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1.
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. 相似文献
2.
The use of coenzyme Q 10 (CoQ 10) 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 CoQ 10 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 CoQ 10 by microbial fermentation. In this report, the effect of a coupled fermentation–extraction process on CoQ 10 production by newly isolated Sphingomonas sp. ZUTEO3 was evaluated. It was found that the CoQ 10 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 CoQ 10 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 CoQ 10 production with even higher yield and lower cost. This is the first report of CoQ 10 production by Sphingomonas sp. using a coupled fermentation–extraction process. 相似文献
3.
By the optimization of nitrogen source for coenzyme Q 10 (ubiquinone, CoQ 10) production in Agrobacterium tumefaciens KCCM 10413 culture, the highest CoQ 10 production was achieved in medium containing corn steep powder (CSP). Components for a stimulatory effect on the production
of CoQ 10 in CSP were screened, and lactate was found to increase dry cell weight (DCW) and the specific CoQ 10 content. In a fed-batch culture of A. tumefaciens, supplementation with 1.5 g of lactate l −1 further improved DCW, the specific CoQ 10 content, and CoQ 10 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 CoQ 10 content in A. tumefaciens culture, probably by accelerating TCA cycle and energy production as reported previously, leading to the increase of CoQ 10 production. 相似文献
4.
The production yield of Coenzyme Q 10 (CoQ 10) 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 CoQ 10 content, CoQ 10 production, and the production yield of CoQ 10 from the sucrose consumed increased, whereas EPS production decreased as maintained sucrose concentration decreased. The
pH-stat fed-batch culture system was adapted for CoQ 10 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 CoQ 10 content, CoQ 10 production, and the product yield of CoQ 10 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 CoQ 10 production results obtained (626.5 mg l −1 of CoQ 10 and 9.25 mg g DCW −1 of specific CoQ 10 content) were similar to those obtained at the laboratory scale. Thus, an efficient and highly competitive process for microbial
CoQ 10 production is available. 相似文献
5.
This report describes the optimization of culture conditions for coenzyme Q 10 (CoQ 10) production by Agrobacterium
tumefaciens KCCM 10413, an identified high-CoQ 10-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 CoQ 10 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 CoQ 10 production of 320 mg l −1 were obtained after 96 h of batch culture, corresponding to a specific CoQ 10 content of 6.61 mg g-DCW −1. In a fed-batch culture of sucrose, the DCW, specific CoQ 10 content, and CoQ 10 production increased to 53.6 g l −1, 8.54 mg g-DCW −1, and 458 mg l −1, respectively. CoQ 10 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. CoQ 10 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 CoQ 10 in A. tumefaciens. 相似文献
6.
For the commercial production of CoQ 10, batch-type fermentations were attempted in a 150-l fermenter using a mutant strain of R. sphaeroides. Optimum temperature and initial aeration rate were found to be 30°C and 2 vvm, respectively. Under optimum fermentation
conditions, the maximum value of specific CoQ 10 content was achieved reproducibly as 6.34 mg/g DCW after 24 h, with 3.02 g/l of DCW. During the fermentation, aeration shift
(from the adequate aeration at the early growth phase to the limited aeration in active cellular metabolism) was a key factor
in CoQ 10 production for scale-up. A higher value of the specific CoQ 10 content (8.12 mg/g DCW) was achieved in fed-batch fermentation and comparable to those produced by the pilot-scale fed-batch
fermentations of A. tumefaciens, which indicated that the mutant strain of R. sphaeroides used in this study was a potential high CoQ 10 producer. This is the first detailed study to demonstrate a pilot-scale production of CoQ 10 using a mutant strain of R. sphaeroides. 相似文献
7.
Summary In the production of coenzyme Q 10 (CoQ 10) by Agrobacterium sp. the culture broth becomes highly viscous. In an attempt to improve the production process, the effects of chemical and physical factors on broth viscosity and CoQ 10 production were studied, using Agrobacterium sp. KY-8593. A particular concentration ratio of sugar to ammonium-nitrogen (NH 4–N) in the medium could effectively enhance CoQ 10 production without increasing broth viscosity. An increase in culture temperature to between 32°C and 34°C lowered broth viscosity without reducing CoQ 10 production. NH 4–N concentration and temperature had a correlative effect on broth viscosity. At a temperature of about 33°C, there was a wide range of NH 4–N concentration which was optimal for both broth viscosity and CoQ 10 production. In optimal conditions with 8% sugar the apparent broth viscosity was reduced to less than 10 pseudo-cP and CoQ 10 production was increased to more than 80 mg/l. 相似文献
8.
Coenzyme Q 10 (CoQ 10), a strong antioxidant, is used extensively in food, cosmetic and medicine industries. A natural producer, Rhodopseudomonas palustris, was engineered to overproduce CoQ 10. For increasing the CoQ 10 content, crtB gene was deleted to block the carotenoid pathway. crtB gene deletion led to 33% improvement of CoQ 10 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 CoQ 10 pathway. To further increase the CoQ 10 content, hopanoid pathway was blocked by deleting shc gene, resulting in R. palustris [ Δshc, ΔcrtB] to produce 4·7 mg g −1 DCW CoQ 10, which was 1·2 times higher than the CoQ 10 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 CoQ 10, 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 CoQ 10 production and provides viable strategies to increase CoQ 10 titer. 相似文献
9.
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. 相似文献
10.
A higher Coenzyme Q 10 (CoQ 10) 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 CoQ 10 production in airlift bioreactor. The fed-batch operation in airlift bioreactor could increase the biomass concentration
and led to the maximum CoQ 10 concentration of 33.91 mg/l measured, but a lower CoQ 10 cell content (3.5 mg CoQ 10/DCW) was observed in the fed-batch operation as compared to the batch operation. To enhance the CoQ 10 content, an aeration-change strategy was proposed in the fed-batch operation of airlift bioreactor. This strategy led to
the maximum CoQ 10 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 CoQ 10 production. 相似文献
11.
Coenzyme Q 10 (CoQ 10) is a quinine consisting of ten units of the isoprenoid side-chain. Because it limits the oxidative attack of free radicals to DNA and lipids, CoQ 10 has been used as an antioxidant for foods, cosmetics and pharmaceuticals. Decaprenyl diphosphate synthase (DPS) is the key enzyme for synthesis of the decaprenyl tail in CoQ 10 with isopentenyl diphosphate. The ddsA gene coding for DPS from Gluconobacter suboxydans was expressed under the control of an Escherichia coli constitutive promoter. Analysis of the cell extract in recombinant E. coli BL21/pACDdsA by high performance liquid chromatography and mass spectrometry showed that CoQ 10 rather than endogenous CoQ 8 was biologically synthesized as the major coenzyme Q. Expression of the ddsA gene with low copy number led to the accumulation of CoQ 10 to 0.97 mg l –1 in batch fermentation. A high cell density (103 g l –1) in fed-batch fermentation of E. coli BL21/pACDdsA increased the CoQ 10 concentration to 25.5 mg l –1 and its productivity to 0.67 mg l –1 h –1, which were 26.0 and 6.9 times higher than the corresponding values for batch fermentation. 相似文献
12.
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. 相似文献
13.
Coenzyme Q 10 (CoQ 10) is a redox molecule critical for the proper function of energy metabolism and antioxidant defenses. Despite its essential role in cellular metabolism, the regulation of CoQ 10 biosynthesis in humans remains mostly unknown. Herein, we determined that PPTC7 is a regulatory protein of CoQ 10 biosynthesis required for human cell survival. We demonstrated by in vitro approaches that PPTC7 is a bona fide protein phosphatase that dephosphorylates the human COQ7. Expression modulation experiments determined that human PPTC7 dictates cellular CoQ 10 content. Using two different approaches ( PPTC7 over-expression and caloric restriction), we demonstrated that PPTC7 facilitates and improves the human cell adaptation to respiratory conditions. Moreover, we determined that the physiological role of PPTC7 takes place in the adaptation to starvation and pro-oxidant conditions, facilitating the induction of mitochondrial metabolism while preventing the accumulation of ROS. Here we unveil the first post-translational mechanism regulating CoQ 10 biosynthesis in humans and propose targeting the induction of PPTC7 activity/expression for the treatment of CoQ 10-related mitochondrial diseases. 相似文献
14.
Mutation of Pseudomonas N842 was carried out to increase CoQ 10 production. The productivity of CoQ 10 was improved considerably by repeated mutation, and the content of CoQ 10 per unit cell of the fifth generation mutant was approximately 6 times that of the wild strain, Pseudomonas N842. CoQ 11, which was hardly detectable in the wild strain, increased significantly by mutation, and the ratio of CoQ 11 to total CoQ exceeded 20% in the fourth generation mutant. Intermittent feeding of glucose to the culture medium during cultivation increased cell yield and CoQ production. When total glucose added was 5 times that of basal medium, cell yield and CoQ formation respectively increased about 3 and 4 times. 相似文献
15.
BackgroundCoenzyme Q 10 (CoQ 10) and its analogs are used therapeutically by virtue of their functions as electron carriers, antioxidant compounds, or both. However, published studies suggest that different ubiquinone analogs may produce divergent effects on oxidative phosphorylation and oxidative stress. Methodology/Principal FindingsTo test these concepts, we have evaluated the effects of CoQ 10, coenzyme Q 2 (CoQ 2), idebenone, and vitamin C on bioenergetics and oxidative stress in human skin fibroblasts with primary CoQ 10 deficiency. A final concentration of 5 µM of each compound was chosen to approximate the plasma concentration of CoQ 10 of patients treated with oral ubiquinone. CoQ 10 supplementation for one week but not for 24 hours doubled ATP levels and ATP/ADP ratio in CoQ 10 deficient fibroblasts therein normalizing the bioenergetics status of the cells. Other compounds did not affect cellular bioenergetics. In COQ2 mutant fibroblasts, increased superoxide anion production and oxidative stress-induced cell death were normalized by all supplements. Conclusions/SignificanceThese results indicate that: 1) pharmacokinetics of CoQ 10 in reaching the mitochondrial respiratory chain is delayed; 2) short-tail ubiquinone analogs cannot replace CoQ 10 in the mitochondrial respiratory chain under conditions of CoQ 10 deficiency; and 3) oxidative stress and cell death can be counteracted by administration of lipophilic or hydrophilic antioxidants. The results of our in vitro experiments suggest that primary CoQ 10 deficiencies should be treated with CoQ 10 supplementation but not with short-tail ubiquinone analogs, such as idebenone or CoQ 2. Complementary administration of antioxidants with high bioavailability should be considered if oxidative stress is present. 相似文献
16.
AbstractEndogenous coenzyme Q 10 (CoQ 10) is a lipid-soluble antioxidant and essential for the electron transport chain. We previously demonstrated that hydrogen peroxide enhanced CoQ 10 levels, whereas disruption of mitochondrial membrane potential by a chemical uncoupler suppressed CoQ 10 levels, in human 143B cells. In this study, we investigated how CoQ 10 levels and expression of two PDSS and eight COQ genes were affected by oligomycin, which inhibited ATP synthesis at Complex V without uncoupling the mitochondria. We confirmed that oligomycin increased the production of reactive oxygen species (ROS) and decreased mitochondria-dependent ATP production in 143B cells. We also demonstrated that CoQ 10 levels were decreased by oligomycin after 42 or 48 h of treatment, but not at earlier time points. Expression of PDSS2 and COQ2–COQ9 were up-regulated after 18-hour oligomycin treatment, and the expression of PPARGC1A ( PGC1-1α) elevated concurrently. Knockdown of PPARGC1A down-regulated the basal mRNA levels of PDSS2 and five COQ genes and suppressed the induction of COQ8 and COQ9 genes by oligomycin, but did not affect CoQ 10 levels under these conditions. N-acetylcysteine suppressed the augmentation of ROS levels and the enhanced expression of COQ2, COQ4, COQ7, and COQ9 induced by oligomycin, but did not modulate the changes in CoQ 10 levels. These results suggested that the condition of mitochondrial dysfunction induced by oligomycin decreased CoQ 10 levels independent of oxidative stress. Up-regulation of PDSS2 and several COQ genes by oligomycin might be regulated by multiple mechanisms, including the signaling pathways mediated by PGC-1α and ROS, but it would not restore CoQ 10 levels. 相似文献
17.
Coenzyme Q 10 (CoQ 10) is a blockbuster nutraceutical molecule which is often used as an oral supplement in the supportive therapy for cardiovascular diseases, cancer and neurodegenerative diseases. It is commercially produced by fermentation process, hence constructing the high yielding CoQ 10 producing strains is a pre-requisite for cost effective production. Paracoccus denitrificans ATCC 19367, a biochemically versatile organism was selected to carry out the studies on CoQ 10 yield improvement. The wild type strain was subjected to iterative rounds of mutagenesis using gamma rays and NTG, followed by selection on various inhibitors like CoQ 10 structural analogues and antibiotics. The screening of mutants were carried out using cane molasses based optimized medium with feeding strategies at shake flask level. In the course of study, the mutant P-87 having marked resistance to gentamicin showed 1.25-fold improvements in specific CoQ 10 content which was highest among all tested mutant strains. P-87 was phenotypically differentiated from the wild type strain on the basis of carbohydrate assimilation and FAME profile. Molecular differentiation technique based on AFLP profile showed intra specific polymorphism between wild type strain and P-87. This study demonstrated the beneficial outcome of induced mutations leading to gentamicin resistance for improvement of CoQ 10 production in P. denitrificans mutant strain P-87. To investigate the cause of gentamicin resistance, rpIF gene from P-87 and wild type was sequenced. No mutations were detected on the rpIF partial sequence of P-87; hence gentamicin resistance in P-87 could not be conferred with rpIF gene. However, detecting the mutations responsible for gentamicin resistance in P-87 and correlating its role in CoQ 10 overproduction is essential. Although only 1.25-fold improvement in specific CoQ 10 content was achieved through mutant P-87, this mutant showed very interesting characteristic, differentiating it from its wild type parent strain P. denitrificans ATCC 19367, which are presented in this paper. Electronic supplementary materialThe online version of this article (doi:10.1007/s12088-014-0506-4) contains supplementary material, which is available to authorized users. 相似文献
18.
The cell growth and CoQ 10 (coenzyme Q 10) formation of Rhizobium radiobacter WSH2601 were investigated in a 7-1 bioreactor under different dissolved oxygen (DO) concentrations. A maximal CoQ 10 content (C/B) of 1.91 mg/g dry cell weight (DCW) and CoQ 10 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 CoQ 10 production, a new DO-stat feeding strategy was proposed, which significantly improved cell growth and CoQ 10 formation. With this strategy, the maximal CoQ 10 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. 相似文献
20.
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-Q 10 (CoQ 10) is a promising enhancer candidate. CoQ 10 is well known as a strong antioxidant in the respiratory chain and is used for healthcare and other applications. Because
CoQ 10 is negligibly water soluble, most studies are limited by low concentrations. We added CoQ 10 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 CoQ 10 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. 相似文献
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