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首页> 外文期刊>Bioprocess and Biosystems Engineering >Production and characteristics of the recombinant extracellular bifunctional endoglucanase of the polyextremophilic bacterium Bacillus halodurans and its applicability in saccharifying agro-residues
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Production and characteristics of the recombinant extracellular bifunctional endoglucanase of the polyextremophilic bacterium Bacillus halodurans and its applicability in saccharifying agro-residues

机译：多嗜性芽孢杆菌嗜碱性芽孢杆菌的重组细胞外双功能内切葡聚糖酶的生产，特性及其在糖化农残中的应用

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摘要

The recombinant alkalistable and moderately thermostable bifunctional endoglucanase gene (BhCell-Xyl) of polyextremophilic bacterium Bacillus halodurans TSLV1 has been expressed in Pichia pastoris under constitutive GAP as well as inducible AOX promoters. A higher titre of recombinant BhCell-Xyl was attained after induction (4.8 U mL(-1)) as compared to that of the constitutive production (2.1 U mL(-1)). The recombinant P. pastoris strains integrated two copies of BhCell-Xyl under AOX and GAP promoters. The pure recombinant BhCell-Xyl is a glycoprotein of 66 kDa, which is optimally active at 60 A degrees C and pH 6.0 and 8.0. Glycosylated BhCell-Xyl exhibits higher thermostability than that of the native enzyme. The analysis of amino acids of BhCell-Xyl revealed that multiple factors are responsible for its thermostability. Kinetics and in silico analysis of the enzyme suggested that BhCell-Xyl has one active site for both endocellulase and endoxylanase activities. The BhCell-Xyl possesses a carbohydrate binding domain and saccharifies lignocellulosic agro-residues to xylo-oligosaccharides and cello-oligosaccharides, suggesting its potential application in generating fermentable sugars from renewable agro-residues for biofuel and fine chemical industries.
机译：多嗜性芽孢杆菌嗜碱性芽孢杆菌TSLV1的重组耐碱和中等耐温双功能内切葡聚糖酶基因（BhCell-Xyl）已在毕赤酵母中以组成型GAP和诱导型AOX启动子表达。与组成型生产（2.1 U mL（-1））相比，诱导后获得了更高的重组BhCell-Xyl滴度（4.8 U mL（-1））。重组巴斯德毕赤酵母菌株在AOX和GAP启动子下整合了两个拷贝的BhCell-Xyl。纯的重组BhCell-Xyl是66 kDa的糖蛋白，在60 A摄氏度和pH 6.0和8.0下具有最佳活性。糖基化的BhCell-Xyl具有比天然酶更高的热稳定性。 BhCell-Xyl氨基酸的分析表明，多种因素是其热稳定性的原因。该酶的动力学和计算机分析表明，BhCell-Xyl具有一个内切酶和内切木聚糖酶活性的活性位点。 BhCell-Xyl具有碳水化合物结合结构域，可将木质纤维素农业残渣糖化为木糖寡糖和纤维寡糖，这表明其在从可再生农业残渣生产可发酵糖用于生物燃料和精细化工行业中的潜在应用。

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