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利用啤酒糟作为底物生产、纯化和表征一种重要的青霉木聚糖酶。

Production, purification, and characterization of a major Penicillium glabrum xylanase using Brewer's spent grain as substrate.

机构信息

Department of Biological Sciences, Midwest State University, Camargo Varela de Sá Street 03, 85040-080 Guarapuava, PR, Brazil.

出版信息

Biomed Res Int. 2013;2013:728735. doi: 10.1155/2013/728735. Epub 2013 May 13.

Abstract

In recent decades, xylanases have been used in many processing industries. This study describes the xylanase production by Penicillium glabrum using brewer's spent grain as substrate. Additionally, this is the first work that reports the purification and characterization of a xylanase using this agroindustrial waste. Optimal production was obtained when P. glabrum was grown in liquid medium in pH 5.5, at 25 °C, under stationary condition for six days. The xylanase from P. glabrum was purified to homogeneity by a rapid and inexpensive procedure, using ammonium sulfate fractionation and molecular exclusion chromatography. SDS-PAGE analysis revealed one band with estimated molecular mass of 18.36 kDa. The optimum activity was observed at 60 °C, in pH 3.0. The enzyme was very stable at 50 °C, and high pH stability was verified from pH 2.5 to 5.0. The ion Mn(2+) and the reducing agents β -mercaptoethanol and DTT enhanced xylanase activity, while the ions Hg(2+), Zn(2+), and Cu(2+) as well as the detergent SDS were strong inhibitors of the enzyme. The use of brewer's spent grain as substrate for xylanase production cannot only add value and decrease the amount of this waste but also reduce the xylanase production cost.

摘要

近几十年来,木聚糖酶已被广泛应用于许多加工业。本研究描述了利用啤酒糟作为底物生产青霉木聚糖酶的过程。此外,这是首次使用这种农业工业废物来纯化和表征木聚糖酶的工作。当青霉在 pH5.5、25°C、静置条件下生长六天时,可获得最佳产量。采用硫酸铵分级沉淀和分子筛层析的快速、廉价方法对青霉木聚糖酶进行了纯化为均相。SDS-PAGE 分析显示,该酶具有约 18.36 kDa 的估计分子质量,有一条带。该酶在 60°C、pH3.0 时具有最佳活性。该酶在 50°C 时非常稳定,并且在 pH2.5 至 5.0 之间验证了高 pH 稳定性。Mn(2+)离子和还原剂 β -巯基乙醇和 DTT 增强了木聚糖酶的活性,而 Hg(2+)、Zn(2+)和 Cu(2+)离子以及去污剂 SDS 则是该酶的强烈抑制剂。利用啤酒糟作为木聚糖酶生产的底物,不仅可以增加其附加值、减少这种废物的数量,还可以降低木聚糖酶的生产成本。

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