RESUMO
Blakeslea trispora is used commercially to produce ß-carotene. Isopentenyl pyrophosphate isomerase (IPI) and geranylgeranyl pyrophosphate synthase (GGPS) are key enzymes in the biosynthesis of carotenoids. The cDNAs of genes ipi and carG were cloned from the fungus and expressed in Escherichia coli. Greater GGPS activity was needed in the engineered E. coli when IPP activity was increased. The introduction of GGPS and IPI increased the ß-carotene content in E. coli from 0.5 to 0.95 mg/g dry wt.
Assuntos
Isomerases de Ligação Dupla Carbono-Carbono/metabolismo , Escherichia coli/genética , Proteínas Fúngicas/metabolismo , Geranil-Geranildifosfato Geranil-Geraniltransferase/metabolismo , Mucorales/enzimologia , Mucorales/genética , beta Caroteno/biossíntese , Isomerases de Ligação Dupla Carbono-Carbono/química , Isomerases de Ligação Dupla Carbono-Carbono/genética , Clonagem Molecular , Biologia Computacional , Eletroforese em Gel de Poliacrilamida , Escherichia coli/enzimologia , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Geranil-Geranildifosfato Geranil-Geraniltransferase/química , Geranil-Geranildifosfato Geranil-Geraniltransferase/genética , Hemiterpenos , Redes e Vias Metabólicas , FilogeniaRESUMO
Coenzyme Q10 (CoQ10) is a vitamin-like substance which plays a crucial role in the respiratory chain ranging from bacteria to humans and in the radical scavenging in human body. In this study, the full-length hmgR gene (encoding 3-hydroxy-3-methyl-glutaryl-CoA reductase, HMG-CoA reductase) was cloned and overexpressed in Schizosaccharomyces pombe. Using the pREPG yeast depressed under the thiamine as the control, CoQ10 contents increased up to 2.68 and 3.09 times when recombinant cells were incubated without and with arachidonic acid, respectively. It demonstrated that arachidonic acid could upregulate the activity of HMG-CoA reductase and that hmgR gene played a significant role in CoQ10 biosynthesis. So, it has an importance to be utilized for fermentation.
