• Title/Summary/Keyword: 키틴분해

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Classification and Characteristics of Chitin/Chitosan Hydrolases (키틴/키토산 가수분해효소의 분류 및 특성)

  • Lee, Han-Seung
    • Journal of Life Science
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    • v.18 no.11
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    • pp.1617-1624
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    • 2008
  • Chitin and chitosan, which is deacetylated form of chitin, are one of the most abundant biomass on the earth. They showed various biological activities including antimicrobial activity, heavy metal chelating, immune system activation, and have very diverse applications in food, pharmaceutical, medicinal, and environmental industry. There have been reported many chitin/chitosan-hydrolyzing enzymes, their structures and genes from three domains, archaea, bacteria, and eukarya. Carbohydrate hydrolyzing enzymes are classified in CAZy (Carbohydrate Active Enzymes) database according to their amino acid sequence similarity. Interestingly, chitinases and chitosanases are classified in various glycosyl hydrolase(GH) families, GH2, GH5, GH7, GH8, GH18, GH19, GH20, GH46, GH48, GH73, GH75, GH80, GH84, and GH85. Here, we review characteristics and structures of chitin/chitosan hydrolyzing enzymes according to glycosyl hydrolase families in order to provide information about gene mining.

Enhancement of chitinolytic activity of by co-expression of endochitinase and chitobiosidase genes (Endochitinase와 Chitobiosidase 유전자의 동시발현에 의한 키틴분해 활성의 증가)

  • Kim, Jungtae;Choi, Shin-Geon
    • Journal of Industrial Technology
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    • v.30 no.B
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    • pp.69-74
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    • 2010
  • Chitinolytic activity was enhanced by coexpression of endo-chitinase gene (chiA) and chitobiosidase gene (chiB) from Serratia marcescens KFRI314 using constitutive expression vector, pHCEIA, in E. coli. Coexpression vector was constructed by inserting ribosome binding site (RBS) into junction between two chitinase genes. SDS-PAGE analyses showed that two chitinase were constitutively expressed while E. coli clones expressing two chitinases simultaneously increased halo size on colloidal chitin plate. Furthermore, the chitinolytic activities were much enhanced in coexpressed clones when degradation patterns of substrate analogues such as 4-MU-(NAG), $4-MU-(NAG)_2$,$4-MU-(NAG)_3$ were used. Consequently, the combined use of endochitinase and chitobiosidase greatly increased overall chitinolytic activities on recombinant E. coli clones.

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Cloning of Serratia marcescens KFRI314 chitinase genes and its role on chitin degradation (Serratia marcescens KFRI314 chitinase 유전자의 클로닝과 키틴분해에 관한 효소의 역할)

  • Kim, Jungtae;Choi, Shin-Geon
    • Journal of Industrial Technology
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    • v.30 no.B
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    • pp.61-68
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    • 2010
  • Three chitinase genes (chiA, chiB, and chiC) were cloned into E. coli by PCR amplification from Serratia marcescens KFRI314. The sizes of cloned chitinase genes were 1692 bp, 1500 bp, and 1443 bp which correspond to 563, 499, and 480 amino acids, respectively. Recombinant chitinases were overexpressed using pHCEIA expression vector and purified to homogenity. The molecular weights of chitinases were about 60kDa, 50 kDa, 52 kDa, respectively. Optimum pHs were around pH 5~6 and optimum temperatures were $45{\sim}50^{\circ}C$ while 90% of enzyme activities were stable up to $50^{\circ}C$. The specific activities of ChiA, ChiB, and ChiC were 233.1, 278.8, $111.3{\mu}mol\;(min)^{-1}\;mg^{-1}$ against colloidal chitin. From experiments using TLC and fluorescent substrate analogues, it was demonstrated that ChiA was endo-chitinase while ChiB and ChiC were chitobiosidase.

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Survey on the Chitinolytic Activity from Some Plants for the Industrial Utilization (공업적 이용을 위한 식물성 키틴분해효소의 탐색)

  • Han, Beom-Ku;Lee, Woo-Jin;You, Tak;Park, In-Ho;Jo, Do-Hyun
    • Applied Biological Chemistry
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    • v.39 no.6
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    • pp.466-471
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    • 1996
  • The survey on the chitinolytic activity of some plants was performed for the purpose of obtaining some reliable and inexpensive sources of chitinase. Rice, soybean for sprouting, kiwi fruit, almond and crude papain were investigated. Rice bran, seed coat of the soybean and the pericarp of kiwi fruit showed a considerable activity, while the bean after the removal of the seed coat, the mixture of rice integument and endosperm, polished rice, and defatted soybean powder didn't have any detectable activity. These crude enzymes have shown to contain both endo- and exochitinase activity. The effects of pH and temperature on the enzyme activity were variable. Furthermore we have observed the chitosanolytic activity from these enzyme Preparations. The rice bran had the highest activity in the enzymatic degradation of chitosan, and seed coat of soybean and the pericarp of kiwi fruit followed. On the basis of the fact that crude papain was not only commercially available but also the most potent in the endochitinase activity and the lowest in the exochitinase activity, we could conclude that crude papain was considered as the most suitable source of the chitinase among plants studied in this paper. In addition, rice bran was worth further investigation from the point of utilizing agricultural by-product.

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Physiological Characteristics of Actinomycetes Isolated from Turfgrass Rhizosphere (잔디 근권에서 분리된 Actinomycetes균주의 생리학적 특성)

  • Lee, Jung Han;Min, Gyu Young;Shim, Gyu Yul;Jeon, Chang Wook;Kwak, Youn-Sig
    • Weed & Turfgrass Science
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    • v.4 no.4
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    • pp.348-359
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    • 2015
  • Total 443 isolates of actinomycetes were isolated from turfgrass rhizosphere as potential biological control agents. The two isolates (S11 and S4) showed highest cellulase activity with compared to the other isolates that exhibited a clear zone of 1.2 mm around the colony on cellulose agar medium. S12 strain appeared the most active chitin degrading, which exhibited a 1.2 mm of clear zone. The highest proteolytic activity on skim milk agar was which exhibited a 7.5 mm of clear zone by S2 strain. S1 strain from the soli showed siderophore production ability, which exhibited a 0.6 mm of large clear zone on chrome azurol S agar. The antifungal activity of the volatile compound producing by 4 selected actinomycetes was investigated that inhibition rate against Rhizoctonia solani AG2-2 and Sclerotinia homoeocarpa. Growth inhibition effect of S8 isolate against S. homoeocarpa was appeared to 94.8%, S2 to 76.9%, S5 to 46.1% and S12 to 43.5%. The significant inhibition effects on mycelial growth of S. homoeocarpa were shown on media with four strains. The inhibition effect was the highest with S8 strain treatment at 94.8%.

Control of Ginseng Damping-Off Disease Using Chitinolytic Bacterial Mixtures (키틴분해미생물을 이용한 인삼 잘록병 방제)

  • Kim, Young Cheol;Chung, Hyun Chae;Bae, Yeoung Seuk;Park, Seur Kee
    • Research in Plant Disease
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    • v.24 no.4
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    • pp.353-358
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    • 2018
  • An effective bioformulation of mixtures of chitin-degrading bacteria has been used successfully to control plant diseases and nematodes. In this study, the bioformulation approach was assessed to control damping-off disease of ginseng. In pot experiments with soils infested with dapming-off pathogens of ginseng, root-drenchings of Chrobacterium sp. C-61, Lysobacterium enzymogenes C-3, and mixture of two bacterial strains grown in chitin minimal medium were signficantly increased emergence of seeds and reduced damping-off disease incidence of seedlings. Efficacy of the bioformulated product depended on the dose and timing of application. In two-year-old ginseng field, the high control efficacies were achieved by soil drenching of two times with an undiluted product or three times with a 10-fold diluted product. In a To-jik nursery (self soil nursery), biocontrol efficacy of the undiluted product against damping-off disease were similar to that of a seed dressing with fungicide, Tolclofos-methyl WP. These results suggest that the bioformulated product containing Chromobacterium sp. C-61 and L. enzymogenes C-3 could be an effective approach to control of ginseng damping-off disease.

Effects of Organic Materials on Insect and Disease Occurrence and Fruit Quality in Pear Orchards (친환경 자재가 배 과원의 병해충 방제 효과 및 과실 품질 특성에 미치는 영향)

  • Choi, Hyun-Sug;Wu, Xiu-Yu;Kim, Wol-Soo;Lee, Youn;Choi, Byoung-Min;Kuk, Yong-In
    • Korean Journal of Organic Agriculture
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    • v.19 no.3
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    • pp.405-416
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    • 2011
  • Organic materials, such as chitin incubated solution (CIS) combined with neem oil (NO), nano silver silica (NSS), and Bordeaux mixture (BDM), were applied with and without agricultural chemicals (AC) (insecticide and fungicide) to investigate scab and mealybug occurrences and fruit qualities on 'Niitaka' pear trees in orchards in 2006. Fruits and leaves grown under CIS+NO without AC had less than 30%, scab occurrence, but CIS+NSS or CIS+BDM without AC had higher scab occurrence. Organic materials with AC decreased the scab to less than 20%. All treatments decreased mealybug occurrences to less than 10%, except for the fruits grown under CIS+BDM without AC. Fruit qualities varied among the treatments. Hunter value a, representing for the redness degree, was higher for fruits treated with CIS+NSS and CIS+BDM without AC than those with AC. Fruits treated with organic materials without AC had greater total phenolic and flavonoid compounds as well as antioxidant capacity in flesh and greater total phenolc compounds and antioxidant capacity in peel than those treated with the AC.

Control of the Root-Knot Nematode (Meloidogyne spp.) on Cucumber by a Liquid Bio-Formulation Containing Chitinolytic Bacteria, Chitin and Their Products (키틴분해세균, 키틴 및 그들의 산물이 함유된 미생물제에 의한 오이의 뿌리혹선충(Meloidogyne spp.) 방제)

  • Ha, Woo Jong;Kim, Young Cheol;Jung, Hyuncha;Park, Seur Kee
    • Research in Plant Disease
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    • v.20 no.2
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    • pp.112-118
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    • 2014
  • A liquid bio-formulation containing chitinolytic bacteria, chitin and their products was assessed for its potential biological control against root-knot nematodes on cucumber. The bio-formulation was prepared by cultures of three chitinolytic bacteria, Chromobacterium sp. strain C-61, Lysobacter engymogenes and Serratia plymuthica in minimal medium supplemented with chitin. Under pot conditions, the bio-formulation showed better growth of cucumber plants, and less root galls and population density of Meloidogyne spp. than control media without the bio-formulation. In a greenhouse, 75-fold diluted bio-formulations were treated instead of water around cucumber plants through hoses for drip irrigation six times at 5-day intervals from the transplanting date. After 30 and 60 days, the treatment provided about 7% and 10% enhancement in the plant height and about 78% and 69% reduction in population density of Meloidogyne spp. in the rhizosphere, respectively. In addition, the experiments showed that the control effects occurred only in the soils contacted with the bio-formulation. Undiluted bio-formulations were drenched three times at 10-day intervals around cucumber plants severely infested with Meloidogyne spp. The treatment showed about 37% plant enhancement without dead plants compared with 37% death in the untreated control, and about 82% nematode reduction. These results suggest that the bio-formulation can be practically used to control the root-knot nematode on cucumber.

Preparation of an Immobilized Enzyme for Enhancing Thermostability of the Crude Proteinase from Fish Intestine (어류 내장 유래 단백질 분해효소로부터 열안정성 개선을 위한 고정화 효소의 제조)

  • 전유진;박표잠;변희국;송병권;김원석;김세권
    • Journal of Life Science
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    • v.8 no.6
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    • pp.627-637
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    • 1998
  • In order to utilize tuna pyloric caeca among fish intestines wasted when treated raw fish in fish processing manufactory, a crude enzyme with high proteolytic activity was extracted and its optimum condition were investigated. An immobilized enzymes also were prepared by adsorption method to enhance thermostability of the crude proteinase. The yield of the crude proteinase was approximately 2.7% on dry basis. The proteolytic activity for casein was 0.54 U/mg protein, for BTEE 1.10 U/mg protein, and for BAEE 2.69 U/mg protein. It was almost similar to that of the commercial trypsin purified. Optimum hydrolysis activity of the crude proteinase was about 80%, as the degree of hydrolysis for casein, at pH 10.0 and 45$^{\circ}C$ for 12 hrs. Also, when the crude proteinase was immobilized on DEAE-Cellulose and chitin, the residual activities remained after 7 days of pre-incubation time were maintained about 90% or more and their thermostabilities were enhanced by about 50%, compared with the native enzyme.

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