the morphological and molecular characterization of the fungal strain, CMSS06 and evaluates its raw starch hydrolyzing ability in four different agricultural substrates (rice bran, banana peel, cassava tubers, and coconut water). The potential use of each agricultural substrate to replace the expensive fermentation media was evaluated with six different fermentation media: rice bran (RB), banana peel (BP), cassava starch (CS), cassava in coconut water (CSCW), cassava in modified coconut water (CMCW), and pure Coconut water (CW). The fungal strain CMSS06 was identified as Thielaviopsis ethacetica by the analysis of the ITS sequences. The T. ethacetica alpha amylase enzyme exhibited maximum alpha amylase activity at 72 h, pH 7.0, and $40^{\circ}C$ on soluble starch. This species resulted in the highest enzyme activity (mU/ml) of 26.06, 10.89, 58.82, 14.2, and 54.67 with the RB, BP, CS, CSCW, and CMCW fermentation media, respectively. The results indicate that CS can be used as a carbon substrate and CMCW can be used to accelerate the fermentation by T. ethacetica. The enzyme was partially purified by 40-60% ammonium sulphate fraction, and it showed total enzyme activity, total protein content, specific activity, purification fold, and a recovery of 2400 mU, 30 mg, 80 mU/mg, 2.7, and 71.1%, respectively. The molecular mass of the T. ethacetica alpha amylase was estimated on SDS-PAGE, and two bands around 50 kDa and 70 kDa were identified. The present study implies that T. ethacetica can produce alpha amylase, and it can be used to hydrolyze raw starch during the fermentation processes.

• Applied Biological Chemistry
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• 제27권1호
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• pp.52-54
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• 1984

Ethanol fermentation from the chemically gelatinized starchy material was examined. The critical concentration of sodium hydroxide solution for gelatinization was dependent on the species of starch; 0.4M for potato and 0.6M for tapioca at room temperature. For alcohol fermention the starchy material was gelatinized by addition of sodium hydroxide solution, neutralized by sulfuric acid, and then yeast was added. The amount of $CO_2$ evolved during ethanol fermentation indicates that non-fermentable material was not produced from the starch by chemical gelatinization. In ethanol fermentation of potato and tapioca starch no significant difference was observed between the thermal and the chemical gelatinization.

• Journal of Microbiology and Biotechnology
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• 제11권1호
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• pp.13-16
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• 2001

Maltose and soluble starch were used as secondary sources of carbon for glucoamylase production by Aspergillus awamori in solid state fermentation. During batch cultivation, maltose above 2.5%(w/w) repressed glucoamylase production, but, by adding either 2.5% (w/w) maltose or 1.25% (w/w) soluble starch to fed-batch cultivations, glucoamylase activity was increased by 15% and 170% over standard medium, respectively. The data showed that maltose is a weak inducer of glucoamylase production in solid stat fermentation.

• Journal of Microbiology and Biotechnology
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• 제4권1호
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• pp.7-12
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• 1994

A yeast strain secreting glucoamylase was transformed with an expression vector (pMS12) containing the promoter of yeast alcohol dehydrogenase I gene ADC1, mouse salivary $\alpha$-amylase cDNA, and a segment of yeast $21\mu m$ plasmid. The transformed strain could produce ethanol from starch (4%, w/v) through a direct one-step process with the conversion efficiency of 93.2%, during 5 days of fermentation, while the original, untransformed strain exhibited a conversion efficiency of 38.1% under the same condition. When the regulatory site of the ADC1 promoter region was removed, the production of ethanol increased to 29~37% in the presence of exogenous 3%(v/v) ethanol in the fermentation medium.

• Mycobiology
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• 제34권3호
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• pp.128-130
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• 2006

The solid waste of sago industry using cassava was fermented by Aspergillus niger, Aspergillus terreus and Rhizopus stolonifer in solid state fermentation. Cassava waste contained 52 per cent starch and 2.9 per cent protein by dry weight. The amylase activity was maintained at a high level and the highest amylase activity was observed on the $8^{th}$ day in R. stolonifer mediated fermentation. R. stolonifer was more efficient than Aspergillus niger and Aspergillus terreus in bioconverting cassava waste into fungal protein (90.24 mg/g) by saccharifying 70% starch and releasing 44.5% reducing sugars in eight days of solid state fermentation.

• Asian-Australasian Journal of Animal Sciences
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• 제2권3호
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• pp.386-387
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• 1989

• 한국미생물·생명공학회지
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• 제27권1호
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• pp.62-69
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• 1999

Clostridium butyricum NCIB 9576 evolved hydrogen gas and produced various organic acids from glucose, lactose, starch, and glycerol. Total amount of hydrogen gas produced from 1 and 2% glucose were 630 and 950ml $H_2$/l-broth, respectively, for the first 24 hrs of incubation and the maximum hydrogen production rates were 42 and 94ml $H_2$/hr/1-broth, respectively. Teh initial pH 6.8 decreased to 4.2~4.5 during the first 12~16 hrs of fermentation when the pH was not controlled, resulting in ceasing the cell growth and hydrogen evolution and in degradation of 82 and 40% glucose after 24hrs of incubation from 1 and 2% glucose, respectively. When pH was controlled to 5.5, glucose was consumed completely and resulted in increasing hydrogen production approximately 38~50% compared to the experiments without the pH control. C. butyricum NCIB 9576 produced hydrogen gas approximately 644, 1,700 and 3,080 ml $H_2$/l-broth with 0.5, 1 and 2% lactose, respectively and the maximum hydrogen production rates were 41, 141 and 179ml $H_2$/hr/l-broth, respectively. All of the lactose added was degraded completely during fermentation even though pH was not controlled. C. butyricum NCIB 9576 produced 183 and 709ml $H_2$/l-broth with 0.1 and 0.5% starch for 48 hrs, respectively, when pH was not controlled. The maximum rates of hydrogen gas production were 43 and 186ml $H_2$/l-broth, respectively and 80~100% of starch added was fermented. Approximately 107ml $H_2$/l-broth was produced using 1% glycerol by C. butyricum NCIB 9576 and the pH was maintained higher than 6.1 during fermentation without pH control. The degradation of glucose, lactose, starch and glycerol by C. butyricum NCIB 9576 were affected by the pH of fermentation broth and the organic acids released during fermentation. The pH of feremtntation broth dropped to 4.2~4.6 after 12~14 hrs incubation when glucose was used as a substrate while pHs were maintained above pH 5 under the same experimental conditions when lactose, starch and glycerol were used. The organic solvents and acids produced during glucose fermentation were mainly ethanol, butyrate, acetate and a little of propionate, while butyrate was the main organic acids during the lactose, starch, and glycerol fermentation by C. butyricum NCIB 9576.

• 한국수소및신에너지학회논문집
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• 제13권4호
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• pp.321-329
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• 2002

Clostridium butyricum, Lactobacillus amylophillus, Lactobacillus amylovorus, Lactobacillus acidophillus, AI-9 produced hydrogen and /or organic acids using glucose, lactose and starch at the anaerobic culture conditions. Cl. butyricum NCIB 9576 evolved 1,700 ml H2/L-culture broth and accumulated butyric acid, acetic acid, propionic acid and ethanol in its culture broth when lactose was used as a carbon source during 24 hrs of fermentation. L. amylovorus ATCC 33620 accumulated lactic and acetic acids and some reducing sugars when starch was used as a carbon source without hydrogen production. Instead of starch as a carbon source, L. amylovorus ATCC 33620 produced lactic acid from algal biomass during fermentation and the acid-heat or freeze-thaw pretreatment of algal biomass accelerate the lactic acid fermentation.