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Production of recombinant porcine colipase secreted by Pichia pastoris and its application to improve dietary fat digestion and growth of postweaning piglets
Author(s) -
Liu FangChueh,
Chen HsiaoLing,
Chong KowitYu,
Hsu ALi,
Chen ChuanMu
Publication year - 2008
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.46
Subject(s) - pichia pastoris , yeast , biology , colipase , signal peptide , recombinant dna , biochemistry , microbiology and biotechnology , gene , enzyme , lipase , pancreatic lipase
Abstract Recombinant porcine pancreatic colipase (pCoL) was produced in the methylotrophic yeast Pichia pastoris. A synthetic yeast secretion cassette was constructed with the constitutive promoter of the glyceraldehyde‐3‐phosphate‐dehydrogenase (GAPDH) gene and the yeast α‐mating factor signal peptide. The pCoL cDNA corresponding in the coding sequence, excluding a 16‐amino acid segment of the native signal sequence, was cloned into the pGAPZαB vector and integrated into the genome of P. pastoris. Yeast transformants were cultured and bioactive pCoL protein was detected in the supernatant at a high‐level of 126.8 mg/L after 3 days of culture. The transformed yeast containing the highest recombinant colipase level (pCoL yeast) and native yeast GS 115 not containing pCoL (non‐pCoL yeast, as a control group) were separately cultured and the supernatants were adsorbed by dried skim milk. In an animal trial, two concentrations of colipase activity (0 vs. 5,000 U/kg in the diet) were blended with the pig corn‐soybean basal diet and fed to weaned piglets for 4 weeks. The pCoL‐administrated test group gained significantly more weight than piglets in the control group when measured at Day 15 (11.84 ± 0.70 vs. 10.59 ± 0.39 kg, P < 0.05), Day 22 (15.84 ± 0.95 vs. 14.32 ± 0.59 kg, P < 0.01), and Day 28 (20.19 ± 1.47 vs. 18.54 ± 0.92 kg, P < 0.01) after weaning. The blood triglyceride (TG) concentrations were significantly increased in the experimental test group that received recombinant colipase on the 28th day of postweaning when compared with that of the control group (32.50 vs. 16.37 mg/dL; P < 0.0001). These experimental data suggest that the use of recombinant porcine colipase as a dietary supplement provides an alternative approach for improving fat digestion and enhancing growth in postweaning piglets.