Enhancement of antibody production by the addition of Coenzyme-Q<Subscript>10</Subscript>

Recently, there has been a growing demand for therapeutic monoclonal antibodies (MAbs) on the global market. Because therapeutic MAbs are more expensive than low-molecular-weight drugs, there have been strong demands to lower their production costs. Therefore, efficient methods to minimize the cost of goods are currently active areas of research. We have screened several enhancers of specific MAb production rate (SPR) using a YB2/0 cell line and found that coenzyme-Q₁₀ (CoQ₁₀) is a promising enhancer candidate. CoQ₁₀ is well known as a strong antioxidant in the respiratory chain and is used for healthcare and other applications. Because CoQ₁₀ is negligibly water soluble, most studies are limited by low concentrations. We added CoQ₁₀ to a culture medium as dispersed nanoparticles at several concentrations (Q-Media) and conducted a fed-batch culture. Although the Q-Media had no effect on cumulative viable cell density, it enhanced SPR by 29%. In addition, the Q-Media had no effect on the binding or cytotoxic activity of MAbs. Q-Media also enhanced SPR with CHO and NS0 cell lines by 30%. These observations suggest that CoQ₁₀ serves as a powerful aid in the production of MAbs by enhancing SPR without changing the characteristics of cell growth, or adversely affecting the quality or biological activity of MAbs.     

Coenzyme Q<Subscript>10</Subscript> production by <Emphasis Type="Italic">Rhodobacter sphaeroides</Emphasis> in stirred tank and in airlift bioreactor

A higher Coenzyme Q₁₀ (CoQ₁₀) concentration of 25.04 mg/l was found in airlift bioreactor than the value of 18.11 mg/l obtained in stirred tank under the aerobic-dark cultivation of Rhodobacter sphaeroides. Aeration rate didn’t show obvious impact to CoQ₁₀ production in airlift bioreactor. The fed-batch operation in airlift bioreactor could increase the biomass concentration and led to the maximum CoQ₁₀ concentration of 33.91 mg/l measured, but a lower CoQ₁₀ cell content (3.5 mg CoQ₁₀/DCW) was observed in the fed-batch operation as compared to the batch operation. To enhance the CoQ₁₀ content, an aeration-change strategy was proposed in the fed-batch operation of airlift bioreactor. This strategy led to the maximum CoQ₁₀ concentration of 45.65 mg/l, a 35% increase as compared to the simple fed-batch operation. The results of this study suggested that a fed-batch operation in airlift bioreactor accompanying aeration-change could be suitable for CoQ₁₀ production.     

Coenzyme Q<Subscript>10</Subscript> production directly from precursors by free and gel-entrapped <Emphasis Type="Italic">Sphingomonas</Emphasis> sp. ZUTE03 in a water-organic solvent,two-phase conversion system

In a water-organic solvent, two-phase conversion system, CoQ₁₀ could be produced directly from solanesol and para-hydroxybenzoic acid (PHB) by free cells of Sphingomonas sp. ZUTE03 and CoQ₁₀ concentration in the organic solvent phase was significantly higher than that in the cell. CoQ₁₀ 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. CoQ₁₀ 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 CoQ₁₀ 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 CoQ₁₀ from the mixed phase of aqueous and organic. The gel-entrapped cells could be re-used to produce CoQ₁₀ by a repeated-batch culture. After 15 repeats, the yield of CoQ₁₀ kept at a high level of more than 40 mg l⁻¹. After 8 h conversion under optimized precursor’s concentration, CoQ₁₀ 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 CoQ₁₀ in a two-phase conversion system by gel-entrapped cells of Sphingomonas sp. ZUTE03.     

Optimization of culture conditions and scale-up to pilot and plant scales for coenzyme Q<Subscript>10</Subscript> production by <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>

This report describes the optimization of culture conditions for coenzyme Q₁₀ (CoQ₁₀) production by Agrobacterium tumefaciens KCCM 10413, an identified high-CoQ₁₀-producing strain (Kim et al., Korean patent. 10-0458818, 2002b). Among the conditions tested, the pH and the dissolved oxygen (DO) levels were the key factors affecting CoQ₁₀ production. When the pH and DO levels were controlled at 7.0 and 0–10%, respectively, a dry cell weight (DCW) of 48.4 g l⁻¹ and a CoQ₁₀ production of 320 mg l⁻¹ were obtained after 96 h of batch culture, corresponding to a specific CoQ₁₀ content of 6.61 mg g-DCW⁻¹. In a fed-batch culture of sucrose, the DCW, specific CoQ₁₀ content, and CoQ₁₀ production increased to 53.6 g l⁻¹, 8.54 mg g-DCW⁻¹, and 458 mg l⁻¹, respectively. CoQ₁₀ production was scaled up from a laboratory scale (5-l fermentor) to a pilot scale (300 l) and a plant scale (5,000 l) using the impeller tip velocity (V _tip) as a scale-up parameter. CoQ₁₀ production at the laboratory scale was similar to those at the pilot and plant scales. This is the first report of pilot- and plant-scale productions of CoQ₁₀ in A. tumefaciens.     

Ca<Superscript>2+</Superscript> increases the specific coenzyme Q<Subscript>10</Subscript> content in <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>

Of various metal ions (Ca²⁺, Cr³⁺, Cu²⁺, Fe²⁺, Mg²⁺, Mn²⁺, Ni²⁺ and Zn²⁺) added to the culture medium of Agrobacterium tumefaciens at 1 mM, only Ca²⁺ increased Coenzyme Q10 (CoQ₁₀) content in cells without the inhibition of cell growth. In a pH-stat fed-batch culture, supplementation with 40 mM of CaCO₃ increased the specific CoQ₁₀ content and oxidative stress by 22.4 and 48%, respectively. Also, the effect of Ca²⁺ on the increase of CoQ₁₀ content was successfully verified in a pilot-scale (300 L) fermentor. In this study, the increased oxidative stress in A. tumefaciens culture by the supplementation of Ca²⁺ is hypothesized to stimulate the increase of specific CoQ₁₀ content in order to protect the membrane against lipid peroxidation. Our results improve the understanding of Ca²⁺ effect on CoQ₁₀ biosynthesis in A. tumefaciens and should contribute to better industrial production of CoQ₁₀ by biological processes.     

Bioenergetic and Antioxidant Properties of Coenzyme Q<Subscript>10</Subscript>: Recent Developments

For a number of years, coenzyme Q (CoQ₁₀ in humans) was known for its key role in mitochondrial bioenergetics; later studies demonstrated its presence in other subcellular fractions and in plasma, and extensively investigated its antioxidant role. These two functions constitute the basis on which research supporting the clinical use of CoQ₁₀ is founded. Also at the inner mitochondrial membrane level, coenzyme Q is recognized as an obligatory co-factor for the function of uncoupling proteins and a modulator of the transition pore. Furthermore, recent data reveal that CoQ₁₀ affects expression of genes involved in human cell signalling, metabolism, and transport and some of the effects of exogenously administered CoQ₁₀ may be due to this property. Coenzyme Q is the only lipid soluble antioxidant synthesized endogenously. In its reduced form, CoQH₂, ubiquinol, inhibits protein and DNA oxidation but it is the effect on lipid peroxidation that has been most deeply studied. Ubiquinol inhibits the peroxidation of cell membrane lipids and also that of lipoprotein lipids present in the circulation. Dietary supplementation with CoQ₁₀ results in increased levels of ubiquinol-10 within circulating lipoproteins and increased resistance of human low-density lipoproteins to the initiation of lipid peroxidation. Moreover, CoQ₁₀ has a direct anti-atherogenic effect, which has been demonstrated in apolipoprotein E-deficient mice fed with a high-fat diet. In this model, supplementation with CoQ₁₀ at pharmacological doses was capable of decreasing the absolute concentration of lipid hydroperoxides in atherosclerotic lesions and of minimizing the size of atherosclerotic lesions in the whole aorta. Whether these protective effects are only due to the antioxidant properties of coenzyme Q remains to be established; recent data point out that CoQ₁₀ could have a direct effect on endothelial function. In patients with stable moderate CHF, oral CoQ₁₀ supplementation was shown to ameliorate cardiac contractility and endothelial dysfunction. Recent data from our laboratory showed a strong correlation between endothelium bound extra cellular SOD (ecSOD) and flow-dependent endothelial-mediated dilation, a functional parameter commonly used as a biomarker of vascular function. The study also highlighted that supplementation with CoQ₁₀ that significantly affects endothelium-bound ecSOD activity. Furthermore, we showed a significant correlation between increase in endothelial bound ecSOD activity and improvement in FMD after CoQ₁₀ supplementation. The effect was more pronounced in patients with low basal values of ecSOD. Finally, we summarize the findings, also from our laboratory, on the implications of CoQ₁₀ in seminal fluid integrity and sperm cell motility.     

Stimulation of insulin release from pancreatic islets by quinones

Coenzyme Q (CoQ₀) and other quinones were shown to be potent insulin secretagogues in the isolated pancreatic islet. The order of potency was CoQ₀benzoquinonehydroquinonemenadione. CoQ₆ and CoQ₁₀ (ubiquinone), duroquinone and durohydroquinone did not stimulate insulin release. CoQ₀'s insulinotropism was enhanced in calcium-free medium and CoQ₀ appeared to stimulate only the second phase of insulin release. CoQ₀ inhibited inositol mono-, bis- and trisphosphate formation. Inhibitors of mitochondrial respiration (rotenone, antimycin A, FCCP and cyanide) and the calcium channel blocker verapamil, did not inhibit CoQ₀-induced insulin release. Dicumarol, an inhibitor of quinone reductase, did not inhibit CoQ₀-induced insulin release, but it did inhibit glucose-induced insulin release suggesting that the enzyme and quinones play a role in glucose-induced insulin release. Quinones may stimulate insulin release by mimicking physiologically-occuring quinones, such as CoQ₁₀, by acting on the plasma membrane or in the cytosol. Exogenous quinones may bypass the quinone reductase reaction, as well as many reactions important for exocytosis.     

Identification of bottlenecks in Escherichia coli engineered for the production of CoQ(10)

In this work, Escherichia coli was engineered to produce a medically valuable cofactor, coenzyme Q₁₀ (CoQ₁₀), 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 CoQ₁₀ precursor para-hydroxybenzoate (PHB) was increased. The production of isoprenoid precursors of CoQ₁₀ 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 CoQ₁₀-producing E. coli strain resulted in an increase in CoQ₁₀ 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 CoQ₁₀ 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.     

Controlling the sucrose concentration increases Coenzyme Q10 production in fed-batch culture of <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>

The production yield of Coenzyme Q₁₀ (CoQ₁₀) from the sucrose consumed by Agrobacterium tumefaciens KCCM 10413 decreased, and high levels of exopolysaccharide (EPS) accumulated after switching from batch culture to fed-batch culture. Therefore, we examined the effect of sucrose concentration on the fermentation profile by A. tumefaciens. In the continuous fed-batch culture with the sucrose concentration maintained constantly at 10, 20, 30, and 40 g l⁻¹, the dry cell weight (DCW), specific CoQ₁₀ content, CoQ₁₀ production, and the production yield of CoQ₁₀ from the sucrose consumed increased, whereas EPS production decreased as maintained sucrose concentration decreased. The pH-stat fed-batch culture system was adapted for CoQ₁₀ production to minimize the concentration of the carbon source and osmotic stress from sucrose. Using the pH-stat fed-batch culture system, the DCW, specific CoQ₁₀ content, CoQ₁₀ production, and the product yield of CoQ₁₀ from the sucrose consumed increased by 22.6, 13.7, 39.3, and 39.3%, respectively, whereas EPS production decreased by 30.7% compared to those of fed-batch culture in the previous report (Ha SJ, Kim SY, Seo JH, Oh DK, Lee JK, Appl Microbiol Biotechnol, 74:974–980, 2007). The pH-stat fed-batch culture system was scaled up to a pilot scale (300 l), and the CoQ₁₀ production results obtained (626.5 mg l⁻¹ of CoQ₁₀ and 9.25 mg g DCW⁻¹ of specific CoQ₁₀ content) were similar to those obtained at the laboratory scale. Thus, an efficient and highly competitive process for microbial CoQ₁₀ production is available.     

INFLUENCE OF pH AND TEMPERATURE ON THE ACTIVITY OF SnO2-BOUND α-AMYLASE: A GENOTOXICITY ASSESSMENT OF SnO2 NANOPARTICLES

Immobilization of biologically important molecules on a myriad of nanosized materials has attracted great attention due to their small size, biocompatibility, higher surface-to-volume ratio, and lower toxicity. These properties make nanoparticles (NPs) a superior matrix over bulk material for the immobilization of enzymes and proteins. In the present study, Bacillus amyloliquefaciens α-amylase was immobilized on SnO₂ nanoparticles by a simple adsorption mechanism. Nanoparticle-adsorbed enzyme retained 90% of the original enzyme activity. Thermal stability of nanosupport was investigated by thermogravimetric and differential thermal analysis. Scanning electron microscopic studies showed that NPs have porous structure for the high-yield immobilization of α-amylase. The genotoxicity of SnO₂-NPs was analyzed by pUC¹⁹ plasmid nicking and comet assay and revealed that no remarkable DNA damage occurred in lymphocytes. The pH-optima was found to be the same for both free and SnO₂-NPs bound enzyme, while the temperature-optimum for NPs-adsorbed α-amylase was 5°C higher than its free counterpart. Immobilized enzyme retained more than 70% enzyme activity even after its eight repeated uses.     

Impact of Carrier Systems on the Interactions of Coenzyme Q<Subscript>10</Subscript> with Model Lipid Membranes

We investigated the influence of carrier systems for different commercially available water-soluble formulations for coenzyme Q₁₀ on structural changes of model lipid membranes formed by 1,2-dipalmitoyl-sn-glycero-3-phosphocholine and by a mixture of phosphatidylcholine and sphingomyelin (2.4:1). Structural changes in the membranes were measured using fluorescence anisotropy, electron paramagnetic resonance, and differential scanning calorimetry. Two fluorophores and two spin probes were used to monitor membrane characteristics close to the water-lipid interface and in the middle of the bilayer of the model lipid membranes. Different water-soluble carrier systems were tested. These data show that different systems can facilitate penetration of CoQ₁₀ in the lipid membranes, where an increase in the lipid order parameter was observed. In addition, water soluble CoQ₁₀ formulations better protect lipids from oxidation in liposome solution. With the exception of the carriers in an emulsified formulation of CoQ₁₀, those in the other samples did not have any significant effects on membrane fluidity.     

Water-soluble formulation of Coenzyme Q10 inhibits Bax-induced destabilization of mitochondria in mammalian cells

Oxidative stress leads to mitochondrial dysfunction, which triggers the opening of the permeability transition pores (PTP) and the release of pro-apoptotic factors causing apoptotic cell death. In a limited number of cell systems, anti-oxidants and free-radical scavengers have been shown to block this response. We have previously reported that coenzyme Q₁₀ (CoQ₁₀), an electron carrier in the mitochondrial respiratory chain, is involved in the reactive oxygen species (ROS) removal and prevention of oxidative stress-induced apoptosis in neuronal cells. However, the mechanism of this protection has not been fully elucidated. In the present study we investigated the effects of CoQ₁₀ on the mitochondrial events characteristic to apoptosis, especially on the function of pro-apoptotic protein Bax. Our results demonstrated that following a brief exposure of two human cell lines (fibroblasts and HEK293 cells) to H₂O₂ the intracellular levels of ROS and the association of Bax with the mitochondria significantly increased and the cells underwent apoptosis. Both of these events, as well as the release of cytochrome c from the mitochondria, were blocked by a 24 h pre-treatment with CoQ₁₀. It is therefore believed that CoQ₁₀ prevented the collapse of the mitochondrial membrane potential in response to the H₂O₂ treatment. Recombinant Bax protein alone caused the ROS generation and release of cytochrome c from isolated mitochondria and, again, CoQ₁₀ inhibited these Bax-induced mitochondrial dysfunctions.     

Increasing coenzyme Q10 yield from Rhodopseudomonas palustris by expressing rate-limiting enzymes and blocking carotenoid and hopanoid pathways

Coenzyme Q₁₀ (CoQ₁₀), a strong antioxidant, is used extensively in food, cosmetic and medicine industries. A natural producer, Rhodopseudomonas palustris, was engineered to overproduce CoQ₁₀. For increasing the CoQ₁₀ content, crtB gene was deleted to block the carotenoid pathway. crtB gene deletion led to 33% improvement of CoQ₁₀ content over the wild type strain. However, it was found that the yield of hopanoids was also increased by competing for the precursors from carotenoid pathway with CoQ₁₀ pathway. To further increase the CoQ₁₀ content, hopanoid pathway was blocked by deleting shc gene, resulting in R. palustris [Δshc, ΔcrtB] to produce 4·7 mg g⁻¹ DCW CoQ₁₀, which was 1·2 times higher than the CoQ₁₀ content in the wild type strain. The common strategy of co-expression of rate-limiting enzymes (DXS, DPS and UbiA) was combined with the pathway blocking method resulted in 8·2 mg g⁻¹ DCW of CoQ₁₀, which was 2·9 times higher than that of wild type strain. The results suggested a synergistic effect among different metabolic engineering strategies. This study demonstrates the potential of R. palustris for CoQ₁₀ production and provides viable strategies to increase CoQ₁₀ titer.     

Plasma coenzyme Q10 concentrations are not decreased in male patients with coronary atherosclerosis

Coenzyme Q₁₀ (CoQ₁₀) is an important mitochondrial electron transfer component and has been postulated to function as a powerful antioxidant protecting LDL from oxidative damage. It could thus reduce the risk of cardiovascular disease. Thus far, beneficial effects of supplementation with CoQ₁₀ have been reported. To study the relation between unsupplemented concentrations of plasma CoQ₁₀ and coronary atherosclerosis, we performed a case-control study among 71 male cases with angiographically documented severe coronary atherosclerosis and 69 healthy male controls free from symptomatic cardiovascular disease and without atherosclerotic plaques in the carotid artery.

Plasma CoQ₁₀ concentrations (mean ± SE) were 0.86 ± 0.04 vs. 0.83 ± 0.04 μmol/l for cases and controls, respectively. The CoQ₁₀/LDL-cholesterol ratio (μmol/mmol) was slightly lower in cases than in controls (0.22 ± 0.01 vs. 0.26 ± 0.03). Differences in CoQ₁₀ concentrations and CoQ₁₀/LDL-cholesterol ratio did not reach significance. The odds ratios (95% confidence interval) for the risk of coronary atherosclerosis calculated per μmol/l increase of CoQ₁₀ was 1.12 (0.28–4.43) after adjustment for age, smoking habits, total cholesterol and diastolic blood pressure.

We conclude that an unsupplemented plasma CoQ₁₀ concentration is not related to risk of coronary atherosclerosis.     

Attenuation of Cocaine and Methamphetamine Neurotoxicity by Coenzyme Q10

The neurotoxic effects of cocaine and methamphetamine (METH) were studied in mice brain with a primary objective to determine the neuroprotective potential of coenzyme Q₁₀ (CoQ₁₀) in drug addiction. Repeated treatment of cocaine or METH induced significant reduction in the striatal dopamine and CoQ₁₀ in mice. Cocaine or METH-treated mice exhibited increased thiobarbituric acid reactive substances (TBARs) in the striatum and cerebral cortex without any significant change in the cerebellum. Complex I immunoreactivity was inhibited in both cocaine and METH-treated mice, whereas tyrosine hydroxylase (TH) immunoreactivity was decreased in METH-treated mice and increased in cocaine-treated mice. Neither cocaine nor METH could induce significant change in α-synuclein expression at the doses and duration we have used in the present study. CoQ₁₀ treatment attenuated cocaine and METH-induced inhibition in the striatal ¹⁸F-DOPA uptake as determined by high-resolution microPET neuroimaging. Hence exogenous administration of CoQ₁₀ may provide neuroprotection in drug addiction.     

Effects of dissolved oxygen concentration and DO-stat feeding strategy on CoQ10 production with Rhizobium radiobacter

The cell growth and CoQ₁₀ (coenzyme Q₁₀) formation of Rhizobium radiobacter WSH2601 were investigated in a 7-1 bioreactor under different dissolved oxygen (DO) concentrations. A maximal CoQ₁₀ content (C/B) of 1.91 mg/g dry cell weight (DCW) and CoQ₁₀ concentration of 32.1 mg/l were obtained at the appropriate DO concentration of 40% (of air saturation). High DO concentration was favourable to the cell growth of Rhizobium radiobacter WSH2601. In order to achieve the maximal yield of CoQ₁₀ production, a new DO-stat feeding strategy was proposed, which significantly improved cell growth and CoQ₁₀ formation. With this strategy, the maximal CoQ₁₀ concentration and DCW reached 51.1 mg/l and 23.9 g/l, respectively, which were 67 and 44.8% higher than those obtained in the batch culture with DO concentration controlled.     

Lactate increases coenzyme Q10 production by Agrobacterium tumefaciens

By the optimization of nitrogen source for coenzyme Q₁₀ (ubiquinone, CoQ₁₀) production in Agrobacterium tumefaciens KCCM 10413 culture, the highest CoQ₁₀ production was achieved in medium containing corn steep powder (CSP). Components for a stimulatory effect on the production of CoQ₁₀ in CSP were screened, and lactate was found to increase dry cell weight (DCW) and the specific CoQ₁₀ content. In a fed-batch culture of A. tumefaciens, supplementation with 1.5 g of lactate l⁻¹ further improved DCW, the specific CoQ₁₀ content, and CoQ₁₀ production by 16.0, 5.8, and 22.8%, respectively. It has been reported that lactate stimulates cell growth and acts as an accelerator driving the tricarboxylic acid (TCA) cycle (Roberto et al. 2002, Biotechnol Let 24:427–431; Matsuoka et al. 1996, Biosci Biotechnol Biochem 60:575–579). In this study, lactate supplementation increased DCW and the specific CoQ₁₀ content in A. tumefaciens culture, probably by accelerating TCA cycle and energy production as reported previously, leading to the increase of CoQ₁₀ production.     

Enhanced production of CoQ<Subscript>10</Subscript> by newly isolated <Emphasis Type="Italic">Sphingomonas</Emphasis> sp. ZUTEO3 with a coupled fermentation–extraction process

The use of coenzyme Q₁₀ (CoQ₁₀) as a complementary therapy in heart failure will increase in proportion to the growth of the ageing population and the expansion of statins consumption. Economical production of CoQ₁₀ by microbes will become more important due to the growing demands of the pharmaceutical industry. Process simplification and integration might be one desirable pathway for economic production of CoQ₁₀ by microbial fermentation. In this report, the effect of a coupled fermentation–extraction process on CoQ₁₀ production by newly isolated Sphingomonas sp. ZUTEO3 was evaluated. It was found that the CoQ₁₀ yield of the coupled process was significantly higher than that of the traditional process. As optimal conditions in our experiment, 2% soybean oil was added to the original culture to enhance cell membrane permeability, and 50 mL hexane was added to the 30 h culture as an extracting solvent for the subsequent coupled fermentation–extraction process. The maximal yield of CoQ₁₀ reached 43.2 mg/L and 32.5 mg/g dry cell weight after 38 h of total fermentation period. The coupled process represents one potential pathway for CoQ₁₀ production with even higher yield and lower cost. This is the first report of CoQ₁₀ production by Sphingomonas sp. using a coupled fermentation–extraction process.     

Update on the application of magnetic fields to microalgal cultures

Coenzyme Q₁₀ (CoQ₁₀) is the main CoQ species in human and is used extensively in food, cosmetic and medicine industries because of its antioxidant properties and its benefit in prophylactic medicine and therapy for a variety of diseases. Among various approaches to increase the production of CoQ₁₀, microbial fermentation is the most effective. As knowledge of the biosynthetic enzymes and regulatory mechanisms modulating CoQ₁₀ production increases, opportunities arise for metabolic engineering of CoQ₁₀ in microbial hosts. In this review, we present various strategies used up to date to improve CoQ₁₀ production and focus on metabolic engineering of CoQ₁₀ overproduction in microbes. General strategies of metabolic engineering include providing sufficient precursors for CoQ₁₀, increasing metabolic fluxes, and expanding storage capacity for CoQ₁₀. Based on these strategies, CoQ₁₀ production has been significantly improved in natural CoQ₁₀ producers, as well as in heterologous hosts.