• Title/Summary/Keyword: Saccharification

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Extraction and Application of Bulk Enzymes and Antimicrobial Substance from Spent Mushroom Substrates

  • Lim, Seon-Hwa;Kwak, A Min;Min, Kyong-Jin;Kim, Sang Su;Kang, Hee Wan
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.19-19
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    • 2014
  • Pleurotus ostreatus, P. eryngii, and Flammulina velutipes are major edible mushrooms that account for over 89% of total mushroom production in Korea. Recently, Agrocybe cylindracea, Hypsizygus marmoreus, and Hericium erinaceu are increasingly being cultivated in mushroom farms. In Korea, the production of edible mushrooms was estimated to be 614,224 ton in 2013. Generally, about 5 kg of mushroom substrate is needed to produce 1 kg of mushroom, and consequently about 25 million tons of spent mushroom substrate (SMS) is produced each year in Korea. Because this massive amount of SMC is unsuitable for reuse in mushroom production, it is either used as garden fertilizer or deposited in landfills, which pollutes the environment. It is reasonably assumed that SMS includes different secondary metabolites and extracellular enzymes produced from mycelia on substrate. Three major groups of enzymes such as cellulases, xylanases, and lignin degrading enzymes are involved in breaking down mushroom substrates. Cellulase and xylanase have been used as the industrial enzymes involving the saccharification of biomass to produce biofuel. In addition, lignin degrading enzymes such as laccases have been used to decolorize the industrial synthetic dyes and remove environmental pollutions such as phenolic compounds. Basidiomycetes produce a large number of biologically active compounds that show antibacterial, antifungal, antiviral, cytotoxic or hallucinogenic activities. However, most previous researches have focused on therapeutics and less on the control of plant diseases. SMS can be considered as an easily available source of active compounds to protect plants from fungal and bacterial infections, helping alleviate the waste disposal problem in the mushroom industry and creating an environmentally friendly method to reduce plant pathogens. We describe extraction of lignocellulytic enzymes and antimicrobial substance from SMSs of different edible mushrooms and their potential applications.

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Isolation and Identification of Cellulomonas fimi, Characteristics of its Cellulase and Conversion of the Sawdust into Ethanol (Cellulomonas fimi의 분리(分離) 및 동정(同定), cellulase 특성(特性)과 톱밥의 Ethanol 전환(轉換))

  • Lee, Chan-Yong;Lee, Ke-Ho
    • 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.

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Optimization of Cellulolytic Enzyme Production for newly isolated Bacillus sp. H9-1 from Herbivore Feces (초식동물 배설물로부터 분리한 Bacillus sp. H9-1의 섬유소 분해효소생산 최적화)

  • Yoon, Young Mi;An, Gi Hong;Kim, Jung Kon;Cha, Young-Lok;Park, Yu Ri;Ahn, Jong-Woong;Moon, Youn-Ho;Ahn, Seung-Hyun;Koo, Bon-Cheol;Park, Kwang-Geun
    • KSBB Journal
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    • v.28 no.1
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    • pp.42-47
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    • 2013
  • This study was performed to find cellulolytic strain of enzymatic saccharification for bioethanol production. Cellulolytic strains were isolated from 59 different feces of herbivores from Seoul Grand Park located in Gwacheon Gyeonggi-Do. The celluloytic strain was selected by congo red staining and DNS method. Among the isolated strains, H9-1 strain isolated from the feces of rabbit has the highest CMCase activity. H9-1 strain was identified as Bacillus sp. based on 16S rDNA gene sequencing. The optimal conditions for CMCase activity by Bacillus sp. H9-1 were at $40^{\circ}C$ and at initial pH 8.

Studies on Enzymic Sources and Method of effective Addition in Fermentation of Yack-Tack-Joo Korean liquors (탁약주 제조에 있어서의 발효원 및 그의 효율적 첨가방법에 관한 연구)

  • 이성범
    • Korean Journal of Microbiology
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    • v.5 no.2
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    • pp.43-54
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    • 1967
  • The characteristics of enzymic sources and its effective uses concerned with brewing of Yack-Tack Joo which is traditional and popular liquors for all Korean have been studied. Results obtained are as follows; 1)Kock Ja (enrich of fungi and yeast produced in Korean brewery) is found to be weak in its liquifying(600U.) and saccharifying activity(1300U.), so that it is useful to conbine two factors effectively for better brewing. 2) The additional ratio of Kok Ja per materials is seems proper at line of 20 percent for better fermentation and the enzymic preparations inoculated of microorganisms in wheat bran is seems proper at 25 percent line. 3) Adding the enzymic preperation in which the strain Rhyzopus had been inoculated to the experimental mash at 5 percent per material, the rate of fermentation was revealed highest degree than those of else. 4) It is not proper to add a single Bun Kok in fermentation, as it produce much acid in mash during brewing. 5) However, the enzymic preparation composed of Asp usami and Rhyzopus sp. produced less acid in brewing. 6) The increasing of temparature in enzymic samples, temparatures of the mixtured Kuk(Kok Ja and enzymic preparation) are higher than those of single addition at the first stage in pre-fermentation, but there are no differences at the late stage of post-fermentation. 7) Amount of amino acids in the plot of enzymic prepation are found much more than those of single use at late stage of post-fermentation. In the plot of single use of Kock Ja, the amount was the most than else, the proteinase activity is strongest more than else. 8) In the brewing of Korean Tack-Yack-Joo, it is desirable less amount of acidity, more amount of amino acid, stronger liquifaction of starch and vigorous saccharification. Thren it was found that the application of two prepations(Kock Ja and Bun kok) is most effective to get moderate quality in Tack-Yack-Joo brewing.

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Comparison of Bio-ethanol Productivity Using Food Wastes by Various Culture Modes (에탄올 발효방법에 따른 음식물류 폐기물의 바이오에탄올 생산성 비교)

  • Kang, Hee-Jeong;Li, Hong-Xian;Kim, Yong-Jin;Kim, Seong-Jun
    • KSBB Journal
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    • v.25 no.5
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    • pp.471-477
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    • 2010
  • In order to improve bio-ethanol productivity by various cultivation methods in this paper, the culture modes using food wastes, such as batch culture, high-cell-density fermentation, SSF (simultaneous saccharification and fermentation) by fill & draw, continuous culture by fill & draw were performed and their productivities were compared. SSFs by fill & draw were performed by continuous decompression using 1 L evaporator system, and by 10 L bioreactor without decompression. In addition, the continuous cultures by fill & draw mode using SFW (saccharafied food wastes) medium were performed by changes of 40% culture broth with intervals of 12 h (0.03 $h^{-1}$), 6 h (0.07 $h^{-1}$), 3 h (0.13 $h^{-1}$). Consequently, productivities of bio-ethanol were 2.52 g/L-h and 1.30 g/L-h in batch culture and high- cell-density fermentation, respectively. The productivities of SSF by fill & draw showed 2.24 g/L-h and 2.03 g/L-h in continuous decompression with 1 L evaporator and 10 L bioreactor without decompression, respectively. Also, the productivities in continuous culture by fill & draw modes showed 2.02 g/L-h, 4.07 g/L-h and 6.25 g/L-h by medium change with intervals of 12 h, 6 h, and 3 h, respectively. In conclusion, the highest ethanol productivity was obtained in the continuous culture mode by fill & draw with dilution rate of 0.13 $h^{-1}$.

Construction of a Thermotolerant Saccharomyces cerevisiae Strain for Bioethanol Production with Reduced Fermentation Time and Saccharifying Enzyme Dose

  • Lim, Ji Sung;Jang, You Ri;Lim, Young Hoon;Kim, Keun
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1401-1405
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    • 2012
  • A thermotolerant Saccharomyces cerevisiae mutant strain, TT6, was constructed after multi-parental hybridization of five mutant strains obtained by UV or NTG treatment of the original strain, S. cerevisiae KV1. When incubated at $40^{\circ}C$ in YPD broth, TT6 began to grow exponentially in 10 h, but KV1 did not show any noticeable growth even after 22 h. The thermotolerant growth of TT6 was confirmed by serial dilution assay at $42^{\circ}C$; TT6 grew at a cell concentration ($10^{-5}$) 10,000 times lower than that of KV1 ($10^{-1}$). Whereas ethanol production from YP containing 23% (w/v) glucose by KV1 decreased with increasing temperature from $30^{\circ}C$ to $36^{\circ}C$, ethanol production by TT6 did not decrease at temperatures up to $37^{\circ}C$. When TT6 was tested for ethanol production at $36^{\circ}C$ by simultaneous saccharification and fermentation (SSF) from 23% corn, 24 h of fermentation time or 50% of the glucoamylase dose was saved when compared with KV1 at $30^{\circ}C$. The ethanol yield from corn by SSF with TT6 at $36^{\circ}C$ was 91.7% of the theoretical yield, whereas that of KV1 at $30^{\circ}C$ was 90.6%.

Identification of Off-Flavor in Sea Mustard and Rice Syrup Sold in the Markets (시판 미역 및 쌀엿의 이취성분(異臭成分) 규명)

  • Kim, Hyeon-Wee;Lee, Yoon-Kyoung;Shim, Gun-Sub;Chang, Youn-Kyung
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.728-732
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    • 1998
  • The purpose of this study was to identify off-flavor compounds in sea mustard and rice syrup sold in the markets. Naphthalenes such as naphthalene, 2-methylnaphthalene, 1-methylnaphthalene, 2,6-dimethylnaphthalene, 1,5-dimethylnaphthalene, 1,8-dimethylnaphthalene, 2,7-dimethylnaphthalene, 1,4,6-trimethylnaphthalene, 2,3,6-trimethylnaphthalene etc., were present in sea mustard, while free fatty acids such as butanoic acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, dodecanoic acid, tetradecanoic acid and 2-furanmethanol, 2-furancarboxaldehyde etc. in rice syrup. The former (naphthalenes) have been supposed to be contaminents from paint of ship and the latter (free fatty acids) derived from deteriorated rice for saccharification. From the results of the samples studied, formation of their off-flavor compounds seems to be related with the condition of storage, the process of production and circulation in the markets.

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Improvement of Cheongju Manufacturing Process Using Gelatinized Rice and Zeolite (팽화미분과 zeolite를 이용한 청주 제조공정의 개선)

  • Seo, Min-Jae;Ryu, Sang-Ryeol
    • Korean Journal of Food Science and Technology
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    • v.34 no.4
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    • pp.610-616
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    • 2002
  • In order to improve a complicated Cheongju manufacturing process, saccharification process with gelatinized rice flour was employed during a Cheongju fermentation. High sugar content without unsaccharified residue appeared to impede the yeast growth and fermentation. To solve this problem, addition of zeolite to the saccharifying solution containing 20% (w/v) sugar and fed-batch system were employed. These adjustments resulted in a increase of yeast viability and 40% time-saving alterations of fermentation. The Cheongju, having 18% (v/v) of ethanol content and fresh and rich flavor, could be made in 12 days. Yeast cells recovered from the fermentation precipitates could be reused up to four times without any adverse effect on cell viability, alcohol production, and flavor of the product. The complicated conventional brewing process of Cheongju can thus be simplified effectively.

Simultaneous Overpexpression of Genes Encoding Cellulose- and Xylan-Degrading Enzymes through High Density Culture of a Recombinant Yeast Cell (재조합 효모 세포의 고농도배양을 통한 섬유소와 자일란 분해효소 유전자의 동시 과발현)

  • Kim, Yeon-Hee;Heo, Sun-Yeon;Kim, Gun-Do;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.46 no.4
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    • pp.390-394
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    • 2018
  • For the coexpression of endoxylanase and endoglucanase genes in yeast Saccharomyces cerevisiae, the genes were separately inserted downstream of the yeast ADH1 promoters, resulting the plasmid pAGX3 (9.83 kb). In the batch culture on YPD medium of the yeast transformant, S. cerevisiae SEY2102/pAGX3, the total activities of the enzymes reached about 7.91 units/ml for endoxylanase and 0.43 units/ml for endoglucanase. In the fed-batch culture with intermittent feeding of yeast extract and glucose, the total activities of 24.9 units/ml for endoxylanase and 0.84 units/ml for endoglucanase were produced which were about 3.1-fold and 2.0-fold increased levels, respectively, compared to those of the batch culture. Most of endoxylanase and endoglucanase activities were found in the extracellular media. This recombinant yeast could be useful for the development of simultaneous saccharification bioprocess of the cellulose and xylan mixture.

Characterization of the Starch Degradation Activity of recombinant glucoamylase from Extremophile Deinococcus geothermalis (극한성 미생물Deinococcus geothermalis 유래 재조합 글루코아밀레이즈의 전분 분해 활성 특징)

  • Jang, Seung-Won;Kwon, Deok-Ho;Park, Jae-Bum;Jung, Jong-Hyun;Ha, Suk-Jin
    • 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.