Bioresources and Bioprocessing ( IF 4.3 ) Pub Date : 2022-11-17 , DOI: 10.1186/s40643-022-00606-3 Lingling Ma 1, 2 , Xiaolin Liang 1, 2 , Shiqin Yu 1, 2, 3, 4 , Jingwen Zhou 1, 2, 3, 4
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Collagen is a biofunctional protein that has been widely used in many fields, including but not limited to biomedical, cosmetics and skin care, food, and novel materials. Recombinant collagen has great potential as an alternative to collagen extracted from animals because it avoids the immune response, and the yield and properties are stable. However, challenges remain in the industrial application of recombinant collagen, including improving the expression yield, reducing the cost of purification for industry and expanding applications. In this study, a cloning and recombination method was used to heterologously express the recombinant human-like collagen (RHLC) in Pichia pastoris GS115 using the pPIC9k expression vector. The RHLC expression titre was 2.33 g/L via a 5-L fermenter, and the purification was completed within 48 h and was 98% pure. The characteristics of RHLC were investigated. Furthermore, potential applications for RHLC were explored, such as basal collagen sponge preparation, forming films with chitosan and production of collagen hydrolysed peptides. RHLC has various potential applications due to its triple helical structure, thermostability, good biocompatibility and film-forming ability.
Graphical Abstract
中文翻译：
重组类人胶原蛋白在毕赤酵母中的表达、表征及应用潜力评价
胶原蛋白是一种生物功能蛋白，已广泛应用于许多领域，包括但不限于生物医学、化妆品和护肤品、食品和新型材料。重组胶原蛋白由于避免了免疫反应，且产量和性质稳定，因此具有作为动物提取胶原蛋白替代品的巨大潜力。然而，重组胶原蛋白的工业应用仍面临挑战，包括提高表达产量、降低工业纯化成本和扩大应用范围。本研究采用克隆重组的方法，利用pPIC9k表达载体在毕赤酵母GS115中异源表达重组类人胶原蛋白（RHLC）。通过5L发酵罐，RHLC表达滴度为2.33g/L，48小时内纯化完成，纯度为98%。研究了RHLC的特性。此外，还探索了 RHLC 的潜在应用，例如基础胶原蛋白海绵的制备、壳聚糖成膜以及胶原蛋白水解肽的生产。 RHLC因其三螺旋结构、热稳定性、良好的生物相容性和成膜能力而具有多种潜在的应用前景。
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