• Microbiology and Biotechnology Letters
• /
• v.24 no.5
• /
• pp.560-566
• /
• 1996

A strain capable of producing thermostable chitinase suitable for chitooligosaccharide production was isolated from high temperature environment and identified as Bacillus licheniformis. The chitinase from Bacillus licheniformis KFB-Cl4 was only induced by addition of colloidal thitin into the basal medium as carbon source, showing the decrease of the chitinase production by supplernental addition of other carbon sources into the medium containing 1.0% colloidal chitin. Among organic and inorganic nitrogen sources, yeast extract was the most effective for the increase of total activity and specific activity, and had high affinity for the enzyme production. The optimum temperature of cell growth and thermostable chitinase production was 55$\circ$C. The optimum culture medium was composed of 1.2% colloidal chitin, 0.15% K$_{2}$HPO$_{4}$, 0.05% KH$_{2}$PO$_{4}$, 0.01% MgSO$_{4}$-7H$_{2}$O, 0.1% yeast extract (pH 6.5). Bacillus licheniformis KFB-C14 produced the thermostable chitinase of 3.89 units per ml culture fluid and 7.4 units per mg protein under rotary shaking at 150 rpm for 40 hr.

• Microbiology and Biotechnology Letters
• /
• v.47 no.1
• /
• pp.96-104
• /
• 2019

Beauveria bassiana, one of the most common entomopathogenic fungi, has been isolated, pre defined and characterized in-house from soil of tea cultivation area. Experiments have been performed to verify the presence of chitinase as intracellular metabolite and its release as extracellular product rendering the spores with biopesticide activity. Although there are many responsible enzymes for the pest killer action of B. bassiana, binding property of chitinase depending on presence as well as absence of serine supplemented in the media has been studied with respect to the production and kinetics. A programmed investigation conclusively indicates that the isolated spore (hyphae) of B. bassiana has been metabolically enriched with the enzyme chitinase in presence of an externally added amino acid serine with its inhibitory kinetics.

• Applied Biological Chemistry
• /
• v.36 no.5
• /
• pp.370-375
• /
• 1993

Five electrophoretic bands of crude enzyme extracted from rice leaves were found to possess both chitinase and ${\beta}-1,3-glucanase$ activities. These $chitinase/{\beta}-1,3-glucanase$ were resolved into acidic and basic fractions of protein by DEAE-cellulose and chitin affinity column chromatography. The optimal pH and temperature for ${\beta}-1,3-glucanase$ activity of two fractions were in the same extent as pH 5 and $60^{\circ}C$, whereas those for chitinase activity differed from one another; pH 3 and $60^{\circ}C$ for the acidic and pH 4 and $50^{\circ}C$ for the basic fraction, respectively. In addition, lysozyme activity was found in both fractions.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.4 no.1
• /
• pp.26-31
• /
• 1999

The advantages of the organism Streptomycs griseus HUT 6037 is that the chitinase and chitosanase using chitinaceouse substrate are capable of hydrolyzing both amorphous and crystalline chitin and chitosan. We attempted to investigate the optimization of induction protocol for high-level production and secretion of chitosanase and the influence of chitin and partially deacetylated chitosan sources (75∼99% deacetylation). The maximum specific activity or chitinase has been found at 5 days cultivation with the 48 hours induction time using colloidal chitin as a carbon source. To investigate characteristic of chitosan activity according to substrate, we used chitosan with various degree of deacetylation as a carbon source and found that this strain accumulates chitosanase in the culture medium using chitosanaceous substrates rather than chitinaceous substrates. The highest chitosanase activity was also presented on 4 days with 99% deacetylated chitosan. The partially 53% deacetylated chitosan can secrete both chitinase and chitosanase which was defined as a soluble chitosan. The specific activities of chitinase and chitosanase were 0.89 at 3 days and 1.33 U/mg protein at 5 days, respectively. It indicate that chitosanase obtained from S. griseus HUT 6037 can hydrolyze GlcNAc-GlcN and GlcN-GlcN linkages by exo-splitting manner. This activity increased with increasing degree of deacetylation of chitosan. It is the first attempted to investigate the effects of chitosanase on various degrees of deacetylations of chitosan by S. griseus HUT 6037. The highest specific activity of chitosanase was obtained with 99% deacetylated chitosan.

• BMB Reports
• /
• v.32 no.6
• /
• pp.541-546
• /
• 1999

When we compared the chitinase activity of various plant sources using colorimetric or active gel-staining assay methods, the specific activity of pumpkin leaves was the highest among the samples we analyzed. The highly active chitinase from pumpkin leaves (designated PL-ChtIII) was purified to homogeneity using affinity chitin gel and HPLC Mono-Q anion-exchange cloumn chromatographies. In contrast to other members of the class III chitinase family, PL-ChtIII showed a strong binding affinity to the regenerated chitin gel column. The apparent molecular weight of PL-ChtIII was estimated to be 29 kDa on SDS-PAGE gel, while its optimum pH and temperature were shown to be pH 6.0 and $60^{\circ}C$, respectively. Analyzing the reaction products of PL-ChtIII with swollen chitin as substrate, the dimer and tetramer of N-acetylglucosamine were produced as major products in the first hour of the enzymatic reaction along with a small amount of monomers and trimers. As the reaction time increased, dimeric N-acetylglucosamine became the predominant form of reaction product.

• The Plant Pathology Journal
• /
• v.22 no.2
• /
• pp.107-114
• /
• 2006

The white cottony stem rot pathogen Sclerotinia scierotiorum was subjected to 70 different isolates of actinomycetes indigenous to Jordan as biological control agents. Forty of them demonstrated chitinase activity on crab shell chitin agay (CCA) media and they were segregated into three groups: 14 highly active, 12 moderately active, and 14 with low activity, with average clearing zones of (4.7-8.3), (3.7-4.3), and (2.3-3.3) mm surrounding colonies on CCA, respectively. Further, these isolates were able to inhibit radial mycelium growth of the pathogen and were categorized into three antagonistic groups: 13 strong, 13 moderate, and 14 weak antagonists, with antibiosis inhibition Bones of (32.0-45.7), (22.7-31.3), and (3.7-22.3) mm, respectively. High levels of chitinase activity of the isolates Ma3 (8.3 mm), Jul (7.7 mm), and Sa8 (7.7 mm) with their antagonistic activity against mycelium growth of 45.7, 44.3, and 40.7 mm were observed, respectively. These isolates exhibited fungicidal activity against sclevotia of S. sclerotiorum. On the other hand, isolates Na5, Aj3, and Aj2 that produced no chitinase showed fungistatic effect only.

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

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

• Korean Journal of Plant Tissue Culture
• /
• v.21 no.6
• /
• pp.357-362
• /
• 1994

Chitinases and $\beta$-1,3-glucanases are believed to be important in defending plane against pathogens. Here, we investigated the expression of chitinase(s) in Arabidopsis thaliana cell suspension culture system in response to ethephon (2-chloroethyl phosphonic acid) which produces ethylene or a microbial elicitor, a bacterial pectin-degrading enzyme, $\beta$-1, 4-endopolygalactronic acid Iyase (PGA Iyase), treatment. Chitinase activity was measured either by radio chemical assay using $^3$H-labeled regenerated chitin as substrate or western blot analysis using antibody raised against tobacro chitinase(S). With 1 mg/mL of ethephon or 100 m units/mL of elicitor treatment, maximum levels of activity were reached after 48h. We also investigated distribution of chitinase activity in seedlings, leaves, and root of A. thaliana and found that root have the highest chitinase activity.

• Fisheries and Aquatic Sciences
• /
• v.6 no.1
• /
• pp.7-12
• /
• 2003

Chitinase and chitobiase produced by Aeromonas sp. J-5003 were purified and characterized. The chitinase was purified to 19.4 folds by gel chromatography and ion-exchange chromatography with the overall yield of $2.2\%$ and the specific activity of 93.1 unit/mg. The purified enzyme showed a single band on SDS-PAGE with MW 54kDa. The optimum pH and temperature of the purified chitinase were 7.0 and $37^{\circ}C$, respectively, and this enzyme stable in the range of pH 6.0 to 10.0 below $37^{\circ}C$. $Mg^{2+},\;Ca^{2+}\;and\;Na^+$ slightly stimulated the chitinase activity. However, $Hg^{2+}\;and\;Fe^{3+}$ inhibited chitinase activity. The chitobiase was purified by Sephacryl HR-l00 gel chromatography and DEAE-Sephadex A-50 ion-exchange chromatography with 33.5 purification folds and $4.3\%$ yield. The purified enzyme showed a single band with MW 63 kDa. The optimum pH and temperature of the purified chitobiase were 7.0 and $37^{\circ}C$, respectively. And this enzyme was stable in the range of pH 6.0 to 9.0 and at the temperature below $37^{\circ}C$. The enzyme activity was increased by $Mn^{2+}$, but it was inhibited by $Ag^+$.