具有高維持皮膚健康能力的人XVII型膠原蛋白片段的鑒定

2024年6月，a江南大學食品合成生物技術教育部工程研究中心和生物技術學院，麗湖路1800號;b江南大學未來食品科學中心;c江南大學江蘇省食品合成生物技術工程研究中心;

d江南大學生物技術學院，e博美生物科技有限公司（a Engineering Research Center of Ministry of Education on Food Synthetic Biotechnology and School of Biotechnology, Jiangnan University, 1800 Lihu Road,b Science Center for Future Foods, Jiangnan University,c Jiangsu Province Engineering Research Center of Food Synthetic Biotechnology, Jiangnan University,d School of Biotechnology, Jiangnan University,

e Bloomage Biotechnology Corporation Limited,）Jingwen Zhou研究團隊在《Synthetic and Systems Biotechnology》上發表論文：

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

“具有高維持皮膚健康能力的人XVII型膠原蛋白片段的鑒定”

Abstract：

Collagen XVII (COL17) is a transmembrane protein that mediates skin homeostasis. Due to expression of full length collagen was hard to achieve in microorganisms, arising the needs for selection of collagen fragments with desired functions for microbial biosynthesis. Here, COL17 fragments (27–33 amino acids) were extracted and replicated 16 times for recombinant expression in Escherichia coli. Five variants were soluble expressed, with the highest yield of 223 mg/L. The fusion tag was removed for biochemical and biophysical characterization. Circular dichroism results suggested one variant (sample-1707) with a triple-helix structure at >37 ?C. Sample-1707 can assemble into nanofiber (width, 5.6 nm) and form hydrogel at 3 mg/mL. Sample-1707 was shown to induce blood clotting and promote osteoblast differentiation. Furthermore, sample-1707 exhibited high capacity to induce mouse hair follicle stem cells differentiation and osteoblast migration, demonstrating a high capacity to induce skin cell regeneration and promote wound healing. A strong hydrogel was prepared from a chitosan and sample-1707 complex with a swelling rate of >30 % higher than simply using chitosan. Fed-batch fermentation of sample-1707 with a 5-L bioreactor obtained a yield of 600 mg/L. These results support the large-scale production of sample-1707 as a biomaterial for use in the skin care industry.

摘要：

膠原XVII (COL17)是一種介導皮膚穩態的跨膜蛋白。由于全長膠原蛋白在微生物中難以實現表達，因此需要選擇具有所需功能的膠原片段進行微生物生物合成。提取COL17片段(27-33個氨基酸)，復制16次，在大腸桿菌中進行重組表達。5個變異均可溶表達，最高產量為223 mg/L。去除融合標簽進行生化和生物物理表征。圓二色性結果表明，在>37°C時具有三螺旋結構的一個變體(樣品-1707)。樣品-1707可以組裝成納米纖維(寬度，5.6 nm)，并在3mg /mL時形成水凝膠。樣本-1707可誘導血液凝固，促進成骨細胞分化。此外，樣品-1707具有誘導小鼠毛囊干細胞分化和成骨細胞遷移的高能力，具有誘導皮膚細胞再生和促進傷口愈合的高能力。殼聚糖與樣品-1707復配制備了一種強水凝膠，其溶脹率比單純使用殼聚糖高30%以上。用5-L生物反應器對樣品-1707進行補料分批發酵，產率為600 mg/L。這些結果支持了樣品-1707作為皮膚護理行業使用的生物材料的大規模生產。

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