잿빛곰팜이병균 Botrytis cinera가 분비하는 Polygalacturonase의 부분정제와 특성

Partial Purification and Properties of Polygalacturonase Produced by Botrytis cinerea

  • 나유진 (경상대학교 자연과학대학 미생물학과) ;
  • 김재원 (경상대학교 자연과학대학 미생물학과) ;
  • 정영륜 (경상대학교 자연과학대학 미생물학과) ;
  • 허남응 (경상대학교 자연과학대학 미생물학과) ;
  • 조광연 (한국화학연구소 스크리닝 안정성센터)
  • 발행 : 1994.06.01

초록

Polygalacturonase (PG) produced by Botrytis cinerea in the culture broth containing citrus pectin as a carbon source was partially purified and characterized. PG was produced on a range of carbon sources such as starch, glycerol, cellobiose, and Na+-PAG with total activities of 34.8, 32.0, 29.2, 27.8 units, respectively. The specific activity was highest with 2316.7 units on Na+-PGA. Proteins of culture filtrate were concentrated with polyethylene glycol and acetone and applied to a hydroxyapatite column. Among three active fractions collected from the column, the reaction containing the highest PG activity was resolved by a Q-sepharose column. The active fraction from the Q-sepharose column was further purified by HPLC Mono Q column. The partially purified enzyme was analyzed by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Among a few protein bands revealed, the amount of the protein of which molecular weight estimated to be 43 kDa coincided with the PG activity. The partially purified PG had optimal temperatures between 35~55$^{\circ}C$ and pH between 4.5~5.5.

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