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近年来,分子生物学方法已成为寻找人类疾病新病原微生物的有效方法,使一些不能正常培养的致病因子得以鉴定。利用代表性差异分析(RDA),基于共有序列的聚合酶链反应(PCR),简称共有序列PCR和cDNA文库的筛选已成功地发现了卡波济肉瘤(KS),惠普尔病,汉坦病毒肺型综合征,非甲非乙型肝炎等疾病的病原体。 相似文献
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研究了蔗糖、KH2PO4、NH4NO3比对烟草细胞生长和CoQ10量的影响,结果表明,当蔗糖浓度为30g/L时,CoQ10总量最高,此时细胞产量、CoQ10含量和总量分别为19.8g/L、414.7μg/g(dwt)、8212μg/L。在上述蔗糖浓度下,当KH2PO4起始浓度为340mg/L、NH4NO3为12时,细胞产量、CoQ10含量和总量分别最高,高于此比例时有利于CoQ10形成
,但不利于细胞生长。 相似文献
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莽草酸生物合成途径的调控 总被引:3,自引:0,他引:3
莽草酸是合成生物碱等许多其它物质的前体.近年来,莽草酸作为临床上惟一有效的抗禽流感药物--达啡的原料而倍受关注.简述了莽草酸生物合成的途径,着重从基因工程角度分析了莽草酸合成过程中的关键酶及其对莽草酸产量的影响,旨在为研究和开发微生物工程菌生产莽草酸提供参考. 相似文献
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透明质酸是由N-乙酰氨基葡萄糖和葡萄糖醛酸组成的双糖单位聚合而成的直链酸性黏多糖,已被广泛应用于药物、化妆品和食品添加剂。微生物发酵法是目前生产透明质酸最有效的方法。生物体内透明质酸的合成途径基本一致,均为Leloir途径。透明质酸合成操纵子由透明质酸合酶基因、尿苷二磷酸葡萄糖脱氢酶基因和尿苷二磷酸葡萄糖焦磷酸化酶基因组成,其表达受Cov S/CovR和Lux S等多种调控系统调控。随着分子生物学技术的迅速发展以及对透明质酸合成相关基因了解的不断深入,人们从提高透明质酸安全性、提高透明质酸产量和调控透明质酸分子质量三个方面出发,通过基因工程手段构建出了高产、安全、一定分子质量范围的透明质酸生产菌株。就有关透明质酸生物合成途径、合成相关基因表达调控及生产菌株分子生物学改造的策略与研究进展进行综述和展望。 相似文献
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21世纪,世界上的各类科学技术和专业学科都有了长足的发展和进步,比如说生物化学学科、免疫学学科等,其中作为代表性研究项目的分子生物学得到了越来越多人的关注,随着现代化设施的更新和计算机技术的发展,分子生物学的相关研究成果被大量运用在人们生活中,促使了社会的长久进步。 相似文献
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沼泽红假单胞菌J001产辅酶Q10摇瓶发酵条件优化 总被引:1,自引:0,他引:1
本试验对沼泽红假单胞菌J001菌株250ml摇瓶发酵辅酶Q10条件进行了优化。结果表明其最佳初始pH值为6.5-7.5,温度为28-31℃,摇床转速100 r min-1,摇瓶装液量为200ml,接种量达10%时可直接进入对数生长期;碳源以NaAc较好,氮源以(NH4)2SO4和NH4Cl较好,磷源用量对考察指标影响不明显;用中心组合设计响应曲面法对碳氮源用量进行了优化,当NaAc浓度为5.39(g l-1),(NH4)2SO4浓度为0.385(g l-1)即碳氮比为14/1时,对菌胞生长最有利;当NaAc浓度为5.70(g l-1),(NH4)2SO4浓度为0.365(g l-1)即碳氮比为15.6/1时,对辅酶Q10产量最有利。 相似文献
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辅酶Q10产生菌的抗性筛选及发酵条件优化 总被引:1,自引:0,他引:1
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。 相似文献
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Hesham N. Mustafa Gehan A. Hegazy Sally A. El Awdan Marawan AbdelBaset 《Tissue & cell》2017,49(3):410-426
Doxorubicin (DOX) is a chemotherapeutic agent used for treatment of different cancers and its clinical usage is hindered by the oxidative injury-related cardiotoxicity. This work aims to declare if the harmful effects of DOX on heart can be alleviated with the use of Coenzyme Q10 (CoQ10) or L-carnitine. The study was performed on seventy two female Wistar albino rats divided into six groups, 12 animals each: Control group; DOX group (10 mg/kg); CoQ10 group (200 mg/kg); L-carnitine group (100 mg/kg); DOX + CoQ10 group; DOX + L-carnitine group. CoQ10 and L-carnitine treatment orally started 5 days before a single dose of 10 mg/kg DOX that injected intraperitoneally (IP) then the treatment continued for 10 days. At the end of the study, serum biochemical parameters of cardiac damage, oxidative stress indices, and histopathological changes were investigated. CoQ10 or L-carnitine showed a noticeable effects in improving cardiac functions evidenced reducing serum enzymes as serum interleukin-1 beta (IL-1 β), tumor necrosis factor alpha (TNF-α), leptin, lactate dehydrogenase (LDH), Cardiotrophin-1, Troponin-I and Troponin-T. Also, alleviate oxidative stress, decrease of cardiac Malondialdehyde (MDA), Nitric oxide (NO) and restoring cardiac reduced glutathione levels to normal levels. Both corrected the cardiac alterations histologically and ultrastructurally. With a visible improvements in α-SMA, vimentin and eNOS immunohistochemical markers. CoQ10 or L-carnitine supplementation improves the functional and structural integrity of the myocardium. 相似文献
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Cluis CP Ekins A Narcross L Jiang H Gold ND Burja AM Martin VJ 《Metabolic engineering》2011,13(6):733-744
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. 相似文献
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Cleren C Yang L Lorenzo B Calingasan NY Schomer A Sireci A Wille EJ Beal MF 《Journal of neurochemistry》2008,104(6):1613-1621
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. 相似文献
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Physical and chemical agents were employed in our laboratories to induce mutation in a variety of microorganisms used for
production of antibioties or enzymes. Improved production strains ofPenicillium chrysogenum (penicillin-producer),Streptomyces griseus (streptomycin-producer),Streptomyces nodosus. (amphotericin B-producer),Streptomyces noursei (nystatin-producer),Streptomyces umbrinus (diumycin-producer),Streptomyces prasinus (prasinomycin-producer),Streptomyces roseochromogenes (steroid-16α-hydroxylase-producer), andArthrobacter simplex (steroid-1-dehydrogenase-producer) were developed by use of mutation selection techniques.
The methods found to be most successful with each species are described. The genealogical relationships within species of
a number of strains ofPenicillium chrysogenum, Streptomyces prasinus, andStreptomyces roseochromogenes are presented. 相似文献
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Hyaluronic acid (HA) production in Streptococcus zooepidemicus competes for the carbon source along with biomass formation, lactate formation (via glycolysis) and pentose phosphate pathway (PPP). In our studies, increase in HA molecular weight was observed by redirecting the carbon flux towards HA biosynthesis pathway by partially inhibiting the glycolytic pathway. Batch bioreactor (1.2 L) studies showed that with the addition of 25 μM sodium iodoacetate, 5 g/L tryptophan and 10 g/L pyruvate, which are glycolytic inhibitors, HA molecular weight increased to 3.2, 3.2 and 3.1 MDa respectively compared to control run (2.4 MDa). Yield coefficients YHA/S and YLA/S showed inverse relationship, indicating competition for glucose between HA and lactic acid formation. Addition of 5 g/L glutamine along with 25 μM sodium iodoacetate also increased the HA concentration to 5.0 g/L from 2.0 g/L in control run. Metabolic flux analysis studies show that concentration and molecular weight of HA is increased by decreasing carbon flux towards glycolysis and PPP and increasing carbon flux towards HA precursor formation. It was observed that specific growth rate of the cells correlated positively to the specific HA production rate and negatively to the molecular weight of HA produced. Addition of antioxidant tannic acid also increased molecular weight to 3.0 MDa. 相似文献
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Shemiakin IG Stepanshina VN Manzeniuk OIu Anisimova VA Panfertsev EA Korobova OV Medvedeva IM Dorozhkova IR 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》2000,(2):6-11
The expediency of using molecular biological methods for the evaluation of M. tuberculosis clinical strains by individual genetic certification of circulating M. tuberculosis strains has been substantiated. Considerable genetic heterogeneity of M. tuberculosis strains isolated from patients in different regions of the Russian Federation has been established; this heterogeneity is due to the presence of differences in the number of copies (5-26) of element IS6110 in M. tuberculosis cells. 相似文献
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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. 相似文献
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高产辅酶Q10结构类似物抗性突变株的选育 总被引:1,自引:0,他引:1
以土壤杆菌(Agrobacteriumsp.)TLY-4为出发菌株,采用70%致死剂量的NTG进行诱变处理,通过筛选抗辅酶Q10结构类似物维生素K3突变株,定向选育到了两株辅酶Q10高产突变株,编号为R-122和R-015,其摇瓶发酵72h时的辅酶Q10产量分别为57.3 mg/L和59.9 mg/L,较出发菌株提高了35.7%和41.6%。通过连续传代实验,表明突变株高产辅酶Q10的遗传性状稳定。实验以有机溶剂DMF和吐温-80共同增溶的方法,解决了维生素K3在培养基中易析出的问题,并确定了平板培养基中维生素K3的最小抑菌浓度为0.15 mg/mL。 相似文献