Enhanced Xylanase Production from Bacillus safensis MABS6 using Sorghum Straw Substrate: Optimization, Characterization, and Biotechnological Applications

Amal Hassan Al-Shawi
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Abstract

The objective of this study was to evaluate the synthesis of xylanase enzyme by Bacillus safensis MABS6 using sorghum straw as a substrate. A comprehensive investigation was conducted to optimize xylanase yield by examining various parameters such as sorghum concentration, inoculum size, culture age, pH, temperature, and agitation speed. Additionally, the impact of nutritional additives, metallic ions, organic solvents, and alkaline H 2 O 2 treatment on xylanase production and activity was explored. Experimental trials were performed with specific parameters, including 4% w/v sorghum concentration, 2% inoculum size, 12 hours of culture age, pH 7, 35°C, and 250 rpm. Further improvements involved the addition of nutritional additives such as gelatin, xylose, and potassium nitrate. The effects of initial conditions, various metallic ions (Ca 2+ , Mn 2+ , and Fe 3+ ), and organic solvents (2-methyl propanol, methanol, and ethanol) on xylanase activity were evaluated. Additionally, saccharification investigations with a 4% alkaline H2O2 treatment assessed the enzymatic hydrolysis of sorghum straw. Optimized conditions resulted in a significant increase, with xylanase production reaching 3.49%. The partially purified xylanase exhibited over 60% relative activity within a pH range of 6 to 9 and demonstrated more than 55% activity between 45°C to 65°C after 1 hour. Certain metallic ions and organic solvents further enhanced xylanase activity. Notably, the xylanase derived from Bacillus safensis MABS6, utilizing sorghum straw, showcased desirable characteristics such as heat stability, alkali-solvent stability, and absence of cellulase activity. Its potential as a biocatalyst makes it valuable for various biotechnological applications, particularly in the efficient enzymatic hydrolysis of sorghum straw.
利用高粱秸秆底物提高 safensis MABS6 杆菌生产木聚糖酶的能力:优化、表征和生物技术应用
本研究的目的是评估 safensis MABS6 杆菌以高粱秸秆为底物合成木聚糖酶的情况。通过对高粱浓度、接种物大小、培养时间、pH 值、温度和搅拌速度等各种参数的研究,对木聚糖酶产量进行了全面的优化。此外,还探讨了营养添加剂、金属离子、有机溶剂和碱性 H 2 O 2 处理对木聚糖酶产量和活性的影响。实验采用了特定的参数,包括 4% w/v 高粱浓度、2% 接种物大小、12 小时培养时间、pH 值 7、35°C 和 250 转/分。进一步的改进包括添加营养添加剂,如明胶、木糖和硝酸钾。评估了初始条件、各种金属离子(Ca 2+ 、Mn 2+ 和 Fe 3+ )和有机溶剂(2-甲基丙醇、甲醇和乙醇)对木聚糖酶活性的影响。此外,用 4% 的碱性 H2O2 处理进行的糖化研究评估了高粱秸秆的酶水解作用。优化后的条件显著提高了木聚糖酶的产量,达到 3.49%。部分纯化的木聚糖酶在 pH 值为 6 至 9 的范围内显示出 60% 以上的相对活性,在 45°C 至 65°C 温度范围内,1 小时后显示出 55% 以上的活性。某些金属离子和有机溶剂进一步提高了木聚糖酶的活性。值得注意的是,利用高粱秸秆从 safensis MABS6 杆菌中提取的木聚糖酶具有热稳定性、碱-溶剂稳定性和无纤维素酶活性等理想特性。其作为生物催化剂的潜力使其在各种生物技术应用中,尤其是在高效酶解高粱秸秆方面具有重要价值。
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