Cai Zhi, Wang Yi, You Yang, Yang Nan, Lu Shanshan, Xue Jianheng, Xing Xiang, Sha Sha, Zhao Lihua
SDU-ANU Joint Science College, Shandong University, Weihai 264209, China.
Marine College, Shandong University, Weihai 264209, China.
Microorganisms. 2024 May 13;12(5):979. doi: 10.3390/microorganisms12050979.
Enzyme-production microorganisms typically occupy a dominant position in composting, where cellulolytic microorganisms actively engage in the breakdown of lignocellulose. Exploring strains with high yields of cellulose-degrading enzymes holds substantial significance for the industrial production of related enzymes and the advancement of clean bioenergy. This study was inclined to screen cellulolytic bacteria, conduct genome analysis, mine cellulase-related genes, and optimize cellulase production. The potential carboxymethylcellulose-hydrolyzing bacterial strain Z2.6 was isolated from the maturation phase of pig manure-based compost with algae residuals as the feedstock and identified as . In the draft genome of strain Z2.6, 31 related cellulolytic genes were annotated by the CAZy database, and further validation by cloning documented the existence of an endo-1,4-β-D-glucanase (EC 3.2.1.4) belonging to the GH5 family and a β-glucosidase (EC 3.2.1.21) belonging to the GH1 family, which are predominant types of cellulases. Through the exploration of ten factors in fermentation medium with Plackett-Burman and Box-Behnken design methodologies, maximum cellulase activity was predicted to reach 2.98 U/mL theoretically. The optimal conditions achieving this response were determined as 1.09% CMC-Na, 2.30% salinity, and 1.23% tryptone. Validation under these specified conditions yielded a cellulose activity of 3.02 U/mL, demonstrating a 3.43-fold degree of optimization. In conclusion, this comprehensive study underscored the significant capabilities of strain Z2.6 in lignocellulolytic saccharification and its potentialities for future in-depth exploration in biomass conversion.
产酶微生物在堆肥过程中通常占据主导地位,其中纤维素分解微生物积极参与木质纤维素的分解。探索高产纤维素降解酶的菌株对于相关酶的工业生产和清洁生物能源的发展具有重要意义。本研究倾向于筛选纤维素分解细菌,进行基因组分析,挖掘纤维素酶相关基因,并优化纤维素酶的生产。从以藻类残渣为原料的猪粪基堆肥成熟期分离出潜在的羧甲基纤维素水解细菌菌株Z2.6,并鉴定为 。在菌株Z2.6的基因组草图中,CAZy数据库注释了31个相关纤维素分解基因,通过克隆进一步验证记录了属于GH5家族的内切-1,4-β-D-葡聚糖酶(EC 3.2.1.4)和属于GH1家族的β-葡萄糖苷酶(EC 3.2.1.21)的存在,它们是纤维素酶的主要类型。通过Plackett-Burman和Box-Behnken设计方法对发酵培养基中的十个因素进行探索,理论上预测最大纤维素酶活性可达2.98 U/mL。实现该响应的最佳条件确定为1.09% CMC-Na、2.30%盐度和1.23%胰蛋白胨。在这些特定条件下的验证产生了3.02 U/mL的纤维素活性,表明优化程度为3.43倍。总之,这项综合研究强调了菌株Z2.6在木质纤维素糖化方面的显著能力及其在生物质转化方面未来深入探索的潜力。