Temperature-Controlled Expression of a Recombinant Human-like Collagen I Peptide in Escherichia coli

Abstract

Collagen is the functional protein of the skin, tendons, ligaments, cartilage, bone, and connective tissue. Due to its extraordinary properties, collagen has a wide range of applications in biomedicine, tissue engineering, food, and cosmetics. In this study, we designed a functional fragment of human type I collagen (rhLCOL-I) and expressed it in Escherichia coli (E. coli) BL21(DE3) PlysS containing a thermal-induced plasmid, pBV-rhLCOL-I. The results indicated that the optimal expression level of the rhLCOL-I reached 36.3% of the total protein at 42 °C, and expressed in soluble form. In a 7 L fermentation, the yield of purified rhLCOL-I was 1.88 g/L. Interestingly, the plasmid, pBV220-rhLCOL-I, was excellently stable during the fermentation process, even in the absence of antibiotics. Functional analyses indicated that rhLCOL-I had the capacity to promote skin cell migration and adhesion in vitro and in vivo. Taken together, we developed a high-level and low-cost approach to produce collagen fragments suitable for medical applications in E. coli.

Identification of a human type XVII collagen fragment with high capacity for maintaining skin health

“…Meanwhile, the transcription level induced by sample-1707 was more than 70 % higher than that of BSA, confirming that sample-1707 can promote fibroblast differentiation. The cell migration test was conducted to further assess wound healing capacity based on MC-3T3 cells [ 17 ]. After 24 h cultivation, the wound gap decreased by > 60 % by treating with the three samples, respectively ( Fig.…”

“…Nonetheless, sample-1707 exhibited much stronger capacity to induce differentiation of HFSCs than COL1, which would be beneficial for repair and regeneration of hair follicles [ 40 ]. In addition, sample-1707 generated faster cell migration than COL1, which is critical for wound healing [ 17 ]. Overall, these results suggested that sample-1707 is a robust biocompatible material.…”

“…For the cell migration assay [ 17 ], 5 × 10 5 MC-3T3 cells were plated and incubated to 95 % confluency. A scratch was made using pipette.…”

Exploring and Engineering Novel Strong Promoters for High-Level Protein Expression in Bacillus subtilis DB104 through Transcriptome Analysis

“…Escherichia coli is the most commonly used host strain for recombinant protein production. It has been successfully used to produce various proteins including collagen peptide [1], cellulose [2], endoglucanase [3], and fucosidase [4]. However, E. coli comes with certain disadvantages, such as a propensity of expressed proteins to aggregate [5,6], its production of endotoxin [7,8], and plasmid instability [9].…”