Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Properties of Chitosanase from Chitinolytic $\beta$-Proteobacterium KNU3

  • Yi, Jae-Hyoung (School of Biotechnology and Bioengineering, Kangwon National University) ;
  • Jang, Hong-Ki (School of Biotechnology and Bioengineering, Kangwon National University) ;
  • Lee, Sang-Jae (ProMediTech, Inc.,) ;
  • Lee, Keun-Eok (School of Biotechnology and Bioengineering, Kangwon National University) ;
  • Choi, Shin-Geon (School of Biotechnology and Bioengineering, Kangwon National University)
  • Published : 2004.04.01
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Abstract

A bacterial strain concurrently producing extracellular chitosanase and chitinase was isolated from soil and identified as a member of the $\beta$-subgroup of Proteobacteria through its 16S rRNA analysis and some biochemical analyses. The newly discovered strain, named as KNU3, had 99% homology of its 16S rRNA sequence with chitinolytic $\beta$-Proteobacterium CTE108. Strain KNU3 produced 34 kDa of chitosanase in addition to two chitinases of 68 kDa and 30 kDa, respectively. The purified chitosanase protein (ChoK) showed activity toward soluble, colloidal, and glycol chitosan, but did not exhibit any activity toward colloidal chitin. The optimum pH and temperature of ChoK were 6.0 and $70^{\circ}C$, respectively. The chitosanase was stable in the pH 4.0 to 8.0 range at $70^{\circ}C$, while enzyme activity was relatively stable at below $45^{\circ}C$. MALDI-TOF MS and N-terminal amino acid sequence analyses indicated that ChoK protein is related to chitosanases from Matsuebacter sp. and Sphingobacterium multivorum. HPLC analysis of chitosan lysates revealed that glucosamine tetramers and hexamers were the major products of hydrolysis.

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References

  1. J. Gen. Microbiol. v.126 Purification and mode of action of a chitosanase from Penicillium islandicum Fenton,D.M.;E.D.Eveleigh
  2. J. Microbiol. Biotechnol. v.13 Cloning and Characterization of a bifunctional cellulase-chitosanase gene from Bacillus licheniformis NBL420 Hong,I.P.;H.K.Jang;S.Y.Lee;S.G.Choi
  3. Appl. Microbiol. Biotechnol. v.38 Purification and characterization of a chitosanase from Streptomyces N174 Isabelle,B.;A.Dupuy;P.Vidal
  4. Application of Chitin and Chitosan Japanase Society for Chitin and Chitosan
  5. J. Microbiol. Biotechnol. v.13 Characterization of endochitosanases-producing Bacillus cereus P16 Jo,Y.Y.;K.J.Jo;Y.L.Jin;W.J.Jung;J.H.Kuk;K.Y.Kim;T.H.Kim;R.D.Park
  6. Biosci. Biotech. Biochem. v.56 Purification and properties of a chitosanase from Pseudomonas sp. H-14 Kazutoshi,Y.;J.Hosokawa;T.Kubo;M.Nishiyama;Y.Koba https://doi.org/10.1271/bbb.56.972
  7. Curr. MIcrobiol. v.40 Properties of chitosanase from Bacillus cereus S1 Kurakake,M.;S.You;K.Nakagawa;M.Sugihara;T.Komaki https://doi.org/10.1007/s002849910002
  8. J. Microbiol. Biotechnol. v.6 Purification and characteristics of chitosanase from Bacillus sp. HW-002 Lee,H.W.;J.W.Choi;D.P.Han;N.W.Lee;S.L.Park;D.H.Yi
  9. Biosci. Biotech. Biochem. v.56 Action pattern of Bacillus sp. No. 7-M chitosanase on partially N-acetylated chitosan Masato,I.;S.Nagae;H.Kawagishi;M.MItsuromi;A.Ohtakara https://doi.org/10.1271/bbb.56.448
  10. J. Gen. Plant Pathol. v.67 In Vitro suppression of mycelial growth of Fusarium oxysporum by extracellular chitosanase of Sphingobacterium multivorum and cloning of the chitosanase gene csnSM1 Matsuda,Y.;Y.Iida;T.Shinogi;K.Kakutani;T.Nonomura;H.Toyoda https://doi.org/10.1007/PL00013039
  11. J. Bacteriol. v.181 Purification, characterization, and gene anlaysis of a chitosanase (choA) from Matsuebacter chitsanotbidus 3001 Park,J.K.;K.Shimono;N.Ochiai;K.Shigeru;M.Kurita;Y.Ohta;K.Tanaka;H.Matsuda;M.Kawamukai
  12. J. Biosci. Bioeng. v.91 The bacterium Burkholderia gladioli strain CHB101 produces two different kinds of chitinases belonging to families 18 and 19 of the glycosyl hydrolases Shimosaka,M.;Y.Fukumori;T.Narita;X.Y.Zhang;N.J.He;R.Kodaira;M.Nogawa;M.Okazaki https://doi.org/10.1263/jbb.91.103
  13. Appl. Microbiol. Biotechnol. v.54 Molecular cloning and characterization of a chitosanase from the chitosanolytic bacterium Burholderia gladioli strain CHB101 Shimosaka,M.;Y.Fukumori;X.Y.Zhang;N.J.He;R.Kodaira;M.Okazaki https://doi.org/10.1007/s002530000388
  14. Bioresource Technol. v.55 Chitosanase-properties and applications: a review Somashekar,D.;R.Joseph https://doi.org/10.1016/0960-8524(95)00144-1
  15. Biosci. Biotech. Biochem. v.67 Novel chitosanase from Streptomyces griseus HUT6037 Tanabe,T.;K.Morinaga;T.Fukamizo;M.Mitsutomi https://doi.org/10.1271/bbb.67.354
  16. J. Bacteriol. v.173 16S ribosomal DNA amplification for phylogenetic study Weisburg,W.G.;S.M.Barns;D.A.Pelletier;D.J.Lane https://doi.org/10.1128/jb.173.2.697-703.1991
  17. Biosci. Biotech. Biochem. v.57 Purification and mode of action of chitosanolytic enzyme from Enterobacter sp. G-1 Yamasaki,Y.;I.Hayashi;Y.Ohta;T.Nakagawa;M.Kawamukai;H.Matsuda https://doi.org/10.1271/bbb.57.444
  18. Methods in Enzymol. v.161 Chitosanase from Bacillus species Yasushi,U.;A.Ohtakara https://doi.org/10.1016/0076-6879(88)61066-4
  19. Appl. Environ. Microbiol. v.66 Thermostable chitosanase from Bacillus sp. Strain CK4: Cloning and expression of the gene and characterization of the enzyme Yoon,H.G.;H.Y.Kim;Y.H.Lim;H.K.Kim;D.H.Shin;B.S.Hong;H.Y.Cho https://doi.org/10.1128/AEM.66.9.3727-3734.2000
  20. Biosci. Biotechnol. Biochem. v.66 Gene cloning and biochemical analysis of thermostable chitosanase (TCH-2) from Bacillus coagulans CK108 Yoon,H.G.;K.H.Lee;H.Y.Kim;H.K.Kim;D.H.Shin;B.S.Hong;H.Y.Cho https://doi.org/10.1271/bbb.66.986
  21. Biosci. Biotech. Biochem. v.56 Purification and properties of chitosanase from Pseudomonas sp. H-14 Yoshihara,K.;J.Hosokawa;T.Kubo;M.Nishiyama;Y.Koba https://doi.org/10.1271/bbb.56.972
  22. Biosci. Biotechnol. Biochem. v.67 Fermentation conditions and properties of a chitosanase Acinetobacter sp. C-17 Zhu,X.F.;X.Y.Wu;Y.Dai https://doi.org/10.1271/bbb.67.284