利用修饰的真核脂肪酸合酶在昆虫细胞中生产聚羟基脂肪酸酯生物聚合物。
Production of a polyhydroxyalkanoate biopolymer in insect cells with a modified eucaryotic fatty acid synthase.
作者信息
Williams M D, Rahn J A, Sherman D H
机构信息
Department of Microbiology and Biological Process Technology Institute, St. Paul, Minnesota 55108-6106, USA.
出版信息
Appl Environ Microbiol. 1996 Jul;62(7):2540-6. doi: 10.1128/aem.62.7.2540-2546.1996.
Abstract
A novel pathway for the synthesis of poly-3-hydroxybutyrate has been engineered by simultaneous delivery of two genes into insect cells (Spodoptera frugiperda) by use of individual baculovirus vectors. This system includes expression of a dehydrase-domain mutant rat fatty acid synthase cDNA and the phbC gene encoding polyhydroxyalkanoate synthase from Alcaligenes eutrophus. The dehydrase-deficient fatty acid synthase provides de novo synthesis of R-(-)-3-hydroxybutyryl-coenzyme A as a premature termination product rather than palmityl-coenzyme A, the normal product of wild-type rat fatty acid synthase. High levels of this mutant multifunctional protein provide a suitable precursor pool of R-(-)-3-hydroxybutyryl-coenzyme A for conversion to poly-3-hydroxybutyrate in insect cells coexpressing the phbC gene product. This strategy for redesigning a poly-3-hydroxybutyrate biosynthetic pathway suggests a new method for generating structurally diverse polyhydroxyalkanoates by metabolic engineering.
摘要
通过利用单个杆状病毒载体将两个基因同时导入昆虫细胞（草地贪夜蛾），设计出了一条合成聚-3-羟基丁酸酯的新途径。该系统包括脱水酶结构域突变型大鼠脂肪酸合酶cDNA的表达以及编码产碱杆菌聚羟基链烷酸酯合酶的phbC基因的表达。缺乏脱水酶的脂肪酸合酶从头合成R-(-)-3-羟基丁酰辅酶A作为提前终止产物，而不是野生型大鼠脂肪酸合酶的正常产物棕榈酰辅酶A。这种突变型多功能蛋白的高水平表达为在共表达phbC基因产物的昆虫细胞中转化为聚-3-羟基丁酸酯提供了合适的R-(-)-3-羟基丁酰辅酶A前体库。这种重新设计聚-3-羟基丁酸酯生物合成途径的策略提示了一种通过代谢工程产生结构多样的聚羟基链烷酸酯的新方法。
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本文引用的文献
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Polyhydroxybutyrate, a biodegradable thermoplastic, produced in transgenic plants.聚羟基丁酸酯，一种可生物降解的热塑性塑料，在转基因植物中生产。
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