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CN107164397A 一种磷脂酶D的基因挖掘及其应用方法 无效 - 驳回

一种磷脂酶D的基因挖掘及其应用方法
摘要：
本发明提供了一种基于基因挖掘方法获得的磷脂酶D及其基因编码序列与应用方法。该磷脂酶D基因全长1,614bp，编码538个氨基酸，以质粒pET28a(+)为表达载体，以大肠杆菌E.coli BL21(DE3)为表达宿主，实现了磷脂酶D的高效表达。该磷脂酶D基因的基因挖掘方法简易便捷，经挖掘获得的目标基因所构建的重组菌株经发酵诱导可以分泌磷脂酶D，该磷脂酶D的最适反应温度为60℃，最适反应pH为7.5，且Ca2+对其酶活具有较好的激活促进作用。该酶能对磷脂进行改性，催化底物磷脂酰胆碱和L‑丝氨酸合成磷脂酰丝氨酸，在食品、医药和保健行业具有良好的应用前景。
摘要（英）：

The invention provides a phospholipase D acquired based on a gene mining method, as well as a gene coding sequence and application method thereof. A phospholipase D gene has a full length of 1,614bp, encodes 538 amino acids, uses plasmid pET28a (+) as an expression vector, and uses E. coli BL21 (DE3) as an expression host, to realize efficient expression of phospholipase D. The phospholipase D gene is simple and convenient in gene mining method; a recombinant strain constructed by the mined target gene can be fermented and induced to excrete the phospholipase D. For the phospholipase D, an optimum reaction temperature is 60 DEG C; an optimum reaction pH is 7.5; and Ca<2+> has good activating and promoting effects on enzyme activity of the phospholipase D; the phospholipase D can modify phospholipids, and can catalyze substrates phosphatidylcholine and L-serine to synthesize phosphatidylserine, thereby having good application prospects in food, medicine and healthcare industries.

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