Journal
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
Volume 41, Issue 11, Pages 1667-1675Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s10295-014-1500-x
Keywords
Chaperone; Co-expression; Escherichia coli; Over-expression; ptsG deletion; RT-qPCR
Categories
Funding
- National Natural Science Foundation of China [21176200]
- Scientific Research Program of Shaanxi Provincial Department of Education, China [2010JC21]
- Shaanxi Biochemical Engineering Key Discipline Program, China
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Human-like collagen (HLC) is a novel biomedical material with promising applications. Usually, insoluble HLC was formed due to over-expression. In order to improve the production of soluble HLC, the effective chaperone proteins and their mediation roles on HLC were clarified. Trigger factor (TF) pathway with low specificity and high binding affinity to nascent chains could increase soluble HLC expression; GroEL-GroES could increase the expression level of HLC by assisting the correct folding of HLC and increase mRNA level of the gene coding for HLC by enhancing mRNA stability. DnaK chaperone system did not work positively on soluble HLC due to the unbalanced ratio of DnaK:DnaJ:GrpE, especially too high GrpE significantly inhibited DnaK-mediated refolding. The production of soluble HLC with co-expression of exogenous TF and GroEL-GroES was increased by 35.3 % in comparison with the highest value 0.26 g/L reported previously.
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