期刊
BIOTECHNOLOGY LETTERS
卷 33, 期 3, 页码 557-563出版社
SPRINGER
DOI: 10.1007/s10529-010-0456-9
关键词
D-Amino acid oxidase; Permeabilization; Pichia pastoris; Rhodotorula gracilis; Stability
资金
- Osterreichische Forschungsforderungsgesellschaft mbH (FFG)
- province of Vienna
- province of Styria
- province of Tyrol
- Steirische Wirtschaftsforderungsgesellschaft mbH (SFG)
- Ingenza Ltd.
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.
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