Optimization of Culture Conditions for toe Production of Chitinase

Chitinase 생성을 위한 배did 조건 최적화

  • 차진명 (조선대학교 공과대학, 화학·고분자공학부) ;
  • 석근영 ((주)환경과 생명 연구소, 조선이공대학 식품공업과) ;
  • 차월석 (조선대학교 공과대학, 화학·고분자공학부)
  • Published : 2001.08.01

Abstract

Chitinase producing microorganism, Serratia marcescens KY, was isolated from seashore mud around Beobseongpo in Chunnam province by selective enrichment culture. As the colloidal chitin concentration increased, chitinase production was increased. But chitinase production with addition of other carbon sources (glucose, fructose, galactose, maltose, sucrose, starch) was decreased. The effect of nitrogen sources on the chitinase production with serratia marcescens KY was as fellows. The opitimum mineral concentration for chitinase production was K$_2$HPO$_4$ 0.2 g/L and MgSO$_4$ 0.20 ∼ 0.25 g/L, respectively. The effect of nitrogen sources on chitinase production by Serratia marcescens KY was increased as follows, tryptone > yeast extract > beef extract > asparagine.

Serratia marcescens KY와 Serratia marcescens ATCC 27117 두 균주 모두 기본 배지에 $K_2$HPO$_4$ 농도를 0.2 g/L 농도 첨가할 경우 최대 chitinase 생성을 나타냈다. 그러나 세포 성장에 따른 균체량은 기본 배지에 $K_2$PHO$_4$ 농도 0.2 g/L 이상에서는 균체량은 약간 감소한다. 1.2% colloidal chitin과 0.2 g/L의 $K_2$PHO$_4$가 포함된 기본 배지에 MgSO$_4$를 0.2-0.25 g/L를 첨가하였을 때 최대의 chitinase 생성을 보이고, 두 균주 모두 K, P, Mg 및 기타 mineral을 영양 요구 인자로서 필요한다. Colloidal chitin을 1.2% 함유하고 있는 기본 배지에 각종 탄소원의 종류에 따른 세포 성장과 chitinase 생성은 colloidal chitin만을 첨가하였을 때가 상대적으로 가장 우수하고, 탄소원을 첨가할 경우 Serratia marcescens는 모든 탄소원에서 chitinase 생성이 억제되었다. 또한 질소원에 따른 세포 성장과 chitinase 생성은 Serratia marcescens KY와 Serratia marcescens ATCC 27117 모두 tryptone이 가장 우수하였고, 2.0 g/L의 질소원 농도까지는 질소원 농도가 증가함에 따라 chitinase 생성은 증가하다가 2.0 g/L 이상의 농도에서는 질소원 농도가 증가함에 따라 chitinase 생성은 감소하였다. 이들 질소원 중 chitinase 생성은 trypotone>yeast extract > beef extract > asparagine 순서로 chitinaserk 생성되므로, Serratia marcescens는 chitinase 생성에 있어 vitamin B군과 같은 질소원을 growth factor로 요구한다.

Keywords

References

  1. J.Biol.Chem. v.244 Extracellular nuclease from Serratia marcescens Nestle,M;W.K.Roberts
  2. Appl.Envir.Microbiol v.63 Purification and characterization of two bifuctional chitinase/lysozymers extracellularly produced by Pseudomonas aeruginosa K-187 in shrimp and crab shell powder medium Wang,S.L.;W.T.Chang
  3. Biosci.Biotech.Biochem v.95 The action of Bacillus circulans WL-12 chitinase on partially N-acetylated chitosan Mitutomi,M.;H.Kidoh;H.Tomita;T.Watanabe
  4. Agric.Biol.Chem v.54 Action pattern of Streptomyces griseus chitinase on partially N-acetylated chitosan Ohtakara,A.;H.Matsunaga;M.Mitsutomi
  5. Appl.Biochem.Biotechnol v.37 Purification and some properties of chitinase from Aspergillus carneus Mohamed,A.Abdel-Naby;Nefsia M.El-Shayeb;A.A.Sherief
  6. J.Biol.Chem. v.257 Endochitinase a mannon-associated enzyme from Saccharomyces cerevisiae Correa,J.U.;N.Elango;I.Polacheek;E.Cabib
  7. J.Bacteriol v.20 Extracelluar enzyme from Myxobacter AL-1 that exhibits both B-1,4-glucanase and chitosanase activities Hedges,A.;R.S.Wolfe
  8. Korean Biochem.J. v.24 Detection of chitinase activity using fluorescence-labeled substrate on polyscrylamide gel Kim,Y.S.;K.B.Lee;R.J.Linhardt
  9. J.Gen.Appl.Microbiol v.32 Purification and characterization of chitinase and chitobiase produced by Aeromonas hydrophita subsp Yabuki,M.;K.Mizushima,T.;Ando,T;Fugi;M.Sbimada;M.Yamasbita
  10. Anal.Biochem. v.127 Serratia marcescens chitinase:one-step purification and use for the determination of chitin Roberts,R.H.;E.Cubib
  11. Can.J.Microbiol v.15 The chitinase of Serraita marcescens Monreal,J.;E.T.Reese
  12. Anal.Chem v.31 Use of dinitrosalicylic acid reagent for determination of reducing sugar Miller,L.
  13. J.Ferment.Technol v.57 Purification and some properties of chitinase from Vibiro sp. Ohtakara,A.;M.Mitsutomi
  14. J.Microbiol v.10 no.2 Isolation and characterization of chitosanase-producing microorganism,Aureobacterium sp. Lee,D.H.;E.L.Lee;K.H.Lee
  15. J.P.Zikakis(ed.) Chitin,chitosan and related enzyme Hadwiger,L.A.;B.Fristensky;R.C.Riggleman
  16. Appl.Envir.Microbiol v.63 no.2 Purification and characterization of two bifuctional chitinase/lysozymers extracellularly produced by Pseudomonas aeruginosa K-187 in shrimp and crab shell powder medium Wang,S.L.;W.T.Chang
  17. Can.J.Microbiol v.38 Purification,properties and partial amino acid sequence chitinase from a marine Alteromonas sp. Hiroshi,T.;Y.Yugio;K.Miyamoto
  18. EMBO J v.5 Isolation and characterization of genes encoding two chitinase enzymes from Serraita marcescenes Jones,J.D.G.;K.L.Grady;T.V.Suslow;J.R.Bedbrook