High-level expression, purification and characterisation of porcine β-defensin 2 in Pichia pastoris and its potential as a cost-efficient growth promoter in porcine feed

Applied Microbiology and Biotechnology

Volumn 98, Issue 12, 2014, Pages 5487-5497

  Peng, Zixin ^a,b   Wang, Anru ^a   Feng, Qiuyue ^a   Wang, Zhaoyue ^b   Ivanova, Iskra Vitanova ^c   He, Xiuping ^b   Zhang, Borun ^b   Song, Weiping ^a  

^a State Key Laboratory of Direct Fed Microbial Engineering   (China)

^b INSTITUTE OF MICROBIOLOGY   (China)

^c UNIVERSITY OF SOFIA   (Bulgaria)

Author keywords

Antimicrobial activity; Expression; Pichia pastoris; Porcine defensin 2; Purification; Stability

Indexed keywords

CONVERGENCE OF NUMERICAL METHODS; MASS SPECTROMETRY; PROTEINS; PURIFICATION; SALMONELLA; SOLVENTS;

ANTI-MICROBIAL ACTIVITY; DEFENSINS; EXPRESSION; GEL EXCLUSION CHROMATOGRAPHY; HIGH THERMAL STABILITY; LACTOBACILLUS PLANTARUM; MINIMAL INHIBITORY CONCENTRATION; PICHIA PASTORIS;

YEAST;

BETA DEFENSIN 2; COMPLEMENTARY DNA; DIMETHYL SULFOXIDE; DITHIOTHREITOL; RECOMBINANT PROTEIN; BETA DEFENSIN;

ALCOHOL; ANTIMICROBIAL ACTIVITY; COST-BENEFIT ANALYSIS; CULTIVATION; GENE EXPRESSION; GROWTH RATE; INHIBITOR; PURIFICATION; RECOMBINATION; STABILIZATION; YEAST;

AMINO ACID SEQUENCE; ANIMAL FOOD; ANTIMICROBIAL ACTIVITY; ARTICLE; BACILLUS SUBTILIS; BACTERICIDAL ACTIVITY; CONTROLLED STUDY; DNA SEQUENCE; ENZYME ACTIVITY; ESCHERICHIA COLI; GASTROINTESTINAL TRACT; GENE SEQUENCE; GROWTH REGULATION; HAEMOPHILUS PARASUIS; LACTOBACILLUS PLANTARUM; MATRIX ASSISTED LASER DESORPTION IONIZATION TIME OF FLIGHT MASS SPECTROMETRY; MINIMUM INHIBITORY CONCENTRATION; NONHUMAN; NUCLEOTIDE SEQUENCE; OXIDATION; PH; PICHIA PASTORIS; PRECIPITATION; PROTEIN ANALYSIS; PROTEIN DEGRADATION; PROTEIN EXPRESSION; PROTEIN PURIFICATION; REDUCTION; SACCHAROMYCES CEREVISIAE; SALMONELLA ENTERICA; STAPHYLOCOCCUS AUREUS; STREPTOCOCCUS SUIS; SWINE; THERMOSTABILITY; ANALYSIS; ANIMAL; BACTERIAL INFECTION; BACTERIUM; CHEMISTRY; DRUG EFFECTS; GENETICS; GROWTH, DEVELOPMENT AND AGING; ISOLATION AND PURIFICATION; METABOLISM; MICROBIAL SENSITIVITY TEST; MICROBIOLOGY; PICHIA; PIG; PROTEIN ENGINEERING; SWINE DISEASE; VETERINARY;

BACILLUS SUBTILIS; LACTOBACILLUS PLANTARUM; PICHIA PASTORIS; SACCHAROMYCES CEREVISIAE; SALMONELLA CHOLERAESUIS; STAPHYLOCOCCUS AUREUS; STREPTOCOCCUS SUIS; SUIDAE; SUS;

ANIMAL FEED; ANIMALS; BACTERIA; BACTERIAL INFECTIONS; BETA-DEFENSINS; MICROBIAL SENSITIVITY TESTS; PICHIA; PROTEIN ENGINEERING; SWINE; SWINE DISEASES;

EID: 84903818046     PISSN: 01757598     EISSN: 14320614     Source Type: Journal    
DOI: 10.1007/s00253-014-5560-7     Document Type: Article

References (40)

1. Almeida MS, Cabral KS, de Medeiros LN, Valente AP, Almeida FC, Kurtenbach E (2001) cDNA cloning and heterologous expression of functional cysteine-rich antifungal protein Psd1 in the yeast Pichia pastoris: overcoming the inefficiency of the STE13 protease. Arch Biochem Biophys 395(2):199-207 (Pubitemid 33078254)
2. Cabral KM, Almeida MS, Valente AP, Almeida FC, Kurtenbach E (2003) Production of the active antifungal Pisum sativum defensin 1 (Psd 1) in Pichia pastoris: overcoming the inefficiency of the STE13 protease. Protein Expr Purif 31:115-122 (Pubitemid 37207649)
3. Chen Z, Wang D, Cong Y, Wang J, Zhu J, Yang J, Hu Z, Hu X, Tan Y, Hu F, Rao X (2011) Recombinant antimicrobial peptide hPAB-β expressed in Pichia pastoris, a potential agent active against methicillin-resistant Staphylococcus aureus. Appl Microbiol Biotechnol 8:281-291
4. Corrales-Garcia LL, Possani LD, Corzo G (2011) Expression systems of human β-defensins: vectors, purification and biological activities. Amino Acids 40:5-13
5. Damaso MC, Almeida MS, Kurtenbach E, Martins OB, Pereira N Jr, Andrade CM, Albano RM (2003) Optimized expression of a thermostable xylanase from Thermomyces lanuginosus in Pichia pastoris. Appl Environ Microbiol 69(10):6064-6072 (Pubitemid 37287580)
6. Dhople V, Krukemeyer A, Ramamoorthy A (2006) The human beta-defensin-3, an antibacterial peptide with multiple biological functions. Biochim Biophys Acta 1758(9):1499-1512 (Pubitemid 44444831)
7. Fujita K, Ichimasa S, Zendo T, Koga S, Yoneyama F, Nakayama J, Sonomoto K (2007) Structural analysis and characterization of lacticin Q, a novel bacteriocin belonging to a new family of unmodified bacteriocins of gram-positive bacteria. Appl Environ Microbiol 73(9):2871-2877. doi:10.1128/AEM.02286-06 (Pubitemid 46718569)
8. Ganz T (2005) Defensins and other antimicrobial peptides: a historical perspective and an update. Comb Chem High Throughput Screen 8:209-217
9. Hsiao CP, Siebert KJ (1999) Modeling the inhibitory effects of organic acids on bacteria. Int J Food Microbiol 47:189-201
10. Hsu KH, Pei C, Yeh JY, Shih CH, Chung YC, Hung LT, Ou BR (2009) Production of bioactive human α-defensin 5 in Pichia Pastoris. J Gen Appl Microbiol 55:395-401
11. Hu H, Yu B, He Q (2011) Expressing of porcine beta-defensin-2 mature peptide in the yeast. Wei Sheng Wu Xue Bao 51:704-709
12. Li L, Wang JX, Zhao XF, Kang CJ, Liu N, Xiang JH, Li FH, Sueda S, Kondo H (2005) High level expression, purification, and characterization of the shrimp antimicrobial peptide, Ch-penaeidin, in Pichia pastoris. Protein Expr Purif 39:144-151 (Pubitemid 40084599)
13. Li CL, Zhao YC, Song XY, Huang XX, Zhao WD (2013) Molecular cloning, expression and characterization of the porcine β defensin 2 in E. coli. Protein Pept Lett 20:715-723
14. Liu J, Cai Y, Wang J, Zhou Q, Yang B, Lu Z, Jiao L, Zhang D, Sui S, Jiang Y, Ying W, Qian X (2007) Phosphoproteome profile of human liver Chang's cell based on 2-DE with fluorescence staining and MALDI-TOF/TOF-MS. Electrophoresis 28(23):4348-4358. doi:10.1002/elps.200600696 (Pubitemid 350259037)
15. Looft T, Johnson TA, Allen HK, Bayles DO, Alt DP, Stedtfeld RD, Sul WJ, Stedtfeld TM, Chai B, Cole JR, Hashsham SA, Tiedje JM, Stanton TB (2012) In-feed antibiotic effects on the swine intestinal microbiome. Proc Natl Acad Sci U S A 109(5):1691-1696
16. Mao R, Teng D, Wang X, Xi D, Zhang Y, Hu X, Yang Y, Wang J (2012) Design, expression, and characterization of a novel targeted plectasin against methicillin-resistant Staphylococcus aureus. Appl Microbiol Biotechnol 97(9):3991-4002. doi:10.1007/s00253-012-4508-2
17. Mccluskey AJ, Poon GM, Gariepy J (2007) A rapid and universal tandem-purification strategy for recombinant proteins. Protein Sci 16:2726-2732 (Pubitemid 350172759)
18. Merchant AA, McConnell EL, Liu F, Ramaswamy C, Kulkami RP, Basit AW, Murdan S (2011) Assessment of gastrointestinal pH, fluid and lymphoid tissue in the guinea pig, rabbit and pig, and implications for their use in drug development. Eur J Pharm Sci 42:3-10
19. Sanchez-Hidalgo M, Martinez-Bueno M, Fernandez-Escamilla AM, Valdivia E, Serrano L, Maqueda M (2008) Effect of replacing glutamic residues upon the biological activity and stability of the circular enterocin AS-48. J Antimicrob Chemother 61(6):1256-1265. doi:10.1093/jac/dkn126 (Pubitemid 351753591)
20. Sang YM, Patil AA, Zhang GL, Ross CR, Blecha F (2006) Bioinformatic and expression analysis of novel porcine beta-defensins. Mamm Genome 17:332-339
21. Schäger H (2006) Tricine-SDS-PAGE. Nat Protoc 1:16-22
22. Schroeder BO, Wu Z, Nuding S, Groscurth S, Marcinowski M, Beisner J, Buchner J, Schaller M, Stange EF, Wehkamp J (2011) Reduction of disulphide bonds unmasks potent antimicrobial activity of human beta-defensin 1. Nature 469(7330):419-423. doi:10.1038/nature09674
23. Song W, Shi Y, Xiao MZ, Lu H, Qu TJ, Li P, Wu G, Tian Y (2009) In vitro bactericidal activity of recombinant human-defensin-3 against pathogenic bacterial strains in human tooth root canal. Int J Antimicrob Agents 33:237-243
24. Su S, Liu P, Zhang H, Li Z, Song Z, Zhang L, Chen S (2011) Proteomic analysis of human age-related nuclear cataracts and normal lens nuclei. Invest Ophthalmol Vis Sci 52(7):4182-4191. doi:10.1167/iovs.10-7094
25. Tian ZG, Dong TT, Yang YL, Teng D, Wang JH (2009) Expression of antimicrobial peptide LH multimers in Escherichia coli C43(DE3). Appl Microbiol Biotechnol 83:143-149
26. Veldhuizen EJA, Hendriks HGCJM, Hogenkamp A, van Dijk A, Gaastra W, Tooten PCJ, Haagsman HP (2006) Differential regulation of porcine beta-defensins 1 and 2 upon Salmonella infection in the intestinal epithelial cell line IPI-2I. Vet Immunol Immunopathol 114:94-102 (Pubitemid 44415996)
27. Veldhuizen EJA, van Dijk A, Tersteeg MHG, Kalkhove SIC, van der Meulen J, Niewold TA, Haagsman HP (2007) Expression of betadefensins pBD-1 and pBD-2 along the small intestinal tract of the pig: lack of upregulation in vivo upon Salmonella typhimurium infection. Mol Immunol 44:276-283 (Pubitemid 44287064)
28. Veldhuizen EJA, Rijnders M, Claassen EA, van Dijk A, Haagsman HP (2008) Porcine beta-defensin 2 displays broad antimicrobial activity against pathogenic intestinal bacteria. Mol Immunol 45:386-394 (Pubitemid 47332639)
29. Veldhuizen EJA, Koomen I, Ultee T, van Dijk A, Haagsman HP (2009) Salmonella serovar specific upregulation of porcine defensins 1 and 2 in a jejunal epithelial cell line. Vet Microbiol 136:69-75
30. Wang A, Wang S, Shen M, Chen F, Zou Z, Ran X, Cheng T, Su Y, Wang J (2009) High level expression and purification of bioactive human α-defensin 5 mature peptide in Pichia pastoris. Appl Microbiol Biotechnol 84:877-884
31. Wehkamp J, Fellermann K, Herrlinger KR, Bevins CL, Stange EF (2005a) Mechanisms of disease: defensins in gastrointestinal diseases. Nat Clin Pract Gastroenterol Hepatol 2:406-415 (Pubitemid 41486093)
32. Wehkamp J, Salzman NH, Porter E, Nuding S, Weichenthal M, Petras RE, Shen B, Schaeffeler E, Schwab M, Linzmeier R, Feathers RW, Chu H, JrH L, Fellermann K, Ganz T, Stange EF, Bevins CL (2005b) Reduced Paneth cell alpha-defensins in ileal Crohn's disease. Proc Natl Acad Sci U S A 102:18129-18134 (Pubitemid 41798513)
33. Yadava A, Ockenhouse CF (2003) Effect of codon optimization on expression levels of a functionally folded malaria vaccine candidate in prokaryotic and eukaryotic expression systems. Infect Immun 71(9):4961-4969 (Pubitemid 37070738)
34. Yang YL, Teng D, Zhang J, Tian ZG, Wang SR, Wang JH (2011) Characterization of recombinant plectasin: solubility, antimicrobial activity and factors that affect its activity. Process Biochem 46:1050-1055
35. Zhang GL, Wu H, Shi JS, Ganz T, Ross CR, Blecha F (1998) Molecular cloning and tissue expression of porcine beta-defensin-1. FEBS Lett 424:37-40 (Pubitemid 28130347)
36. Zhang D, Sun L, Yang L, Liu W(2010) Fusion expression and bioactivity comparison of porcine beta-defensin-2 and porcine interferon-gamma in Pichia pastoris. Sheng Wu Gong Cheng Xue Bao 26:1652-1659
37. Zhang J, Yang YL, Teng D, Tian ZG, Wang SR, Wang JH (2011) Expression of plectasin in Pichia pastoris and its characterization as a new antimicrobial peptide against Staphyloccocus and Streptococcus. Protein Expr Purif 78:189-196
38. Zhang Y, Teng D, Mao R, Wang X, Xi D, Hu X, Wang J (2014) High expression of a plectasin-derived peptide NZ2114 in Pichia pastoris and its pharmacodynamics, postantibiotic and synergy against Staphylococcus aureus. Appl Microbiol Biotechnol 98(2):681-694. doi:10.1007/s00253-013-4881-2
39. Zhu YG, Johnson TA, Su JQ, Qiao M, Guo GX, Stedtfeld RD, Hashsham SA, Tiedje JM (2013) Diverse and abundant antibiotic resistance genes in Chinese swine farms. Proc Natl Acad Sci U S A 110(9):3435-3440
40. Zilbauer M, Dorrell N, Boughan PK, Harris A, Wren BW, Klein NJ, Bajaj-Elliott M (2005) Intestinal innate immunity to Campylobacter jejuni results in induction of bactericidal human beta-defensins 2 and 3. Infect Immun 73(11):7281-7289 (Pubitemid 41587627)