Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Production and Purification of Polygalacturonase from Rhizopus sp.

Rhizopus속이 생성하는 Polygalacturonase의 생산 및 정제

  • Chung, Yung-Gun (Dept. of Food Science and Technology, Yeungnam University) ;
  • Cho, Young-Je (Dept. of Food Science and Technology, Yeungnam University) ;
  • Kwon, Oh-Jin (Dept. of Food Science and Technology, Yeungnam University) ;
  • Choi, Cheong (Dept. of Food Science and Technology, Yeungnam University)
  • 정영건 (영남대학교 식품가공학과) ;
  • 조영제 (영남대학교 식품가공학과) ;
  • 권오진 (영남대학교 식품가공학과) ;
  • 최청 (영남대학교 식품가공학과)
  • Published : 1992.04.01
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Abstract

Rhizopus oryzae CJ-2114 was selected for its strong polygalacturonase activity among various strains of mold found in soil. It was found that the production of polygalacturonase reached to maximum when the wheat bran medium containing 1% albumin, 1% sorbitol and 0.2% (NH$_4$)$_2$C$_2$O$_4$was cultured for 96 hrs at 3$0^{\circ}C$. Polygalacturonase was purified 11.13 fold from Rhizopus oryzae CJ-2114. The purification procedures include ammonium sulfate treatment, gel filtration on Sephadex G-75, G-150 and DEAE-cellulose ion exchange chromatography. Yield of the enzyme purification was 40.3% .Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. When the purified enzyme was applied to SDS-poly-acrylamide gel electrophoresis, the molecular weight was estimated to be 47,000. The amino acid composition indicated relatively high contents of glutamic acid and glyrine.

Rhizopus oryzae CJ-2114의 polygalacturonase 생성을 위한 최적조건은 수분이 60% 함유된 밀기울 배지에 1% albumin, 0.2% (NH$_4$)$_2$C$_2$O$_4$, 1% sorbitol을 첨가하여 96시간 배양시 최대활성을 나타내었으며, Sep-hadex G-75 및 G-150을 사용한 gel filtration과 DEAE-cellulose에 의한 이온교환 크로마토그라피를 통하여 이 효소를 11.13배 정제할 수 있었고, 수율은 40.3% 였다. 정제효소는 polyacrylamide gel 전기 영동에 의하여 단일밴드로 확인되었으며 분자량은 SDS-polyacryl-amide 전기영동에 의하여 47,000정도로 측정되었다. 효소의 결정구조는 표면이 거친 기둥모양을 형성하고 있었으며 아미노산 조상은 17종류로써 glutamic acid 함량이 198.74mg/g enzyme로 가장 많았다.

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