• Title/Summary/Keyword: Ethanol production from starch

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Genome Analysis and Optimization of Caproic Acid Production of Clostridium butyricum GD1-1 Isolated from the Pit Mud of Nongxiangxing Baijiu

  • Min Li;Tao Li;Jia Zheng;Zongwei Qiao;Kaizheng Zhang;Huibo Luo;Wei Zou
    • Journal of Microbiology and Biotechnology
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    • v.33 no.10
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    • pp.1337-1350
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    • 2023
  • Caproic acid is a precursor substance for the synthesis of ethyl caproate, the main flavor substance of nongxiangxing baijiu liquor. In this study, Clostridium butyricum GD1-1, a strain with high caproic acid concentration (3.86 g/l), was isolated from the storage pit mud of nongxiangxing baijiu for sequencing and analysis. The strain's genome was 3,840,048 bp in length with 4,050 open reading frames. In addition, virulence factor annotation analysis showed C. butyricum GD1-1 to be safe at the genetic level. However, the annotation results using the Kyoto Encyclopedia of Genes and Genomes Automatic Annotation Server predicted a deficiency in the strain's synthesis of alanine, methionine, and biotin. These results were confirmed by essential nutrient factor validation experiments. Furthermore, the optimized medium conditions for caproic acid concentration by strain GD1-1 were (g/l): glucose 30, NaCl 5, yeast extract 10, peptone 10, beef paste 10, sodium acetate 11, L-cysteine 0.6, biotin 0.004, starch 2, and 2.0% ethanol. The optimized fermentation conditions for caproic acid production by C. butyricum GD1-1 on a single-factor basis were: 5% inoculum volume, 35℃, pH 7, and 90% loading volume. Under optimal conditions, the caproic acid concentration of strain GD1-1 reached 5.42 g/l, which was 1.40 times higher than the initial concentration. C. butyricum GD1-1 could be further used in caproic acid production, NXXB pit mud strengthening and maintenance, and artificial pit mud preparation.

Isolation and Characterization of Cyclodextrin Glycosyl Transferase Producing Alkalophilic Bacillus sp. (Cyclodextrin glycosyltransferase를 생산하는 호알칼리성 Bacillus속 미생물)

  • 유주현;정용준;이정수
    • Microbiology and Biotechnology Letters
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    • v.17 no.2
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    • pp.148-153
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    • 1989
  • A strain of alkalophilic Bacillus sp. YC-335 has been isolated from soil. The strain was capable of producing large amount of cyclodextrin glycosyl transferase (CGTase) in the culture broth. The preferable medium composition has been determined to be as follows : 1.5% soluble starch, 5% corn steep liquor, 0.1% $K_2$HPO$_4$, 0.02%mgSO$_4$.7$H_2O$, 1% CaCO$_3$and 1% Na$_2$CO$_3$(pH 10.3). The highest enzyme production was observed after 48 hours of cultivation at 31$^{\circ}C$. The optimum pH and temperature for the activity of crude enzyme were 6.0 and 5$0^{\circ}C$, respectively. The enzyme was stable between pH 5 and 9, and upto 5$0^{\circ}C$. The enzyme converted starch into $\alpha$-, $\beta$- and ${\gamma}$-CD in the relative amounts of 1:10:1.5, respectively.

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Fermentation Characteristics of Takju Prepared with Lotus Leaf (연잎을 첨가한 탁주의 발효 특성)

  • Yoo, Ha-Na;Chung, Chang-Ho
    • Korean journal of food and cookery science
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    • v.27 no.5
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    • pp.577-587
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    • 2011
  • In this study, lotus leaf was added to a Takju (Korean milky rice wine) preparation to evaluate its effects on physicochemical and fermentation properties. For rapid saccharification Takju was stored at 25$^{\circ}C$ for 2 days and then temperature was switched to 18$^{\circ}C$ to mature the wine for 21 days. From the start of fermentation to 2-days, the pH of Takju decreased rapidly and acidity increased. Maltose, a byproduct of starch saccharification, increased for 2 days then rapidly decreased. Glucose fluctuated for 7 days and of the 21-days fermentation. The ethanol production rate was highest during the first 7 days, then slowed. Total viable yeast and lactic acid bacterial counts increased rapidly for 2 days and then decreased gradually thereafter. Leuconostoc spp. grew rapidly for 1 day and sharply disappeared with decreasing pH. DPPH radical scavenging activity was significantly higher for Takju prepared with lotus leaf than without. The overall acceptance of Takju tended to increase when it was prepared with lotus leaf.

Liquefaction and Saccharification of Tapioca Starch for Fuel Ethanol Production (연료용 알콜 생산을 위한 타피오카 전분의 액화 및 당화)

  • 김기호;박성훈
    • KSBB Journal
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    • v.10 no.3
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    • pp.304-316
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    • 1995
  • For fuel alcohol production, enzymatic liquefaction and saccharification of tapioca starch by ${\alpha}$-amylase and glucoamylase were studied. The thermophilic ${\alpha}$-amylase Termamyl produced from Bacillus licheniformis gave a better liquefaction than the relalively low temperature enzyme BAN from B. subtilis. Oplimal temperature and pH with Termamyl were $90∼95^{\circ}C$ and 5.8, respectively. Minimal amount of Termamyl 240uc for a satisfactory liquefaction for a two-hour reaction was about 0.0125% (v/w) with respect to the mass of tapioca used. For saccharification experiments two enzymes, Novo AMG and Do-I1 enzymes were compared. The enzymatic activity of each enzyme was a little different depending on the substrate used and the latter was found to have a significant amount of ${\alpha}$-amylase activity. With Novo AMG optimal temperature was about $58^{\circ}C$ The pH optimum was 4.3 with maltose, however, with tapioca, no difference was observed between pH 4.3 and 5.7 which is a natural, unadjusted pH of liquefied tapioca. For 85% of completion of saccharification, it was necessary to use 0.0625% (v/w) of Novo AMG 400L for tapioca and to run the reaction for more than 10 hr, Packed volume of solid particles in tapioca slurry remained at around 30% during liquefaction and saccharification. This indicates that the removal of the solid particle before fermentation is not economically feasible at all, even though the solid particles make it very difficult to operate the bioreactor in a continuous mode with cell-recycle.

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Alcohol Fermentation at High Temperature and the Strain-specific Characteristics Required to Endow the Thermotolerance of Sacchromyces cerevisiae KNU5377

  • Paik, Sang-Kyoo;Park, In-Su;Kim, Il-Sup;Kang, Kyung-Hee;Yu, Choon-Bal;Rhee, In-Koo;Jin, In-Gnyol
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2005.06a
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    • pp.154-164
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    • 2005
  • Saccharomyces cerevisiae KNU5377 is a thermotolerant strain, which can ferment ethanol from wasted papers and starch at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. This strain showed alcohol fermentation ability to convert wasted papers 200 g (w/v) to ethanol 8.4% (v/v) at 40$^{\circ}C$, meaning that 8.4% ethanol is acceptable enough to ferment in the industrial economy. As well, all kinds of starch that are using in the industry were converted into ethanol at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. Hyperthermic cell killing kinetics and differential scanning calorimetry (DSC) revealed that exponentially growing cells of this yeast strain KNU5377 were more thermotolerant than those of S. cerevisiae ATCC24858 used as a control. This intrinsic thermotolernace did not result from the stability of entire cellular components but possibly from that of a particular target. Heat shock induced similar results in whole cell DSC profiles of both strains and the accumulation of trehalose in the cells of both strains, but the trehalose contents in the strain KNU5377 were 2.6 fold higher than that in the control strain. On the contrary to the trehalose level, the neutral trehalase activity in the KNU5377 cells was not changed after the heat shock. This result made a conclusion that though the trehalose may stabilize cellular components, the surplus of trehalose in KNU5377 strain was not essential for stabilization of whole cellular components. A constitutively thermotolerant yeast, S. cerevisiae KNU5377, was compared with a relatively thermosensitive control, S. cerevisiae ATCC24858, by assaying the fluidity and proton ATPase on the plasma membrane. Anisotropic values (r) of both strains were slightly increased by elevating the incubation temperatures from 25$^{\circ}C$ to 37$^{\circ}C$ when they were aerobically cultured for 12 hours in the YPD media, implying the membrane fluidity was decreased. While the temperature was elevated up to 40$^{\circ}C$, the fluidity was not changed in the KNU5377 cell, but rather increased in the control. This result implies that the plasma membrane of the KNU5377 cell can be characterized into the more stabilized state than control. Besides, heat shock decreased the fluidity in the control strain, but not in the KNU5377 strain. This means also there's a stabilization of the plasma membrane in the KNU5377 cell. Furthermore, the proton ATPase assay indicated the KNU5377 cell kept a relatively more stabilized glucose metabolism at high temperature than the control cell. Therefore, the results were concluded that the stabilization of plasma membrane and growth at high temperature for the KNU5377 cell. Genome wide transcription analysis showed that the heat shock responses were very complex and combinatory in the KNU5377 cell. Induced by the heat shock, a number of genes were related with the ubiquitin mediated proteolysis, metallothionein (prevent ROS production from copper), hsp27 (88-fold induced remarkably, preventing the protein aggregation and denaturation), oxidative stress response (to remove the hydrogen peroxide), and etc.

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Optimization of Thickness and Maturation Period of Andong-Soju Nuruk for Fermentation of Andong-Soju (안동소주 누룩 제조를 위한 누룩 두께 및 누룩 띄움시간의 최적화)

  • Bae, Kyung-Hwa;Ryu, Hee-Young;Kwun, In-Sook;Kwon, Chong-Suk;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.231-237
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    • 2007
  • To support the fermentational superiority of Korean nuruks and maintain the various domestic nuruks, the optimal nuruk production of Andong-Soju, which was designated as an intangible cultural asset of Gyungsangbukdo province from 1987, was investigated. Different thickness of nuruks ($2.2{\sim}5.5\;cm$) were manufactured based on traditional Andong-Soju nuruk method, while the size of round form of nuruk was set to 23 cm. During the 3 weeks maturation, changes of water content, weight, pH, brix, the amount of reducing sugar, sac-charifying activity, viable cell and major microorganisms were determined, Also, ethanol fermentation abilities of the manufactured nuruks were evaluated using 20% glucose medium or 16% starch medium, respectively. Our results indicated that the production of high quality of Andong-Soju nuruk needs $4.0{\sim}5.5\;cm$ thickness and 3 weeks maturation without extraneous yeast addition. These results would be applied to production of homogeneous, and high quality of Andong-Soju nuruk.

Anti-diabetic Effects of CCCA, CMESS, and Cordycepin from Cordyceps militaris and the Immune Responses in Streptozotocin-induced Diabetic Mice

  • Yun, Yun-Ha;Han, Shin-Ha;Lee, Seung-Jeong;Ko, Sung-Kwon;Lee, Chong-Kil;Ha, Nam-Joo;Kim, Kyung-Jae
    • Natural Product Sciences
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    • v.9 no.4
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    • pp.291-298
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    • 2003
  • Anti-diabetic effect of various fractions of Cordyceps militaris (CM), CCCA (crude cordycepin containing adenosine), CMESS (ethanol soluble supernatant), and cordycepin were evaluated in streptozotocin (STZ) induced diabetic mice, CMESS showed potent inhibitory activity of 34.7% in starch-loaded mice (2 g/kg) while acarbose as a positive standard exhibited 37.8% of inhibition rate. After 3 days administration (50 mg/kg), cordycepin (0.2 mg/kg), and acarbose (10 mg/kg) dramatically reduced blood glucose level (inhibition ratio: 46.9%, 48.4% and 37.5% respectively). CCCA that has high contents of cordycepin (0.656 mg/4 mg) did not influence on reducing blood glucose level. The proliferation of splenocytes and peritoneal macrophages derived from STZ-induced diabetic mice administered samples were evaluated out by addition of mitogens to see the stability of the usage of these herbal medicines. Proliferation of T-lymphocyte was significantly decreased; while NO production was increased more than two fold to STZ control in the cordycepin-administered group. Changes of serum enzyme levels of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) were also evaluated. Cordycepin administered group was appeared to acarbose. We conclude that CMESS and cordycepin may be useful tools in the control of blood glucose level in diabetes and promising new drug as an anti-hyperglycemic agent without defects of immune responses and other side effects.

Optimization of Culture Conditions for the Bioconversion of Vitamin $D_3\;to\;1{\alpha}$,25-Dihydroxyvitamin $D_3$ Using Pseudonocardia autotrophica ID9302

  • Kang, Dae-Jung;Lee, Hong-Sub;Park, Joon-Tae;Bang, Ji-Sun;Hong, Soon-Kwang;Kim, Tae-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.408-413
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    • 2006
  • We assessed the ability of a Pseudonocardia sp. from soil samples to bioconvert vitamin $D_3$. The optimal culture conditions for the bioconversion of vitamin $D_3$ to active $1{\alpha}$,25-dihydroxyvitamin $D_3$ were investigated by varying the carbon and nitrogen sources, the metal salt concentrations, the initial pH, and the temperature. Microbial transformations were carried out with the addition of vitamin $D_3$ dissolved in ethanol. They were sampled by extraction with methanol-dichloromethane and the samples were examined by HPLC. Optimum culture conditions were found to be 0.4% yeast extract, 1% glucose, 3% starch, 1% fish meal, 0.2% NaCl, 0.01% $K_2HPO_4$, 0.2% $CaCO_3$, 0.01% NaF, and pH 7.0 at $28^{\circ}C$. The optimal timing of the addition of vitamin $D_3$ for the production of calcitriol by Pseudonocardia autotrophica ID9302 was concurrent with the inoculation of seed culture broth. Maximum calcitriol productivity and the yield of bioconversion reached a value of 10.4mg/L and 10.4% respectively on the 7th day in a 75L fementer jar under the above conditions.

Studies on the Amylase Production by Bacteria (세균(細菌)에 의(依)한 Amylase생산(生産)에 관한 연구(硏究))

  • Park, Yoon-Joong
    • Applied Biological Chemistry
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    • v.13 no.2
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    • pp.153-170
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    • 1970
  • 1. Isolation and identification of amylase-producing bacteria. The powerful strain A-12 and S-8 were respectively isolated from air and soil after screening a large number of amylase-producing bacteria. Their bacterial characteristics have been investigated and it has been found that all characteristics of strain A-12 and S-8 are similar to Bac. subtilis of Bergey's manual except for the acid formation from a few carbohydrates and the citrate utilization, i.e., the strain A-12 shows negative in the citrate utilization, and the acid formation from arabinose and xylose, S-8 shows negative in the acid formation from xylose. 2. Amylase production by Liquid cultures with solid materials. Several conditions for amylase production by strain A-12 in stationary cultures have been studied. The results obtained are as follows. (1) The optimum conditions are:temperature $35^{\circ}C$, initial pH 6.5 to 7.0 and incubation time 3 to 4 days. (2) The amylase production is not affected by the preservation period of the stock cultures. (3) Among the various solid material, the defatted soy bean is found to be the best for t1e amylase production. However, the alkali treatment of the defatted soy bean gives no effect contrary to the cage of defatted rape seed. The addition of soluble starch to the alkali extract of defatted soy bean shows the increased amylase production. (4) Up to 1% addition of ethanol to carbon dificient media gives the improved amylase production, whereas the above effect is not found in the case of carbon rich media. (5) The amylase production can be increased 2.5 times when 10% of defatted soy bean is admixed to cheaply available wheat bran. (6) The excellent effect is found for amylase production when 20% of wheat bran is admixed to defatted dry milk which is a poor medium. The activity is found to be $D^{40^{\circ}}_{30'}$ 7,000(L.S.V. 1,800) in 10% medium. (7) No significant effect is observed due to the addition of various inorganic salts. 3. Amylase production by solid cultures. Several conditions for amylase production by strain A-12 in wheat bran cultures have been studied and the results obtained are as follows. (1) The optimum conditions: are temperature $33^{\circ}C$, incubation lime 2 days, water content added 150 to 175% and the thickness of the medium 1.5cm, The activity is found to be $D^{40^{\circ}}_{30'}$ 36,000(L.S.V. 15,000) (2) No significant effect is found in the case of the additions of various organic and inorganic substances.

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A New Sweetpotato Cultivar for Use of Bioethanol 'Daeyumi' (바이오에탄올용 고구마 신품종 '대유미')

  • Lee, Joon-Seol;Ahn, Young-Sup;Chung, Mi-Nam;Kim, Hag-Sin;Jeong, Kwang-Ho;Bang, Jin-Ki;Song, Yeon-Sang;Shim, Hyeong-Kwon;Han, Seon-Kyeong;Suh, Sae-Jung
    • Korean Journal of Breeding Science
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    • v.42 no.6
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    • pp.674-678
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    • 2010
  • A new sweetpotato variety, 'Daeyumi', was developed by Bioenergy Crop Research Center, National Institute of Crop Science (NICS), RDA in 2008. This variety was obtained from the cross between 'Jinhongmi' and 'Xusju 18' in 2000. The seedling and line selections were performed from 2001 to 2003, preliminary and advanced yield trials were carried out from 2004 to 2005, and the regional yield trials were conducted at six locations from 2006 to 2008. 'Daeyumi' has cordate leaf, green vine and petiole, elliptic storage root, red skin and yellow flesh color of storage root. This variety is also resistant to Fusarium wilt and nematode. The starch value was 25.9%, ethanol yield was 418 L/Ton, which was 7% higher than that of 'Yulmi' variety, and the total sugar content was 2.47 g/100g, dry weight. 'Daeyumi's initial temperature of starch gelatinization was lower, 76.2$^{\circ}C$, and the retrogradation process was earlier than 'Yulmi'. The average yield of storage root was 27.8 ton/ha in the regional yield trials, which was 36% higher than that of 'Yulmi' variety. Number of storage roots over 50 gram per plant was 3.0, and the average weight of one storage root was 152 gram. This variety can be used for the production of bioethanol and starch processing.