EXPRESSION OF L-N-CARBAMOYLASE GENE IN PICHIA PASTORIS
- VernacularTitle:N-氨甲酰基氨基酸水解酶在毕赤酵母中的表达
- Author:
Wei-Cai ZHANG
;
Ying-Li LI
;
Yan-Ming ZHANG
;
Bing-Bing DENG
;
Liu-Yu HUANG
;
- Publication Type:Journal Article
- Keywords:
L-N-Carbamoylase, Pichia pastaris, Gene expression
- From:
Microbiology
1992;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
N-carbamoylase is a part of hydantoinase operon which can transform N-carbamoylamino acid to corresponding ammo acids. The L-N-carbamoylase of Arthrobacter BT801, codied by the HyuC gene, is the rate-limiting and the only stereoselective enzyme. HyuC DNA fragment was amplified by PCR from the plasmid of pUC18-169. The target fragment was introduced into pPIC3. 5K plasmid to construct the pPIC3. 5K-hyuC expressing vector which was then transduced into Pichia pastoris GS115 cells after being linearized by BglⅡ digestion. Multi-copies insertion transformants were screened on G418 plates. The recombinant protein was proved to have biological activity of hydrolyzing N-carbamoylphenylalanine into phenylalanine through enzyme activity determination.
