coenzyme-q10 and 4-hydroxybenzoic-acid

Coenzyme Q (ubiquinone or CoQ) is an essential lipid that plays a role in mitochondrial respiratory electron transport and serves as an important antioxidant. In human and yeast cells, CoQ synthesis derives from aromatic ring precursors and the isoprene biosynthetic pathway.

Topics: Ataxia; Genes, Fungal; Genome, Human; Humans; Mitochondrial Diseases; Mitochondrial Proteins; Models, Biological; Muscle Weakness; Mutation; Parabens; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; Ubiquinone

Topics: Chromatography, Liquid; NAD; NADH, NADPH Oxidoreductases; Schizosaccharomyces; Schizosaccharomyces pombe Proteins; Tandem Mass Spectrometry; Ubiquinone

Coenzyme Q10 (CoQ10) plays an indispensable role in ATP generation through oxidative phosphorylation and helps in scavenging superoxides generated during electron transfer reactions. It finds extensive applications specifically related to oxidative damage and metabolic dysfunctions. This article reports the use of a statistical approach to optimize the concentration of key variables for the enhanced production of CoQ10 by Rhodotorula glutinis in a lab-scale fermenter. The culture conditions that promote optimum growth and CoQ10 production were optimized and the interaction of significant variables para-hydroxybenzoic acid (PHB, 819.34 mg/L) and soybean oil (7.78% [v/v]) was studied using response surface methodology (RSM). CoQ10 production increased considerably from 10 mg/L (in control) to 39.2 mg/L in batch mode with RSM-optimized precursor concentration. In the fed-batch mode, PHB and soybean oil feeding strategy enhanced CoQ10 production to 78.2 mg/L.

Topics: Analysis of Variance; Batch Cell Culture Techniques; Bioreactors; Biostatistics; Biotechnology; Models, Theoretical; Parabens; Rhodotorula; Soybean Oil; Ubiquinone

A three-phase fluidized bed reactor (TPFBR) was designed to evaluate the potential of CoQ(10) production by gel-entrapped Sphingomonas sp. ZUTE03 via a conversion-extract coupled process. In the reactor, the CoQ(10) yield reached 46.99 mg/L after 8 h of conversion; a high-level yield of about 45 mg/L was maintained even after 15 repetitions (8 h/batch). To fully utilize the residual precursor (para-hydroxybenzoic acid, PHB) in the aqueous phase, the organic phase was replaced with new solution containing 70 mg/L solanesol for each 8 h batch. The CoQ(10) yield of each batch was maintained at a level of about 43 mg/L until the PHB ran out. When solid solanesol was fed to the organic phase for every 8 h batch, CoQ(10) could accumulate and reach a yield of 171.52 mg/L. When solid solanesol and PHB were fed to the conversion system after every 8 h batch, the CoQ(10) yield reached 441.65 mg/L in the organic phase after 20 repetitions, suggesting that the conversion-extract coupled process could enhance CoQ(10) production in the TPFBR.

Topics: Bioreactors; Biotechnology; Cells, Immobilized; Culture Media; Parabens; Sphingomonas; Terpenes; Ubiquinone

In a water-organic solvent, two-phase conversion system, CoQ(10) could be produced directly from solanesol and para-hydroxybenzoic acid (PHB) by free cells of Sphingomonas sp. ZUTE03 and CoQ(10) concentration in the organic solvent phase was significantly higher than that in the cell. CoQ(10) yield reached a maximal value of 60.8 mg l(-1) in the organic phase and 40.6 mg g(-1)-DCW after 8 h. CoQ(10) also could be produced by gel-entrapped cells in the two-phase conversion system. Soybean oil and hexane were found to be key substances for CoQ(10) production by gel-entrapped cells of Sphingomonas sp. ZUTE03. Soybean oil might improve the release of CoQ10 from the gel-entrapped cells while hexane was the suitable solvent to extract CoQ(10) from the mixed phase of aqueous and organic. The gel-entrapped cells could be re-used to produce CoQ(10) by a repeated-batch culture. After 15 repeats, the yield of CoQ(10) kept at a high level of more than 40 mg l(-1). After 8 h conversion under optimized precursor's concentration, CoQ(10) yield of gel-trapped cells reached 52.2 mg l(-1) with a molar conversion rate of 91% and 89.6% (on PHB and solanesol, respectively). This is the first report on enhanced production of CoQ(10) in a two-phase conversion system by gel-entrapped cells of Sphingomonas sp. ZUTE03.

Topics: Cells, Immobilized; Chemical Fractionation; Industrial Microbiology; Parabens; Sphingomonas; Terpenes; Ubiquinone

In this work, Escherichia coli was engineered to produce a medically valuable cofactor, coenzyme Q(10) (CoQ(10)), by removing the endogenous octaprenyl diphosphate synthase gene and functionally replacing it with a decaprenyl diphosphate synthase gene from Sphingomonas baekryungensis. In addition, by over-expressing genes coding for rate-limiting enzymes of the aromatic pathway, biosynthesis of the CoQ(10) precursor para-hydroxybenzoate (PHB) was increased. The production of isoprenoid precursors of CoQ(10) was also improved by the heterologous expression of a synthetic mevalonate operon, which permits the conversion of exogenously supplied mevalonate to farnesyl diphosphate. The over-expression of these precursors in the CoQ(10)-producing E. coli strain resulted in an increase in CoQ(10) content, as well as in the accumulation of an intermediate of the ubiquinone pathway, decaprenylphenol (10P-Ph). In addition, the over-expression of a PHB decaprenyl transferase (UbiA) encoded by a gene from Erythrobacter sp. NAP1 was introduced to direct the flux of DPP and PHB towards the ubiquinone pathway. This further increased CoQ(10) content in engineered E. coli, but decreased the accumulation of 10P-Ph. Finally, we report that the combined over-production of isoprenoid precursors and over-expression of UbiA results in the decaprenylation of para-aminobenzoate, a biosynthetic precursor of folate, which is structurally similar to PHB.

Topics: 4-Aminobenzoic Acid; Alkyl and Aryl Transferases; Dimethylallyltranstransferase; Escherichia coli; Gene Deletion; Genetic Engineering; Mevalonic Acid; Parabens; Phenols; Polyisoprenyl Phosphates; Promoter Regions, Genetic; Sesquiterpenes; Sphingomonadaceae; Sphingomonas; Terpenes; Ubiquinone; Up-Regulation