title: Effect of cooperation of chaperones and gene dosage on the expression of porcine PGLYRP-1 in Pichia pastoris.
abstract: 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.
authors.affiliation: Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University, Wushan Road, Tianhe district, Guangzhou, Guangdong, 510642, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Yang
authors.affiliation: Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University, Wushan Road, Tianhe district, Guangzhou, Guangdong, 510642, China.
authors.forename: Zhipeng
authors.initials: Z
authors.lastname: Lu
authors.affiliation: Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University, Wushan Road, Tianhe district, Guangzhou, Guangdong, 510642, China.
authors.forename: Jiawei
authors.initials: J
authors.lastname: Chen
authors.affiliation: Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University, Wushan Road, Tianhe district, Guangzhou, Guangdong, 510642, China.
authors.forename: Pinpin
authors.initials: P
authors.lastname: Chu
authors.affiliation: Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University, Wushan Road, Tianhe district, Guangzhou, Guangdong, 510642, China.
authors.forename: Qingmei
authors.initials: Q
authors.lastname: Cheng
authors.affiliation: Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University, Wushan Road, Tianhe district, Guangzhou, Guangdong, 510642, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Liu
authors.affiliation: Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University, Wushan Road, Tianhe district, Guangzhou, Guangdong, 510642, China.
authors.forename: Feiping
authors.initials: F
authors.lastname: Ming
authors.affiliation: Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University, Wushan Road, Tianhe district, Guangzhou, Guangdong, 510642, China.
authors.forename: Chaoyuan
authors.initials: C
authors.lastname: Huang
authors.affiliation: Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University, Wushan Road, Tianhe district, Guangzhou, Guangdong, 510642, China.
authors.forename: Anji
authors.initials: A
authors.lastname: Xiao
authors.affiliation: Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University, Wushan Road, Tianhe district, Guangzhou, Guangdong, 510642, China.
authors.forename: Haiming
authors.initials: H
authors.lastname: Cai
authors.affiliation: Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University, Wushan Road, Tianhe district, Guangzhou, Guangdong, 510642, China. lhzhang@scau.edu.cn.
authors.forename: Linghua
authors.initials: L
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002352:Carrier Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D018628:Gene Dosage; D015870:Gene Expression; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D010843:Pichia / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D019704:Protein Disulfide-Isomerases / Q000235:genetics / Q000378:metabolism; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis; D013211:Staphylococcus aureus / Q000187:drug effects; D013552:Swine; D014170:Transformation, Genetic; D056811:Unfolded Protein Response
chemical_list: D000900:Anti-Bacterial Agents; D002352:Carrier Proteins; D005656:Fungal Proteins; D018840:HSP70 Heat-Shock Proteins; C060153:KAR2 protein, yeast; D018832:Molecular Chaperones; D011994:Recombinant Proteins; C118897:peptidoglycan recognition protein; D019704:Protein Disulfide-Isomerases
keywords: BiP; Dosage; Expression; PDI; Peptidoglycan recognition protein; Pichia pastoris