Abstract <p>The limitations associated with animal-derived collagen, such as the risk of zoonotic pathogen transmission and batch variability, have expedited the development of recombinant alternatives. Nonetheless, achieving an optimal balance between the bioactivity of recombinant collagen and production efficiency to ensure superior techno-economic performance remains a significant challenge in the field. In this study, we engineered a novel recombinant humanized collagen, designated as SynthCol1, by incorporating a 9-mer repeat sequence from the human type I collagen α1 chain (G674–A736) that includes integrin-binding motifs (GFPGER/GMPGER). This design strategy effectively addressed the critical challenges of soluble expression and production yield, resulting in a high-producing strain. SynthCol1 was expressed at high titers (15.3&#xa0;g/L) in a 500 L bioreactor using <i>Pichia pastoris</i> GS115 and was purified to greater than 95% homogeneity. Furthermore, functional assays demonstrated its capability to enhance cell adhesion. In a model of full-thickness human skin damaged by UVA exposure, SynthCol1 demonstrated significant efficacy in promoting tissue repair through structural reconstitution of the basement membrane, barrier regeneration and modulation of the inflammatory microenvironment. These results substantiate a strategic approach in the design of potent recombinant collagens, positioning SynthCol1 as a versatile and scalable biomaterial platform with substantial potential for therapeutic and cosmetic applications.</p> Key points <p><UnorderedList Mark="Bullet"> <ItemContent> <p><i>The study engineered a novel recombinant humanized type I collagen with high yield</i></p> </ItemContent> <ItemContent> <p><i>SynthCol1 was designed with enhanced bioactivity via rational design</i></p> </ItemContent> <ItemContent> <p><i>SynthCol1 was demonstrated to be effective in skin repair and photoprotection</i></p> </ItemContent> </UnorderedList></p>