Production of a polyhydroxyalkanoate biopolymer in insect cells with a modified eucaryotic fatty acid synthase.
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.
Williams
M D
MD
Department of Microbiology and Biological Process Technology Institute, St. Paul, Minnesota 55108-6106, USA.
Rahn
J A
JA
Sherman
D H
DH
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001704:Biopolymers / Q000096:biosynthesis* / Q000737:chemistry; D002460:Cell Line; D003594:Cytoplasmic Granules / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D064429:Fatty Acid Synthases / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005818:Genetic Engineering; D051381:Rats; D018411:Spodoptera
D001704:Biopolymers; D018076:DNA, Complementary; D064429:Fatty Acid Synthases