Effect of cooperation of chaperones and gene dosage on the expression of porcine PGLYRP-1 in Pichia pastoris

• Published in: Applied microbiology and biotechnology Vol. 100; no. 12; pp. 5453 - 5465
• Main Authors: Yang, Jun, Lu, Zhipeng, Chen, Jiawei, Chu, Pinpin, Cheng, Qingmei, Liu, Jie, Ming, Feiping, Huang, Chaoyuan, Xiao, Anji, Cai, Haiming, Zhang, Linghua
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.06.2016; Springer; Springer Nature B.V
• Subjects: Animals; Anti-Bacterial Agents - pharmacology; Antimicrobial agents; Applied Genetics and Molecular Biotechnology; Ascomycota; Bacteria; Biomedical and Life Sciences; Biotechnology; Carrier Proteins - biosynthesis; Carrier Proteins - chemistry; Carrier Proteins - genetics; Carrier Proteins - pharmacology; Cooperation; Escherichia coli - drug effects; Fungal Proteins - genetics; Fungal Proteins - metabolism; Gene Dosage; Gene Expression; Genetic aspects; Genetic engineering; Gram-positive bacteria; Health aspects; HSP70 Heat-Shock Proteins - genetics; HSP70 Heat-Shock Proteins - metabolism; Immune system; Life Sciences; Mammals; Microbial Genetics and Genomics; Microbiology; Molecular chaperones; Molecular Chaperones - genetics; Molecular Chaperones - metabolism; Observations; Optimization; Peptides; Pharmaceutical industry; Pichia - chemistry; Pichia - genetics; Pichia - metabolism; Pichia pastoris; Plasmids; Protein Disulfide-Isomerases - genetics; Protein Disulfide-Isomerases - metabolism; Protein Folding; Proteins; Recombinant Proteins - biosynthesis; Staphylococcus aureus - drug effects; Studies; Swine; Transformation, Genetic; Unfolded Protein Response; BiP; PDI; Peptidoglycan recognition protein; Dosage; Expression; Pichia pastoris
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-016-7372-4

Mammalian peptidoglycan recognition proteins (PGLYRPs) are highly conserved pattern-recognition molecules of the innate immune system with considerable bactericidal activity, which manifest their potential values for the application to food and pharmaceutical industry. However, the effective expression of porcine PGLYRP-1 in Pichia pastoris has not been reported so far. In this study, expression in P. pastoris was explored as an efficient way to produce functional porcine PGLYRP-1. Cooperation of chaperones co-expression and gene dosage (including protein disulfide isomerase ( PDI )/binding protein ( BiP ) and pglyrp - 1 ) were used to enhance functional expression of antimicrobial protein in P. pastoris . Overexpression of PDI was certainly able to increase secretion level of PGLYRP-1 protein because the increase in secreted PGLYRP-1 secretion was correlated with the copy numbers of PDI in high copy pglyrp - 1 clones. However, co-expression of BiP was proved to be detrimental to PGLYRP-1 secretion. In addition, we also found that excessive expression of PDI and/or BiP could decrease the mRNA expression of pglyrp - 1 gene. This showed that PDI and BiP as the target genes of unfolded protein response (UPR) might regulate the transcription of the target protein. These data demonstrated for the first time that the combination of chaperones and gene dosages could improve the yield of PGLYRP-1, which could facilitate the application to food and pharmaceutical industry.