Abstract
A β-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 60°C, and the half-lives were 1,737, 330, 34, and 1 hr at 50, 55, 60, and 65°C, respectively. Its activity was inhibited by 47% by 5 mM Ni2+. The enzyme exhibited hydrolytic activity for p-nitrophenyl-β-D-glucopyranoside (pNP-Glu), p-nitrophenyl-β-D-cellobioside, p-nitrophenyl-β-D-xyloside, and cellobiose, however, no activity was observed for p-nitrophenyl-β-D-lactopyranoside, p-nitrophenyl-β-D-galactopyranoside, carboxymetyl cellulose, xylan, and cellulose, indicating that the enzyme was a β-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 β-glucosidases. Non-competitive inhibition of the enzyme by both glucose (Ki = 8.9 mM) and glucono-δ-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 β-glucosidase by glucose and glucono-δ-lactone.
| Original language | English |
|---|---|
| Pages (from-to) | 173-180 |
| Number of pages | 8 |
| Journal | Mycobiology |
| Volume | 40 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - 2012 Dec 1 |
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Keywords
- β-Glucosidase
- Cellulolytic fungi
- Characterization
- Identification
- Purification
ASJC Scopus subject areas
- Plant Science
- Microbiology
- Infectious Diseases
Cite this
Biochemical characterization of an extracellular β-glucosidase from the fungus, Penicillium italicum, Isolated from rotten citrus peel. / Park, Ah Reum; Hong, Joo Hee; Kim, Jae-Jin; Yoon, Jeong Jun.
In: Mycobiology, Vol. 40, No. 3, 01.12.2012, p. 173-180.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Biochemical characterization of an extracellular β-glucosidase from the fungus, Penicillium italicum, Isolated from rotten citrus peel
AU - Park, Ah Reum
AU - Hong, Joo Hee
AU - Kim, Jae-Jin
AU - Yoon, Jeong Jun
PY - 2012/12/1
Y1 - 2012/12/1
N2 - A β-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 60°C, and the half-lives were 1,737, 330, 34, and 1 hr at 50, 55, 60, and 65°C, respectively. Its activity was inhibited by 47% by 5 mM Ni2+. The enzyme exhibited hydrolytic activity for p-nitrophenyl-β-D-glucopyranoside (pNP-Glu), p-nitrophenyl-β-D-cellobioside, p-nitrophenyl-β-D-xyloside, and cellobiose, however, no activity was observed for p-nitrophenyl-β-D-lactopyranoside, p-nitrophenyl-β-D-galactopyranoside, carboxymetyl cellulose, xylan, and cellulose, indicating that the enzyme was a β-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 β-glucosidases. Non-competitive inhibition of the enzyme by both glucose (Ki = 8.9 mM) and glucono-δ-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 β-glucosidase by glucose and glucono-δ-lactone.
AB - A β-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 60°C, and the half-lives were 1,737, 330, 34, and 1 hr at 50, 55, 60, and 65°C, respectively. Its activity was inhibited by 47% by 5 mM Ni2+. The enzyme exhibited hydrolytic activity for p-nitrophenyl-β-D-glucopyranoside (pNP-Glu), p-nitrophenyl-β-D-cellobioside, p-nitrophenyl-β-D-xyloside, and cellobiose, however, no activity was observed for p-nitrophenyl-β-D-lactopyranoside, p-nitrophenyl-β-D-galactopyranoside, carboxymetyl cellulose, xylan, and cellulose, indicating that the enzyme was a β-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 β-glucosidases. Non-competitive inhibition of the enzyme by both glucose (Ki = 8.9 mM) and glucono-δ-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 β-glucosidase by glucose and glucono-δ-lactone.
KW - β-Glucosidase
KW - Cellulolytic fungi
KW - Characterization
KW - Identification
KW - Purification
UR - http://www.scopus.com/inward/record.url?scp=84875185625&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84875185625&partnerID=8YFLogxK
U2 - 10.5941/MYCO.2012.40.3.173
DO - 10.5941/MYCO.2012.40.3.173
M3 - Article
C2 - 23115510
AN - SCOPUS:84875185625
VL - 40
SP - 173
EP - 180
JO - Mycobiology
JF - Mycobiology
SN - 1229-8093
IS - 3
ER -