Molecular Characterization, Production and Optimization of Cellulase Producer and its Industrial Applications

Nimisha Dharmesh Patel, Dharmesh Amrutbhai Patel
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Abstract

Background: Plant cell walls are composed primarily of cellulose, making it the most common organic polymer on Earth. The utilization of microorganisms to convert cellulose into valuable products has gained significant attention worldwide. This study aims to isolate novel cellulase-producing microorganisms and investigate the influence of dietary and physicochemical factors on cellulase production, as well as evaluate the potential applications of a selected isolated strain. Materials and Methods: The Bacillus amyloliquefaciens strain was identified as the most efficient producer of cellulase among various cellulase-producing bacterial strains isolated during the research. The morphological and biochemical characteristics of this strain were characterized. To determine the optimal conditions for cellulase production, various parameters such as raw substrates, carbon sources, nitrogen sources, temperature, inoculum age, inoculum size, agitation rate, incubation duration, and pH were investigated. Results: The study found that sugarcane bagasse, molasses, and paper pulp exhibited the highest cellulase production capacity, with respective enzyme activities of 0.97, 0.98, and 0.88 IU/mL. Maximum cellulase production occurred at a temperature of 37°C, an inoculum size of 2.5%, an inoculum age of 24 hours, an agitation rate of 120 rpm, an incubation time of 48 hours, and a pH of 7 with 1% peptone. The Bacillus amyloliquefaciens isolate was successfully applied to convert lignocellulosic waste into ethanol and fermentable sugars through saccharification and fermentation of cellulose, demonstrating its significant effectiveness. Conclusion: Bacillus amyloliquefaciens strain isolated in this study proved to be a highly efficient producer of cellulase. The optimized production media led to improved enzyme production, and the isolate showed promising results in the industrial application of converting cellulosic waste into ethanol. The 16s rRNA sequencing confirmed the identity of Bacillus amyloliquefaciens , which was deposited under the accession number MN081796 in the NCBI database. In this study we discussed the possible low-cost, enzymatic pretreatment methods of lignocellulosic material in order to use it as an efficient raw material for biofuel production. These findings contribute to the understanding and potential utilization of cellulose producing microorganisms in various biotechnological applications.
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纤维素酶生成物的分子表征、生产与优化及其工业应用
背景:植物细胞壁主要由纤维素组成,使其成为地球上最常见的有机聚合物。利用微生物将纤维素转化为有价值的产品已经引起了全世界的广泛关注。本研究旨在分离新型纤维素酶生产微生物,研究膳食和理化因素对纤维素酶生产的影响,并评价筛选出的分离菌株的潜在应用价值。材料与方法:在本研究分离的多种产纤维素酶菌株中,解淀粉芽孢杆菌是产纤维素酶效率最高的菌株。对该菌株的形态和生化特征进行了表征。为了确定纤维素酶生产的最佳条件,研究了原料底物、碳源、氮源、温度、接种年龄、接种量、搅拌速率、孵育时间和pH等参数。结果:研究发现蔗渣、糖蜜和纸浆的纤维素酶生产能力最高,酶活性分别为0.97、0.98和0.88 IU/mL。在温度为37℃,接种量为2.5%,接种年龄为24小时,搅拌速度为120转/分,孵育时间为48小时,pH为7,蛋白胨含量为1%的条件下,纤维素酶产量最大。解淀粉芽孢杆菌分离物通过对纤维素的糖化和发酵,成功地将木质纤维素废物转化为乙醇和可发酵糖,证明了其显著的有效性。结论:本研究分离得到的解淀粉芽孢杆菌是一株高效产纤维素酶的菌株。优化后的生产培养基提高了酶的产量,该分离物在将纤维素废料转化为乙醇的工业应用中显示出良好的效果。16s rRNA测序证实了该解淀粉芽孢杆菌的身份,该菌株已在NCBI数据库中保存,登录号为MN081796。在这项研究中,我们讨论了可能的低成本,木质纤维素材料的酶预处理方法,以便将其用作生物燃料生产的有效原料。这些发现有助于对产纤维素微生物在各种生物技术应用中的认识和潜在利用。
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