原创力文档high level expression of bikunin in pichia pastoris by fusion of human serum albumin在毕赤酵母属高水平表达bikunin pastoris人类血清白蛋白的融合.pdfVIP
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high level expression of bikunin in pichia pastoris by fusion of human serum albumin在毕赤酵母属高水平表达bikunin pastoris人类血清白蛋白的融合
Gou et al. AMB Express 2012, 2:14
/content/2/1/14
ORIGINAL Open Access
High level expression of bikunin in Pichia pastoris
by fusion of human serum albumin
1 2 1 2 1 1 1
Xing-Hua Gou , Yu-Ying Liu , Qi-Lei Chen , Jian-Jun Tang , Da-Yu Liu , Liang Zou , Xiao-Yong Wu and
Wei Wang1,3*
Abstract
Bikunin is a proteoglycan exhibiting broad-spectrum inhibitory activity against serine proteases and could
potentially suppress tumor cell invasion and metastasis. Here, we have successfully expressed recombinant human
bikunin (rh-bikunin) in Pichia pastoris and also established the purification procedure. Different fusion genes of h-
UTI and domain I, domain I and domain II, domain I, domain II and domain III of human serum albumin (HSA)
were inserted into expression vector pPICZaA. After expressed in shake flask, rh-bikunin was produced in an 30-L
fermenter and purified by affinity chromatography and cation exchange chromatography. The final expression
levels were 200 mg/L and we got totally 1.08 g (3650 IU/mg) of active purified rh-bikunin (purity is 98%) from 20 L
of fermentation broth. The rh-bikunin consists of unique form with molecular masses of 25 kDa, and has the same
N-terminals sequence as human native bikunin. This study provided a new method for high level expression of
active rh-bikunin by using HSA as fusion parter.
Keywords: Human bikunin, Fusion expression, Human serum albumin, Pichia pastoris
Introduction 2005]; [Qing xia 2005]; [Zhang, Liu et al. 2011]). The
Bikunin, also being called urinary trypsin inhibitor bikunin has many advantages such as evident effect in
(UTI),
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