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来自龙舌兰芽孢杆菌CGX5-1的一种新型1,3-1,4-β-葡聚糖酶基因的特性分析

Characterization of a new 1,3-1,4-β-glucanase gene from Bacillus tequilensis CGX5-1.

作者信息

Wang Jinjing, Niu Chengtuo, Liu Xiaoling, Chen Xi, Li Qi

机构信息

Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, Jiangsu, China.

出版信息

Appl Biochem Biotechnol. 2014 Jun;173(3):826-37. doi: 10.1007/s12010-014-0900-8. Epub 2014 Apr 12.

Abstract

1,3-1,4-β-Glucanase received great interest due to its application in brewing and feed industries. Application of 1,3-1,4-β-glucanase in brewing industry helps make up for the defect that plant-derived β-glucanases are heat-sensitive. A new strain, CGX5-1, exhibited remarkable 1,3-1,4-β-glucanase, was isolated from Asian giant hornet nest and identified Bacillus tequilensis. Moreover, a new 1,3-1,4-β-glucanase gene from B. tequilensis was cloned and measured to be 720 bp encoding 239 amino acids, with a predicted molecular weight of 26.9 kDa. After expressed in Escherichia coli BL21, active recombinant enzyme of 24 kDa was detected in the supernatant of cell culture, with the activity of 2,978.2 U/mL. The new enzyme was stable in the pH 5.0-7.5 with the highest activity measured at pH 6.0. Moreover, it is thermostable within 45 to 60 °C. The property of the new recombinant enzyme makes this enzyme a broad prospect in brewing industry. Moreover, this is the first report on 1,3-1,4-β-glucanase produced by B. tequilensis.

摘要

1,3-1,4-β-葡聚糖酶因其在酿造和饲料行业的应用而备受关注。1,3-1,4-β-葡聚糖酶在酿造工业中的应用有助于弥补植物来源的β-葡聚糖酶对热敏感的缺陷。从亚洲大黄蜂巢中分离出一株表现出显著1,3-1,4-β-葡聚糖酶活性的新菌株CGX5-1,并鉴定为特基拉芽孢杆菌。此外,克隆了来自特基拉芽孢杆菌的一个新的1,3-1,4-β-葡聚糖酶基因,测得其长度为720 bp,编码239个氨基酸,预测分子量为26.9 kDa。在大肠杆菌BL21中表达后,在细胞培养上清液中检测到活性为24 kDa的重组酶,活性为2978.2 U/mL。该新酶在pH 5.0-7.5范围内稳定,在pH 6.0时活性最高。此外,它在45至60°C范围内具有热稳定性。这种新重组酶的特性使其在酿造工业中具有广阔的应用前景。此外,这是关于特基拉芽孢杆菌产生1,3-1,4-β-葡聚糖酶的首次报道。

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