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1.
为进一步明确利迪链菌素生物合成时从初级代谢到次级代谢的转变,运用ESI-MS和PCA分析,对利迪链霉菌AS 4.2501生产利迪链菌素不同发酵阶段的细胞内外代谢物进行代谢物组研究。结果表明,利用代谢物组学的方法能区分开不同发酵阶段的代谢产物,特别是根据第三主成分(PC3)可明显将细胞进入次级代谢与初级代谢阶段区分开,结合文献和MS/MS分析,推断代谢转变过程的生物标志物主要源于磷脂和蛋白类化合物。  相似文献   

2.
杨祖明  王颖  姚明东  肖文海 《化工进展》2019,38(5):2402-2412
菌种进化工程是绿色生物制造的重要策略,利用高效的高通量筛选方法和技术可以快速地获得理想的实用菌株。针对菌种进化工程中的高通量筛选方法,本文重点综述了基于颜色或荧光、基于细胞生长、基于生物传感器以及基于液滴微流体平台等4个方面的高通量筛选技术的重要进展,同时也介绍了各种高通量筛选技术的应用范围和特点,为研究人员从不同进化文库中获得生理特性或者代谢能力显著提高的目标菌株提供了理论指导,极大地提高进化文库的筛选效率,降低了菌株筛选的时间和成本。最后展望了人工智能、合成生物学以及生物信息学的发展对高通量筛选技术的重要影响,以期提高高通量筛选技术的精度、效率和应用范围,进而加速菌种进化过程和工业化进程。  相似文献   

3.
通过对组合催化研究的关键因素--催化剂高通量筛选技术的评价分析认为:在对已有的催化体系和催化剂优化的同时,如何高校研制开发新筛选检测技术,诸如反应器设计和检测器的选择,是组合催化研究的主导思想,对此的深刻理解和运用将增强实验室的创新能力。  相似文献   

4.
于淑晶  边强  王满意  张晓  李宝聚  赵卫光 《农药》2012,51(8):550-553,564
高通量筛选技术是发现创新药物的重要技术手段之一,在医药研究领域已经得到了广泛的应用,但在农药研究领域还处于初始阶段。综述了近年来高通量筛选技术在农用杀菌剂创制中的应用,并展望了其发展前景。  相似文献   

5.
生物催化技术是工业可持续发展最有希望的技术,生物催化剂有着很好的应用前景。介绍了高通量筛选生物催化剂的常用方法。  相似文献   

6.
高通量筛选与组合合成技术相结合是发现创新药物的重要技术手段之一。这极大地提高了化合物库的建立速度及对目标分子、活性物质以及先导药物的筛选速度,在农药创制过程中可以缩短新农药分子的研发周期,提高成功率。  相似文献   

7.
目的:为快速评估填料及筛选工艺条件,探究高通量筛选技术在双特异性抗体的亲和层析工艺开发中的应用。方法:采用高通量微孔滤板进行工艺开发,利用Langmuir等温吸附曲线评估8种商业Protein A填料的结合能力,筛选出高载量的2种填料后对5种洗脱条件和7种淋洗条件进行筛选,并在层析柱上对确定的工艺进行验证。结果:筛选出的BioCap rProtein A PC填料以60 g/L的载量,在pH=3.6的洗脱溶液和含0.5 mol/L NaCl的淋洗2的工艺下,柱层析的收率(亲和收率和中和收率分别为93.6%和100.0%)及产品质量(纯度95.5%,宿主细胞蛋白含量1 282 ng/mg)均满足工艺要求。结论:高通量筛选技术可快速、高效地从多种亲和填料中筛选出最佳填料以及从多种工艺参数中筛选出适合的工艺条件,缩短了实验周期,提高了实验效率。  相似文献   

8.
介绍了近几年来用于高通量筛选技术的反应器和检测器,对高通量筛选技术在烯烃聚合领域中的应用情况进行了综述,并对其在烯烃聚合领域的应用前景做了展望.  相似文献   

9.
将脱硫菌的筛选与新一代高通量测序技术相结合,探索脱硫菌筛选的新方法,为生物甲烷的高值化利用提供技术支持。实验将富含硫细菌的环境土样及经过培养基富集培养后的液样分别进行16S rDNA V6区的高通量测序,并分析了物种组成和丰度、Alpha多样性和菌群结构。结果表明,高通量测序的平均数据利用率达99.177%,测序量能够充分反映样品在该区域细菌的群落组成和结构。取自环境的土壤样品物种组成丰富,含有Pseudomonadales(假单胞菌目)、Rhizobiales(根瘤菌目)、Desulfuromonadales(除硫单孢菌目)、Desulfobacterales(脱硫杆菌目)、Acidithiobacillales(酸硫杆状菌目)等脱硫菌目。经过培养基富集后,菌群多样性显著降低,主要组成菌为Pseudomonadales(假单胞菌目)和Rhizobiales(根瘤菌目)。通过高通量测序技术,能够在筛选脱硫菌之前就了解实验样品的菌群结构和组成,为有针对性地设计筛选脱硫菌实验提供了一种新的方法。  相似文献   

10.
简要介绍了邻近闪烁分析技术及由其发展而来的Alphascreen分析技术 ,对其中使用的荧光微球的要求、特点作了介绍 ,并按荧光物质在微球中所处的位置对荧光微球的制备方法作了较为详细的讨论。  相似文献   

11.
To achieve higher antibiotic streptolydigin productivity through metabolic regulation, propionate was fed during the fermentation of Streptomyces lydicus AS 4.2501. The effects of propionate feeding on streptolydigin production and intracellular fluxes were investigated. The highest streptolydigin production yield of 95.10mg·L^-1 was obtained when 2mmol·L^-1 of sodium propionate was added at 60h of cultivation into shake-flask culture. This yield is 23.06% higher when compared to that of a batch culture without propionate supplementation. It was also found that when propionate was added, much more organic acids were excreted. Metabolic flux analysis was performed and it demonstrated that the carbon fluxes of the pentose phosphate pathway and the anaplerotic reaction were significantly increased after propionate feeding. The carbon flux from pyruvate to acetyl-CoA was determined to be 24.7, which was 12.27% higher than that in the batch culture. This study indicated that the glucose-6-phosphate and pyruvate nodes were potential bottlenecks for increasing streptolydigin productivity. Potential targets and strategies that could be manipulated through genetic and process engineering to increase the production of streptolydigin were also suggested.  相似文献   

12.
The effects of nitrogen sources on streptolydigin production and distribution of secondary metabolites were investigated for flask cultured S.lydicus AS 4.2501.When peptone,asparamide,and glutamic acid were ex- amined as the nitrogen source,respectively,liquid chromatography-mass spectrometry(LC-MS)and photodiode array(PDA)analyses revealed the formation of two analogues of streptolydigin in the fermentation broth.When soybean meal was used as the source of nitrogen,three analogues of streptolydigin were detected.The use of am- monium sulfate as a source of nitrogen resulted in a lower pH value of the fermentation system,thus inhibiting streptolydigin biosynthesis and changing the metabolic profiling.Among the nitrogen sources that were made use of,glutamic acid was most favorable to the formation of streptolydigin.Simultaneously,this study also showed that the changing nitrogen sources resulted in altering the production and relative ratios of streptolydigin and its analogues.  相似文献   

13.
    
The ubiquitylation machinery regulates several fundamental biological processes from protein homeostasis to a wide variety of cellular signaling pathways. As a consequence, its dysregulation is linked to diseases including cancer, neurodegeneration, and autoimmunity. With this review, we aim to highlight the therapeutic potential of targeting E3 ligases, with a special focus on an emerging class of RING ligases, named tri-partite motif (TRIM) proteins, whose role as targets for drug development is currently gaining pharmaceutical attention. TRIM proteins exert their catalytic activity as scaffolds involved in many protein–protein interactions, whose multidomains and adapter-like nature make their druggability very challenging. Herein, we give an overview of the current understanding of this class of single polypeptide RING E3 ligases and discuss potential targeting options.  相似文献   

14.
杨平  衣雪竹 《广州化工》2015,(6):135-136,147
为建立一种工业杀菌剂高通量筛选方法,采用96孔板培养法,以大肠杆菌作为待测菌,利用显色剂显色反应对杀菌剂的抑菌能力进行评价。结果显示,采用1~4 d培养菌液,稀释至10%或菌液吸光度值A=0.038,显色剂TTC(2,3,5-氯化三苯基四氮唑)加入量20 mg/kg,显色18~24 h可得到与试管法一致的结果。该方法操作简捷,微量省力,结果易观察,成本低,可用于工业杀菌剂的大量筛选和应用评估。  相似文献   

15.
为了评价由新型助剂制成的草甘膦水剂的生物活性,研究建立了用空心莲子草评价草甘膦水剂的室内生测方法.以空心莲子草为生测靶标,采朋喷雾法,培养介质为土壤,以地上部分鲜重抑制率为评价指标来筛选适合旱地使用的草甘膦水剂;以空心莲子草为生测靶标,采用喷雾法,培养介质为清水,以总鲜重抑制率为评价指标来筛选适合湿地或水域使用的草甘膦水剂.  相似文献   

16.
    
Drosophila melanogaster (Drosophila) models of cancer are emerging as powerful tools to investigate the basic mechanisms underlying tumour progression and identify novel therapeutics. Rapid and inexpensive, it is possible to carry out genetic and drug screens at a far larger scale than in vertebrate organisms. Such whole-organism-based drug screens permits assessment of drug absorption and toxicity, reducing the possibility of false positives. Activating mutations in the Wnt and Ras signalling pathways are common in many epithelial cancers, and when driven in the adult Drosophila midgut, it induces aggressive intestinal tumour-like outgrowths that recapitulate many aspects of human colorectal cancer (CRC). Here we have taken a Drosophila CRC model in which tumourous cells are marked with both GFP and luciferase reporter genes, and developed novel high-throughput assays for quantifying tumour burden. Leveraging these assays, we find that the Drosophila CRC model responds rapidly to treatment with standard CRC-drugs, opening the door to future rapid genetic and drug screens.  相似文献   

17.
    
The industrial production of chemicals by microorganisms usually requires improvements to the enzymes, pathways, and strain that go beyond the capacity of innate enzymes. To achieve these phenotypes and overcome our limited capacity to de novo design these parts, directed and adaptive evolutionary approaches are used to explore new functions. This review highlights the recent advances in both sequence diversity generation and selection strategies from traditional in vitro mutagenesis to novel in vivo continuous evolution applications. The focus here is on comparison of the different gene diversity methods in an attempt to distinguish the best strategy for protein or strain engineering for a given goal. Furthermore, the important role that screening and selection can play in advancing directed and adapted evolution is discussed. © 2018 Society of Chemical Industry  相似文献   

18.
以螺旋链霉菌25-1为出发菌株,分别采用紫外线(UV)、Co60射线、甲基磺酸乙酯(EMS)、氯化锂(LiCl)、UV+LiCl、亚硝基胍(NTG)+UV进行诱变筛选,比较各方法的致死率、正变率、最大提高幅度及对传代稳定性的影响,结果发现UV、Co60射线、UV+LiCl 3种方法对螺旋链霉茵的诱变效果较好,化学诱变剂...  相似文献   

19.
    
High-throughput virtual screening (HTVS) is, in conjunction with rapid advances in computer hardware, becoming a staple in drug design research campaigns and cheminformatics. In this context, virtual compound library design becomes crucial as it generally constitutes the first step where quality filtered databases are essential for the efficient downstream research. Therefore, multiple filters for compound library design were devised and reported in the scientific literature. We collected the most common filters in medicinal chemistry (PAINS, REOS, Aggregators, van de Waterbeemd, Oprea, Fichert, Ghose, Mozzicconacci, Muegge, Egan, Murcko, Veber, Ro3, Ro4, and Ro5) to facilitate their open access use and compared them. Then, we implemented these filters in the open platform Konstanz Information Miner (KNIME) as a freely accessible and simple workflow compatible with small or large compound databases for the benefit of the readers and for the help in the early drug design steps.  相似文献   

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