辅酶Q10产生菌的抗性筛选及发酵条件优化

以根癌土壤杆菌(Agrobacterium tumefaciens)WSHAT12为出发菌株,通过硫酸二乙酯诱变,获得遗传稳定性好的抗L-乙硫氨酸(Eth)突变株WSH-E01,通过进一步的诱变处理,获得L-乙硫氨酸和维生素K3(VK3)双抗性突变株WSH-V01,以双抗性突变株WSH-V01为出发菌株,再进行诱变处理,获得一株X-gal利用能力提高的突变株WSH-X01,与出发菌株WSHAT12相比,突变株WSH-X01的辅酶Q10产量提高幅度达50.6%,同时,对突变株WSH-E01的发酵条件进行优化。出发菌株WSHAT12、突变株WSH-E01、WSH-V01和WSH-X01在优化后的发酵条件下辅酶Q10产量分别达到23.1mg/L、26.8mg/L、29.5mg/L和34.8mg/L。     

微波结合紫外诱变选育辅酶Q_(10)高产菌株

以根瘤土壤杆菌LNUB335为出发菌株,以维生素K3和NaN3双抗性为筛选标记,在根瘤土壤杆菌中首次利用紫外线及微波联合诱变处理,获得1株生产性能比LNUB335显著提高的突变株ARN007,其CoQ10产量为12.01mg/L,较出发菌株提高68.67%,每克干细胞含CoQ102.46mg,较出发菌株提高38.20%。通过传代实验证明该突变株的遗传性稳定,可作为进一步研究的实验菌株。     

发酵生产辅酶Q10高产菌株的选育

以实验室保存的类球红细菌(Rhodobacter sphaeroides)JDW61为出发菌株,考察了紫外、紫外结合氯化锂和亚硝基胍对菌株产生辅酶Q10能力的诱变效应,并结合辅酶Q10的合成途径设计了快速筛选辅酶Q10高产菌株的模型,获得一株辅酶Q10产量提高的突变株CP222,该菌株摇瓶发酵的辅酶Q10产量为276.14mg·L-1,较出发菌株提高了190%,并且遗传性能稳定。     

微生物发酵生产辅酶Q10的研究进展

利用微生物发酵生产辅酶Q1 0 产物活性好 ,并可通过规模放大提高生产能力 ,因而颇受国内外学者的关注。文章综述菌种的选择和遗传改造 ,发酵条件的优化及其分离纯化     

微生物法高产辅酶Q10的研究进展

辅酶Q10是呼吸链上的一种电子传递体,具有抗氧化功能.微生物法生产辅酶Q10具有产物活性高、原料成本低并可以通过规模放大提高生产能力等优点.综述了微生物法生产辅酶Q10的生产菌种,以及能够提高辅酶Q10产量的各种不同的方法策略.     

半导体激光和紫外线对辅酶Q10产生菌的复合诱变效应

采用紫外线、半导体激光及紫外线与半导体激光复合作用的方法,诱变产辅酶Q10红酵母菌SY-3,以提高辅酶Q10的产量。结果表明,紫外线和半导体激光单独作用,诱变效果不佳,而二者的复合作用却能产生很好的效果。用紫外照射120 s再经半导体激光辐射8 min,得到一株叠氮钠和维生素K3双抗性突变株,产辅酶Q10的量达到157.7 mg/L,比原始菌株提高了88.1%,并具有良好的遗传稳定性。     

辅酶Q10和维生素E配伍的稳定性和食用安全性研究

目的：研究辅酶Q10和维生素E配伍的稳定性和食用安全性。方法：采用高效液相色谱法,对经过加速破坏的辅酶Q10和维生素E配伍的样品进行含量分析和安全性毒理学评价。结果：辅酶Q10和维生素E在24个月内未发生明显化学反应,辅料也未对其稳定性产生影响,毒理学试验未发现安全性问题。结论：辅酶Q10和维生素E配伍稳定,并且食用安全。     

辅酶Q10的生理作用及临床应用

辅酶Q10是线粒体电子传递链中的一种重要辅酶,参与细胞氧化磷酸化及ATP生成过程。辅酶Q10是细胞代谢呼吸激活剂和免疫增强剂,具有抗氧化和自由基清除功能。辅酶Q10药物的临床应用主要在心血管疾病、高血压、神经系统疾病和免疫系统疾病方面。     

代谢工程改造大肠杆菌生产辅酶Q10

辅酶Q10(CoQ10)是一种脂溶性抗氧化剂,具有提高人体免疫力、延缓衰老和增强人体活力等功能,广泛应用于制药行业和化妆品行业。微生物发酵法能可持续性生产辅酶Q10,具有越来越多的商业价值。本研究首先将来自类球红细菌的十聚异戊二烯焦磷酸合成酶基因(dps)整合到大肠杆菌ATCC 8739染色体上,敲除内源的八聚异戊二烯焦磷酸合成酶基因(ispB),使内源的辅酶Q8合成途径被辅酶Q10合成途径取代,得到稳定生产辅酶Q10的菌株GD-14,其辅酶Q10产量达0.68 mg/L,单位细胞含量达0.54 mg/g DCW。随后用多个固定强度调控元件在染色体上对MEP途径的关键基因dxs和idi基因以及ubiCA基因进行组合调控,将辅酶Q10单位细胞含量提高2.46倍(从0.54到1.87 mg/g)。进一步引入运动发酵单胞菌Zymomonas mobilis的Glf转运蛋白代替自身的磷酸烯醇式丙酮酸:碳水化合物磷酸转移酶系统(PTS),使辅酶Q10产量进一步提高16%。最后,对高产菌株GD-51进行分批补料发酵,辅酶Q10产量达433 mg/L,单位细胞含量达11.7 mg/g DCW。这是目前为止文献报道的大肠杆菌产辅酶Q10最高菌株。     

辅酶Q10的生物合成及其高表达研究进展

辅酶Q10(CoQ10)不仅是呼吸链上的电子传递体,同时也具有抗氧化功能。目前全球市场上的CoQ10正处于一种供不应求的状态。我们简要论述了CoQ10的结构、性质、功能及其生物合成过程,同时概括总结了现阶段为提高CoQ10产量而采用的新型技术手段。     

提高微生物发酵辅酶Q10产量的研究进展

辅酶Q10具有很高的保健和临床应用价值,开发潜力巨大。主要从菌种筛选、发酵条件优化以及提取方法改进三方面介绍了如何提高微生物发酵辅酶Q10产量的一些研究进展。     

Therapeutic effects of coenzyme Q10 (CoQ10) and reduced CoQ10 in the MPTP model of Parkinsonism

Coenzyme Q₁₀ (CoQ₁₀) is a promising agent for neuroprotection in neurodegenerative diseases. We tested the effects of various doses of two formulations of CoQ₁₀ in food and found that administration in the diet resulted in significant protection against loss of dopamine (DA), which was accompanied by a marked increase in plasma concentrations of CoQ₁₀. We further investigated the neuroprotective effects of CoQ₁₀, reduced CoQ₁₀ (ubiquinol), and CoQ₁₀ emulsions in the (MPTP) model of Parkinson's disease (PD). We found neuroprotection against MPTP induced loss of DA using both CoQ₁₀, and reduced CoQ₁₀, which produced the largest increases in plasma concentrations. Lastly, we administered CoQ₁₀ in the diet to test its effects in a chronic MPTP model induced by administration of MPTP by Alzet pump for 1 month. We found neuroprotective effects against DA depletion, loss of tyrosine hydroxylase neurons and induction of alpha-synuclein inclusions in the substantia nigra pars compacta. The finding that CoQ₁₀ is effective in a chronic dosing model of MPTP toxicity, is of particular interest, as this may be more relevant to PD. These results provide further evidence that administration of CoQ₁₀ is a promising therapeutic strategy for the treatment of PD.     

Response surface methodology for optimization and characterization of limonene-based coenzyme Q10 self-nanoemulsified capsule dosage form

The aim of this study was to systematically obtain a model of factors that would yield an optimized self-nanoemulsified capsule dosage form (SNCDF) of a highly lipophilic model compound, Coenzyme Q10 (CoQ). Independent variables such as amount of R-(+)-limonene (X ₁), surfactant (X ₂), and cosurfactant (X ₃), were optimized using a 3-factor, 3-level Box-Behnken statistical design. The dependent variables selected were cumulative percentage of drug released after 5 minutes (Y ₁) with constraints on drug release in 15 minutes (Y ₂), turbidity (Y ₃), particle size (Y ₄), and zeta potential (Y ₅). A mathematical relationship obtained,Y ₁=78.503+6.058X ₁ +13.738X ₂+5.986X ₃−25.831X ₁ ² +9.12X ₁X₂−26.03X ₁X₃−38.67X ₂ ² +11.02X ₂X₃−15.55X ₃ ³ (r ²=0.97), explained the main and quadratic effects, and the interaction of factors that affected the drug release. Response surface methodology (RSM) predicted the levels of factorsX ₁,X ₂, andX ₃ (0.0344, 0.216, and 0.240, respectively), for a maximized response ofY ₁ with constraints of >90% release onY ₂. The observed and predicted values ofY ₁ were in close agreement. In conclusion, the Box-Behnken experimental design allowed us to obtain SNCDF with rapid (>90%) drug release within 5 minutes with desirable properties of low turbidity and particle size.     

Quantitative determination of coenzyme Q10 in human blood for clinical studies

A quantitative method for the determination of coenzyme Q10 (CoQ10) in human blood has been devised which allows recovery of essentially 100% of the CoQ10. The use of whole blood rather than plasma includes the CoQ10 in white cells. The method utilizes TLC instead of saponification to fractionate lipid impurities, because CoQ10 is sensitive to saponification, and utilizes CoQ11 as an internal standard which is advantageous over CoQ9 and a synthetic quinone. The final step of HPLC frequently reveals a peak with a retention time like that of CoQ9 which, being less than that of CoQ10, can be near other peaks of impurities.     

Mitochondrial membrane depolarization and the selective death of dopaminergic neurons by rotenone: protective effect of coenzyme Q10

Chronic exposure to the pesticide rotenone induces a selective degeneration of nigrostriatal dopaminergic neurons and reproduces the features of Parkinson's disease in experimental animals. This action is thought to be relevant to its inhibition of the mitochondrial complex I, but the precise mechanism of this suppression in selective neuronal death is still elusive. Here we investigate the mechanism of dopaminergic neuronal death mediated by rotenone in primary rat mesencephalic neurons. Low concentrations of rotenone (5-10 nM) induce the selective death of dopaminergic neurons without significant toxic effects on other mesencephalic cells. This cell death was coincident with apoptotic events including capsase-3 activation, DNA fragmentation, and mitochondrial membrane depolarization. Pretreatment with coenzyme Q10, the electron transporter in the mitochondrial respiratory chain, remarkably reduced apoptosis as well as the mitochondrial depolarization induced by rotenone, but other free radical scavengers such as N-acetylcysteine, glutathione, and vitamin C did not. Furthermore, the selective neurotoxicity of rotenone was mimicked by the mitochondrial protonophore carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP), a cyanide analog that effectively collapses a mitochondrial membrane potential. These data suggest that mitochondrial depolarization may play a crucial role in rotenone-induced selective apoptosis in rat primary dopaminergic neurons.     

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.     

快速提取类球红细菌中辅酶Q10的方法研究

目的：建立一种从类球红细菌中快速分离纯化辅酶Q10的方法。方法：对影响超声提取辅酶Q10的各因素,包括提取试剂、超声频率、循环次数及工作时间的最佳条件进行正交试验,比较超声破碎法与碱醇皂化法提取辅酶Q10的差异。结果：在超声提取中,提取试剂和循环次数对辅酶Q10提取效果具有显著性影响;在超声频率0.5s、丙酮提取3min、循环3次的条件下提取的辅酶Q10的含量比碱醇皂化法提高了近6倍。结论：超声破碎法是一种简单、迅速、高效的提取辅酶Q10方法。