A β-glucosidase gene (bsbgl1a) from Bacillus sp. CGMCC 1.16541 was expressed in Escherichia coli BL21 and subsequently characterized. The amino acid sequence shared 83.64% identity with β-glucosidase (WP_066390903.1) from Fictibacillus phosphorivorans. The recombinant β-glucosidase (BsBgl1A) had a molecular weight of 52.2 kDa and could hydrolyze cellobiose, cellotriose, cellotetrose, p-nitrophenyl-β-d-glucopyranoside (pNPG), and p-nitrophenyl-β-d-xylopyranoside (pNPX). Optimal activity for BsBgl1A was recorded at 45 °C with a pH between 5.6 and 7.6, and 100% of its activity was maintained after a 24 h incubation between pH 4 and 9. Kinetic characterization revealed an enzymatic turnover (Kcat) of 616 ± 2 s⁻¹ (with cellobiose) and 3.5 ± 0.1 s⁻¹ (with p-nitrophenyl-β-d-glucopyranoside). Interestingly, the recombinant enzyme showed cupric ion (Cu²⁺), sodium dodecyl sulfate (SDS) and alcohol tolerance at 10 mM for Cu²⁺ and 10% for both SDS and alcohol. Additionally, BsBgl1A had high tolerance for glucose (Ki = 2095 mM), which is an extremely desirable feature for industrial applications. Following the addition of BsBgl1A (0.05 mg/ml) to a commercial cellulase reaction system, glucose yields from sugarcane bagasse increased 100% after 1 day at 45 °C. This work identifies a Cu²⁺, SDS, alcohol, and glucose tolerant GH1 β-glucosidase with potential applications in the hydrolysis of cellulose for the bioenergy industry.

来自芽孢杆菌属的β-葡萄糖苷酶基因（bsbgl1a）。CGMCC 1.16541在大肠杆菌BL21中表达，并随后进行了表征。该氨基酸序列与来自磷光杆菌的β-葡糖苷酶（WP_066390903.1）具有83.64％的同一性。重组β葡糖苷酶（BsBgl1A）具有52.2 kDa的分子量，并可以水解纤维二糖，纤维三糖，cellotetrose，对硝基苯基β- d吡喃葡萄糖苷（p NPG），和对硝基苯基β- d -xylopyranoside（pNPX）。BsBgl1A的最佳活性记录在45°C，pH在5.6和7.6之间，并且在pH 4和9之间孵育24小时后，其BsBgl1A的活性保持了100％。动力学表征显示酶转化率（Kcat）为616±2 s ^-1（具有纤维二糖）和3.5±0.1 s ^-1（具有对硝基苯基-β - d-吡喃葡萄糖苷）。有趣的是，该重组酶表现出的铜离子（铜²⁺），十二烷基硫酸钠（SDS）和酒精耐受性以10mM对Cu ²⁺SDS和酒精含量均为10％。此外，BsBgl1A对葡萄糖具有很高的耐受性（Ki = 2095 mM），这是工业应用中极为需要的功能。向商业纤维素酶反应系统中添加BsBgl1A（0.05 mg / ml）之后，在45°C下放置1天后，甘蔗渣中的葡萄糖产率提高了100％。这项工作确定了Cu 2 ⁺，SDS，酒精和葡萄糖耐性GH1β-葡萄糖苷酶在生物能源行业纤维素水解中的潜在应用。