YAKHAK HOEJI (약학회지)

The Pharmaceutical Society of Korea (대한약학회)
권호 표지

Thermostability and Resistance to Proteolysis of L-Asparaginase Purified from Strepfomyces lincolnensis M-20

Strepfomyces lincolnensis M-20 균주로 부터 분리, 정제된 L-Asparaginase의 열안정성과 단백 가수 분해 효소에 대한 저항성

  • 김경자 (순천향대 자연과학대학 생명공학과)
  • Published : 2007.06.30
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Abstract

Thermostable asparaginase was purified to homogeneity from mesophilic Strepfomyces lincolnensis M-20 by 30${\sim}$70% ammonium sulfate precipitation and asparagine-Sepharose CL 6B affinity column chromatography, The apparent molecular mass of L-asparaginase by SDS-PAGE was found to be 47 kDa, whereas by its mobility on Sephacryl S-300 column was around 180 kDa, indicating that the enzyme at the native stage acts as tetramer, The purified enzyme showed a single band on acrylamide gel electrophoresis. The optimum pH and temperature were pH 9.5 and 55${\circ}$C, respectively. Chemical modification experiments of purified asparagines implied the existence cystein residue located at or near active site. Purified asparaginase retained the 85% of the initial activity after incubation at 90${\circ}$C for 30 min. A correlation between themostability and resistance to proteolysis of commercial asparaginase and purified asparaginase from Strepfomyces lincolnensis M-20 was investigated. Purified thermostable asparaginase was resistant to trypsin and chymotrypsin treatment, while the commercial asparaginase was not themostable and was susceptible to proteolytic treatment with trypsin and chymotrypsin.

Keywords

References

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