Fhl1p protein, a positive transcription factor in Pichia pastoris, enhances the expression of recombinant proteins
Abstract Background The methylotrophic yeast Pichia pastoris is well-known for the production of a broad spectrum of functional types of heterologous proteins including enzymes, antigens, engineered antibody fragments, and next gen protein scaffolds and many transcription factors are utilized to add...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-11-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-019-1256-0 |
| _version_ | 1818966101382397952 |
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| author | Xueyun Zheng Yimin Zhang Xinying Zhang Cheng Li Xiaoxiao Liu Ying Lin Shuli Liang |
| author_facet | Xueyun Zheng Yimin Zhang Xinying Zhang Cheng Li Xiaoxiao Liu Ying Lin Shuli Liang |
| author_sort | Xueyun Zheng |
| collection | DOAJ |
| description | Abstract Background The methylotrophic yeast Pichia pastoris is well-known for the production of a broad spectrum of functional types of heterologous proteins including enzymes, antigens, engineered antibody fragments, and next gen protein scaffolds and many transcription factors are utilized to address the burden caused by the high expression of heterologous proteins. In this article, a novel P. pastoris transcription factor currently annotated as Fhl1p, an activator of ribosome biosynthesis processing, was investigated for promoting the expression of the recombinant proteins. Results The function of Fhl1p of P. pastoris for improving the expression of recombinant proteins was verified in strains expressing phytase, pectinase and mRFP, showing that the productivity was increased by 20–35%. RNA-Seq was used to study the Fhl1p regulation mechanism in detail, confirming Fhl1p involved in the regulation of rRNA processing genes, ribosomal small/large subunit biogenesis genes, Golgi vesicle transport genes, etc., which contributed to boosting the expression of foreign proteins. The overexpressed Fhl1p strain exhibited increases in the polysome and monosome levels, showing improved translation activities. Conclusion This study illustrated that the transcription factor Fhl1p could effectively enhance recombinant protein expression in P. pastoris. Furthermore, we provided the evidence that overexpressed Fhl1p was related to more active translation state. |
| first_indexed | 2024-12-20T13:27:33Z |
| format | Article |
| id | doaj.art-c8e8fd693d444d04b510df4036a30803 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-20T13:27:33Z |
| publishDate | 2019-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-c8e8fd693d444d04b510df4036a308032022-12-21T19:39:13ZengBMCMicrobial Cell Factories1475-28592019-11-0118111210.1186/s12934-019-1256-0Fhl1p protein, a positive transcription factor in Pichia pastoris, enhances the expression of recombinant proteinsXueyun Zheng0Yimin Zhang1Xinying Zhang2Cheng Li3Xiaoxiao Liu4Ying Lin5Shuli Liang6Guangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, South China University of TechnologyGuangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, South China University of TechnologyGuangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, South China University of TechnologyGuangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, South China University of TechnologyGuangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, South China University of TechnologyGuangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, South China University of TechnologyGuangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, South China University of TechnologyAbstract Background The methylotrophic yeast Pichia pastoris is well-known for the production of a broad spectrum of functional types of heterologous proteins including enzymes, antigens, engineered antibody fragments, and next gen protein scaffolds and many transcription factors are utilized to address the burden caused by the high expression of heterologous proteins. In this article, a novel P. pastoris transcription factor currently annotated as Fhl1p, an activator of ribosome biosynthesis processing, was investigated for promoting the expression of the recombinant proteins. Results The function of Fhl1p of P. pastoris for improving the expression of recombinant proteins was verified in strains expressing phytase, pectinase and mRFP, showing that the productivity was increased by 20–35%. RNA-Seq was used to study the Fhl1p regulation mechanism in detail, confirming Fhl1p involved in the regulation of rRNA processing genes, ribosomal small/large subunit biogenesis genes, Golgi vesicle transport genes, etc., which contributed to boosting the expression of foreign proteins. The overexpressed Fhl1p strain exhibited increases in the polysome and monosome levels, showing improved translation activities. Conclusion This study illustrated that the transcription factor Fhl1p could effectively enhance recombinant protein expression in P. pastoris. Furthermore, we provided the evidence that overexpressed Fhl1p was related to more active translation state.https://doi.org/10.1186/s12934-019-1256-0Pichia pastorisFhl1pTranscription factorEnhanced protein productionTranslation |
| spellingShingle | Xueyun Zheng Yimin Zhang Xinying Zhang Cheng Li Xiaoxiao Liu Ying Lin Shuli Liang Fhl1p protein, a positive transcription factor in Pichia pastoris, enhances the expression of recombinant proteins Microbial Cell Factories Pichia pastoris Fhl1p Transcription factor Enhanced protein production Translation |
| title | Fhl1p protein, a positive transcription factor in Pichia pastoris, enhances the expression of recombinant proteins |
| title_full | Fhl1p protein, a positive transcription factor in Pichia pastoris, enhances the expression of recombinant proteins |
| title_fullStr | Fhl1p protein, a positive transcription factor in Pichia pastoris, enhances the expression of recombinant proteins |
| title_full_unstemmed | Fhl1p protein, a positive transcription factor in Pichia pastoris, enhances the expression of recombinant proteins |
| title_short | Fhl1p protein, a positive transcription factor in Pichia pastoris, enhances the expression of recombinant proteins |
| title_sort | fhl1p protein a positive transcription factor in pichia pastoris enhances the expression of recombinant proteins |
| topic | Pichia pastoris Fhl1p Transcription factor Enhanced protein production Translation |
| url | https://doi.org/10.1186/s12934-019-1256-0 |
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