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首页> 外文期刊>Catalysis science & technology >Discovery of new levansucrase enzymes with interesting properties and improved catalytic activity to produce levan and fructooligosaccharides
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Discovery of new levansucrase enzymes with interesting properties and improved catalytic activity to produce levan and fructooligosaccharides

机译：新发现的levansucrase酶有趣的属性和改善催化生产果聚糖和活动

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摘要

Levansucrases ( EC 2.4.1.10, LS) are of high interest for the synthesis of novel prebiotic fructooligosaccharides. Hindered by the limited availability of LS enzymes and a propensity for performing the hydrolysis of sucrose rather than the transfructosylation reaction, a desire to find new LSs was fueled. Genome mining was employed to explore their biodiversity using 26 characterized LSs as a reference set for a sequence driven- approach leading to a collection of 32 enzymes representative of the biodiversity for which the gene was cloned and over- expressed in E. coli. These enzymes underwent an initial screening process based upon total activity, transfructosylation activity and levan forming ability which narrowed the candidates to 10 potential enzymes. These LS enzymes were found to have high levan production ( 33 +/- 1 g levan per mg protein) and able to produce very large polymers ( 6986 kDa). The LS from Paraburkholderia graminis had a very high natural thermal stability with a half- life of 291 minutes at 50 degrees C. The full kinetic parameters of the top candidates were characterized for the enzymes with the highest potential. Enzymes with higher catalytic efficiency and activity for transfructosylation over hydrolysis were identified. The acceptor specificity of these new levansucrase enzymes was briefly explored. It showed wide specificity for each of the selected enzymes.
机译：Levansucrases (EC 2.4.1.10 LS)高对小说的合成生命起源以前的fructooligosaccharides。LS的酶和倾向蔗糖的水解而不是表演transfructosylation反应,渴望找到新的LSs推动。使用26个用来探索生物多样性LSs特征作为参考的序列驱动——方法导致一个集合32酶生物多样性的代表基因的克隆和表达在大肠杆菌。筛选过程基于总活动,transfructosylation活动和果聚糖形成能力,缩小候选人10潜在的酶。有很高的果聚糖生产(33 + / - 1 g果聚糖/毫克蛋白),能够产生非常大的聚合物(6986 kDa)。Paraburkholderia茎有很高的自然热稳定性的一半——291年的生命分钟50度c .完整的动能参数的热门候选人最高的酶的特征的潜力。transfructosylation效率和活动在确定了水解。这些新levansucrase酶的特异性简要探讨。每个选定的酶。

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