Expression of monomeric insulin precursor in Pichia pastoris and its conversion into monomeric B27 Lys destripeptide insulin by tryptic hydrolysis

	Expression of monomeric insulin precursor in Pichia pastoris and its conversion into monomeric B27 Lys destripeptide insulin by tryptic hydrolysis
	Ding, JG; Feu, J; Cui, DF; Zhang, YS
刊名	ACTA BIOCHIMICA ET BIOPHYSICA SINICA
	2005
卷号	37 期号:4 页码:234-240
关键词	monomeric B27 Lys destripeptide insulin tryptic hydrolysis Pichia pastoris
通讯作者	Cui, DF (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China.,dfcui@sibs.ac.cn ; yszhang@sibs.ac.cn
英文摘要	Monomeric B27 Lys destripeptide insulin (1327 Lys DTrI) was designed and produced from its precursor expressed in Pichia pastoris through tryptic hydrolysis instead of the less efficient tryptic transpeptidation. The monomeric B27 Lys DTrI precursor (MIP) was purified from a cultured medium of P. pastoris by a combination of hydrophobic, size-exclusion, and ion-exchange chromatography. The purified MIP was converted, by tryptic hydrolysis, to B27 Lys DTrI, which was then purified by ion-exchange chromatography to homogeneity as assessed by native gel electrophoresis, HPLC, amino acid composition, and electrospray mass-spectrometric analysis. B27 Lys DTrI exhibited superior monomeric properties in size-exclusion chromatography. The yield of MIP was 200 mg per liter of culture, and the overall yield of purified B27 Lys DTrI from the crude MIP was 70%. The in vivo biological activity of B27 Lys DTrI as determined by the mouse convulsion assay was 21 U/mg, identical to that obtained by semisynthesis.
学科主题	Biochemistry & Molecular Biology; Biophysics
类目[WOS]	Biochemistry & Molecular Biology ; Biophysics
关键词[WOS]	SECRETORY EXPRESSION ; YEAST ; ASSOCIATION
收录类别	SCI
语种	英语
WOS记录号	WOS:000228545300004
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1894]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Ding, JG,Feu, J,Cui, DF,et al. Expression of monomeric insulin precursor in Pichia pastoris and its conversion into monomeric B27 Lys destripeptide insulin by tryptic hydrolysis[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2005,37(4):234-240.
APA	Ding, JG,Feu, J,Cui, DF,&Zhang, YS.(2005).Expression of monomeric insulin precursor in Pichia pastoris and its conversion into monomeric B27 Lys destripeptide insulin by tryptic hydrolysis.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,37(4),234-240.
MLA	Ding, JG,et al."Expression of monomeric insulin precursor in Pichia pastoris and its conversion into monomeric B27 Lys destripeptide insulin by tryptic hydrolysis".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 37.4(2005):234-240.