title: Truncation of a mannanase from Trichoderma harzianum improves its enzymatic properties and expression efficiency in Trichoderma reesei.
abstract: To obtain high expression efficiency of a mannanase gene, ThMan5A, cloned from Trichoderma harzianum MGQ2, both the full-length gene and a truncated gene (ThMan5AΔCBM) that contains only the catalytic domain, were expressed in Trichoderma reesei QM9414 using the strong constitutive promoter of the gene encoding pyruvate decarboxylase (pdc), and purified to homogeneity, respectively. We found that truncation of the gene improved its expression efficiency as well as the enzymatic properties of the encoded protein. The recombinant strain expressing ThMan5AΔCBM produced 2,460 ± 45.1 U/ml of mannanase activity in the culture supernatant; 2.3-fold higher than when expressing the full-length ThMan5A gene. In addition, the truncated mannanase had superior thermostability compared with the full-length enzyme and retained 100 % of its activity after incubation at 60 °C for 48 h. Our results clearly show that the truncated ThMan5A enzyme exhibited improved characteristics both in expression efficiency and in its thermal stability. These characteristics suggest that ThMan5AΔCBM has potential applications in the food, feed, paper, and pulp industries.
authors.affiliation: College of Life Science, Shenzhen Key Laboratory of Microbial Genetic Engineering, Shenzhen University, Shenzhen, 518060, Guangdong, People's Republic of China, wangjuan@szu.edu.cn.
authors.forename: Juan
authors.initials: J
authors.lastname: Wang
authors.forename: Desheng
authors.initials: D
authors.lastname: Zeng
authors.forename: Gang
authors.initials: G
authors.lastname: Liu
authors.forename: Shaowen
authors.initials: S
authors.lastname: Wang
authors.forename: Shaowen
authors.initials: S
authors.lastname: Yu
mesh_terms: D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D013379:Substrate Specificity; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics; D044902:beta-Mannosidase / Q000235:genetics* / Q000378:metabolism*
chemical_list: D044902:beta-Mannosidase