Zhihang Zhang , Yangyang Li , Zhenmin Liu , Shixiu Cui , Xianhao Xu , Yanfeng Liu , Jianghua Li , Guocheng Du , Xueqin Lv , Long Liu
{"title":"在 Komagataella phaffii 中高效分泌表达人乳骨化蛋白","authors":"Zhihang Zhang , Yangyang Li , Zhenmin Liu , Shixiu Cui , Xianhao Xu , Yanfeng Liu , Jianghua Li , Guocheng Du , Xueqin Lv , Long Liu","doi":"10.1016/j.fufo.2024.100393","DOIUrl":null,"url":null,"abstract":"<div><p>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 <em>Komagataella phaffii</em> for the recombinant expression of human OPN (hOPN). The results of host cell screening revealed that X33 cells were more suitable than other <em>K. phaffii</em> 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 P<em><sub>AOX1</sub></em> 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.</p></div>","PeriodicalId":34474,"journal":{"name":"Future Foods","volume":"10 ","pages":"Article 100393"},"PeriodicalIF":7.2000,"publicationDate":"2024-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2666833524000996/pdfft?md5=f012303d8de0808c5a3bf01f3df767f3&pid=1-s2.0-S2666833524000996-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Efficient secretory expression of human milk Osteopontin in Komagataella phaffii\",\"authors\":\"Zhihang Zhang , Yangyang Li , Zhenmin Liu , Shixiu Cui , Xianhao Xu , Yanfeng Liu , Jianghua Li , Guocheng Du , Xueqin Lv , Long Liu\",\"doi\":\"10.1016/j.fufo.2024.100393\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>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 <em>Komagataella phaffii</em> for the recombinant expression of human OPN (hOPN). The results of host cell screening revealed that X33 cells were more suitable than other <em>K. phaffii</em> 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 P<em><sub>AOX1</sub></em> 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.</p></div>\",\"PeriodicalId\":34474,\"journal\":{\"name\":\"Future Foods\",\"volume\":\"10 \",\"pages\":\"Article 100393\"},\"PeriodicalIF\":7.2000,\"publicationDate\":\"2024-06-08\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2666833524000996/pdfft?md5=f012303d8de0808c5a3bf01f3df767f3&pid=1-s2.0-S2666833524000996-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Future Foods\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2666833524000996\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"FOOD SCIENCE & TECHNOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Future Foods","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2666833524000996","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"FOOD SCIENCE & TECHNOLOGY","Score":null,"Total":0}
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.
Future FoodsAgricultural 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:
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Emerging Sources Citation Index (ESCI)
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