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排序方式: 共有127条查询结果,搜索用时 15 毫秒
1.
Dr. Qingyun Tang Assist. Prof. Dr. Ioannis V. Pavlidis Dr. Christoffel P. S. Badenhorst Prof. Dr. Uwe T. Bornscheuer 《Chembiochem : a European journal of chemical biology》2021,22(16):2584-2590
Halide methyltransferases (HMTs) enable the enzymatic synthesis of S-adenosyl-l -methionine (SAM) from S-adenosyl-l -homocysteine (SAH) and methyl iodide. Characterisation of a range of naturally occurring HMTs and subsequent protein engineering led to HMT variants capable of synthesising ethyl, propyl, and allyl analogues of SAM. Notably, HMTs do not depend on chemical synthesis of methionine analogues, as required by methionine adenosyltransferases (MATs). However, at the moment MATs have a much broader substrate scope than the HMTs. Herein we provide an overview of the discovery and engineering of promiscuous HMTs and how these strategies will pave the way towards a toolbox of HMT variants for versatile chemo- and regioselective biocatalytic alkylations. 相似文献
2.
Increase of empty pool-activated Ca2+ influx using an intracellular Ca2+ chelating agent 总被引:1,自引:0,他引:1
We demonstrate here that stimulated 45Ca2+ influx in A7r5 vascular smooth muscle cells induced either by receptor activation with [Arg]8 vasopressin or by the SR-Ca(2+)-ATPase inhibitor thapsigargin was increased more than threefold if cells were preloaded with the intracellular calcium chelator BAPTA (1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid). The increased influx is probably due to an attenuation of negative feedback of Ca2+ on its own entry accompanied by increased Ca2+ storage capacity of BAPTA-loaded cells leading to diminished cellular Ca2+ release. We propose that BAPTA preloading could be a useful approach to investigate receptor-induced Ca2+ influx. 相似文献
3.
Protein Engineering of the Progesterone Hydroxylating P450‐Monooxygenase CYP17A1 Alters Its Regioselectivity 下载免费PDF全文
Lisa K. Morlock Sascha Grobe Dr. Kathleen Balke Dr. Stephan Mauersberger Dr. Dominique Böttcher Prof. Dr. Uwe T. Bornscheuer 《Chembiochem : a European journal of chemical biology》2018,19(18):1954-1958
The CYP171 enzyme is known to catalyse a key step in the steroidogenesis of mammals. The substrates progesterone and pregnenolone are first hydroxylated at the C17 position, and this is followed by cleavage of the C17?C20 bond to yield important precursors for glucosteroids and androgens. In this study, we focused on the reaction of the bovine CYP17A1 enzyme with progesterone as a substrate. On the basis of a created homology model, active‐site residues were identified and systematically mutated to alanine. In whole‐cell biotransformations, the importance of the N202, R239, G297 and E305 residues for substrate conversion was confirmed. Additionally, mutation of the L206, V366 and V483 residues enhanced the formation of the 16α‐hydroxyprogesterone side product up to 40 % of the total product formation. Furthermore, residue L105 was found not to be involved in this side activity, which contradicts a previous study with the human enzyme. 相似文献
4.
Markus A. Durban Uwe T. Bornscheuer 《European Journal of Lipid Science and Technology》2007,109(5):469-473
Phospholipases C (PLC, EC 3.1.4.3) are enzymes that specifically hydrolyze the C‐O‐P bond in phospholipids, yielding sn‐1,2(2,3)‐diacylglycerides and the phosphate residue bearing the corresponding headgroup. The biochemical characterization of PLC requires methods for the reliable determination of their activity. Here, an assay is described in which the phosphate residue released by the PLC is cleaved with an alkaline phosphatase. The phosphate formed is then extracted with n‐butanol and quantified as phosphomolybate complex. The applicability of this method is demonstrated for a concentration range from 10 nM to 10 mM for a range of phospholipids bearing different headgroups in an aqueous and a two‐phase system. The method has the additional advantage that the crude enzyme can be used without the need for purification. 相似文献
5.
Conversion of a Mono‐ and Diacylglycerol Lipase into a Triacylglycerol Lipase by Protein Engineering 下载免费PDF全文
Dr. Dongming Lan Dr. Grzegorz Maria Popowicz Dr. Ioannis V. Pavlidis Pengfei Zhou Prof. Dr. Uwe T. Bornscheuer Prof. Dr. Yonghua Wang 《Chembiochem : a European journal of chemical biology》2015,16(10):1431-1434
Despite the fact that most lipases are believed to be active against triacylglycerides, there is a small group of lipases that are active only on mono‐ and diacylglycerides. The reason for this difference in substrate scope is not clear. We tried to identify the reasons for this in the lipase from Malassezia globosa. By protein engineering, and with only one mutation, we managed to convert this enzyme into a typical triacylglycerol lipase (the wild‐type lipase does not accept triacylglycerides). The variant Q282L accepts a broad spectrum of triacylglycerides, although the catalytic behavior is altered to some extent. From in silico analysis it seems that specific hydrophobic interactions are key to the altered substrate specificity. 相似文献
6.
Mohamed M. Soumanou Uwe T. Bornscheuer Ulrich Menge Rolf D. Schmid 《Journal of the American Oil Chemists' Society》1997,74(4):427-433
Structured triglycerides (ST) that contain medium- and long-chain fatty acids were synthesized by lipase-catalyzed interesterification
between tricaprylin and peanut oil. To select appropriate enzymes, we investigated nine commercial lipase preparations for
their ability to hydrolyze pure triglycerides as well as natural oils. Three microbial lipases from Rhizomucor miehei (RML), Candida sp. (CSL), and Chromobacterium viscosum (CVL) gave good results, and immobilized preparations were used in the interesterification. RML gave the highest yields of
ST (73%, 40°C), although its hydrolytic activity toward triolein was low. As the temperature was raised to 50°C, the yield
of ST increased to 79%. After 120 h reaction time, remaining activities were high for CSL (71%), moderate for CVL (48%), and
low for RML (20%).
Parts of this paper were presented as a poster at the Biochemical Engineering Conference IX, May 1995, Davos, Switzerland. 相似文献
7.
8.
Sirirung Wongsakul Poonsuk Prasertsan Uwe T. Bornscheuer Aran H‐Kittikun 《European Journal of Lipid Science and Technology》2003,105(2):68-73
Commercial immobilized lipases were used for the synthesis of 2‐monoglycerides (2‐MG) by alcoholysis of palm and tuna oils with ethanol in organic solvents. Several parameters were studied, i.e., the type of immobilized lipases, water activity, type of solvents and temperatures. The optimum conditions for alcoholysis of tuna oil were at a water activity of 0.43 and a temperature of 60 °C in methyl‐tert‐butyl ether for ~12 h. Although immobilized lipase preparations from Pseudomonas sp. and Candida antarctica fraction B are not 1, 3‐regiospecific enzymes, they were considered to be more suitable for the production of 2‐MG by the alcoholysis of tuna oil than the 1, 3‐regiospecific lipases (Lipozyme RM IM from Rhizomucor miehei and lipase D from Rhizopus delemar). With Pseudomonas sp. lipase a yield of up to 81% 2‐MG containing 80% PUFA (poly‐unsaturated fatty acids) from tuna oil was achieved. The optimum conditions for alcoholysis of palm oil were similar as these of tuna oil alcoholysis. However, lipase D immobilized on Accurel EP100 was used as catalyst at 40 °C with shorter reaction times (<12 h). This lead to a yield of ~60% 2‐MG containing 55.0‐55.7% oleic acid and 18.7‐21.0% linoleic acid. 相似文献
9.
Yannik Brack Dr. Chenghai Sun Prof. Dr. Dong Yi Prof. Dr. Uwe T. Bornscheuer 《Chembiochem : a European journal of chemical biology》2022,23(10):e202200062
p-Coumaric acid (p-CA) is a key precursor for the biosynthesis of flavonoids. Tyrosine ammonia lyases (TALs) specifically catalyze the synthesis of p-CA from l -tyrosine, which is a convenient enzymatic pathway. To explore novel and highly active TALs, a phylogenetic tree-building approach was conducted including 875 putative TALs and 46 putative phenylalanine/tyrosine ammonia lyases (PTALs). Among them, 5 TALs and 3 PTALs were successfully characterized and found to exhibit the proposed enzymatic activity. The TAL from Chryseobacterium luteum sp. nov (TALclu) has the highest affinity (Km =0.019 mm ) and conversion efficiency (kcat/Km= 1631 s−1 ⋅ mm −1) towards l -tyrosine. The reaction conditions for two purified enzymes and their E. coli recombinant cells were optimized and p-CA yields of 2.03 g/L after 8 hours by TALclu and 2.35 g/L after 24 h by TAL from Rivularia sp. PCC 7116 (TALrpc) in whole cells were achieved. These TALs are thus candidates for the construction of whole-cell systems to produce the flavonoid precursor p-CA. 相似文献
10.
Przemyslaw Krause Lutz Hilterhaus Georg Fieg Andreas Liese Uwe Bornscheuer 《European Journal of Lipid Science and Technology》2009,111(2):194-201
The esterification of benzoic acid with n‐hexanol, n‐octanol, 2‐ethyl hexanol and n‐decanol was investigated in detail. An analysis of the reaction kinetics of esterification in the presence of different commercially available chemical catalysts was carried out. The effects of catalyst type and loading on the reaction rate were studied. Although the considered reaction is bimolecular, it showed a first‐order behavior, and a linear dependence with respect to the catalyst concentration was observed. Hence, a new approach is presented to describe the reaction kinetics accurately over a wide range. The application of biotechnological synthesis applying different enzymes as catalysts offers an interesting alternative besides chemical synthesis. Especially an esterase from Bacillus subtilis immobilized on Sepabeads EC‐EP showed high stability and was applied for 2 days in the synthesis of hexyl benzoate. Nevertheless, the chemical reaction route remains superior with respect to the catalyst activities under the applied conditions, which were 25 kU/g for the chemical reaction and 0.7 kU/g for the best enzymatic conversion. 相似文献