Porduction and Enzymatic Characteristics of ${\alpha}-Galactosidase$ from Aspergillus niger

Aspergillus niger에 의한 ${\alpha}-Galactosidase$의 생산 및 효소적 특성

  • Chun, Hyang-Sook (Department of Food & Nutrition, Ewha Woman's University) ;
  • Lee, Su-Rae (Department of Food & Nutrition, Ewha Woman's University)
  • 전향숙 (이화여자대학교 식품영양학과) ;
  • 이서래 (이화여자대학교 식품영양학과)
  • Published : 1988.02.01

Abstract

${\alpha}$-Galactosidase from Aspergillus niger as a possible enzyme for removal of flatulence factors in soybean foods was produced the highest in 120 hours in either Czapeck-Dox liquid medium or wheat bran solid medium. The most efficient carbon and nitrogen sources in Czapeck-Dox medium were raffinose and sodium nitrate, respectively, whereas the addition of the sources showed negative effects in wheat bran. pH optima for enzyme activity and stability were 4.0-5.0 and 3.5-6.5, respectively, and optimum temperature for stability was $40-50^{\circ}C$. Upon reaction on p-nitrophenyl-${\alpha}$-D-galactoside, Michaelis constant was 0.42 mM and maximum velocity was 152 ${\mu}moles$ substrate/minute/kg solid medium. Mercuric chloride acted as a strong noncompetitive inhibitor and p-chloromercuribenzoate, even in low concentration, acted as a competitive inhibitor. Crude ${\alpha}$-galactosidase hydrolyzed raffinose and stachyose completely, giving spots of monosaccharides only on thin-layer chromatogram.

대두(大豆)중에 존제하는 장내(腸內) 가스 발생인자인 raffinose와 stachyose의 제거에 이용될 수 있는 Aspergillus niger의 ${\alpha}$-galactosidase는 Czapeck-Dox 액체배지와 밀기울 배지에서 5일간 배양하였을 때 효소생성이 제일 높았다. Czapeck-Dox 액체배양에서 raffinose와 $NaNO_3$가 효소생산에 가장 효과적인 탄소원과 질소원으로 나타났으며 밀기울 배양에서는 질소원 또는 탄소원의 첨가효과가 없었다. 생성된 조효소(組酵素)용액의 작용 최적 pH는 4.0-5.0, 안정도 최적 pH는 3.5-6.5, 최적온도는 $40-50^{\circ}C$ 였다. 합성기질인 p-nitrophenyl-${\alpha}$-D-galactoside에 대한 Michaelis상수는 0.42mM, 최대반응속도는 152${\mu}moles$ substrate/min/kg (고체배지)이었다. $HgCl_2$는 강력한 비경쟁 저해제로 작용하였고 p-chloromercuribenzoate는 낮은 농도에서 경쟁 저해제로 작용하였다. 조효소 용액은 raffinose와 stachyose를 완전 분해하여 단당류 만을 생성하였다.

Keywords