title: Influence of Randomly Inserted Feruloyl Esterase A on β-Glucosidase Activity in Trichoderma reesei.
abstract: As a well-known industrial fungus for cellulase production, the strain RUT-C30 of Trichoderma reesei was selected to produce the feruloyl esterase A (FAEA) by a random integration protocol. The strong promoter of cellobiohydrolase 1 (cbh1) gene was used to drive the expression of FAEA. Using double-joint PCR protocol, Pcbh1-faeA-TtrpC expression cassette was successfully constructed and co-transformed into RUT C30 strain of T. reesei. One transformant with high feruloyl esterase yield (3.44 ± 0.16 IU/mL) was obtained through plate screening and named TrfaeA1. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of fermentation supernatant from transformant TrfaeA1 showed a distinct protein band appearing at the position of about 34 kDa, indicating that faeA gene has been successfully expressed in T. reesei. Compared with that in original RUT C30 strain, β-glucosidase production in transformant TrfaeA1 was significantly increased by about 86.4%, reaching 63.2 IU/mL due to the random insertion of faeA. Moreover, the total secretion protein and filter paper activities of the transformant TrfaeA1 were also improved by up to 5.5 and 4.3%, respectively. The present results indicated that the random insertion strategy could be an effective and feasible method to improve and optimize the cellulase system of filamentous fungi.
authors.affiliation: Department of Bioengineering, Qilu University of Technology, Shandong Provincial Key Laboratory of Microbial Engineering, Jinan, 250353, People's Republic of China.
authors.forename: YunHua
authors.initials: Y
authors.lastname: Hou
authors.affiliation: Department of Bioengineering, Qilu University of Technology, Shandong Provincial Key Laboratory of Microbial Engineering, Jinan, 250353, People's Republic of China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Pan
authors.affiliation: Department of Bioengineering, Qilu University of Technology, Shandong Provincial Key Laboratory of Microbial Engineering, Jinan, 250353, People's Republic of China.
authors.forename: MengJie
authors.initials: M
authors.lastname: Yan
authors.affiliation: Department of Bioengineering, Qilu University of Technology, Shandong Provincial Key Laboratory of Microbial Engineering, Jinan, 250353, People's Republic of China.
authors.forename: Huan
authors.initials: H
authors.lastname: He
authors.affiliation: Department of Bioengineering, Qilu University of Technology, Shandong Provincial Key Laboratory of Microbial Engineering, Jinan, 250353, People's Republic of China. yangqinzheng@sina.com.
authors.forename: QinZheng
authors.initials: Q
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University, Jinan, Shandong, 250100, China.
authors.forename: YaoHua
authors.initials: Y
authors.lastname: Zhong
mesh_terms: D002265:Carboxylic Ester Hydrolases / Q000096:biosynthesis / Q000235:genetics; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics; D014242:Trichoderma / Q000201:enzymology / Q000235:genetics; D001617:beta-Glucosidase / Q000096:biosynthesis / Q000235:genetics
chemical_list: D005656:Fungal Proteins; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase; D001617:beta-Glucosidase
keywords: Feruloyl esterase; Random insertion strategy; Trichoderma reesei; β-glucosidase