التفاصيل البيبلوغرافية
| العنوان: |
High-level expression and characterization of carboxypeptidase Y from Saccharomyces cerevisiae in Pichia pastoris GS115. |
| المؤلفون: |
Yu, Xianhong, Zhai, Chao, Zhong, Xing, Tang, Wei, Wang, Xiaojuan, Yang, Hu, Chen, Wanping, Ma, Lixin |
| المصدر: |
Biotechnology Letters; Jan2015, Vol. 37 Issue 1, p161-167, 7p |
| مصطلحات موضوعية: |
CARBOXYPEPTIDASES, SACCHAROMYCES cerevisiae, PICHIA pastoris, GENE expression, AMINO acid sequence, ENZYMES |
| مستخلص: |
Carboxypeptidase Y is widely used in peptide sequencing and mass spectrometry. PRC1 coding for proteinase C from Saccharomyces cerevisiae was expressed in Pichia pastoris GS115 as procarboxypeptidase Y with a yield of ~605 mg/l in shake-flasks after 168 h induction with 1 % (v/v) methanol. This precursor of carboxypeptidase Y was cleaved by endogenous proteinases of P. pastoris and released into the fermentation broth as active carboxypeptidase Y within 2 weeks at 10 °C, which facilitated the preparation of mature carboxypeptidase Y. The recombinant enzyme was purified. It was optimally active at 30 °C and pH 6.0, with an optimal activity of ~305 U/mg using benzyloxycarbonyl- l-phenylalanyl- l-leucine as substrate. This is the first report about high-level expression and activation of carboxypeptidase Y in P. pastoris. [ABSTRACT FROM AUTHOR] |
|
Copyright of Biotechnology Letters is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) |
| قاعدة البيانات: |
Complementary Index |
