工业微生物在渗透胁迫下的应激反应及保护措施

发酵工业作为生物技术产业的重要组成部分,在我国工业结构中占据了极大比重,而在工业发酵后期菌体代谢物、中和剂以及补料物的累积使微生物受到极大的渗透胁迫,严重影响了细胞生长及目标产物代谢,致使发酵产量与效率偏低。本文主要针对高渗胁迫下微生物的细胞结构、应答途径、基因、蛋白、代谢、分裂机制进行综述与分析,并以微生物菌种特性结合工业发酵技术为改良思路,从菌种改良、外源添加保护剂、改良中和剂、去除渗透抑制因子、膜过滤技术等方面找寻潜在渗透保护措施,以期为发酵行业生产力水平的提升、节能减排降耗提供参考。     

微生物菌种改良筛选新技术研究进展

从自然界筛选出来的微生物菌株往往活性比较低,耐受性比较差,不能满足工业化生产的要求.而采用新颖的诱变技术,如微波、离子注入、原生质体诱变和原生质体融合、基因工程技术等能有效改良菌种,提高菌种的活性,降低生产成本,满足工业化生产的要求.同时,构建一种简单易行的改良路线和快速有效的筛选模式是菌种改良选育的关键.     

加快微生物油脂研究为生物柴油产业提供廉价原料

当前国内外致力于发展生物柴油,因其性能优良,成为化石柴油的替代品。由于以植物油脂生产生物柴油原料成本占总成本的70％-85％,所以亟待开发廉价油脂资源。微生物油脂主要是微生物利用碳水化合物合成的甘油脂,其脂肪酸组成和植物油相近。产油微生物具有资源丰富、油脂含量高、碳源利用谱广等特点,开发潜力大。然而,目前微生物油脂生产成本偏高,研究工作仍以富含多不饱和脂肪酸的高附加值菌油为目标。随着现代分子生物学和生物化工技术的发展,对产油微生物菌种筛选、改良、代谢调控和发酵工程的研究日趋深入,将降低微生物油脂生产成本,为未来生物柴油产业提供廉价原料。     

微生物原生质体融合育种技术及其应用

工业微生物菌种选育在发酵工业中占有重要地位。微生物原生质体融合(microbial protoplast fusion)技术具有重组频率高、受结合型或致育型限制小以及遗传物质传递完整等优点,是微生物育种最常用的方法之一。结合相关研究进展,分析了原生质体融合技术的组成,包括制备、再生、融合的影响因素以及融合子的筛选方法,重点评述了原生质体融合技术应用在微生物育种中的最新进展,以及微生物原生质体融合技术的发展前景。     

基因组改组及其在木质纤维素水解液乙醇发酵菌种选育中的应用展望

能高效代谢木质纤维素水解液中的可发酵糖、同时可耐受/分解发酵抑制剂的菌种, 是利用木质纤维素为原料生产燃料乙醇技术的关键。基因组改组技术是近些年发展起来的一项新型育种技术, 该技术已运用于食品和医药行业菌种的改良。本文综述了基因组改组技术的原理、方法、特点、及其运用, 并对其在木质纤维素水解液乙醇发酵菌种选育方面的应用进行了展望。     

发酵工业菌种的迭代创制

生物发酵是以微生物菌种为生物催化剂,以淀粉糖、生物质等可再生资源为原料发酵生产各种食品、化学品、燃料、材料等物质的生产过程,具有绿色、低碳和可持续等特征。我国拥有全球规模最大的生物发酵产业,尤其氨基酸、维生素等传统发酵产品占全球市场份额的60%–80%。发展生物发酵产业对于我国实现“碳中和、碳达峰”的战略目标和生物经济发展具有重要的意义。微生物工业菌种是生物发酵产业的核心,直接影响原料路线、产品种类和生产成本。创新发酵工业菌种,提升其原料转化利用效率,提高产物生产水平,拓展产品种类,是生物发酵产业高质量发展的关键。近年来,合成生物学、系统生物学等学科的发展,进一步加深了研究者对微生物底盘细胞生理代谢机制的理解,加速了基因编辑等菌种设计创制使能技术的发展,为发酵工业菌种改造提升提供了新动能。本文选取了具有代表性的大宗氨基酸、B族维生素、柠檬酸、燃料乙醇等发酵产业,从其工业微生物底盘的基础研究和技术开发角度,综述发酵工业菌种改造提升的最新进展,并展望人工智能、自动化与生命科学交叉融合将对工业菌种迭代产生的重要影响。     

有关微生物菌种选育技术的探索

所谓微生物菌种选育技术指的是,有机运用以遗传学为代表的相关原理,对目标微生物菌种展开相应筛选及其改造,从而达成所需产物大幅增产的一门技术。鉴于此,本文首先对传统微生物菌种选育技术进行了简要介绍,然后针对基因组改组技术进行了重点讨论和研究,以便同行参考。     

利用紫外诱变选育辅酶q10菌种的研究

微生物育种指的是运用遗传学、诱变、杂交等原理,对负有特定作用的微生物菌种开展筛选、改造,从而得到符合某种需要的技术。本文对微生物菌种选育技术中的紫外诱变来选育辅酶q10高产菌株进行了一系列的试验和研究。。     

蛋白质组学技术在工业发酵梭菌研究中的应用

介绍了目前应用较广泛的蛋白质组学技术的原理、应用及优缺点;总结了发酵工业中常用的梭菌属细菌;重点阐述了蛋白质组学技术在工业发酵梭状芽孢杆菌研究中的应用,为工业发酵菌种的改良和发酵工艺的优化提供理论依据。最后讨论了今后工业发酵菌种蛋白质组学研究的重点和方向。     

混菌发酵对白酒液态发酵效率和风味物质的影响

<正>我国白酒发酵属于典型的自然发酵过程,其特点是在开放的生产环境中,多种不同微生物共同发酵,相互作用,最终形成具有独特风格的白酒。因此,认识微生物群体发酵机制的关键之一是认识微生物之间的相互作用。研究微生物之间的相互作用对于白酒酿造机制的认识,以及酿造技术发展具有重要作用。发酵体系中微生物相互作用关系是白酒功能微生物研究的关键,以往研究多集中于白酒微生物菌群结构及单菌种功能。而选择不同的微生物组合进行发酵,不仅是阐明微生物之间相互作用的常用研究策略,     

The influence of metabolic network structures and energy requirements on xanthan gum yields

The metabolic network of Xanthomonas campestris is complex since a number of cyclic pathways are present making simple stoichiometric yield predictions difficult. The influence of certain pathway configurations and the resulting variations in flux have been examined as regards the maximum yield potential of this bacteria for xanthan gum production. These predictions have been compared with experimental results showing that the strain employed is functioning close to its theoretical maximum as regards yield criteria. The major constraint imposed on the network concerns energy availability which has a more pronounced effect on yield than carbon precursor supply. This can be attributed to the relatively high maintenance requirements determined experimentally and incorporated into the model. While some of this overall energy burden will undoubtedly be associated with incompressible metabolic requirements such as sugar uptake and xanthan efflux mechanisms, future strain improvement strategies will need to attack other non-essential energy-consuming reactions, if yields are to be further increased.     

Chromosome mapping in lactic acid bacteria

The chromosome structure of lactic acid bacteria has been investigated only recently. The development of pulsed-field gel electrophoresis (PFGE) combined with other DNA-based techniques enables whole-genome analysis of any bacterium, and has allowed rapid progress to be made in the knowledge of the lactic acid bacteria genome. Lactic acid bacteria possess one of the smallest eubacterial chromosomes. Depending on the species, the genome sizes range from 1.1 to 2.6 Mb. Combined physical and genetic maps of several species are already available or close to being achieved. Knowledge of the genomic structure of these organisms will serve as a basis for future genetic studies. Macrorestriction fingerprinting by PFGE is already one of the major tools for strain differentiation, identification of individual strains, and the detection of strain lineages. The genome data resulting from these studies will be of general application strain improvement.     

Technical guide for genetic advancement of underdeveloped and intractable Clostridium

In recent years, the genus Clostridium has risen to the forefront of both medical biotechnology and industrial biotechnology owing to its potential in applications as diverse as anticancer therapy and production of commodity chemicals and biofuels. The prevalence of hyper-virulent strains of C. difficile within medical institutions has also led to a global epidemic that demands a more thorough understanding of clostridial genetics, physiology, and pathogenicity. Unfortunately, Clostridium suffers from a lack of sophisticated genetic tools and techniques which has hindered the biotechnological exploitation of this important bacterial genus. This review provides a comprehensive summary of biotechnological progress made in clostridial genetic tool development, while also aiming to serve as a technical guide for the advancement of underdeveloped clostridial strains, including recalcitrant species, novel environmental samples, and non-type strains. Relevant strain engineering techniques, from genome sequencing and establishment of a gene transfer methodology through to deployment of advanced genome editing procedures, are discussed in detail to provide a blueprint for future clostridial strain construction endeavors. It is expected that a more thorough and rounded-out genetic toolkit available for use in the clostridia will bring about the construction of superior bioprocessing strains and a more complete understanding of clostridial genetics, physiology, and pathogenicity.     

红枣活性成分及其生物活性研究进展

红枣中的活性成分以多糖、黄酮类、环核苷酸类、多酚类、五环三萜类、生物碱为主,具有抗氧化、免疫调节及抗肿瘤、保护肝脏、降血糖、抗炎等多种生物活性。本文综述了红枣中活性成分及生物活性的研究进展,并对红枣产业的发展进行展望,为红枣中活性成分的开发与利用提供科学依据。     

Improvement of Rhizobium inoculants by mutation, genetic engineering and formulation

The use of mutation and genetic engineering techniques have resulted in Rhizobium strains with improved characteristics. The latter approach can provide improvement and new traits not achievable previously. Enhanced commercial Rhizobium inoculants, however, still rely solely on traditional approaches including formulation improvement, mutation, and strain selection. The lack of contribution to the commercial product lines by genetic engineering is mostly due to the time delay and financial burdens cast by the regulatory policies rather than a lack of progress in the application of modern genetic technologies. Such constraints may lessen in time as the public becomes more educated about the technologies and the subsequent political pressure decreases.     

Recent advances and state-of-the-art strategies in strain and process engineering for biobutanol production by Clostridium acetobutylicum

Butanol as an advanced biofuel has gained great attention due to its environmental benefits and superior properties compared to ethanol. However, the cost of biobutanol production via conventional acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum is not economically competitive, which has hampered its industrial application. The strain performance and downstream process greatly impact the economics of biobutanol production. Although various engineered strains with carefully orchestrated metabolic and sporulation-specific pathways have been developed, none of them is ideal for industrial biobutanol production. For further strain improvement, it is necessary to develop advanced genome editing tools and a deep understanding of cellular functioning of genes in metabolic and regulatory pathways. Processes with integrated product recovery can increase fermentation productivity by continuously removing inhibitory products while generating butanol (ABE) in a concentrated solution. In this review, we provide an overview of recent advances in C. acetobutylicum strain engineering and process development focusing on in situ product recovery. With deep understanding of systematic cellular bioinformatics, the exploration of state-of-the-art genome editing tools such as CRISPR-Cas for targeted gene knock-out and knock-in would play a vital role in Clostridium cell engineering for biobutanol production. Developing advanced hybrid separation processes for in situ butanol recovery, which will be discussed with a detailed comparison of advantages and disadvantages of various recovery techniques, is also imperative to the economical development of biobutanol.     

Knockout of pgdS and ggt genes improves γ‐PGA yield in B. subtilis

One of the emerging biopolymers that are currently under active investigation is bacterial poly(γ‐glutamic acid) (γ‐PGA). However, before its full industrial exploitation, a substantial increase in microbial productivity is required. γ‐PGA obtained from the Bacillus subtilis laboratory strain 168 offers the advantage of a producer characterized by a well defined genetic framework and simple manipulation techniques. In this strain, the knockout of genes for the major γ‐PGA degrading enzymes, pgdS and ggt, leads to a considerable improvement in polymer yield, which attains levels analogous to the top wild γ‐PGA producer strains. This study highlights the convenience of using the laboratory strain of B. subtilis over wild isolates in designing strain improvement strategies aimed at increasing γ‐PGA productivity. Biotechnol. Bioeng. 2013; 110: 2006–2012. © 2013 Wiley Periodicals, Inc.     

Exploiting biological complexity for strain improvement through systems biology

Cellular complexity makes it difficult to build a complete understanding of cellular function but also offers innumerable possibilities for modifying the cellular machinery to achieve a specific purpose. The exploitation of cellular complexity for strain improvement has been a challenging goal for applied biological research because it requires the coordinated understanding of multiple cellular processes. It is therefore pursued most efficiently in the framework of systems biology. Progress in strain improvement will depend not only on advances in technologies for high-throughput measurements but, more importantly, on the development of theoretical methods that increase the information content of these measurements and, as such, facilitate the elucidation of mechanisms and the identification of genetic targets for modification.