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一种高效葡聚糖分支酶的酶学性质及其在淀粉改性中的潜在应用。

Enzymatic properties of an efficient glucan branching enzyme and its potential application in starch modification.

作者信息

Chengyao Xia, Yan Qiao, Chaonan Dong, Xiaopei Chen, Yanxin Wang, Ding Li, Xianfeng Ye, Jian Han, Yan Huang, Zhongli Cui, Zhoukun Li

机构信息

Key Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture, College of Life Science, Nanjing Agricultural University, Nanjing, 210095, PR China.

Institute of Veterinary Immunology &Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, PR China.

出版信息

Protein Expr Purif. 2021 Feb;178:105779. doi: 10.1016/j.pep.2020.105779. Epub 2020 Oct 25.

Abstract

Glucan branching enzymes (GBEs, EC 2.4.1.18) catalyze the formation of α-1,6-linked branch in starch, which is important for the starch modification with prospective properties. In this study, the aqGBE gene encoding an efficient glucan branching enzyme was cloned from Aquabacterium sp. strain A7-Y and successfully expressed in Escherichia coli BL21 (DE3). The specific activity of the purified recombinant enzyme rAqGBE was 2850 U/mg with potato starch as the optimal substrate, and the K and V values of rAqGBE were 1.18 mg/mL and 588.2 μmol/min/mg, respectively. Enzymological characterization showed that rAqGBE exhibits its optimal activity under the condition of 40 °C and pH 7.0, respectively, which is independent of calcium ions. Otherwise, rAqGBE-treated potato starch showed different chain length distribution compared with control, the numbers of short chains (degree of polymerization, DP < 7) and long chains (DP > 25) increased from 4.5% to 9.6% and 6.1%-15.7% after enzymatic treatment, respectively. In starch anti-ageing assay, with minimum usage of 0.8 mg rAqGBE per g starch, the rAqGBE-treated potato starch exhibited reduced retrogradation properties. Our results indicate that the branching enzyme AqGBE may therefore be a promising tool for the enzymatic modification of starch.

摘要

葡聚糖分支酶(GBEs,EC 2.4.1.18)催化淀粉中α-1,6-连接分支的形成,这对于具有预期特性的淀粉改性很重要。在本研究中,从水生细菌菌株A7-Y中克隆了编码高效葡聚糖分支酶的aqGBE基因,并在大肠杆菌BL21(DE3)中成功表达。纯化的重组酶rAqGBE以马铃薯淀粉为最佳底物时的比活性为2850 U/mg,rAqGBE的K和V值分别为1.18 mg/mL和588.2 μmol/min/mg。酶学特性表明,rAqGBE分别在40°C和pH 7.0的条件下表现出最佳活性,且该活性与钙离子无关。此外,与对照相比,经rAqGBE处理的马铃薯淀粉显示出不同的链长分布,酶处理后短链(聚合度,DP < 7)和长链(DP > 25)的数量分别从4.5%增加到9.6%和从6.1%增加到15.7%。在淀粉抗老化试验中,每克淀粉最低使用0.8 mg rAqGBE时,经rAqGBE处理的马铃薯淀粉表现出降低的回生特性。我们的结果表明,分支酶AqGBE因此可能是一种用于淀粉酶促改性的有前途的工具。

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