• Korean Journal Plant Pathology
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• v.11 no.1
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• pp.47-52
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• 1995

One hundred and thirty bacterial isolates with high chitinolytic activity on chitin agar media were isolated and identified. Most of the isolates were Aeromonas hydrophila (110 isolates), and the others were Serratia marcescens (11 isolates), Aeromonas caviae (3 isolates), Chromobacterium violaceum strain C-61 (2 isolates), Chromobacterium violaceum strain C-72 (1 isolate) and unknown species (3 isolates). Among them, C. violaceum strain C-61 had highest chitinolytic activity and fungal growth inhibition on PDA. This bacterium also inhibited the growth of Rhizoctonia solani, Sclerotinia scelrotiorum, Phytophthora capsici and Pythium ultimum, but it didn't inhibit the growth of Fusarium oxysprum and Fusarium solani. C. violaceum strain C-61 suppressed damping-off of eggplant caused by R. solani. Populations of the chitinolytic bacteria such as Aeromonas hydrophila, Serratia marcescens, Aeromonas caviae, Chromobacterium violaceum strain C-61 and Chromobacterium violaceum strain C-72 introduced into R. solani-infested soil were continuously decreased until 20 days after treatment, but their populations except A. caviae were not changed significantly and maintained over 5$\times$104 CFU per g of soil thereafter.

• Mycobiology
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• v.50 no.4
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• pp.244-253
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• 2022

Trichoderma fungi have been intensively studied for mycoparasitism, and the latter is closely related to their cell-wall degrading enzymes including chitinase. Here, we studied marine-derived Trichoderma spp., isolated from distinct sources and locations, for chitinolytic and antifungal activity. Based on morphological and phylogenetic analyses, two strains designated GJ-Sp1 and TOP-Co8 (isolated from a marine sponge and a marine alga, respectively) were identified as Trichoderma bissettii. This species has recently been identified as a closely related species to Trichoderma longibrachiatum. The extracellular crude enzymes of GJ-Sp1 and TOP-Co8 showed activities of chitobiosidase and b-N-acetylglucosaminidase (exochitinase) and chitotriosidase (endochitinase). The optimum chitinolytic activity of the crude enzymes was observed at 50 ℃, pH 5.0, 0-0.5% NaCl concentrations, and the activities were stable at temperatures ranging from 10 to 40 ℃ for 2 h. Moreover, the crude enzymes showed inhibitory activity against hyphal growth of two filamentous fungi Aspergillus flavus and Aspergillus niger. To the best of our knowledge, this is the first report of the chitinolytic and antifungal activity of T. bissettii.

• Microbiology and Biotechnology Letters
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• v.25 no.2
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• pp.151-158
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• 1997

For the production of potent chitinolytic enzyme from bacteria, screening was carried out. Of 100 samples from soil, fresh water and sea water collected from the Kyung-gi area, 7 strains of chitinolytic bacteria were isolated. Among them, Aeromonas hydrophila 5-3K showed the highest chitinolytic activity. Culture conditions of Aeromonas hydrophila for the production of chitinolytic enzyme were inverstigated and lytic enzyme was fractionated by the use of ammonium sulfate and Sephadex G-100. Maximum production of chitinolytic enzyme was obtained at pH 7.0 and 30$\circ$C with chitin concentration between 0.2% and 1.0%. Conditions for the enzyme production were optimized including fermentor cultivation. The chitinolytic system of Aeromonas hydrophila 5-3K was composed of two enzymes, chitinase and chitobiase.

• Fisheries and Aquatic Sciences
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• v.2 no.1
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• pp.105-111
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• 1999

In order to produce functional chitin oligosaccharides, a chitinolytic bacterium was newly screened from the viscera of Korean bony fishes, and identified as Bacillus sp. LJ-25. For the production of chitinolytic enzymes, $1.0\%$ nutrient broth and $0.3\%$ colloidal chitin were used as nitrogen and carbon source, respectively. The optimal temperature, initial pH and concentration of NaCl for the enzyme production by Bacillus sp. LJ-25 were $30^{\circ}C$ 6.5-7.0 and $1.0\%$, respectively. The enzyme activity of Bacillus sp. LJ-25 increased until the incubation time of 168 hr, followed by a decrease in activity.

• 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.

• Korean Journal of Veterinary Research
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• v.43 no.1
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• pp.145-150
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• 2003

To determine effectively the chitinolytic activity of rCHT1 from the hard tick H. longicornis expressed in baculovirus-mediated Spodoptera frugtperda (Sf) 9 cells, a simple and sensitive assay system was established in solid phase using agarose gel containing ethylene glycol chitin as substrate. The various factors affecting the efficacy of the assay were also investigated. The effects of various temperature, dosages of proteins, pH of media and time courses of reaction were examined to verify the sensitivity of assay for chitinolytic activity of rCHT1 protein. It was found that the optimal reactive conditions were $37^{\circ}C$ of temperature, 12 to 15 hours of reactive times, $0.1{\mu}g$ of protein concentration and pH 5 to 7 of media. Using the assay system designed, the functional activities of H. longicornis rCHT1l protein could be evaluated simply and sensitively.

• The Korean Journal of Pesticide Science
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• v.19 no.4
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• pp.402-410
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• 2015

The purpose of this study was to estimate the chitinolytic and antifungal activity of Actinomycetes sp.isolated from waste mushroom media. In five kinds of waste mushroom media, Sinyeong mushroom and Yangsongi were the order of the population density of actinomycetes. Totally 91 chitinolytic isolates of Actinomycetes sp. were obtained from waste mushroom media. The isolates were categorized into 3 groups based on chitinolytic activity and antagonisms against Phytophthora capsici, Rhizoctonia solani, Sclerotinia sclerotiorum, Collectotrichum gloeosporioides, and Cladosporium cucumerinum in vitro. CA-23 was selected as a representative isolate of a group showing strong chitinolytic and antagonistic activities to all of the plant pathogens, while AA-65 was selected as a representative isolate showing no chitinolytic activities but strong antagonistic activities to the pathogens. CA-23 and AA-65 were highly effective on control of Phytophthora blight of hot-pepper, powdery mildew and scab of cucumber in a greenhouse tests. Among the isolates tested, CA-23 showed highest control efficacy, while AA-65 not only effectively controlled the diseases but also consistently increased plant growth and yield. Although the isolates are similarly affected on suppression of plant pathogens, the isolates could be differ from each other in modes of action. Further studies on mechanisms and practical applications are being progressed.

• BMB Reports
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• v.43 no.11
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• pp.726-731
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• 2010

We report the tissue-specific distribution of chitinolytic activity in Korean ginseng root and characterize two 31-kDa chitinolytic enzymes. These two enzymes (SBF1 and SBF2) were purified 70- and 81-fold with yields of 0.75 and 1.25%, respectively, and exhibited optimal pH and temperature ranges of 5.0-5.5 and 40-$50^{\circ}C$. With [$^3H$]-chitin as a substrate, $K_m$ and $V_{max}$ values of SBF1 were 4.6 mM and 220 mmol/mg-protein/h, respectively, while those of SBF2 were 7.14 mM and 287 mmol/mg-protein/h. The purified enzymes showed markedly less activity with p-nitrophenyl-N-acetylglucosaminide and fluorescent 4-methylumbelliferyl glycosides of D-N-acetylglucosamine oligomers than with [$^3H$]-chitin. End-product inhibition of both enzymes demonstrated that both are endochitinases with different N-acetylglucosaminidase activity. Furthermore, the $NH_2$-terminal sequence of SBF1 showed a high degree of homology with other plant chitinases whereas the $NH_2$-terminal amino acid of SBF2 was blocked.

• Journal of fish pathology
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• v.21 no.3
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• pp.247-257
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• 2008

To understand the activity of non-specific defence factors in cultured Sebastes, the antibacterial effect of the serum, skin mucus and homogenate of various organs from cultured oblong rockfish (Sebastes oblongus) and rockfish(Sebastes schlegeli) against pathogenic bacteria, Aeromonas hydrophila, Edwardsiella tarda, Vibrio anguillarum, and Streptococcus sp. was compared with that of flounder(Paralichthys olivaceus) and seabass(Leteolabrax japonicus). And the activities of proteolytic enzyme, chitinolytic enzyme and haemolycin as non-specific defence factor were investigated on the oblong rockfish and rockfish. Samples from oblong rockfish showed the highest antibacterial activity by lysoplate assay on agar plate mixed with pathogens, followed in descending order by rockfish, seabass, and flounder. Turbidimetric assay was carried to evaluate the lysozyme activity of fish samples against lyophilized cells of Micrococcus lysodeiktikus. The serum, kidney, liver, stomach, intestine and eyeball of oblong rockfish and the mucus and gill of rockfish appeared to have the highest lysozyme activity among the fish strains investigated. All samples except skin mucus, liver, and eyeball of oblong rockfish and rockfish showed proteolytic enzyme activity. Chitinolytic enzyme activity was showed in random sampling and haemolytic activity was remarkable in oblong rockfish. Therefore, Sebastes strain was proved to have effective defense mechanisms based on the antibacterial activities, and lysozyme, proteolytic enzyme, chitinolytic enzyme, and haemolycin were considered to act as the non-specific defence factor of Sebastes.

• Journal of Radiation Industry
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• v.4 no.1
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• pp.65-71
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• 2010

Two chitinase producing strains CHI2 and CHI4 were isolated from soybean rhizosphere soil. Both the strains belonged to Lysobacter enzymogenes as indicated by 16S rDNA sequence analysis. Though strain CHI2 and CHI4 produced extracellular chitinase, they differ in their chitinolytic activity. CHI4 produced approximately three times the higher amounts of enzyme than that of CHI2 under specified conditions. CHI2 produced $535.67U\;l^{-1}$ of chitinase after 48 h incubation with a specific activity of $3.91U\;mg^{-1}$ of protein while strain CHI4 produced $1584.13U\;l^{-1}$ of chitinase with a specific activity of $10.88U\;mg^{-1}$ protein. SDS-PAGE analysis indicated that the molecular weight of chitinase enzyme was approximately 45 kDa. A faint band with a molecular weight of 55 kDa reveals the possibility for the presence of another kind of chitin binding protein. Mutant library was developed by exposing the isolates to gamma rays at their $LD_{99}$ value (0.23 kGy). Totally, 11 mutants of CHI2 and CHI4 are reported to have enhanced chitinase activity. Several leaky mutant clones with decreased enzyme activity and a defective mutant (CHI2-M16) with complete loss of chitinase activity were also identified. CHI4-M18, CHI4-M8 and CHI4-M29 showed 78.8, 41.5, and 31.9% increased chitinase activity over wild type CHI4.