High-level expression of Rhodotorula gracilis d-amino acid oxidase in Pichia pastoris

被引：12

作者：

Abad, Sandra ^{[1
]}

Nahalka, Jozef ^{[1
,2
]}

Winkler, Margit ^{[1
]}

Bergler, Gabriele ^{[1
]}

Speight, Robert ^{[4
]}

Glieder, Anton ^{[1
,3
]}

Nidetzky, Bernd ^{[1
,2
]}

机构：

[1] Austrian Ctr Ind Biotechnol, A-8010 Graz, Austria

[2] Graz Univ Technol, Inst Biotechnol & Biochem Engn, A-8010 Graz, Austria

[3] Graz Univ Technol, Inst Mol Biotechnol, A-8010 Graz, Austria

[4] Ingenza Ltd, Roslin EH25 9PP, Midlothian, Scotland

来源：

BIOTECHNOLOGY LETTERS | 2011年 / 33卷 / 03期

关键词：

D-Amino acid oxidase; Permeabilization; Pichia pastoris; Rhodotorula gracilis; Stability; TRIGONOPSIS-VARIABILIS; CATALYTIC-PROPERTIES; CEPHALOSPORIN-C; STABILIZATION; IMMOBILIZATION; MECHANISM; PROTEIN; ENZYME;

D O I：

10.1007/s10529-010-0456-9

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：

By combining gene design and heterologous over-expression of Rhodotorula gracilis d-amino acid oxidase (RgDAO) in Pichia pastoris, enzyme production was enhanced by one order of magnitude compared to literature benchmarks, giving 350 kUnits/l of fed-batch bioreactor culture with a productivity of 3.1 kUnits/l h. P. pastoris cells permeabilized by freeze-drying and incubation in 2-propanol (10% v/v) produce a highly active (1.6 kUnits/g dry matter) and stable oxidase preparation. Critical bottlenecks in the development of an RgDAO catalyst for industrial applications have been eliminated.

引用

页码：557 / 563

页数：7

共 26 条

[1] Stepwise engineering of a Pichia pastoris D-amino acid oxidase whole cell catalyst [J].