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从甘蔗渣中分离出的ITV-01所产α-葡萄糖苷酶的制备及其生化特性研究

Production and biochemical characterization of α-glucosidase from ITV-01 isolated from sugar cane bagasse.

作者信息

Del Moral S, Barradas-Dermitz D M, Aguilar-Uscanga M G

机构信息

Instituto de Biotecnología. Universidad del Papaloapan, Circuito Central 200. Tuxtepec, 68400 Oaxaca, Mexico.

Unidad de Desarrollo en Alimentos (UNIDA), Tecnológico Nacional de México, Instituto Tecnológico de Veracruz, Av. Miguel Ángel de Quevedo, 2779, Col. Formando Hogar, 91850 H. Veracruz, Veracruz Mexico.

出版信息

3 Biotech. 2018 Jan;8(1):7. doi: 10.1007/s13205-017-1029-6. Epub 2017 Dec 11.

Abstract

ITV-01 presents amylolytic activity, identified as α-glucosidase, an enzyme that only produces α-d-glucose from soluble starch and that presents transglucosylase activity on α-d-glucopyranosyl-(1-4)-α-d-glucopyranose (maltose) (200 gL). Biochemical characterization was performed on ITV-01 α-glucosidase; its optimum parameters were pH 4.3, temperature 80 °C but stable at 40 °C, with an energy of activation (Ea) 176.25 kJ mol. Using soluble starch as the substrate, and were 5 mg mL and 1000 U mg, respectively. As α-glucosidase is not a metalloenzyme, calcium and EDTA did not have any effect on its activity. The molecular weight was estimated by SDS-PAGE to be about 75 kDa. It was also active in methanol and ethanol. When ammonium sulfate (AS) and yeast extract (YE) nitrogen sources and calcium effect were evaluated, the greatest activity occurred using YE and calcium, as opposed to AS media where no activity was detected. The results obtained showed that this enzyme has industrial application potential in the processes to produce either ethanol or malto-oligosaccharides from α-d-glucopyranosyl-(1-4)-α-d-glucopyranose (maltose).

摘要

ITV-01具有淀粉分解活性,被鉴定为α-葡萄糖苷酶,该酶仅能从可溶性淀粉产生α-D-葡萄糖,并且对α-D-吡喃葡萄糖基-(1-4)-α-D-吡喃葡萄糖(麦芽糖)(200 g/L)具有转葡萄糖苷酶活性。对ITV-01α-葡萄糖苷酶进行了生化特性分析;其最佳参数为pH 4.3、温度80℃,但在40℃稳定,活化能(Ea)为176.25 kJ/mol。以可溶性淀粉为底物时, 和 分别为5 mg/mL和1000 U/mg。由于α-葡萄糖苷酶不是金属酶,钙和乙二胺四乙酸对其活性没有任何影响。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)估计其分子量约为75 kDa。它在甲醇和乙醇中也具有活性。当评估硫酸铵(AS)和酵母提取物(YE)氮源以及钙的影响时,使用YE和钙时活性最高,而在AS培养基中未检测到活性。所得结果表明,该酶在从α-D-吡喃葡萄糖基-(1-4)-α-D-吡喃葡萄糖(麦芽糖)生产乙醇或麦芽低聚糖的过程中具有工业应用潜力。

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Application of microbial α-amylase in industry - A review.微生物 α-淀粉酶在工业中的应用——综述。
Braz J Microbiol. 2010 Oct;41(4):850-61. doi: 10.1590/S1517-83822010000400004. Epub 2010 Dec 1.

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