天然活性先导化合物生物转化研究进展

天然活性先导化合物生物转化是利用生物催化剂(如：酶、微生物、动植物细胞)将加入到生物反应系统中的天然活性先导化合物进行特异性的分子结构修饰以获得高效、低毒新化合物的方法。该方法可以有效地提高已知的天然活性先导化合物的活性、降低毒副作用、改善水溶性和生物利用度,也可以用来生产具有重要应用价值的微量天然活性先导化合物,同时可用于药物代谢机制的研究。国内外学者已经针对甾体、醌类、黄酮类、萜类等化合物开展了天然活性先导化合物生物转化研究,筛选出一批有重要应用价值的生物转化反应类型,但针对天然活性先导化合物生物转化的机制、生物转化过程工程以及生物转化产物活性等方面的研究较少。将现代生命科学技术(如：生物催化剂的定向改造、高通量筛选、组合生物转化、非水相生物转化)引入天然活性先导化合物生物转化研究中,必将推进天然活性先导化合物的快速发展。     

甾体C11β—羟基生物转化菌株新月弯孢霉的筛选和鉴定

文章对甾体C11β－羟某化生物转化菌株－新月弯孢霉的生长和生物转化特征进行了研究。经过反复筛选,最终选出了一株氧化可的松转化率达30％的最优菌株。文中描述了新月弯孢霉菌体生长过程中,菌体干重与PH的变化情况,并通过正交实验确定了该菌株生长的最适培养基配方。经过研究,对新月弯孢霉菌株的生长和生物转化特性及其进行甾体C11β－羟基化生物转化过程有了初步的了解。     

二氧化碳生物转化制甲烷技术研究进展

二氧化碳减量化与转化是当前业界关注及着手解决的重要问题,将二氧化碳作为资源转化为甲烷,有利于环境与社会的可持续发展。本文在分析二氧化碳转化为甲烷技术的基础上,重点介绍了国内外二氧化碳生物转化的研究与进展;总结了二氧化碳生物转化途径及其影响因素,分析了氢营养型、甲基营养型生物转化甲烷机理和生物转化能量来源;探讨了不同产甲烷菌微生物电合成产甲烷和氢气研究进展,总结了微生物电合成法、光合作用法和厌氧消化法等二氧化碳生物转化技术在反应器设计、电极材料选择、工艺条件优化及试验结果评估等方面取得的进展及存在的问题。重点就微生物电合成法的未来研究提出了增强微生物活性、提升氢气利用率、加快高效电极开发、提高能量效率、加强工业废气试验研究和强化光能转化等研究重点和发展方向,同时加强计算机模拟等交叉学科协同创新是促进二氧化碳生物转化技术进步的新方向。     

纤维素制酒精的分散、耦合、并行系统

生物转化是纤维素制酒精的有效途径。以强化纤维素酒精生物转化过程,降低转化成本为目的,在对该过程深入分析的基础上,提出了纤维素制酒精的分散、耦合、并行系统并阐述了其设计原则与解决方案,有普遍意义。     

甾类化合物微生物转化与分解代谢机制研究进展

甾类化合物具有重要的生理医药作用,市场需求巨大。甾类化合物及其关键甾类药物通过微生物转化制备工艺较化学合成法具有区域立体选择性、减少合成步骤、缩短生产周期、提高收率以及生态友好等优点逐步被应用,然而甾类物质微生物分解代谢机制还有待进一步深入探索研究并确定。本文从甾类化合物结构种类与主要来源、生理功能、微生物转化与分解代谢机制的研究等方面进行了归纳,着重解析甾类化合物分解代谢过程关键酶系及其分子作用机制,为甾药化合物生产菌种改造与工程菌构建,以及微生物转化工业化生产工艺的开发提供参考。     

有机硅橡胶固定化细胞进行的生物转化

有机硅橡胶固定化细胞进行的生物转化潘冰峰,戴学倩,冯青,李祖义（中国科学院上海有机化学研究所,２０００３２）关键词硅橡胶;白地霉;固定化细胞;生物转化近年来,有机化学领域的一个重要进展是用酶或微生物进行生物转化反应。其中研究和应用较多的是碳基还原,特...     

两种光合细菌生物转化槲寄生培养液菌体中几种同工酶的变化

采用聚丙烯酰胺凝胶电泳法对两种光合细菌的生物转化槲寄生培养液中菌体的蛋白质和几种同工酶进行研究,并以纯光合细菌培养液中菌体作对照。结果表明,光合细菌生物转化槲寄生过程中,两种光合细菌的蛋白质、酯酶同工酶和过氧化物酶同工酶均发生改变,某些蛋白质、酯酶和过氧化物酶的合成受到抑制,并有新的蛋白质、酯酶和过氧化物酶生成;超氧化物歧化酶的表达未明显改变。由此可见,槲寄生能诱导光合细菌合成新的酯酶和过氧化物酶,这些诱导酶可能参与了槲寄生的生物转化。为光合细菌生物转化槲寄生转化机理的研究及槲寄生在抗肿瘤领域的进一步应用奠定了基础。     

工业生物技术的过程科学研究概况及发展趋势

简述了工业生物技术领域在过程科学方面的研究现状及发展趋势,从"细胞群体效应及过程放大原理"、"工业生物过程物质和能量传递与生物转化规律"、"工业生物过程优化新方法"等3个方面介绍了我国工业生物技术过程科学的重点进展和在国际上的地位,最后从"生物原料高效转化"、"生物转化过程物质和能量协调和匹配"、"生物过程强化"、"生物过程系统集成"等方面提出了工业生物技术在过程科学研究方面未来的战略方向。     

生物催化与生物转化研究进展

由于生物催化过程具有高效、高选择性、条件温和、环境友好等优点,因此成为可持续发展过程中替代和拓展传统有机化学合成的重要方法。近两年的进展集中于新生物催化剂的发现和改造,以及将生物催化和生物转化应用于工业过程的探索,包括开发新的反应体系,新的固定化方法等。可以预见,在医药中间体等高附加值化工产品的生产过程中,生物催化和生物转化的应用将呈现加速增长趋势。     

生物转化中药化学成分的研究进展

生物转化是实现中药现代化的重要途径之一,阐明了生物转化中药化学成分的研究内容和应用范围,讨论了目前存在的问题和解决办法,并对生物转化中药化学成分的发展趋势进行了展望。     

甘油歧化为1,3-丙二醇的代谢及关键酶研究进展

微生物发酵生产1,3-丙二醇因对环境友好而成为研究热点。通过对发酵菌种、代谢途径、调节子和关键酶的分析,阐述了微生物转化甘油为1,3-丙二醇的分子机理。尤其对还原途径的限速酶-甘油脱水酶的分子结构及再激活因子进行了详细分析,为菌种的遗传改造提供了理论依据。     

甘油歧化为1,3-丙二醇的代谢及关键酶研究进展

微生物发酵生产1,3-丙二醇因对环境友好而成为研究热点。通过对发酵菌种、代谢途径、调节子和关键酶的分析,阐述了微生物转化甘油为1,3-丙二醇的分子机理。尤其对还原途径的限速酶-甘油脱水酶的分子结构及再激活因子进行了详细分析,为菌种的遗传改造提供了理论依据。     

聚羟基丁酸路径在克雷伯氏菌中的构建

以生物柴油的副产物甘油生产高附加值的1,3-丙二醇,现已成为提升生物柴油产业链经济性的重要途径,而中间代谢产物3-羟基丙醛积累造成细胞死亡,发酵异常终止是生物法生产1,3-丙二醇过程中的关键问题。不同于传统的降低3-羟基丙醛积累的思路,本文从增强克雷伯氏菌对3-羟基丙醛的抗逆性出发,改善克雷伯氏菌1,3-丙二醇的生产性能,首次将聚羟基丁酸路径引入克雷伯氏菌中,构建了新型基因工程菌,并对其1,3-丙二醇发酵性能及聚羟基丁酸代谢进行了初步的研究。经IPTG诱导,工程菌中检测到聚羟基丁酸,其含量随IPTG浓度增加而增大。优化的IPTG浓度为0.5 mmol/L。初始甘油50 g/L时,野生菌可正常发酵生产1,3-丙二醇,1,3-丙二醇浓度达到22.1 g/L,其质量得率为46.4%。当初始甘油达到70 g/L时,由于高浓度3-HPA积累,野生菌发酵终止,而工程菌可正常发酵生产1,3-丙二醇,PDO产量可达31.3 g/L,其质量得率为43.9%。同时检测到聚羟基丁酸积累。研究结果有助于加深对克雷伯氏菌1,3-丙二醇代谢机理的认识,为克雷伯氏菌的进一步优化提供了新的思路。     

Glycerol and propanediols degradation by Desulfovibrio alcoholovorans in pure culture in the presence of sulfate, or in syntrophic association with Methanospirillum hungatei

Abstract In a mineral medium containing sulfate as terminal electron acceptor, the sulfate-reducing bacterium Desulfovibrio alcoholovorans oxidized stoichiometrically 1 mol glycerol to 1 mol acetate and 1 mol 1,3-propanediol to 1 mol acetate with the concomitant reduction of 0.75 and 1 mol sulfate, respectively; 1 mol 1,2-propanediol was degraded to 0.8 mol acetate and 0.1 mol proprionate, with the reduction of approximately 1 mol sulfate. The maximum specific growth rates (μ_max in h⁻¹) were 0.22, 0.086 and 0.09 with glycerol, 1,3-propanediol and 1,2-propanediol, respectively. The growth yields were 12.7 g, 11.1 g and 7.2 g dry weight/mol 1,3-propanediol, glycerol and 1,2-propanediol degraded, respectively. The growth yields and maximum specific growth rates of the H₂-transferring associations were also calculated. In the absense of sulfate, all these reduced substrates were degraded to acids and methane when D. alcoholovorans was cocultured with Methanospirillum hungatei . Changes in the metabolic pathway were observed in the degradation of 1,2- and 1,3-propanediol. The metabolic efficiency of D. alcoholovorans to degrade glycerol, 1.2- and 1,3-propanediol is discussed.     

甘油转化生产1,3-丙二醇发酵液中甘油含量的测定

对文献介绍滴定法测定甘油的方法进行了改进,使之能够用于1,3－丙二醇发酵液中甘油含量测定。实验表明,化学滴定法测定结果具有较好的准确性和重复性,与酶法和变色酸比色法相比,测定结果接近,化学滴定法测定发酵液中甘油含量是一个较为经济简便的方法。     

Construction of stress-induced metabolic pathway from glucose to 1,3-propanediol in <Emphasis Type="Italic">Escherichia coli</Emphasis>

1,3-Propanediol is an important chemical widely used in polymer production, but its availability is being restricted owing to its expensive synthesis. The aim of this study was to engineer an Escherichia coli strain that can produce 1,3-propanediol directly from glucose. We successfully constructed a stress-induced metabolic pathway from glucose to 1,3-propanediol in recombinant E. coli by the expression of gpd1 and gpp2 genes from Saccharomyces cerevisiae and dha operon from Klebsiella pneumoniae, respectively. Batch cultivation of the recombinant E. coli showed that 12.1 g/L 1,3-propanediol was accumulated in the culture without using any inducer.     

Expression of 1,3-propanediol oxidoreductase and its isoenzyme in Klebsiella pneumoniae for bioconversion of glycerol into 1,3-propanediol

In the Klebsiella pneumoniae reduction pathway for 1,3-propanediol (1,3-PD) synthesis, glycerol is first dehydrated to 3-hydroxypropionaldehyde (3-HPA) and then reduced to 1,3-PD with NADH consumption. Rapid conversion of 3-HPA to 1,3-PD is one of the ways to improve the yield of 1,3-PD from glycerol and to avoid 3-HPA accumulation, which depends on enzyme activity of the reaction and the amount of reducing equivalents available from the oxidative pathway of glycerol. In the present study, the yqhD gene, encoding 3-propanediol oxidoreductase isoenzyme from Escherichia coli and the dhaT gene, encoding 3-propanediol oxidoreductase from K. pneumoniae were expressed individually and co-expressed in K. pneumoniae using the double tac promoter expression plasmid pEtac-dhaT-tac-yqhD. The three resultant recombinant strains (K. pneumoniae/pEtac-yqhD, K. pneumoniae/pEtac-dhaT, and K. pneumoniae/pEtac-dhaT-tac-yqhD) were used for fermentation studies. Experimental results showed that the peak values for 3-HPA production in broth of the three recombinant strains were less than 25% of that of the parent strain. Expression of dhaT reduced formation of by-products (ethanol and lactic acid) and increased molar yield of 1,3-PD slightly, while expression of yqhD did not enhance molar yield of 1,3-PD, but increased ethanol concentration in broth as NADPH participation in transforming 3-HPA to 1,3-PD allowed more cellular NADH to be used to produce ethanol. Co-expression of both genes therefore decreased by-products and increased the molar yield of 1,3-PD by 11.8%, by catalyzing 3-HPA conversion to 1,3-propanediol using two cofactors (NADH and NADPH). These results have important implications for further studies involving use of YqhD and DhaT for bioconversion of glycerol into 1,3-PD.     

Coordination of metabolic pathways: Enhanced carbon conservation in 1,3-propanediol production by coupling with optically pure lactate biosynthesis

Metabolic engineering has emerged as a powerful tool for bioproduction of both fine and bulk chemicals. The natural coordination among different metabolic pathways contributes to the complexity of metabolic modification, which hampers the development of biorefineries. Herein, the coordination between the oxidative and reductive branches of glycerol metabolism was rearranged in Klebsiella oxytoca to improve the 1,3-propanediol production. After deliberating on the product value, carbon conservation, redox balance, biological compatibility and downstream processing, the lactate-producing pathway was chosen for coupling with the 1,3-propanediol-producing pathway. Then, the other pathways of 2,3-butanediol, ethanol, acetate, and succinate were blocked in sequence, leading to improved d-lactate biosynthesis, which as return drove the 1,3-propanediol production. Meanwhile, efficient co-production of 1,3-propanediol and l-lactate was also achieved by replacing ldhD with ldhL from Bacillus coagulans. The engineered strains PDL-5 and PLL co-produced over 70 g/L 1,3-propanediol and over 100 g/L optically pure d-lactate and l-lactate, respectively, with high conversion yields of over 0.95 mol/mol from glycerol.     

Complex metabolic network of 1,3-propanediol transport mechanisms and its system identification via biological robustness

The bioconversion of glycerol to 1,3-propanediol (1,3-PD) by Klebsiella pneumoniae (K. pneumoniae) can be characterized by an intricate metabolic network of interactions among biochemical fluxes, metabolic compounds, key enzymes and genetic regulation. Since there are some uncertain factors in the fermentation, especially the transport mechanisms of 1,3-PD across cell membrane, the metabolic network contains multiple possible metabolic systems. Considering the genetic regulation of dha regulon and inhibition of 3-hydroxypropionaldehyde to the growth of cells, we establish a 14-dimensional nonlinear hybrid dynamical system aiming to determine the most possible metabolic system and the corresponding optimal parameter. The existence, uniqueness and continuity of solutions are discussed. Taking the robustness index of the intracellular substances together as a performance index, a system identification model is proposed, in which 1,395 continuous variables and 90 discrete variables are involved. The identification problem is decomposed into two subproblems and a parallel particle swarm optimization procedure is constructed to solve them. Numerical results show that it is most possible that 1,3-PD passes the cell membrane by active transport coupled with passive diffusion.     

异源表达木糖异构酶基因对克雷伯氏菌合成1,3-丙二醇的影响

【目的】提高克雷伯氏菌胞内还原力以强化1,3-丙二醇合成。【方法】将来源于大肠杆菌的木糖异构酶基因在克雷伯氏菌中异源表达,构建重组菌。研究重组菌添加不同浓度木糖为辅底物与甘油共发酵过程中代谢产物和NADH的变化规律。【结果】与对照菌相比,重组菌细胞内还原力NADH提高了0.1?0.3倍,1,3-丙二醇产量达到23.31 g/L,提高20%,1,3-丙二醇转化率从0.60 mol/mol提高到0.73 mol/mol。【结论】木糖异构酶基因的表达强化了木糖代谢途径,经磷酸戊糖途径积累大量还原力,促进了1,3-丙二醇的生成。