Protoplast Fusion Between Aspergillus oryzae and Aspergillus shirousamii

Aspergillus oryzae와 Aspergillus shirousamii간의 원형질체의 융합

  • Shin, Dong-Bun (Department of Food Science and Technology, Kyungsung University) ;
  • Ryu, Beung-Ho (Department of Food Science and Technology, Kyungsung University) ;
  • Jin, Seung-Heun (Department of Food Science and Technology, Kyungsung University)
  • 신동분 (경성대학교 식품공학과) ;
  • 류병호 (경성대학교 식품공학과) ;
  • 진성현 (경성대학교 식품공학과)
  • Published : 1993.08.01

Abstract

This study mainly designed to high quality of mirin production by using protopast fusion. In order to enhance the acid carboxypeptidase (ACPase) activity by the method of protoplast fusion. In order to enhance the acid carboxypeptidase (ACPase) activity by the method of protopalst fusion, the mutants, Aspergillus oryzae 9-12 and Aspergillus shirosamii IFO 6082-60 were selected by mutation among various mutants. Protoplast of Aspergillus oryzae 9-12 and Aspergillus shirousamii IFO 6082-60 were formed effectively by incubation of the mixtures of chitinase (10mg/ml), cellulase (10mg/ml) and zymolase 20T (5mg/ml). For protopalst fusion, the mixture of two mutant were fused to effective under the optimum conditions by solutions containing 30% PEG 6,000, 0.01M $CaCl_2\;2H_2O$, 0.6M KCl and 0.05M glycine. Fusion frequency was 0.71% and fusant, F-50 appeared ACPase activity of 20,800 unit/g which has 1.5 times higher than that of each mutants.

본 연구는 미림 제조시 미림의 품질을 높이기 위한 방법의 하나로 세포 원형질체의 융합체 의해 ACPase 활성이 높은 균주를 얻으려고 시도하였다. 돌연변이 방법으로 acid carboxypeptidase(ACPase) 활성이 높은 A. oryzae 9-12와 A. shirousamii IFO 6082-60을 선별하여 사용하였다. A. oryzae 9-12와 A. shirousamii IFO 608260의 원형질체의 형성조건은 chitinase (10mg/ml), cellulase(10mg/ml) 및 zymolase(20T(5mg/ml)등의 효소 혼합용액에서 $30^{\circ}C$에서 4시간 반응시켰을 때 가장 높았으며, pH6.0일 때, 안정제로서는 0.6M KCl 및 무기 염류로서는 0.01M $CaCl_2\;2H_2O$가 가장 높았고, 30% PEG 6,000이 가장 좋았다. 이때의 융합빈도는 0.71%였다. 융합주의 ACPase의 활성은 F-50이 20,800unit/g으로 변이주보다 1.4배 높았다.

Keywords