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来自于……的果胶甲酯酶的组成型和细胞外表达

Constitutive and extracellular expression of pectin methylesterase from in .

作者信息

Acar Melek, Unver Yagmur

机构信息

Department of Molecular Biology and Genetics, Atatürk University, 25240 Erzurum, Turkey.

出版信息

3 Biotech. 2022 Sep;12(9):219. doi: 10.1007/s13205-022-03291-3. Epub 2022 Aug 10.

Abstract

UNLABELLED

Pectin methylesterase (PME) which is widely used in the cosmetic, food and pharmaceutical industries catalyses the hydrolysis of the methyl ester of pectin to yield methanol and free carboxyl groups. This study was performed to produce active pectin methylesterase (PME) extracellularly from in Firstly, pGKBα was constructed for the secretion of heterologous protein. After it was cloned in  cells and the sequence was affirmed, PME gene was inserted into pGKBα. So, pGKBα-PME carried the PME gene in correct position was cloned in  cells. Then, X-33 cells were transformed with linearized pGKBα-PME and six transformants were cultivated for recombinant PME production. It was observed that one of them had a high-capacity secretion of active PME. The molecular mass of extracellular PME enzyme was found to be about 59 kDa. The PME enzyme from was produced by for the first time in this study. This recombinant enzyme might be produced in a large scale and also purified from the culture medium. Then, the purified enzyme might be used for clarification and increasing yield of juice in food industrial applications.

SUPPLEMENTARY INFORMATION

The online version contains supplementary material available at 10.1007/s13205-022-03291-3.

摘要

未标记

果胶甲酯酶(PME)广泛应用于化妆品、食品和制药行业,催化果胶甲酯水解生成甲醇和游离羧基。本研究旨在从[具体来源未提及]中胞外生产活性果胶甲酯酶(PME)。首先,构建用于分泌异源蛋白的pGKBα。将其克隆到[具体细胞未提及]细胞中并确认序列后,将PME基因插入pGKBα。于是,携带正确位置PME基因的pGKBα - PME被克隆到[具体细胞未提及]细胞中。然后,用线性化的pGKBα - PME转化X - 33细胞,并培养六个转化体以生产重组PME。观察到其中一个具有高容量分泌活性PME的能力。发现胞外PME酶的分子量约为59 kDa。本研究首次由[具体来源未提及]生产出这种PME酶。这种重组酶可能会大规模生产,并从培养基中纯化。然后,纯化后的酶可用于食品工业应用中果汁的澄清和提高产量。

补充信息

在线版本包含可在10.1007/s13205 - 022 - 03291 - 3获取的补充材料。

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