• KSBB Journal
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• v.13 no.2
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• pp.147-154
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• 1998

Enzymatic hydrolysis using an immobilized enzyme was carried out to produce chitosan oligosaccharides (COSs) from chitosan effectively. Chitosanase was immobilized on eight different carriers by physical adsorption. The enzyme immobilized on chitin had higher activity than those immobilized on the other carriers in spite of its lower adsorption. The activity of chitin-immobilized enzyme was more than 90% of the original activity. Optimal temperature of the immobilized enzyme increased by about $15^{\circ}C$ and its thermostability was excellent in relatively wide range of temperature. But its effects of pH did not improve compared to the free enzyme. The immobilized enzyme produced 153 mg/g chitosan of the reducing sugar for 3hrs of hydrolytic incubation time. The total content of higher oligomers, tetramer to hexamer, among amount of total COSs obtained for 2hrs was more than 90%. In kinetic parameters for both enzymes, immobilized enzyme showed lower affinity for substrate and reaction rate than free enzyme, however, no reduction of the rate for high substrate concentrations. Consequently, chitin-immobilized could effectively hydrolyse chitosan and produce the higher COSs without activity decrease in comparison with the free enzyme.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.96-97
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• 2000

최근 기능성 생리활성물질로서 각광을 받고있는 chitooligosaccharide의 제조방법은 염산, 황산 등을 이용한 화학적 분해법과 효소를 이용하는 생물학적 방법을 들 수 있다. 화학적 분해법은 비용을 적게들여 쉽게 저분자 올리고당을 만들 수 있으나, 올리고당의 수율이 낮고 저중합도의 올리고당이 많이 생성되며, 원료의 불안전성으로 인해 인체에 유해한 부반응 물질을 생성시키는 문제점이 보고되고 있다(Choi. et al.,1997). (중략)

• Korean Journal of Plant Tissue Culture
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• v.22 no.2
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• pp.95-99
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• 1995

We have previously established a system for plant regeneration through somatic embryogenesis and Agrobacterium-mediated transformation of Korean ginseng. In this study to produce a fungus-resistant plant, we introduced a bean chitinase gene into ginseng using the transformation system. A binary vector pChi/748 was constructed by introducing the bean basic chitinase gene into EcoRI site of pGA748 which carries the CaMV 35S promoter governing the introduced gene and neomycin phosphotransferase II(NPT-II)gene as a positive selection marker. Cotyledonary explants were cocultured with A. tumefaciens strain LBA4404 harboring the binary vertor pChi/748 for 48 h, and transferred to MS medium supplemented with l mg/L2,4-D,0.1mg/L kinetin, 100 mg/L kanamycin, and 500mg/L carbenicillin. Kanamycin-resistant calli were formed on the cut surface of cotyledonary explants after one month of culture, and subsequently they gave rise to somatic embryos. Upon transfer onto medium containing 1 mg/L each of BA and GA$_3$, most of them converted to plantlets after 5 weeks of culture. The genomic DNA of eight kanamycin-resistant regenerants was subjected to polymerase chain reaction (PCR) using two specific 21-mer oligonucleotides derived from the chitinase gene. PCR-Southern blot analysis confirmed that the chitinase gene was incorporated into six out of the eight regenerants..

• Applied Biological Chemistry
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• v.35 no.3
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• pp.191-195
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• 1992

In order to elucidate the plant-microorganism relationship, we purified an ethylene-inducible, basic 30 KD endochitinase from bean leaves and studied its antifungal activity by a hyphal extension-inhibition assay. The purified chitinase was effective in the inhibition of hyphal growth of Aspergillus fumigatus, Botrytis cinerea, Fusarium oxysporum, Rhizoctonia solani, while microbial chitinases of Serratia marcescens and Streptomyces griceus, egg white lysozyme and papya protease didn't affect hyphal growth of the fungi. The chitinase degraded the cell walls of Micrococcus lysodeikticus, suggesting the lysozyme activity of the chitinase. We discussed the implication of the bifunctional chitinase/lysozyme activities of the protein with hydrolysis of chitin in the rapidly extending hyphae of the fungi.

• Applied Biological Chemistry
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• v.37 no.4
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• pp.255-258
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• 1994

Using the enzyme activity staining, we studied the induction and distribution of chitinase isozymes, pathogenesis-related proteins, in intercellular fluids of bean and soybean leaves under stress conditions. The chitinase in intercellular fluids was barely detected in healthy plant leaves. By treatment of ethylene, pathogen (Fusarium oxysporum), or wounding, only 34 kD intercellular endochitinase was induced in bean leaves, while 30 kD and 36 kD intercellular endochitinases were induced in soybean leaves.

• Korean Journal of Soil Science and Fertilizer
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• v.39 no.1
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• pp.15-20
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• 2006

Biological control by chitinolytic microorganisms is being evaluated as management options for soilborne diseases. Forty kilograms of chitin compost (CTC) and control compost (CC) were amended on tomato plots ($15m{\times}0.5m$) 7 d before transplanting to evaluate enzymatic activities and the control of Fusarium wilt. Samples were taken on day 1, 3, 5, and 7, the day 1 corresponded to the 66 d after transplanting, the day on which the initial wilting symptoms occurred in plants of CC treated plots. The chitinase activity in soil of CTC was always higher compared to the control. Pathogenesis related (PR) protein (chitinase, ${\beta}$-1, 3-glucanase and peroxidase) activities in tomato roots in CC increased every day and showed marked differences compared to CTC. Wilting symptoms (96 d after transplanting) were reduced by 25% in CTC compared to the control. Protection of tomato plant may be correlated with the high levels of soil enzyme activities resulting from the chitin compost.

• Applied Chemistry for Engineering
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• v.19 no.5
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• pp.457-465
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• 2008

Development of various medical systems was accomplished through the progress of biotechnological method for therapy of human diseases. Furthermore, drug delivery systems have been investigated to carry the bioactive materials such as drug or gene in the body effectively. The most important thing in this system is to develop biomedical polymers having biocompatibility, biodegradability, and non-toxicity. Chitosan, a natural polymer, has been importantly considered as biomedical materials due to its good biocompatibility and various bio-active characteristics. Since the property of chitosan is differently explained according to the crystalline structures of chitin, the study for structural analysis of chitin has to proceed to apply as a biomaterial. From this point of view, this article introduced the analysis of crystalline structural of chitin, general property of chitosan and potential characteristics of low molecular weight water-soluble chitosan (LMWSC) as a biomaterials. Furthermore, chemical modification of LMWSC using various functional groups was also performed to enhance its bioavailability and emphasize their potential as drug delivery carriers (DDS).

• Microbiology and Biotechnology Letters
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• v.33 no.1
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• pp.9-15
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• 2005

Chitin deacetylase (CDA; EC 3.5.1.41) catalyzes the hydrolysis of N-acetamide bonds of chitin, converting it to chitosan. Chitosan has several applications in areas such as biomedicine, food ingredients, cosmetics, pharmaceuticals, and agriculture. In this paper, occurrence, assay and purification protocols, enzymatic characteristics, substrate specificity, and mode of action of microbial CDAs have been described. Several lines of evidence have substantiated the biological roles involved in cell wall formation and plant-pathogen interactions for fungal CDAs. The gene structure of CDAs has been compared with other family 4 carbohydrate esterases which deacetylate a wide variety of acetylated poly/oligo-saccharides. The use of CDAs for the conversion of chitin to chitosan, in contrast to the presently used chemical procedure, offers the possibility of a controlled, non-degradable process, resulting in the production of well-defined chitosan oligomers and polymers. Insect pathogen that can secrete high levels of chitin-metab­olizing enzymes including CDA can be a possible alternative for new pest management tools.

• KSBB Journal
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• v.28 no.4
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• pp.217-224
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• 2013

A bacterium CGH18 exhibiting antioxidizing and chitin-degrading activities in the colloidal chitin culture medium was isolated from salt-fermented crab. This strain was identified as Bacillus idriensis based on 16S rDNA sequence homology search. Its crude extract was partitioned between n-BuOH and $H_2O$. The organic layer was further partitioned between $CH_2$ $Cl_2$ and $H_2O$. Antioxidant activities of crude extract and its solvent fractions were evaluated using five different bioassay methods, including the degree of occurrence of intracellular reactive oxygen species (ROS), peroxynitrite scavenging (ONOO), and oxidative damage of genomic DNA. All fractions exhibited significant antioxidant activity in bioassay systems used.

• The Korean Journal of Mycology
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• v.31 no.3
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• pp.168-174
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• 2003

In this study, Cordyceps militaris was grown in a liquid medium containing colloidal chitin. A chitinase was purified from the supernatant or cultured medium by ammonium sulfate fractionation, DEAE-Sephadex A-25 and Sephadex G-50 column chromatography. Optimum temperature and pH of this enzyme were $35^{\circ}C$ and 5.5, respectively. The molecular weight of the chitinase was estimated to be 48.5 kDa by SDS-PAGE and its Km value was 0.57 mM. The activity of this enzyme was inhibited by $Cu^{2+},\;Mn^{2+},\;Hg^{2+},\;Zn^{2+},\;CO_{3}^{2-},\;SO_4^{2-},\;CN^-,\;ion,\;and\;OCN^-$ maleic anhydride, acetic anhydride or N-bromo succinimide, especially strongly inhibited by sodium cyanate for 84.0 percentage. But its activity wag slightly stimulated by $Mg^{2+}\;and\;K^+$ ion, respectively. The products formed during hydrolysis of the hexa-N-acetylchitohexaose with this enzyme were N,N'-diacetylchitobiose and N,N',N'-triacetylchitotriose. These results imply that this purified enzyme may be an endo-chitinase.