Cloning, Expression and Enzymatic Characterization of Pectin Methyl Esterase from Populus trichocarpa and Its Application

Processes Pub Date : 2024-07-18 DOI:10.3390/pr12071511
Yanjiao Feng, Lifen Huang, Yue Zeng, Yiyuan Zhang, Wei Liu, Gang He
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Abstract

The pectin methyl esterase gene from Populus trichocarpa (PtPME) was successfully cloned through PCR amplification and subsequently inserted into the expressing vector pMAL-c5e for successful expression in Escherichia coli BL21 (DE3). Initially, we determined the primary enzymatic properties of PtPME, a pectin methyl esterase derived from Populus trichocarpa. Notably, this enzyme exhibits a higher affinity towards citrus pectin, with an esterification degree exceeding 60%. Furthermore, this enzyme’s optimal reaction temperature and pH were found to be 30 °C and 8, respectively. Importantly, its exceptional stability under neutral conditions highlights its potential application in the industrial production of low-ester pectin.
杨树果胶甲基酯酶的克隆、表达和酶学特征及其应用
通过 PCR 扩增成功克隆了杨树果胶甲酯酶(PtPME)基因,随后将其插入表达载体 pMAL-c5e 中,并在大肠杆菌 BL21 (DE3) 中成功表达。最初,我们确定了 PtPME 的主要酶学特性,它是一种源自毛白杨的果胶甲酯酶。值得注意的是,这种酶对柑橘果胶具有较高的亲和力,酯化度超过 60%。此外,这种酶的最佳反应温度和 pH 值分别为 30 °C 和 8。重要的是,这种酶在中性条件下具有极高的稳定性,因此有望应用于低酯果胶的工业生产。
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