Expression and characterization of sweet potato invertase in Pichia pastoris

被引：15

作者：

Huang, WC ^{[1
]}

Wang, AY ^{[1
]}

Wang, LT ^{[1
]}

Sung, HY ^{[1
]}

机构：

[1] Natl Taiwan Univ, Dept Agr Chem, Taipei 10764, Taiwan

来源：

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY | 2003年 / 51卷 / 05期

关键词：

vacuolar invertase; cDNA; sweet potato (Ipomoea batatas); recombinant invertase; Pichia pastoris; glycoprotein;

D O I：

10.1021/jf026032i

中图分类号：

S [农业科学];

学科分类号：

09 ;

摘要：

An invertase cDNA (Ibbetafruct1) was cloned from sweet potato leaves and characterized. The deduced amino acid sequence of the Ibbetafruct1-encoded protein was closely related to vacuolar invertases and included the WECVD catalytic domain characteristic of them. An expression plasmid containing the coding region of Ibbetafruct1 under the control of the alcohol oxidase promoter was used to transform the methylotrophic yeast Pichia pastoris. The biochemical properties for the expressed recombinant enzyme, which was determined to be the acid beta-fructofuranosidase with an acidic pl value (5.1), were similar to those of vacuolar invertases purified from sweet potato. Periodic acid/Schiff staining and Con A-Sepharose gel-binding experiments revealed the recombinant invertase to be a glycoprotein containing glucose and/or mannose residues. Furthermore, the carbohydrate moiety appears to be a key determinant of the enzyme's sucrose hydrolysis activity, substrate affinity, and thermal stability.

引用

页码：1494 / 1499

页数：6

共 36 条

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