• Title/Summary/Keyword: chitin pretreatment

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Isolation and Purification of Chitin from Shrimp Shells by Protease Pretreatment (Protease의 전처리에 의한 새우껍질로부터 키틴의 분리와 정제)

  • Ryu, Beung-Ho;Lee, Sang-Hoon
    • Korean Journal of Food Science and Technology
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    • v.27 no.1
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    • pp.6-10
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    • 1995
  • Chitin was prepared from Solenocera prominentis by deproteinization pretreatment of Neutrase. The optimal enzyme concentration of neutrase, pH, and temperature on deproteinization were 3.0 mg/ml, pH 6.0 and $50^{\circ}C$ as indicated by the minimum protein remaining on the chitin. The residual protein, the degree of deacetylation, Ca and P content in chitin prepared from Solenocera prominentis were similar with commercial chitin. The molecular weight was $1.2{\times}10^{8}$ dalton and the yield of chitin was 25.8%.

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Detection of Zymogenic ChsC Activity in Vegetative Hyphae of Aspergillus nidulans. (Aspergillus nidulans 영양균사에서 효소전구체형 ChsC 활성의 검출)

  • 박범찬;박윤희;박희문
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.178-182
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    • 2004
  • In the vegetative hyphae of Aspergillus nidulans, a zymogenic form of the class I chitin synthase activity was successfully measured by the assay condition for Saccharomyces cerevisiae class I chitin synthase, Chsl. The class I chitin synthase activity of the A. nidulans chsC wild type strain was increased about six-fold by trypsin-pretreatment, but that of the chsC disruption strain revealed no increase. Interestingly enough, level of the class I chitin synthase activity of the chsC disruption strain was almost the same as that of the chsC wild type without trypsin-pretreatment. These results indicated that the A. nidulans ChsC activity could be measured by account-ing the class I chitin synthase activity without the trypsin-pretreatment as an internal control. Consistence to the expression pattern of the chsC revealed by northern blot analysis, the activity of ChsC was increased upon reaching the culture time for acquiring developmental competence. Our results shown here also supported the previous report suggesting the possible involvement of ChsC in vegetative hyphal growth of A. nidulans.

Enzymatic Hydrolysis of Pretreated Chitin by Aspergillus carneus Chitinase

  • Mohamed, Abdel-Naby;Kwon, Dae-Young
    • Journal of Microbiology and Biotechnology
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    • v.2 no.3
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    • pp.197-203
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    • 1992
  • Studies of the pretreatment of chitin and its subsequent hydrolysis by Aspergillus carneus chitinase are reported. Ball milling was found to be the most effective way among the pretreatment methods tested. Data are presented describing the effect of enzyme and substrate concentrations on the rate and extent of the hydrolysis process. It was found that the successive addition of enzyme improved the saccharification yield. Significant product inhibition of the chitinase was observed when N-acetylglucosamine concentration was 3.6% or higher. Adsorption of enzymes to the substrate occurred during a 24 hr hydrolysis period. An initial rapid and extensive adsorption occurred, followed by gradual desorption which increased during the time of reaction. Intermediate removal of the hydrolyzate and continuation of the hydrolysis by adsorbed enzyme on the residual chitin was also investigated. A total of 75.4 g/l reducing sugars, corresponding to 69.2% saccharificaton yield (as N-acetylglucosamine) was obtained. In addition an increase in the amount of recoverable enzymes was observed under these conditions. Evidence presented here suggests that the technique, whereby the free enzymes in the recovered hydrolyzate are re-adsorbed onto the new substrate, may provide a means of recirculating the dissolved enzymes.

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Effective Production of N-Acetyl-$\beta$-glucosamine by Serratia marcescens Using Chitinadceous Waste

  • Kim, Kwang;A. Louise Creagh;Charles A. Haynes
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.2
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    • pp.71-77
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    • 1998
  • The strain of Serratia marcescens QM B1466 produces selectively large amount of chitinolytic enzymes (about 1mg/L medium). Enzymatic hydrolysis of chitin to N-acetyl-${\beta}$-D-glucosamine (NAG) was performed with a system consisting of two hydrolases (chitinase and chitobiase) produced by optimization of a microbial host consuming chitin particles. For the development of Large-scale biological process for the production of NAG from chitinaceous waste, the selection and optimization of a microbial host, particle size of crab/shrimp chitin sources and initial induction time using chitin as a sole carbon source on chitinase/chitobiase production and NAG production were examined. Crab-shell chitin(1.5%) treated by dilute acid and , ball-milled with a normal diameter less than 250m gave the highest chitinase activity over a 7 days culture. Crude chitinase/ chitobiase solution obtained in a 10 L fed-batch fermentation showed a maximum activities of 23.6 U/mL and 5.1 U/mL, respectively with a feeding time of 3 hrs, near pH 8.5 at 30$^{\circ}C$.

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Novel Heterogeneous Carbohydrase Reaction Systems for the Direct Conversion of Insoluble Carbohydrates: Reaction Characteristics and their Applications

  • Lee, Yong-Hyun;Park, Dong-Chan
    • Journal of Microbiology and Biotechnology
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    • v.9 no.1
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    • pp.1-8
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    • 1999
  • Most carbohydrates exist in nature in an insoluble state, which reduces their susceptibility towards various carbohydrases. Accordingly, they require intensive pretreatment for structural modification to enhance an enzyme reaction. The direct conversion of insoluble carbohydrates has distinct advantages for special types of reaction, especially exo-type carbohydrase; however, its application is limited due to structural constraints. This paper introduces two novel heterogeneous enzyme reaction systems for direct conversion of insoluble carbohydrates; one is an attrition coupled enzyme reaction system containing attrition-milling media for enhancing the enzyme reaction, and the other is a heterogeneous enzyme reaction system using extruded starch as an insoluble substrate. The direct conversion of typically insoluble carbohydrates, including cellulose, starch, and chitin with their corresponding carbohydrases, including cellulase, amylase, chitinase, and cyclodextrin glucanotransferase, was carried out using two proposed enzyme reaction systems. The conceptual features of the systems, their reaction characteristics and mechanism, and the industrial applications of the various carbohydrates are analyzed in this review.

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Natural Dyeing of Chitosan Nonwoven Fabric (키토산 부직포의 천연염색)

  • Kim Jong-Jun;Kwon Min-Soo;Jeon Dong-Won
    • The Research Journal of the Costume Culture
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    • v.12 no.6 s.53
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    • pp.999-1009
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    • 2004
  • Chitin is a derived product from the shell of shrimp or crab. Chitosan, a deacetylated product of chitin, has widely been used in the biomedical sector, food industry, and textile industry. Chitosan exhibits fiber-forming property under certain conditions. Nonwoven fabrics made of chitosan fibers may have diverse applications in the industry. Previous studies have revealed that the dye uptake properties of natural dyestuffs improved by the chitosan pretreatment on the fabric specimens. In this case, fabric specimen is coated with acidic salt form of chitosan, which is different from the pure chitosan, since the coating process employes coating with the acidic solution of the chitosan and subesquent drying. In this study, chitosan nonwoven fabric samples were prepared from chitosan sample having deacetylation degree of $100\%$ and molecular weight of 650,000. Chitosan nonwoven fabrics maintain the form of $-NH_2$ end-group. These in turn exhibit higher dye uptake ability than the fabrics coated with chitosan acidic solutions do.

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Intein-mediated expression of Trichoderma reesei Cellobiohydrolase I Cellulose Binding Domain in E. coli (Intein을 이용한 대장균에서의 Trichoderma reesei 유래의 Cellobiohydrolase I 섬유소 결합 도메인의 발현)

  • Choi, Shin-Geon
    • Journal of Industrial Technology
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    • v.36
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    • pp.33-37
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    • 2016
  • Cellulose binding domains (CBDs) of cellulases are thought to assist in the hydrolysis of insoluble crystalline cellulose. To gain sufficient amount of CBDs, the self-cleavable intein tag was used for expression and purification of Trichoderma reesei cellobiohydrolase I CBD in E. coli. Synthetic CBD genes, CBD or linker-CBD were cloned into expression vector pTYB11. Recombinant CBDs were successfully purified by intein mediated purification with an affinity chitin-binding domain. The final yields of recombinant CBD and linker-CBD were 3.2 mg/L and 1.4 mg/L, respectively. The functional bindings of recombinant CBDs were confirmed by Avicel binding experiments. The simple and easy purification method using self-cleavable intein tag can be further used in pretreatment of crystalline cellulose or characterization of engineered CBDs.

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Eliminatory Effect of Mixture including Hot Water Extract of Dendropanax morbifera Lev. on Alcohol-induced Blood Alcohol Concentration and Hangover in Rat (흰쥐에서 황칠나무 열수 추출물을 포함한 혼합물의 혈중 알코올 농도와 숙취 해소 효과)

  • Na, Ju-ryun;Kim, Eun;Park, Soyi;Lee, Kihoon;Jeong, Eui-seon;Kim, Jinseok;Kim, Youngjae;Kim, Sunoh
    • Journal of Chitin and Chitosan
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    • v.23 no.4
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    • pp.267-276
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    • 2018
  • This study was performed to investigate the ameliorating effect of a hangover beverage mixture (SBJ) that contains Dendropanax morbifera Lev. and several medicinal plant extracts, on hepatoprotection and alcohol-metabolizing enzymes in alcohol-induced hangover in both in vitro and in vivo models. In human hepatoma cell line, HepG2, 300 mM of ethanol-induced hepatotoxicity was significantly improved by pretreatment of SBJ by dose-dependent manner. In the in vivo study, administration of alcohol to rats raised to the concentration of blood alcohol and lactate dehydrogenase (LDH). Blood alcohol and LDH levels in SBJ-treated rats significantly decreased at 0.5 h and 8 h after acute ethanol administration (40%, 4.6 g/kg body weight) as compared to alcohol-treated rats. Hepatic alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activity were significantly higher in SBJ-treated rats than in alcohol-treated rats. SBJ supplementation reduced formation of malondialdehyde (MDA), and inhibited reductions of hepatic superoxide dismutase (SOD), hepatic glutathione (GSH), glutathione-S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx) levels, compared with rats administered alcohol. Plasma catalase (CAT), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels showed unaltered resulted in all experimental groups compared with the control group. These results suggest that SBJ exhibit hepatoprotective properties by enhancing ADH, ALDH activity and stimulating the antioxidant defense system in alcohol-induced hangover.