在 Komagataella phaffii 中高效分泌表达人乳骨化蛋白

IF 7.2 Q1 FOOD SCIENCE & TECHNOLOGY
Zhihang Zhang , Yangyang Li , Zhenmin Liu , Shixiu Cui , Xianhao Xu , Yanfeng Liu , Jianghua Li , Guocheng Du , Xueqin Lv , Long Liu
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引用次数: 0

摘要

骨营养蛋白(OPN)是母乳中的一种具有生物活性的乳蛋白,在食品和医药领域有着广泛的应用。OPN 主要从新鲜牛乳中提取,但牛乳中 OPN 的浓度极低。为了解决这些问题,本研究采用 Komagataella phaffii 重组表达人 OPN(hOPN)。宿主细胞筛选结果显示,X33 细胞比其他 K. phaffii 宿主更适合表达 hOPN,其滴度为 340.5 μg/L。随后观察到,使用 PAOX1 启动子和杂交信号肽 PROSCW10-α,XPSA01 菌株的 hOPN 细胞外产量显著增加到 8.02 mg/L。同时,转录因子、翻译因子和分子伴侣的共同表达使细胞外 hOPN 的产量比 XPSA01 提高了 2.94 倍,滴度达到 23.6 mg/L。最后,结合上述策略得到了重组菌株 XPSA01-CP,它在摇瓶发酵和 3 升生物反应器中的滴度分别达到了 35.6 和 128.5 mg/L hOPN。据我们所知,这是迄今为止报道的细胞外产量最高的 hOPN。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Efficient secretory expression of human milk Osteopontin in Komagataella phaffii

Efficient secretory expression of human milk Osteopontin in Komagataella phaffii

Osteopontin (OPN), a bioactive milk protein in breast milk, has diverse applications in food and medicine. OPN is predominantly extracted from fresh bovine milk; however, the concentration of OPN in bovine milk is exceedingly low. To address these issues, in the present study, we used Komagataella phaffii for the recombinant expression of human OPN (hOPN). The results of host cell screening revealed that X33 cells were more suitable than other K. phaffii hosts for hOPN expression, yielding a titer of 340.5 μg/L. Subsequently, it was observed that the extracellular production of hOPN significantly increased to 8.02 mg/L from strain XPSA01 using the PAOX1 promoter and the hybrid signal peptide PROSCW10-α. Meanwhile, the co-expression of transcription factors, translation factors, and molecular chaperones resulted in a 2.94-fold enhancement in extracellular hOPN production compared with that in XPSA01, resulting in a titer of 23.6 mg/L. Finally, the above strategies were combined to obtain the recombinant strain XPSA01-CP, which achieved titers of 35.6 and 128.5 mg/L hOPN in shake-flask fermentation and 3-L bioreactor, respectively. To the best of our knowledge, this is the highest extracellular yield of hOPN ever reported.

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来源期刊
Future Foods
Future Foods Agricultural and Biological Sciences-Food Science
CiteScore
8.60
自引率
0.00%
发文量
97
审稿时长
15 weeks
期刊介绍: Future Foods is a specialized journal that is dedicated to tackling the challenges posed by climate change and the need for sustainability in the realm of food production. The journal recognizes the imperative to transform current food manufacturing and consumption practices to meet the dietary needs of a burgeoning global population while simultaneously curbing environmental degradation. The mission of Future Foods is to disseminate research that aligns with the goal of fostering the development of innovative technologies and alternative food sources to establish more sustainable food systems. The journal is committed to publishing high-quality, peer-reviewed articles that contribute to the advancement of sustainable food practices. Abstracting and indexing: Scopus Directory of Open Access Journals (DOAJ) Emerging Sources Citation Index (ESCI) SCImago Journal Rank (SJR) SNIP
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