Overproduction, Purification and Characterization of Adenylate Deaminase from Aspergillus oryzae

被引：3

作者：

Li, Shubo ^{[1
]}

Qian, Yi ^{[2
]}

Liang, Yunlong ^{[2
]}

Chen, Xinkuan ^{[2
]}

Zhao, Mouming ^{[1
]}

Guo, Yuan ^{[3
]}

Pang, Zongwen ^{[2
,4
]}

机构：

[1] Guangxi Univ, Coll Light Ind & Food Engn, Nanning 530004, Peoples R China

[2] Guangxi Univ, Coll Life Sci & Technol, Nanning 530004, Peoples R China

[3] Guangxi Acad Sci, Natl Engn Res Ctr Nonfood Biorefinery, Nanning 530004, Peoples R China

[4] Guangxi Univ, Coll Life Sci & Technol, Nanning 530005, Peoples R China

来源：

APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY | 2016年 / 180卷 / 08期

关键词：

Adenylate deaminase; Aspergillus oryzae; Purification; Catalytic properties; Inosine-5-monophosphate; MUSCLE AMP-DEAMINASE; PROTEIN; ACID;

D O I：

10.1007/s12010-016-2192-7

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Adenylate deaminase (AMPD, EC 3.5.4.6) is an aminohydrolase that widely used in the food and medicine industries. In this study, the gene encoding Aspergillus oryzae AMPD was cloned and expressed in Escherichia coli. Induction with 0.75 mM isopropyl beta-d-l-thiogalactopyranoside resulted in an enzyme activity of 1773.9 U/mL. Recombinant AMPD was purified to electrophoretic homogeneity using nickel affinity chromatography, and its molecular weight was calculated as 78.6 kDa. Purified AMPD exhibited maximal activity at 35 A degrees C, pH 6.0 and 30 mM K+, with apparent K (m) and V (max) values of 2.7 x 10(-4) M and 77.5 mu mol/mg/min under these conditions. HPLC revealed that recombinant AMPD could effectively catalyse the synthesis of inosine-5'-monophosphate (IMP) with minimal by-products, indicating high specificity and suggesting that it could prove useful for IMP production.

引用

页码：1635 / 1643

页数：9

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