Department of Microbiology, Guru Nanak Dev University, Amritsar 143005, Punjab, India.
Center for Structural and Functional Genomics, Concordia University, 7141 Sherbrooke Street West, Montreal, Quebec H4B 1R6, Canada.
Bioresour Technol. 2022 May;351:127039. doi: 10.1016/j.biortech.2022.127039. Epub 2022 Mar 19.
The objective of this study was to develop thermophilic fungus Rasamsonia emersonii using integrated system biology tools (genomics, proteomics and transcriptional analysis) in combination with classical strain breeding approaches. Developed hyper cellulolytic mutant strain M36 showed endoglucanase (476.35 U/ml), β-glucosidase (70.54 U/ml), cellobiohydrolase (15.17 U/ml), FPase (4.89 U/ml) and xylanase (485.21 U/ml) on cellulose/gram flour based production medium. Comparison of the expression profile at proteome and transcriptional level of the developed strain and wild type parent gave detailed insight into the up-regulation of different CAZymes including glycosyl hydrolases (GH5, GH6, GH7, GH3, GH10) and auxiliary enzymes (lytic polysaccharide monooxygenase, swollenin) at system level. Furthermore, the potential of lignocellulolytic enzyme produced by the developed strain and custom designed cocktail spiked with heterologously expressed lytic polysaccharide monooxygenase from Mycothermus thermophiloides were analyzed for the hydrolysis of biorefinery relevant unwashed pretreated rice straw slurry (PRAJ and IOCL) @17% substrate loading rate.
本研究的目的是利用整合系统生物学工具（基因组学、蛋白质组学和转录分析）结合经典的菌株选育方法，开发嗜热真菌 Rasamsonia emersonii。开发的高纤维素酶突变株 M36 在基于纤维素/麸皮的生产培养基上表现出内切葡聚糖酶（476.35 U/ml）、β-葡萄糖苷酶（70.54 U/ml）、纤维二糖水解酶（15.17 U/ml）、FPase（4.89 U/ml）和木聚糖酶（485.21 U/ml）。比较开发菌株和野生型亲本在蛋白质组和转录水平的表达谱，深入了解不同 CAZymes 的上调，包括糖苷水解酶（GH5、GH6、GH7、GH3、GH10）和辅助酶（溶菌多糖单加氧酶、肿胀素）在系统水平上。此外，还分析了开发菌株和定制设计的鸡尾酒产生的木质纤维素酶的潜力，该鸡尾酒中含有异源表达的来自嗜热真菌 Mycothermus thermophiloides 的溶菌多糖单加氧酶，用于水解生物精炼厂相关的未洗涤预处理水稻秸秆浆料（PRAJ 和 IOCL）@17%的底物加载率。
