Browse > Article
http://dx.doi.org/10.4489/KJM.2003.31.3.168

Purification and Characterization of a Chitinase in Culture Media of Cordyceps militaris(Linn.) Link.  

Lee, Kang-Hyeob (Department of Chemistry, Dongguk University)
Min, Tae-Jin (Department of Chemistry, Dongguk University)
Publication Information
The Korean Journal of Mycology / v.31, no.3, 2003 , pp. 168-174 More about this Journal
Abstract
In this study, Cordyceps militaris was grown in a liquid medium containing colloidal chitin. A chitinase was purified from the supernatant or cultured medium by ammonium sulfate fractionation, DEAE-Sephadex A-25 and Sephadex G-50 column chromatography. Optimum temperature and pH of this enzyme were $35^{\circ}C$ and 5.5, respectively. The molecular weight of the chitinase was estimated to be 48.5 kDa by SDS-PAGE and its Km value was 0.57 mM. The activity of this enzyme was inhibited by $Cu^{2+},\;Mn^{2+},\;Hg^{2+},\;Zn^{2+},\;CO_{3}^{2-},\;SO_4^{2-},\;CN^-,\;ion,\;and\;OCN^-$ maleic anhydride, acetic anhydride or N-bromo succinimide, especially strongly inhibited by sodium cyanate for 84.0 percentage. But its activity wag slightly stimulated by $Mg^{2+}\;and\;K^+$ ion, respectively. The products formed during hydrolysis of the hexa-N-acetylchitohexaose with this enzyme were N,N'-diacetylchitobiose and N,N',N'-triacetylchitotriose. These results imply that this purified enzyme may be an endo-chitinase.
Keywords
Characterization; Cordyceps militaris; Endo-chitinase;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Chitinase from Phaseolus vulgaris leave /
[ Boller, T.;Gehri, A.;Mauch, F.;Vogeli, U. ] / Methods Enzynwl.   DOI
2 Chitinase from Verticillium albo-atum /
[ Pego, G.F. ] / Methods Enzymol.   DOI
3 Chitinase is an inducible enzyme in Beauveria bassiana /
[ Smith, R.J.;Grula, E.A. ] / J. lnvertebr. Pathol.   DOI
4 Biochemistry of chitinase;Chitin and Chitinases /
[ Koga, D.;Mitsutomi, M.;Kono, M.;Matsumiya, M.;Jolles, P.(ed.);Muzzarelli, R.A.(ed.) ] /
5 A submicrodetermination of glucose /
[ Park, J.T.;Johnson, M.J. ] / J. Biol. Chem.
6 Maleylation of amino groups /
[ Butler, P.J.G.;Hartley, B.S. ] / Methods Enzymol.   DOI
7 Powdered chitin agar as a selective medium for enumeration of actinomycetes in water and soil /
[ Hsu, S.C.;Lockwood, J.L. ] / Appl. Microbiol.
8 Chitinase /
[ Jeuniaux, C. ] / Methods Enzymol.   DOI
9 Physiological aspects of chitin catabolism in marine bacteria /
[ Keyhani, N.O.;Roseman, S. ] / Biochim. Biophys. Acta.   DOI   ScienceOn
10 An endochitinase from wheat germ: Activity on nascent and preformed chitin /
[ Molano, J.;Polacheck, I.;Duran, A.;Cabib, E. ] / J. BioI. Chem.
11 Acetylation /
[ Riordan, J.F.;Vallee, B.L. ] / Methods Enzymol.   DOI
12 Cloning and expression of a Streptomyces plicatus chitinase (chitinase-63) in Escherichia coli. /
[ Robbins, P.W.;Albright, C.;Benfield, B. ] / J. Biol. Chem.
13 Serratia marcescens chitinase: One-step purification and use for the determination of chitin /
[ Roberts, R.L.;Cabib, E. ] / Anal. Biochem.   DOI   ScienceOn
14 Dissolution of fungal cell walls by a streptomycete chitinase and <TEX>${\beta}-(1-3)$</TEX> glucanase /
[ Skujins, J.J.;Potgieter, H.J.;Alexander, M. ] / Arch. Biochem. Biophys.   DOI   ScienceOn
15 Requirement for sequential enzymatic activities for penetration of the integument of the com earworm (Heliothis zea) /
[ Smith, R.J.;Pekrul, S.;Grula, E.A. ] / J. lnvertebr. Pathol.   DOI
16 Tryptophan involvement in binding sites of proteins and in enzyme-inhibitor complexes as determined by oxidation with N-bromosuccinimide /
[ Spande, T.F.;Witkop, B. ] / Methods Enzymol.   DOI
17 Modification of proteins with cyanate /
[ Stark, G.R. ] / Methods Enzymol.   DOI
18 Cutiledegrading enzymes of entomopathogenic fungi: Regulation of production of chitinolytic enzymes /
[ Leger, R.J.;Cooper, R.M.;Charnley, A.K. ] / J. Gen. Microbiol.   DOI
19 Studies on distribution and utilization of Cardyceps militaris and C. nutans /
[ Sung, J.M.;Kim, C.H.;Yang, K.J.;Lee, H.K.;Kim, Y.S. ] / Kor. J. Mycol.
20 Distribution and taxonomy of entomopathogenic fungal species from Korea /
[ Sung, J.M.;Lee, H.K.;Choi, Y.S.;Kim, Y.Y.;Kim, S.H.;Sung, G.H. ] / Kor. J. Mycol.
21 Classification of Cordyceps species based on protein anding pattern /
[ Sung, J.M.;Lee, H.K.;Yoo, Y.J.;Choi, Y.S.;Kim, S.H.;Kim, Y.O.;Sung, G.H. ] / Kor. J. Mycol.
22 Chitinase in some Basidiomycetes /
[ Tracey, M.V. ] / Biochem. J.
23 Regulation of chitinase synthesis in Trichoderma harzianum /
[ Ulhoa, C.J.;Peberdy, J.F. ] / J. Gen. Microbiol.   DOI
24 Identification of glutamic acid204 and aspartic acid 200 in chitinase Al of Bacillus circulans WL-12 as essential residues for chitinase activity /
[ Watanabe, T.;Kobori, K.;Miyashita, K.;Fujii, T.;Sakai, H.;Uchida, M.;Tanaka, H. ] / J. BioI. Chem.
25 Substrate distortion to a boat conformation at subsite -1 is critical in the machnism of family 18 chitinase /
[ Brameld, K.A.;Goddard, W.A. III, ] / J. Am. Chem. Soc.   DOI   ScienceOn
26 Chitinase from Neurospora crass /
[ Begovich, A.A. ] / Methods Enzymol.   DOI
27 Partial purification and characterisation of two extracellular N-acety1-D-g1ucosaminidases produced by the entomopathogenic fungus /
[ Bidochka, M.J.;Khachatourians, G.G. ] / Beauveria bassiana. Can. J. Microbiol.   DOI   ScienceOn
28 Physical methods for the determination of chitin structure and conformation /
[ Blackwell, J. ] / Methods Enzymol.   DOI
29 The condition of production of artificial fruiting body of Cordyceps militaris /
[ Choi, I.Y.;Choi, J.S.;Lee, W.H.;Yu, Y.J.;Joung, G.T.;Ju, I.O.;Choi, Y.K. ] / Kor. J. Mycol.
30 The molecular biology of chitin digestion /
[ Cohen-Kupies, R.;Chet, I. ] / Environ. Biotech.   DOI   ScienceOn
31 Chitinolytic enzymes of the gastric mucosa of Perodicticus potto (Primate prosimian): Purification and enzyme specifificity /
[ Cornelius, C.;Dandrifosse, G.;Jeuniaux, C.H. ] / Int. J. Biochem.   DOI   ScienceOn
32 Levels of chitinolytic activity during development of three entomopathogenic fungi /
[ Coudron, T.A.;Kroha, M.J.;Ignoffo, C.M. ] / Comp. Biochem. Physiol.   DOI   ScienceOn
33 Production of microbial chitinases-A revisit /
[ Felse, P.A.;Panda, T. ] / Bioprocess Engineering   DOI
34 Biological control of insect pests by entomogenous fungi /
[ Ferron, P. ] / Ann. Rev. Entomol.   DOI   ScienceOn
35 Ecology of entomopathogenic fungi.;Handbook of Applied Mycology 2 /
[ Glare, T.R.;Milner, R.J.;Arora, D.K.(ed.);Ajello, L.(ed.);Muketji, K.G.(ed.) ] /
36 Characterization and application of chitinases from Trichiderma harzianum-a review /
[ Gokul, B.;Lee, J.H.;Song, K.B.;Rhee, S.K.;Kim, C.H.;Panda, T. ] / Bioprocess Engineering   DOI
37 Gel electrophoresis of protein /
[ Hames, B.D.;Rickwood, D. ] /
38 Purification and characterization of chitinase produced by Strptomyces erythraeus /
[ Hara, S.;Yamamura, Y.;Fujii, Y.;Mega, T.;Ikenaka, T. ] / J. Biochem.
39 Induction and purification of Beauveria bassiana chitinolytic enzymes /
[ Havukkala, I.;Mitarnura, C.;Hara, S.;Hiratae, K.;Nishizawa, Y.;Hibi, T. ] / J. Invertebr. Pathol.   DOI   ScienceOn
40 Classification of chitinases modules;Chitin and Chitinases /
[ Henrissat, B.;Jolles, P.(ed.);Muzzarelli, R.A.(ed.) ] /
41 Cuticle-degrading enzymes of entomopathogenic fungi: Cuticle degradation in vitro by enzymes from entomopathogens /
[ Leger, R.J.;Cooper, R.M.;Charnley, A.K. ] / J. lnvertebr. Pathol.   DOI
42 Regulation and cloning of microbial chitinase gene /
[ Felse, P.A.;Panda, T. ] / Appl. Microbiol. Biotechnol.   DOI
43 Viscosmetric assay for chitinase /
[ Ohtakara, A.;Mitsutomi, M. ] / Methods Enzymol.   DOI