Purification and characterization of a beta-glucosidase from Aspergillus niger

被引:37
作者
Rashid, MH [1 ]
Siddiqui, KS [1 ]
机构
[1] NATL INST BIOTECHNOL & GENET ENGN, FAISALABAD, PAKISTAN
关键词
Cellobiose; Ammonium Sulfate; Wheat Bran; Ammonium Sulfate Precipitation; Kallar Grass;
D O I
10.1007/BF02815462
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The high-molar mass form of beta-glucosidase from Aspergillus niger strain NIAB280 was purified to homogeneity with a 46-fold increase in purification by a combination of ammonium sulfate precipitation, hydrophobic interaction, ion-exchange and gel-filtration chromatography. The native and subunit molar mass was 330 and 110 kDa, respectively. The pH and temperature optima were 4.6-5.3 and 70 degrees C, respectively. The K-m and k(cat) for 4-nitrophenyl beta-D-glucopyranoside at 40 degrees C and pH 5 wee 1.11 mmol/L and 4000/min, respectively. The enzyme was activated by low and inhibited by high concentrations of NaCl. Ammonium sulfate inhibited the enzyme. Thermolysin periodically inhibited and activated the enzyme during the course of reaction and after 150 min of proteinase treatment only 10 % activity was lost with concomitant degradation of the enzyme into ten low-molar-mass active bands. When subjected to 0-9 mol/L transverse urea-gradient-PAGE for 105 min at 12 degrees C, the nonpurified beta-glucosidase showed two major bands which denatured at 4 and 8 mol/L urea, respectively, with half-lives of 73 min.
引用
收藏
页码:544 / 550
页数:7
相关论文
共 20 条
[1]  
[Anonymous], [No title captured]
[2]  
BENNETT JW, 1992, ASPERGILLUS BIOL IND, P175
[3]  
COUGHLAN MP, 1988, METHOD ENZYMOL, V160, P135
[4]  
DESHPANDE V, 1988, METHOD ENZYMOL, V160, P415
[5]  
ESEN A, 1993, ACS SYM SER, V533, P1
[6]  
ESEN A, 1993, ACS SYM SER, V533, P214
[7]  
GOLDENBERG DP., 1989, PROTEIN STRUCTURE PR, P225
[8]   ISOLATION AND CHARACTERIZATION OF 2 FORMS OF BETA-D-GLUCOSIDASE FROM ASPERGILLUS-NIGER [J].
HIMMEL, ME ;
ADNEY, WS ;
FOX, JW ;
MITCHELL, DJ ;
BAKER, JO .
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, 1993, 39 :213-225
[9]   ISOLATION AND CHARACTERIZATION OF BETA-GLUCOSIDASES FROM ASPERGILLUS-NIDULANS MUTANT USDB-1183 [J].
HOH, YK ;
YEOH, HH ;
TAN, TK .
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY, 1993, 9 (05) :555-558
[10]  
KVESITADZE GI, 1990, BIOORG KHIM+, V16, P881