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莱茵衣藻酰基辅酶A合成酶cDNA克隆及其酵母表达

DOI: 10.13560/j.cnki.biotech.bull.1985.2015.09.016, PP. 119-124

Keywords: 莱茵衣藻,酰基辅酶A合成酶,酵母YB525,进化树,脂肪酸

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Abstract:

旨在预测并克隆莱茵衣藻酰基辅酶A合成酶cDNA(cracs),分析其在酵母中的功能。RT-PCR克隆cracs序列,ClustalW和MEGA6.0软件分别分析其编码蛋白保守序列和进化树,表达并分析其在酵母YB525中的底物偏好性。结果表明,首次在莱茵衣藻中克隆获得一个cracs,测序表明其序列大小为2004bp,编码667个氨基酸,编码蛋白crACS的预测分子量为72.3kD,包含酰基辅酶A合成酶的两个保守区:AMP-binding区和FACS区。进化树比对显示,crACS与拟南芥的长链酰基辅酶A合成酶LACSs具有较高的同源性。酵母表达显示cracs编码蛋白能互补酵母YB525LACS的缺陷表型,活化并优先利用C16:n>1和C14:n>0。莱茵衣藻cracs编码蛋白可活化外源脂肪酸,属于酰基辅酶A合成酶家族。

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