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 p...
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Elsevier
2024-12-01
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| Series: | Future Foods |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666833524000996 |
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| author | Zhihang Zhang Yangyang Li Zhenmin Liu Shixiu Cui Xianhao Xu Yanfeng Liu Jianghua Li Guocheng Du Xueqin Lv Long Liu |
| author_facet | Zhihang Zhang Yangyang Li Zhenmin Liu Shixiu Cui Xianhao Xu Yanfeng Liu Jianghua Li Guocheng Du Xueqin Lv Long Liu |
| author_sort | Zhihang Zhang |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-d17ef2d75ec147b6a1dc82cd160e122d |
| institution | DOAJ |
| issn | 2666-8335 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Future Foods |
| spelling | doaj-art-d17ef2d75ec147b6a1dc82cd160e122d2025-08-20T02:48:57ZengElsevierFuture Foods2666-83352024-12-011010039310.1016/j.fufo.2024.100393Efficient secretory expression of human milk Osteopontin in Komagataella phaffiiZhihang Zhang0Yangyang Li1Zhenmin Liu2Shixiu Cui3Xianhao Xu4Yanfeng Liu5Jianghua Li6Guocheng Du7Xueqin Lv8Long Liu9Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, PR China; Science Center for Future Foods, Jiangnan University, Wuxi 214122, PR China; Food Laboratory of Zhongyuan, Jiangnan University, Wuxi 214122, PR ChinaKey Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, PR China; Science Center for Future Foods, Jiangnan University, Wuxi 214122, PR ChinaState Key Laboratory of Dairy Biotechnology, Shanghai Engineering Research Center of Dairy Biotechnology, Dairy Research Institute, Bright Dairy & Food Co., Ltd, Shanghai 200436, PR ChinaScience Center for Future Foods, Jiangnan University, Wuxi 214122, PR China; Jiaxing Institute of Future Food, Jiaxing 314015, PR ChinaKey Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, PR China; Science Center for Future Foods, Jiangnan University, Wuxi 214122, PR ChinaKey Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, PR China; Science Center for Future Foods, Jiangnan University, Wuxi 214122, PR ChinaKey Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, PR China; Science Center for Future Foods, Jiangnan University, Wuxi 214122, PR China; Corresponding author.Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, PR China; Science Center for Future Foods, Jiangnan University, Wuxi 214122, PR ChinaKey Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, PR China; Science Center for Future Foods, Jiangnan University, Wuxi 214122, PR ChinaKey Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, PR China; Science Center for Future Foods, Jiangnan University, Wuxi 214122, PR ChinaOsteopontin (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.http://www.sciencedirect.com/science/article/pii/S2666833524000996Komagataella phaffiiOsteopontinHybrid signal peptideMolecular chaperoneSecretory expression |
| spellingShingle | Zhihang Zhang Yangyang Li Zhenmin Liu Shixiu Cui Xianhao Xu Yanfeng Liu Jianghua Li Guocheng Du Xueqin Lv Long Liu Efficient secretory expression of human milk Osteopontin in Komagataella phaffii Future Foods Komagataella phaffii Osteopontin Hybrid signal peptide Molecular chaperone Secretory expression |
| title | Efficient secretory expression of human milk Osteopontin in Komagataella phaffii |
| title_full | Efficient secretory expression of human milk Osteopontin in Komagataella phaffii |
| title_fullStr | Efficient secretory expression of human milk Osteopontin in Komagataella phaffii |
| title_full_unstemmed | Efficient secretory expression of human milk Osteopontin in Komagataella phaffii |
| title_short | Efficient secretory expression of human milk Osteopontin in Komagataella phaffii |
| title_sort | efficient secretory expression of human milk osteopontin in komagataella phaffii |
| topic | Komagataella phaffii Osteopontin Hybrid signal peptide Molecular chaperone Secretory expression |
| url | http://www.sciencedirect.com/science/article/pii/S2666833524000996 |
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