• Journal of Microbiology and Biotechnology
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• v.6 no.6
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• pp.439-444
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• 1996

A bacterial strain, designated as WY22, producing extracellular chitinase was isolated from the soil around the Youngduck area, after enrichment culture in a medium containing $1{\%}$ (w/v) wet colloidal chitin as a sole carbon source. The isolate was identified as a strain of Bacillus sp. based on its morphological and physiological characteristics. It was observed that Bacillus sp. WY22 could inhibit the growth of Fusarium oxysporum with hyphal extention-inhibition assay on potato dextrose agar plate supplemented with $1{\%}$ collidal chitin. Optimum culture conditions of Bacillus sp. WY22 were examined for chitinase production in a chitin medium. High level production of chitinase was observed not only in the chitin medium but in a medium supplemented with $1{\%}$ N-glucosamine or lactose instead of chitin. The optimum concentrations of colloidal chitin and yeast extract were 3.0 and $0.5{\%}$, and the optimum culture conditions for initial pH of medium and temperature were 7.0 and $30^{\circ}C$, respectively, for the production of chitinase.

• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.839-846
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• 1999

An extracellular chitinase-producing bacterial strain induced by colloidal chitin was isolated from sea sand and was identified to be a member of the genus Cytophaga. The chitinase was purified successively by 30-60% ammonium sulfate fractionation, and DEAE-Bio gel A column, Octyl-Sepharose CL-4B column, and DEAE-Bio gel A column chromatographies. The enzyme had a molecular mass of 59.75 kDa, and the amino terminal amino acid sequence was ATPNAPVISW MPTDXXLQNXS. The enzyme acted better on colloidal chitin as a substrate than on chitosan. For colloidal chitin and chitosan (Degree of Acetylation, 15-25%), $K_{cat}$ values were 0.60U/mg and 0.08U/mg, respectively. HPLC analysis of the enzymatic reaction products showed that the chitinase produced mostly N-acetyl-D-glucosarnine and di-N-acetylchitobiose. The optimum temperature and pH for the enzyme were $50^{\circ}C$ and 4.0, respectively. N-Bromosuccinimide and $Hg^{2+}$ inhibited the chitinase activity as much as 90%, and $Sb^{3+}$, diethylpyrocarbonate, and $Ag^{+}$ inhibited it by 50-70%.

• Korean Journal of Microbiology
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• v.38 no.4
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• pp.224-224
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• 2002

Chitin-degrading marine bacterial strain 98CJ11027 was isolated from bryozoa from the coastal area of Cheju Island, Korea, and identified as a member of the genus Vibrio. The molecular mass of the main extracellular chitinase (chitinase I), purified from strain 98CJ11027, was estimated to be 98 kDa. The optimal condition for chitinase I activity is pH 6.0 and 45℃. The activity was inhibited by $Fe^+2$ and$Cu^+2$. Chitinase I displayed the hydrolysis type of chitobiosidase and catalyzed reversed hydrolysis leading to the synthesis of tetraacetylchitotetraose.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1597-1602
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• 2010

Chitinase is one of the most important mycolytic enzymes with industrial significance. This enzyme is produced by a number of organisms including bacteria. In this study, we describe the optimization of media components with increased production of chitinase for the selected bacteria, Stenotrophomonas maltophilia, isolated from soil. Different components of the defined media responsible for influencing chitinase secretion by the bacterial isolate were screened using Plackett-Burman experimental design and were further optimized by Box-Behnken factorial design of response surface methodology in liquid culture. Maximum chitinase production was predicted in medium containing 4.94 g/l chitin, 5.56 g/l maltose, 0.62 g/l yeast extract, 1.33 g/l $KH_2PO_4$, and 0.65 g/l $MgSO_4{\cdot}7H_2O$ using response surface plots and the point prediction tool of the DESIGN EXPERT 7.1.6 (Stat-Ease, USA) software.

• Microbiology and Biotechnology Letters
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• v.26 no.6
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• pp.515-522
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• 1998

Plant root rotting fungi, Fusarium solani are suppressed their growth by the chitinase which is produced from the antagonistic soil bacteria. The chitinase producable antagonistic bacterium Pseudomonas sp. 3098 was selected as a powerful biocontrol agent of F. solani from ginseng rhizosphere. The antagonistic Pseudomonas sp. 3098 was able to produce a large amount of extracellular chitinase which is key enzyme in the decomposition of fusarial hypal walls. The chitinase was purified from cultural filtrate of Pseudomonas sp. 3098 by the procedure of ammonium sulfate precipitation, anion exchange chromatography, gel filtration on Bio-Gel P-100, and 1st and 2nd hydroxyapatite chromatography. The molecular mass of the purified enzyme was ca. 45 kDa on SDS-FAGE. The optimal pH and temperature for the activity of purified chitinase were 5.0 and 45$^{\circ}C$, respectively. The enzyme was stable in pH range of 5.0 to 9.0 up to 5$0^{\circ}C$ The enzyme was significantly inhibited by metal compounds such as FeCl$_2$, AgNO$_3$ and HgCl$_2$, and was slightly inhibited by p-CMB, iodoacetic acid, urea, 2,4-DNP and EDTA. The enzyme had ability of digestion on colloidal chitin and chitin from shrimp shell, but could not digest chitosan and chitin from crab shell. Km value of the enzyme was 0.11% on colloidal chitin, and the maximum hydrolysis rate of the enzyme was 34% on colloidal chitin.

• Microbiology and Biotechnology Letters
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• v.18 no.4
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• pp.437-441
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• 1990

For the genetic development of more powerful antagonistic Pseudomom - YPL-1 as a biocontxol agent against soilborne plant pathogenic Fuaarium solani causing root rot of many important crops, mutants improving the productivity of chitinase were obtained by mutation with UV radiation or NTG treatment, P. stutzeri YPL-M26 (UV mutant) and P. stutzeri YPL-MI78 (NTG mutant) could improve the productivity of chitinase by 2.5 and 2.0 times, and its antifungal activity by 1.7 and 1.5 times, respectively. The antifungal mechanism of P. stutzeri YPL-M26 was caused by lysis of the fungal cell wall by hydrolytic enzymes such as chitinase. The antifungal activity of crude chitinase of P. stutzeri YPLM26 on the mycelial growth of F. solani was observed to be much higher than that of the original strain. The enzymes produced by P. stutzeri YPL-M26 were the same as the original strain in enzymatic properties such as optimal pH and temperature.

• The Plant Pathology Journal
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• v.28 no.4
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• pp.439-445
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• 2012

The chitinase producing Lysobacter enzymogenes C-3 has previously been shown to suppress plant pathogens in vitro and in the field, but little is known of the regulation of chitinase production, or its role in antimicrobial activity and biocontrol. In this study, we isolated and characterized chitinase-defective mutants by screening the transposon mutants of L. enzymogenes C-3. These mutations disrupted genes involved in diverse functions: glucose-galactose transpoter (gluP), disulfide bond formation protein B (dsbB), Clp protease (clp), and polyamine synthase (speD). The chitinase production of the SpeD mutant was restored by the addition of exogenous spermidine or spermine to the bacterial cultures. The speD and clp mutants lost in vitro antifungal activities against plant fungal pathogens. However, the gluP and dsbB mutants showed similar antifungal activities to that of the wild-type. The growth of the mutants in nutrient rich conditions containing chitin was similar with that of the wild-type. However, growth of the speD and gluP mutants was defective in chitin minimal medium, but was observed no growth retardation in the clp and dsbB mutant on chitin minimal medium. In this study, we identified the four genes might be involved and play different role in the production of extracellular chitinase and antifungal activity in L. enzymogenes C-3.

• Korean Journal of Food Science and Technology
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• v.35 no.6
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• pp.1188-1192
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• 2003

Chitinase producing bacteria, Arthrobacter nicotianae CH4 and A. nicotianae CH13, were isolated from small crabs by an enrichment culture using chitin as the sole carbon source. Crude chitinases from the two isolated strains, A. nicotianae CH4 and A. nicotianae CH13, were stable in the pH range of $3.0{\sim}9.0$ and in the temperature range of $20{\sim}60^{\circ}C$. The reducing sugar $(GlcNAc)_1$, or $(GlcNAc)_4$, corresponding to over 98% of the enzyme reaction products, was obtained. The production of functional $(GlcNAc)_1$ and $(GlcNAc)_4$ from A. nicotianae CH13 and A. nicotianae CH4, respectively, from the chitinases was useful. The chitinase system of A. nicotianae CH13 was supposed to be endo- and exo-chitinase, and N-acetylglucosaminidase.

• Korean Journal of Soil Science and Fertilizer
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• v.36 no.4
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• pp.247-255
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• 2003

A bacterium, KJA-118 showing a strong chitinase activity, was isolated and identified as Bacillus cereus. The strain produced maximum level of chitinase, when grown aerobically at $30^{\circ}C$ for 4 days in basal broth containing 1% colloidal chitin in the initial pH adjusted to 6.0. Among various carbon sources such as crab shell powder, chitin powder, colloidal chitin, and R. solani mycelium, maximum chitinase activity was found in culture broth supplemented with R. solani mycelium. When KJA-118 was incubated with R. solani, the cell wall of the fungus was found to be completely destroyed. SDS-PAGE and active staining results revealed that KJA-118 produced three isoforms of chitinase with molecular weights of 68 kDa, 47 kDa, and 37 kDa. When the suspension of KJA-118 was treated to cucumber seedlings, reducing rate of damping-off caused by R. solani was about 28.1%.

• Applied Biological Chemistry
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• v.37 no.1
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• pp.43-48
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• 1994

A basic inducible protein, ICG, containing chitinase and ${\beta}-1,3-glucanase$ activity concomittantly was purified from cell suspension culture media of rice after the treatment of chitooligosaccharides. The isolated ICG enzyme gave a single band on native and SDS polyacrylamide gel electrophoresis and its molecular weight was estimated to be 52.53 kd. The optimal temperature and optimal pH of both enzyme activities in ICG were $60^{\circ}C$, pH 6.0 for chitinase activity and $37^{\circ}C$, pH 4.0 for ${\beta}-1,3-glucanase$ activity. $K_M$ and $V_{max}$ values for chitinase were 0.474 mM. 2.997 nM/min., and those for ${\beta}-1,3-glucanase$ were 1.004 mM 0.739 nM/min. respectively. TLC analysis of the chitooligosaccharide hydrolysates with ICG enzyme indicated that ICG acts as endochitinase.