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

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Formation and Fusion of Protoplasts from the Cellulolytic Fungi, Aspergillus niger MAN-831 and Aspergillus wentii MAW-538

Cellulase를 생산하는 Aspergillus niger MAN-831과 Aspergillus wentii MAW-538의 원형질체 형성 및 융합

  • Published : 1995.12.01
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Abstract

For the effective utilization of cellulosic biomass, conidial protoplast fusion between Aspergillus niger MAN-831(${\beta}-glucosidase$) and A. wentii MAW-538(CMCase and avicelase), which produced potently cellulolytic enzymes was carried out. Optimal conditions for formation and regeneration of protoplast were conidiospore age-5 dyuas. $2-DG-30\mu\textrm{g}/ml$, preincubation time-4 hours, osmotic stabilizer-0.7M KCl, novozyme(7mg/ml)＋driselase(2.5mg/ml) and reaction time of enzyme-5 hours. Optimal conditions for protoplast fusion were obtained by treatment of protoplasts with 15mM CaCl2 and 25% polyethylene glycol 4000(pH 6~7) as fusogenic agent at $36^{\circ}C$ for 25~30 minutes. The frequency was then $7.94{\times}10^{-4}$. CMCase, avicelase and ${\beta}-glucosidase$ activity of fusant F-208 strain was 1.5, 1.3, 1.2 times higher than those of parental strains, respectively.

섬유성 biomass의 효율적인 이용을 위하여 CMCase, avicelase 분비력이 높은 균주(A. wentii MAW-598)와 ${\beta}-glucosidase$ (A. niger MAN-831) 분비력이 우수한 균주의 포자를 이용한 원형질세 형성.재생조건 및 융합을 실시하였다. 원형질체 형성 및 재생의 최적조건은 5일 성숙한 포자를 $30\mu\textrm{g}/ml$ 농도의 2-DG로 4시간 전배양하여 0.7M KCI 삼투압 안정제, novozyme(7.5mg/ml)+driselase(2.5mg/ml) 혼합효소로 5시간 반응시키는 것이 효과적이었다. 원형질체 융합 조건은 $CaCl_2$ 15mM, 25% PEG 4000, pH 5-7, $36^{\circ}C$에서 25~30분간 처리하였을때 효과적이었으며, 융합빈도는 $7.94\;{\times}\;10^{-4}$로 나타났다. FA-208 융합주는 친주에 비하여 각각 1.5, 1.3, 1.2배 효소활성의 증가를 나타내었다.

Keywords

References

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