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.     

酵母发酵生产辅酶Q10提取方法研究进展

辅酶Q10是存在于哺乳动物中,能与酶蛋白形成复合物以发挥酶学活性的有机小分子化合物,目前广泛应用于医药、日化、保健、食品等不同领域。辅酶Q10来源丰富,其中酵母是其工业生产的主要来源之一。广受关注的酵母发酵生产辅酶Q10的提取分离手段不断革新,产量不断增加,处理方式更加环保,应用日渐拓宽。本文就近年来国内外酵母发酵生产辅酶Q10的提取分离方法进行了综述,包括酵母菌种的类型与优化、辅酶Q10提取检测方法、工业生产放大工艺以及与产品质量相关的各个影响因素,并对酵母发酵生产辅酶Q10的前景进行了展望。     

基因改造Rhodobacter sphaeroides提高

[目的]通过敲除类球红细菌2.4.1基因组中八氢番茄素合成酶基因crtB,让异戊二烯前体更多流向辅酶Q10的合成.引入大肠杆菌编码的分支酸裂解酶基因ubiC和4-羟苯甲酸转移酶基因ubiA,提高4-羟苯甲酸的合成和与聚异戊二烯的连接,从而提高类球红细菌的辅酶Q10产量.[方法]以自杀型质粒pSUP202为载体,构建包含crtB基因上游2.5 kb片段,壮观霉素抗性基因,ubiC、ubiA基因和crtB基因下游2.5 kb片段的基因置换质粒,利用结合转移方法转入类球红细菌2.4.1中,利用抗性机制筛选双交换突变株,RT-PCR方法检测引入的ubiC和ubiA基因转录.用HPLC方法测定出发菌株和基因改造菌株的辅酶Q10产量.[结果]成功构建出基因置换质粒,筛选出发生基因置换的突变株,RT-PCR证实了外源基因的转录,并且突变株辅酶Q10的产量比出发菌株提高40％.[结论]大肠杆菌的ubiC和ubiA基因能够利用自身启动子在类球红细菌中表达,利用基因改造的方法能成功提高类球红细菌的辅酶Q10产量.     

The thermotropic properties of coenzyme Q10 and its lower homologues

The thermotropic properties of coenzymes Q₁₀, Q₉, Q₈, and Q₇ have been examined by differential scanning calorimetry and wide-angle X-ray diffraction. Typical scanning calorimetry cooling curves of coenzyme Q from the liquid state exhibit a single exothermic phase transition into a crystalline state at a temperature that decreases as the length of the polyisoprenoid side-chain substituent decreases. Upon subsequent heating, the molecules undergo a series of thermal events which precede the main crystalline-to-liquid endothermic phase transition. The temperature of these transitions increases with increasing chain length. The crystallization phase transition temperature depends markedly on the rate at which the sample is cooled and increases with decreasing scan rate; the temperature of the melting endotherm is not markedly affected by the scan rate. Detailed calorimetric studies of coenzyme Q₁₀ indicate that two crystalline states are formed, one at relatively high cooling rates to low temperatures and the other when preparations are cooled slowly from the liquid state to relatively high temperatures. Heating the crystalline phase formed by rapid cooling causes its transformation into the phase observed by cooling slowly. X-ray diffraction analysis confirmed the existence of these two crystal phases in coenzymes Q₉ and Q₁₀ and the transformation from the rapidly crystallized form to the more ordered form associated with slower cooling rates. At body temperature (310 K) under equilibrium conditions coenzyme Q₁₀ exists in an ordered crystalline phase; the implications of the thermotropic behavior of coenzyme Q₁₀ on mitochondrial functionin vitro andin vivo are discussed.     

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.