• Mycobiology
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• 제50권4호
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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.

• BMB Reports
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• 제43권11호
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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.

• 한국미생물·생명공학회지
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• 제25권2호
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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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• 제2권1호
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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.

• Fisheries and Aquatic Sciences
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• 제6권1호
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• pp.1-6
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• 2003

A chitinolytic enzyme-producing bacterium was isolated from sea water on the coast of Busan. The bacterium was identified as Aeromonas sp. based on its morphological, cultural and biochemical characteristics and designated Aeromonas sp. J-5003. The strain produced two chitinoloytic enzymes: chitinase and chitobiase. The optimum culture conditions of the strain for production of chitinoloytic enzymes were investigated. For the production of chitinase, the major components of medium were colloidal chitin $0.5\%$, glucose $0.2\%$, yeast extract $0.25\%$ and peptone $0.25\%$ while for the production of chitobiase, they were colloidal chitin $0.5\%$, galactose and tryptone $0.2\%$. The optimum cultural temperature and initial pH for the production of chitinase and chitobiase were $30^{\circ}C$ and pH 7.0, respectively.

• 한국식물병리학회지
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• 제11권3호
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• pp.258-264
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• 1995

Production and some properties of chitinolytic enzymes were investigated by 80% ammonium sulfate precipitates (crude enzymes) from culture supernatant of antagonistic bacteria, Chromobacterium violaceum strain C-61 and strain C-72, Aeromonas hydrophila, Aeromonas caviae, and Serratia marcescens. The maximum production of chitinase was obtained from the 3-day culture at 28$^{\circ}C$ in C. violaceum stains, the 6-day culture in S. marcescens, and the 2-day culture in A. hydrophila and A. caviae. In the optimum culture periods, chitinase activity of C. violaceum strains C-61 was 1.5, 5.5, 12.0 and 11.3 times higher than those of strain C-72, S. marcescens, A. hydrophila and A. caviae, respectively. However, N,N'-diacetylchitobiase activity was 3.2 times higher in S. marcescens than in C. violaceum strain C-61, and that of Aeromonas spp.was very low. On gels containing glycol chitin, chitinase of C. violaceum strains showed four isoforms of 54-, 52-, 50- and 37-kDa, whereas there were four isoforms of 58-, 52-, 48- and 38-kDa in S. arcescens, three isoforms of 70-, 58- and 54-kDa in A. hydrophila and six isoforms of 90-, 79-, 71-, 63-, 58- and 38-kDa in A. caviae. The chitinase of C. violaceum strain C-61 was most active at pH 7.0 and at 5$0^{\circ}C$ and was stable in ranges of pH 5.0~10.0 for 2 hours and of 0~5$0^{\circ}C$ for 30 min.

• 생명과학회지
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• 제9권4호
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• pp.341-347
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• 1999

Insects use chitinolytic enzyme to digest chitin in the exoskelton during the molting process. We have isolated and sequenced a chitinase-encoding cDNA from the silkworm, Bombyx mandarina, compared its sequenced with genes encoding chitinolytic enzymes from other sources. The insert DNA in the clone is 2,675 nucleotides long with an open reading frame of 1,695 uncletides that encodes a protein of 565 amino acids with a molecuar weight of 63.4 kDa. The 3' -untranslated region of 889 nucleotides is AT-rich and contains two putative polyadenylation signals. The N-terminal sequence of the encoded protein contains numerous hydrophobic residues characteristic of a leader peptide. The amino acid alignment revealed that the endo-$\beta$-N-acetylglucosaminidase had 83% and 97% homology with M. sexta and B. mori, respectively. The deduced amino acid had two highly conserved region at the amino acid residues 97-111 and 139-148 that were related to the existing chitinase.

• 한국식물병리학회지
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• 제11권4호
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• pp.304-311
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• 1995

To determine whether chitinolytic enzymes from Chromobacterium violaceum C-61 play an important role in the suppression of Rhizoctonia damping-off, Tn5 insertion mutants deficient in chitinolytic activity (Chi a- mutants) were selected and their chitinolytic and disease suppression were compared with those of the parental strain. Four Chi a- mutants selected from about 2,000 transconjugants did not inhibit mycelial growth of Rhizoctonia solani on nutrient agar-potato dextrose agar (BA-PDA) and their abilities to suppress Rhizoctonia damping-off were much lower than the parental strain. However, population density in the eggplant rhizosphere did not differ significantly between the parental strain and four Chi a- mutants. The crude enzyme of the parental strain inhibited growth of R. solani on NA-PDA and its chitinase activity was much higher than that of Chi a- mutants. But the N,N' -diacetylchitobiase activity between these isolates were not significantly different. The chitinase of Chi a- mutants was defective in 2 isoforms of 52- and 37-kDa among four isoforms of 54-, 52-, 50- and 37-kDa. A Tn5 element was inserted into one site of 10 kb EcoRI fragment of chromosomal DNA in three Chi- mutants, C61-C1, -C2, and -C3. In C61-C4 mutant, a Tn5 element was inserted into two sites of 10 kb and 4.4 kb EcoRI fragments. These results suggest that the chitinase of C. violaceum C-61 play an important role in the suppression of Rhizoctonia damping-off of cucumber and eggplant.

• KSBB Journal
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• 제11권6호
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• pp.696-703
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• 1996

S. marcescens QM 81466 균주는 chitin 분해 효소(1mg/Lmedium)를 선택적으로 높게 생성시킬 수 있는 균주로서, chitin을 N-acetyl-$\beta$-D-glucosa­m mine(NAG)으로 효소적 가수분해를 할 때 chitinase와 chitobiase의 두 가지 가수분해 효소계를 구 성시킨다. 본 연구에서는 이 균주의 chitinase/chitobiase 생성을 위한 chitin 입자크기에 대한 최적화와, 회분 발효계에서 이 균주의 세포 밀도 배양에 따른 두 효소 생성의 변화를 조사하여 NAG 생산성의 증대를 시도하였다. 아울러. chitin과 CM­ chitin이 chitinase/chito biase 생성비 와 NAG 생성 에 미치는 영향을 검토하였는데, CM-chitin을 colloidal 및 결정성 chitin 대신에 사용했을 때, chitinase 활성을 약 7~10U/mL 증가시켰다. 이 경우에 있어서, chitinase/chitobiase의 비는 9:1로 서 NAG의 생성량이 3.0g/L로서 높게 나타났다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제3권2호
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• pp.71-77
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• 1998

The strain of Serratia marcescens QM B1466 produces selectively large amount of chitinolytic enzymes (about 1mg/L medium). Enzymatic hydrolysis of chitin to N-acetyl-${\beta}$-D-glucosamine (NAG) was performed with a system consisting of two hydrolases (chitinase and chitobiase) produced by optimization of a microbial host consuming chitin particles. For the development of Large-scale biological process for the production of NAG from chitinaceous waste, the selection and optimization of a microbial host, particle size of crab/shrimp chitin sources and initial induction time using chitin as a sole carbon source on chitinase/chitobiase production and NAG production were examined. Crab-shell chitin(1.5%) treated by dilute acid and , ball-milled with a normal diameter less than 250m gave the highest chitinase activity over a 7 days culture. Crude chitinase/ chitobiase solution obtained in a 10 L fed-batch fermentation showed a maximum activities of 23.6 U/mL and 5.1 U/mL, respectively with a feeding time of 3 hrs, near pH 8.5 at 30$^{\circ}C$.