International Journal of Horticulture and Floriculture

International Journal of Horticulture and Floriculture ISSN 2756-3790 Vol. 9 (2), pp. 001-006, February, 2021. © International Scholars Journals

Full Length Research Paper

Expression of recombinant human collagen peptide in Escherichia coli and effects of the recombinant protein on UVA-irradiated HaCaT Cells

Ming Wu, Zhen zhen Xu and Guang Chen*

College of Life Sciences, Jilin Agricultural University, Changchun 130000, China.

Accepted 17 October, 2020

Abstract

In this study, we reported the cloning and over- expression of a gene encoding human collagen peptide (CP6) in Escherichia coli and the establishment of a purification protocol for obtaining the recombinant protein. The collagen peptide (CP6) had a molecular weight of about 46 kDa, and CP6 expression comprised approximately 10% of the total bacterial protein expression. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed that most of the collagen peptide (CP6) existed in the soluble fractions. Purified collagen peptide (CP6) was highly soluble. By using high-performance liquid chromatography (HPLC), a CP6 yield of approximately 11.4 mg/L of Luria-Bertani (LB) broth was obtained. What’s more we performed assays with 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and annexin V-fluorescein isothiocyanate/propidium iodide (annexin V-FITC/PI) to investigate the cytoprotective effects of CP6 on the proliferation of UVA-damaged human keratinocyte cell line cells. The results of this study showed that CP6 could prevent UVA-induced DNA damage in cells of the spontaneously immortalized human keratinocyte cell line HaCaT.

Key words: Human collagen peptide, Escherichia coli, HaCaT, UVA.