Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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Purification and characterization of an alkaline protease produced by a xanthomonas sp. YL-37

  • Published : 1995.06.01
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Abstract

The alkaline protease of Xanthomonas sp. YL-37 has been purified, and the properties of the enzyme investigated. The alkaline protease of Xanthomonas sp. YL-37 was purified form crude enzyme by ammonium sulfate fractionation, CM-cellulose ion exchange chromatography, and Sephadex G-100 gel filtration. Through the series of chromatographies, the enzyme was purified to homogenecity with specific activity of 4.23 fold higher than that of the crude broth. The molecular weight of the purified protease has been estimated to be 62 KDa on SDS-polyacrylamide gel electrophoresis. The optimal pH and temperature for alkaline protease activity were 11.0 and 50.deg.C, respectively. The enzyme was stable between pH 5.0 and 10.0 and up to 50.deg.C. Enzyme activity was lost up to 50% on heating at 70.deg.C for 30 minutes. The activity of alkaline protease was inhibited by Cu$\^$2+/, Zn$\^$2+/, Hg$\^$2+/, PMSF, and activated by Mn$\^$2+/ and Ca$\^$2+/. The $K_{m}$ value for casein as a substrate was 4.0 mg/ml.

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References

  1. J. Biochem. v.91 Estimation of molecular weight of proteins by Sephadex gel filtration Andrews,P.
  2. Kor. J. Appl. Microbiol. Bioeng. v.17 Purification and characterization of thermotolerable alkaline protease by alkalophilic Bacillus sp. No. 8-116 Bae,M.;P.Y.Park
  3. Anal. Biochem. v.72 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Bradford,M.M.
  4. Annu. Rev. Biochem. v.29 Proteolytic enzymes Hatley,B.S.
  5. Argic. Biol. Chem. v.35 Production of alkaline enzymes by alkalophilic microorganisms. Part I. Akaline protease produced by Bacillus No. 221 Horikoshi,K.;T.Akiba
  6. Alakalophilic microorganisms: A new microbial world Horikoshi,K.;T.Akiba
  7. Argic. Biol. Chem. v.53 Alkaline cellulase for laundry detergents: Production by Bacillus sp. KSM-615 and enzymatic properties Ito,S.;S.Shikata;K.Ozaki;S.Kawai;K.Okamoto;S.Inous;T.Satoh
  8. Kor. J. Appl. Microbiol. Biotech. v.17 Purification and properties of extracellular protease from Streptomyces rimosus Kim,K.M.;T.K.Lee;H.C.Yang
  9. Kor. J. Appl. Microbiol. Biotech. v.18 Properties of alkaline protease produced by an alkalophilic Bacillus sp. Kim,T.H.;S.H.Park;D.S.Lee;T.K.Kwon;J.K.Kim;S.D.Hong
  10. Nature v.227 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmli,U.K.
  11. Kor. J. Appl. Microbiol. Biotech. v.22 Production and characterization of an alkaline protease from an isolate, Xanthomonas sp. YL.-37 Lee,C.H.;T.J.Kwon;S.M.Kang;H.H.Suh;G.S.Kwon;H.M.Oh;B.D.Yoon
  12. J. Mol. Biol. v.76 The relationship of serine protease activity to RNA polymerase modification and sporulation in Bacillus subtilis Leighton,T.J.;R.H.Doi;R.A.J.Warren;R.A.Kelln
  13. J. Am. Chem. Soc. v.56 The determination of enzyme dissociation constants Lineweaver,J.;D.Burk.
  14. J. Biotech. v.24 Purification of an alkaline serine protease produced by a psychroatrophic Bacillus sp. Margesin,R.;N.Palma;F.Knauseder;F.Schinner
  15. Appl. Environ. Microbiol. v.53 Purification and properties of an extracellular protease produced by the entomopathogenic fungus Beauveria bassiana Michel,J.B.;G.G.Khachatourians
  16. Argic. Biol. Chem. v.55 Purification and properties of a novel surface-active agent and alkaline-resistant protease from Bacillus sp. Y. Shimogaki,H.;K.Takeuchi;T.Nishimo;M.Ohdera;T.Kudo;K.Ohba;M.Iwana;M.Irie
  17. Annu. Rev. Biochem. v.44 Chemical studies of enzyme activities Sigman,D.S.;G.Mooser
  18. J. Appl. Bact. v.69 Purification and characterization of two types of alkaline serine proteases produced by an alkalophilic actinomycete Tsujibo,H.;K.Miyamoto;T.Hesegawa;Y.Inamori