Production of bioactive human beta-defensin-3 in Escherichia coli by soluble fusion expression

Biotechnology Letters
Lei HuangPeilin Cen

Abstract

A codon optimized mature human beta-defensin-3 gene (smHBD3) was synthesized and fused with TrxA to construct pET32-smHBD3 vector, which was transformed into E. coli BL21(DE3) and cultured in MBL medium. The volumetric productivity of fusion protein reached 0.99 g fusion protein l(-1), i.e. 0.21 g mature HBD3 l(-1). Ninety-six percentage of the fusion protein was in a soluble form and constituted about 45% of the total soluble protein. After cell disruption, the soluble fusion protein was separated by affinity chromatography and cleaved by enterokinase, and then the mature HBD3 was purified by cationic ion exchange chromatography. The overall recovery ratio of HBD3 was 43%. The purified mature HBD3 demonstrated antimicrobial activity against E. coli.

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Citations

Aug 28, 2012·Inflammation Research : Official Journal of the European Histamine Research Society ... [et Al.]·Annekatrin CoordesAnton J Kroesen
Apr 18, 2009·Applied Microbiology and Biotechnology·Lei HuangRongrong Jiang
Nov 21, 2007·Applied Biochemistry and Biotechnology·Lei HuangPeilin Cen
Nov 3, 2010·Microbial Cell Factories·Elena García-Fruitós
Jul 4, 2008·Protein Expression and Purification·Lei HuangSusanna Su Jan Leong
Jul 7, 2009·Biotechnology and Applied Biochemistry·Yifeng Li
Feb 6, 2008·Protein Expression and Purification·Feng XuBin Yao
Sep 13, 2014·Biotechnology Letters·Omar González-OrtegaRuth Elena Soria-Guerra
May 19, 2017·FEMS Microbiology Reviews·María Eugenia Pachón-IbáñezJavier Sánchez-Céspedes
Feb 4, 2009·Journal of Peptide Science : an Official Publication of the European Peptide Society·Verica VidovicSebastiaan Werten
May 2, 2020·Expert Review of Clinical Pharmacology·Kamila Botelho Sampaio de OliveiraSimoni Campos Dias
Nov 13, 2019·Protein Expression and Purification·Ligia Luz Corrales-GarcíaGerardo Corzo
Jul 4, 2012·International Journal of Medical Microbiology : IJMM·Ean-jeong SeoTobias A Oelschlaeger
Mar 21, 2009·Journal of Agricultural and Food Chemistry·Krisna Prak, Shigeru Utsumi
Sep 19, 2006·Biochimica Et Biophysica Acta·Vishnu DhopleAyyalusamy Ramamoorthy
Oct 2, 2012·Peptides·Nádia Skorupa ParachinOctávio Luiz Franco

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