Department of Life Sciences, National Chung Hsing University, Taichung, Taiwan.
J Agric Food Chem. 2010 Mar 24;58(6):3322-9. doi: 10.1021/jf903439q.
The aim of the study was to use Pichia pastoris to express a recombinant porcine lipase gene (pLip). The expression-secretion cassette was constructed using the P. pastoris GAPDH (glyceraldehyde-3-phosphate-dehydrogenase) promoter and an 89-residue prepro-alpha-factor secretion signal fused to the AOX1 terminator (the pGAPZalphaA vector). A total of 1,408 bp of pancreatic lipase cDNA was produced, which was located from the position of 4-nt upstream of ATG to 1408-nt inside the intact coding region of the pLip sequence. In an animal trial, three concentrations of recombinant lipase activity (0, 5,000 and 10,000 U/kg) were blended with the basal diet and fed to weaned piglets for six weeks. During the experimental period, the growth performance (bodyweight, feed intake, and feed efficiency) of the test groups was superior to that of the control group (p < 0.05). Furthermore, the group fed the diet blended with 10,000 U/kg of recombinant lipase showed significant (p < 0.05) increases in blood triglyceride (TG) concentration on the seventh day postweaning. These results suggested that the porcine lipase protein yielded by transformed yeast cells may improve fat digestibility and enhance the growth performance in postweaning piglets.
本研究旨在利用巴斯德毕赤酵母表达重组猪脂肪酶基因（pLip）。表达分泌盒由毕赤酵母 GAPDH（甘油醛-3-磷酸脱氢酶）启动子和 89 个残基的前导肽-α因子信号融合到 AOX1 终止子（pGAPZalphaA 载体）构建而成。总共产生了 1408bp 的胰腺脂肪酶 cDNA，其位置从 ATG 上游的 4 个核苷酸到 pLip 序列完整编码区内部的 1408 个核苷酸。在动物试验中，三种浓度的重组脂肪酶活性（0、5000 和 10000U/kg）与基础日粮混合，喂养断奶仔猪 6 周。在实验期间，试验组的生长性能（体重、采食量和饲料效率）优于对照组（p<0.05）。此外，在断奶后第 7 天，喂食含有 10000U/kg 重组脂肪酶的日粮的组，血液甘油三酯（TG）浓度显著增加（p<0.05）。这些结果表明，转化酵母细胞产生的猪脂肪酶蛋白可能提高脂肪消化率，并增强断奶仔猪的生长性能。
