The major recombinant capsid protein L1 of human papillomavirus (HPV) is widely used to produce HPV prophylactic vaccines. However, the quality of soluble and active expression of L1 in Escherichia coli was below the required amount. Coexpression with the chaperonin GroEL/ES enhanced L1 expression. Overexpressing GroEL/ES increased the soluble expression level of glutathione S-transferase-fused L1 (GST-L1) by approximately similar to 3 fold. The yield of HPV type 16 L1 pentamer (L1-p) was similar to 2 fold higher than that in a single expression system after purification through size-exclusion chromatograph. The expression and purification conditions were then optimized. The yield of L1-p was enhanced by similar to 5 fold, and those of HPV types 18 and 58 L1-p increased by -3 and -2 folds, respectively, compared with that in the single expression system. Coexpressing the mono-site mutant HPV16 L1 L469A with GroEL/ES increased L1-p yield by similar to 7 fold compared with strains expressing the wild-type L1 gene. L1-p was then characterized using circular dichroism spectra, UV vis cloud point, dynamic light scattering and transmission electron microscope analyses. Results indicated that the conformation and biological characteristics of L1-p were identical to that of native L1. Hence, overexpressing chaperonin in E. coli can increase the expression level of GST-L1 and L1-p production after purification. This finding may contribute to the development of a platform for prophylactic HPV vaccines. (C) 2015 Published by Elsevier Inc.
基金：
Natural Science Foundation of ChinaNational Natural Science Foundation of China [91027027, 21373101, 20934002]; Innovation Program of State Key Laboratory for Supramolecular Structure and Materials, Jilin UniversityJilin University
语种：
外文
被引次数：
WOS：
PubmedID：
中科院(CAS)分区：
出版当年[2016]版：
大类|4 区生物
小类|4 区生化研究方法4 区生化与分子生物学4 区生物工程与应用微生物
最新[2023]版：
大类|4 区生物学
小类|4 区生化研究方法4 区生化与分子生物学4 区生物工程与应用微生物
JCR分区：
出版当年[2016]版：
Q4BIOTECHNOLOGY & APPLIED MICROBIOLOGYQ4BIOCHEMISTRY & MOLECULAR BIOLOGYQ4BIOCHEMICAL RESEARCH METHODS
最新[2023]版：
Q4BIOCHEMICAL RESEARCH METHODSQ4BIOCHEMISTRY & MOLECULAR BIOLOGYQ4BIOTECHNOLOGY & APPLIED MICROBIOLOGY
第一作者机构：[1]State Key Laboratory of Supramolecular Structure and Materials, Jilin University, No. 2699, Qianjin Street, Changchun, 130012, China
通讯作者：
推荐引用方式(GB/T 7714)：
Pan Dong,Zha Xiao,Yu Xianghui,et al.Enhanced expression of soluble human papillomavirus L1 through coexpression of molecular chaperonin in Escherichia coli[J].PROTEIN EXPRESSION AND PURIFICATION.2016,120:92-98.doi:10.1016/j.pep.2015.12.016.
APA：
Pan, Dong,Zha, Xiao,Yu, Xianghui&Wu, Yuqing.(2016).Enhanced expression of soluble human papillomavirus L1 through coexpression of molecular chaperonin in Escherichia coli.PROTEIN EXPRESSION AND PURIFICATION,120,
MLA：
Pan, Dong,et al."Enhanced expression of soluble human papillomavirus L1 through coexpression of molecular chaperonin in Escherichia coli".PROTEIN EXPRESSION AND PURIFICATION 120.(2016):92-98
