Biochemical Characterization of an Extracellular beta-Glucosidase from the Fungus, Penicillium italicum, Isolated from Rotten Citrus Peel.
- Author:
Ah Reum PARK
1
;
Joo Hee HONG
;
Jae Jin KIM
;
Jeong Jun YOON
Author Information
1. Green Materials Technology Center, Chungcheong Regional Division, Korea Institute of Industrial Technology (KITECH), Cheonan 331-825, Korea. jjyoon@kitech.re.kr
- Publication Type:Original Article
- Keywords:
beta-Glucosidase;
Cellulolytic fungi;
Characterization;
Identification;
Purification
- MeSH:
beta-Glucosidase;
Cellobiose;
Cellulases;
Cellulose;
Chromatography;
Citrus;
Fungi;
Glucose;
Glucosides;
Hydrogen-Ion Concentration;
Penicillium
- From:Mycobiology
2012;40(3):173-180
- CountryRepublic of Korea
- Language:English
-
Abstract:
A beta-glucosidase from Penicillium italicum was purified with a specific activity of 61.8 U/mg, using a chromatography system. The native form of the enzyme was an 88.5-kDa tetramer with a molecular mass of 354 kDa. Optimum activity was observed at pH 4.5 and 60degrees C, and the half-lives were 1,737, 330, 34, and 1 hr at 50, 55, 60, and 65degrees C, respectively. Its activity was inhibited by 47% by 5 mM Ni2+. The enzyme exhibited hydrolytic activity for p-nitrophenyl-beta-D-glucopyranoside (pNP-Glu), p-nitrophenyl-beta-D-cellobioside, p-nitrophenyl-beta-D-xyloside, and cellobiose, however, no activity was observed for p-nitrophenyl-beta-D-lactopyranoside, p-nitrophenyl-beta-D-galactopyranoside, carboxymetyl cellulose, xylan, and cellulose, indicating that the enzyme was a beta-glucosidase. The kcat/Km (s-1 mM-1) values for pNP-Glu and cellobiose were 15,770.4 mM and 6,361.4 mM, respectively. These values were the highest reported for beta-glucosidases. Non-competitive inhibition of the enzyme by both glucose (Ki = 8.9 mM) and glucono-delta-lactone (Ki = 11.3 mM) was observed when pNP-Glu was used as the substrate. This is the first report of non-competitive inhibition of beta-glucosidase by glucose and glucono-delta-lactone.
