CLONING/SEQUENCING/EXPRESSION OF HUMAN BSA LIPASE CDNA

胆盐激活脂肪酶(BSAL)存在于一种有限的乳汁中。 哺乳动物的数量：人类、大猩猩、狗和猫。这个 酵素在婴幼儿饮食中起着重要的消化作用 因此，会产生显著的数量差异 母乳喂养的婴儿和配方奶喂养的婴儿之间的体重增加；在这方面 关于这一点，重要的是要注意到牛乳中不存在Bsal。 在IB阶段的研究中，我们计划克隆、测序和表达 人BSAL基因并为第二阶段提供了框架 研究中将生产重组人BSAL 婴儿配方奶粉中的大量补充剂。 通过克隆获得人Bsal基因的表达 转化成适合于高水平生产的表达载体 酿酒酵母。表达水平将是 酶联免疫吸附试验和直接测定酶 活动。重组蛋白将被纯化并 其特点是在物理和化学上与 纯化的人乳酶。

Bile salt-activated lipase (BSAL) is found in the milk of a limited number of mammals: humans, gorillas, dogs, and cats. The enzyme plays an important role in the infant diet in the digestion of lipids and, as such, effects a striking quantitative difference between breast-fed and formula-fed infants in weight gain; in this regard, it is important to note that BSAL is absent in bovine milk. In Phase IB research, we plan to clone, sequence and express the cDNA for human BSAL and provide a framework for Phase II research wherein recombinant human BSAL will be produced in large quantities for supplementation in infant formulas. Expression of the human BSAL cDNA will be achieved by cloning into an expression vector suitable for production at high levels in Saccharomyces cerevisiae. Levels of expression will be ascertained by ELISA and direct measurements of enzymatic activity. The recombinant protein will be purified and characterized for its physical and chemical similarity to the purified human milk enzyme.