The biocompatibility of collagen extracted from tissue poses a challenge, and the thermal stability and triple helix level of the reported recombinant collagen have been found to be suboptimal.Therefore, Pichia pastoris GS115 was chosen as the heterologous expression strain for inducing the expression of a fragment from the collagen domain of human type I collagen α1 chain.The T4 fibritin promoter sequence was added at the C-terminus.The impact of T4 fibritin on the thermal stability and triple helix level of collagen was investigated through circular dichroism thermal denaturation experiments.The bioactivity of the recombinant collagen was evaluated by cell proliferation-toxicity assay and free radical scavenging rate analysis.Results demonstrated that inclusion of T4 fibritin in recombinant collagen 1A1C increased its thermal denaturation temperature by 4 ℃, with subsequent regeneration after denaturation.Both human-like collagen 1A1 and 1A1C showed good biocompatibility.At 5 μg/mL, human-like collagen 1A1C could promote the proliferation of fibroblasts significantly.Moreover, at 3 mg/mL, the scavenging rates of human-like collagen free radicals measured by two methods were 19.8% and 40.0%, respectively.This study successfully expressed biocompatible recombinant collagen using genetic engineering techniques.The thermal denaturation temperature of collagen was increased to 39.9 ℃ by fusion of terminal peptide, which was higher than the human body temperature of 37 ℃, and have great potential for various applications.
LIU Di
,
GUO Junhui
,
LI Qichang
,
YIN Qiaoling
,
CI Peisong
,
FAN Lihong
,
XIE Hao
. Study on expression, purification, and properties of recombinant type I human-like collagen[J]. Food and Fermentation Industries, 2025
, 51(6)
: 127
-133
.
DOI: 10.13995/j.cnki.11-1802/ts.039247
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