排序方式: 共有56条查询结果,搜索用时 937 毫秒
1.
Summary The retention, selectivity and elution order of fluorescent 1,N6-etheno derivatives of diadenosine polyphosphates and their enzymatic degradation products on octadecyl and phenyl-bonded
silica columns have been studied as a function of mobile phase pH, ionic strength and organic modifier content. Good separations
of the compounds of interest were achieved using mobile phases of around 0.1M potassium phosphate buffers at neutral pH containing
approximately 10% methanol or 4% acetonitrile for C18 columns and 5% methanol or 1.5% acetonitrile for phenyl columns. The data obtained were used to establish isocratic assays
for diadenosine polyphosphate cleaving activities from chromaffin cells using Di(1,N6-ethenoadenosine) polyphosphates as fluorogenic substrate analogues followed by fluorescence detection. 相似文献
2.
3.
4.
5.
《Journal of carbohydrate chemistry》2013,32(4-6):529-547
Some attempts toward the synthesis of novel inhibitors of glycosyl transferases are described. More successfully, the synthesis of an activated cyclopropacyclohexene and an amide and an amine of a cyclopropa‐fused pyranose are described. None of these three novel compounds proved to be a significant inhibitor of a retaining α‐glucosidase from barley. 相似文献
6.
7.
《化学:亚洲杂志》2017,12(19):2539-2543
Enzymes normally lose their activities under extreme conditions due to the dissociation of their active tertiary structure. If an enzyme could maintain its catalytic activity under non‐physiological or denaturing conditions, it might be used in more applications in the pharmaceutical and chemical industries. Recently, we reported a coiled‐coil six‐helical bundle (6HB) structure as a scaffold for designing artificial hydrolytic enzymes. Here, intermolecular isopeptide bonds were incorporated to enhance the stability and activity of such biomolecules under denaturing conditions. These isopeptide bridge‐tethered 6HB enzymes showed exceptional stability against unfolding and retained or even had increased catalytic activity for a model hydrolysis reaction under thermal and chemical denaturing conditions. Thus, isopeptide bond‐tethering represents an efficient route to construct ultrastable artificial hydrolases, with promising potential to maintain biocatalysis under extreme conditions. 相似文献
8.
A Mechanism‐Based Approach to Screening Metagenomic Libraries for Discovery of Unconventional Glycosidases
下载免费PDF全文
![点击此处可从《Angewandte Chemie (International ed. in English)》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Seyed Amirhossein Nasseri Dr. Leo Betschart Daria Opaleva Dr. Peter Rahfeld Prof. Stephen G. Withers 《Angewandte Chemie (International ed. in English)》2018,57(35):11359-11364
Functional metagenomics has opened new opportunities for enzyme discovery. To exploit the full potential of this new tool, the design of selective screens is essential, especially when searching for rare enzymes. To identify novel glycosidases that employ cleavage strategies other than the conventional Koshland mechanisms, a suitable screen was needed. Focusing on the unsaturated glucuronidases (UGLs), it was found that use of simple aryl glycoside substrates did not allow sufficient discrimination against β‐glucuronidases, which are widespread in bacteria. While conventional glycosidases cannot generally hydrolyze thioglycosides efficiently, UGLs follow a distinct mechanism that allows them to do so. Thus, fluorogenic thioglycoside substrates featuring thiol‐based self‐immolative linkers were synthesized and assessed as selective substrates. The generality of the approach was validated with another family of unconventional glycosidases, the GH4 enzymes. Finally, the utility of these substrates was tested by screening a small metagenomic library. 相似文献
9.
Timothy W. Bumpus Prof. Dr. Jeremy M. Baskin 《Angewandte Chemie (International ed. in English)》2016,55(42):13155-13158
Phosphatidic acid (PA) is a potent lipid secondary messenger, the synthesis of which is tightly regulated in both space and time. Established tools for detecting PA involve ex vivo analysis and do not provide information on the subcellular locations where this lipid is synthesized. Here, a chemoenzymatic strategy for imaging sites of cellular PA synthesis by phospholipase D (PLD) enzymes is reported. PLDs were found to be able to catalyze phospholipid head‐group exchange with alkynols to generate alkyne‐labeled PA analogues within cells. Subsequent fluorophore tagging through Cu‐catalyzed azide–alkyne cycloaddition enabled both visualization by fluorescence microscopy and quantification by HPLC. Our studies revealed several intracellular sites of PLD‐mediated PA synthesis. We envision applications of this approach to dissect PA‐dependent signaling pathways, image PLD activity in disease, and remodel intracellular membranes with new functionality. 相似文献
10.
蛋白质是生命体的重要组成部分,其中生物酶在生命体系中发挥至关重要的作用。蛋白质分子设计是研究生物酶结构与功能关系的重要手段。本文综述了基于蛋白质骨架的人工水解酶的理性设计与功能研究进展,包括对天然蛋白的重新利用和重新改造,基于3-股螺旋、4-股螺旋或锌指蛋白的分子设计,以及血红蛋白(如细胞色素b5和肌红蛋白突变体)水解酶催化活性的调控等,阐明了人工水解酶分子设计的基本思路与研究方法,为合理构建人工水解酶或其他生物酶提供了重要的信息。人工水解酶的理性设计进展,不但深化我们对天然酶结构-性质-反应-功能关系的认识,而且还提升我们创造具有优越功能的人工生物酶的能力。 相似文献