PMID- 32737719 OWN - NLM STAT- MEDLINE DCOM- 20210727 LR - 20210727 IS - 1573-6776 (Electronic) IS - 0141-5492 (Linking) VI - 42 IP - 12 DP - 2020 Dec TI - Overexpression of the regulatory subunit of protein kinase A increases heterologous protein expression in Pichia pastoris. PG - 2685-2692 LID - 10.1007/s10529-020-02977-z [doi] AB - OBJECTIVE: Translational regulation plays an important role in protein synthesis. Our goal was to screen translation-related factors to improve heterologous protein expression in Pichia pastoris. RESULTS: Twenty-eight translation-related factors were overexpressed in P. pastoris GS115 expressing enhanced green fluorescent protein (eGFP). The results showed that overexpression of Bcy1, the regulatory subunit of protein kinase A (PKA), significantly increased both eGFP expression and cell biomass by 20% under methanol induction for 120 h. Additionally, overexpression of Bcy1 elevated the growth rate by 18% and increased production of the industrial enzyme Phytase (Phy) by 26%. Transcriptome analysis indicated that the overall expression of ribosomal protein genes was significantly downregulated and that postdiauxic shift genes and stress response element genes were upregulated. CONCLUSIONS: Bcy1 regulates ribosome protein genes, postdiauxic shift genes and stress response element genes, leading to improved cell growth and heterologous protein expression. This study provides a convenient and universal factor for heterologous protein production. FAU - Liao, Xihao AU - Liao X AUID- ORCID: 0000-0002-7395-4708 AD - Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. AD - Guangdong Research Center of Industrial Enzyme and Green Manufacturing Technology, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. FAU - Lin, Wenyang AU - Lin W AD - Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. AD - Guangdong Research Center of Industrial Enzyme and Green Manufacturing Technology, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. FAU - Chen, Nanzhu AU - Chen N AD - Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. AD - Guangdong Research Center of Industrial Enzyme and Green Manufacturing Technology, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. FAU - Li, Lu AU - Li L AD - Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. AD - Guangdong Research Center of Industrial Enzyme and Green Manufacturing Technology, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. FAU - Huang, Dafu AU - Huang D AD - Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. AD - Guangdong Research Center of Industrial Enzyme and Green Manufacturing Technology, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. FAU - Zhong, Bingxu AU - Zhong B AD - Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. AD - Guangdong Research Center of Industrial Enzyme and Green Manufacturing Technology, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. FAU - Chen, Liang AU - Chen L AD - Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. AD - Guangdong Research Center of Industrial Enzyme and Green Manufacturing Technology, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. FAU - Liang, Shuli AU - Liang S AD - Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. AD - Guangdong Research Center of Industrial Enzyme and Green Manufacturing Technology, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. FAU - Lin, Ying AU - Lin Y AD - Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. feylin@scut.edu.cn. AD - Guangdong Research Center of Industrial Enzyme and Green Manufacturing Technology, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China. feylin@scut.edu.cn. LA - eng GR - 31470159/National Natural Science Foundation of China/ PT - Journal Article DEP - 20200731 PL - Netherlands TA - Biotechnol Lett JT - Biotechnology letters JID - 8008051 RN - 0 (Protein Subunits) RN - 0 (Recombinant Proteins) RN - EC 2.7.11.11 (Cyclic AMP-Dependent Protein Kinases) RN - EC 3.1.3.26 (6-Phytase) RN - Y4S76JWI15 (Methanol) RN - Komagataella pastoris SB - IM MH - 6-Phytase/genetics MH - Cyclic AMP-Dependent Protein Kinases/*genetics MH - Gene Expression Profiling MH - Gene Expression Regulation, Enzymologic/genetics MH - Methanol MH - Protein Biosynthesis/*genetics MH - Protein Subunits/*genetics MH - Recombinant Proteins MH - Saccharomycetales/enzymology/*genetics OTO - NOTNLM OT - Bcy1 OT - Heterologous protein OT - Pichia pastoris OT - Translation-related factor EDAT- 2020/08/02 06:00 MHDA- 2021/07/28 06:00 CRDT- 2020/08/02 06:00 PHST- 2020/02/09 00:00 [received] PHST- 2020/07/27 00:00 [accepted] PHST- 2020/08/02 06:00 [pubmed] PHST- 2021/07/28 06:00 [medline] PHST- 2020/08/02 06:00 [entrez] AID - 10.1007/s10529-020-02977-z [pii] AID - 10.1007/s10529-020-02977-z [doi] PST - ppublish SO - Biotechnol Lett. 2020 Dec;42(12):2685-2692. doi: 10.1007/s10529-020-02977-z. Epub 2020 Jul 31.