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|標題:||Production of Recombinant Porcine Colipase Secreted by Pichia Pastoris and Its Application to Improve Dietary Fat Digestion and Growth of Postweaning Piglets||作者:||Liu, F.C.
|關鍵字:||postweaning piglet;recombinant colipase;triglyceride;Picha pastoris;high-level expression;pancreatic lipase;methylotrophic yeast;coconut;oil;bile-salts;gene;pigs;purification;temperature;proteins||Project:||Biotechnology Progress||期刊/報告no：:||Biotechnology Progress, Volume 24, Issue 6, Page(s) 1333-1341.||摘要:||
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 alpha 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 pGAPZ alpha B 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 skin? 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 pCo-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 oil 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.
|Appears in Collections:||生命科學系所|
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