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Co-expression of Beta-Glucosidase and Laccase in Trichoderma reesei by Random Insertion with Enhanced Filter Paper Activity.
- Source :
-
Molecular biotechnology [Mol Biotechnol] 2017 Aug; Vol. 59 (8), pp. 353-364. - Publication Year :
- 2017
-
Abstract
- Trichoderma reesei strain Rut-C30 was modified with enhanced beta-glycosidase (BGL) activity to balance the cellulase system and generated laccase (LAC) protein for lignin degradation. Initially, the binary plasmid p1300-w1 was constructed to express T. reesei bgl2 under the control of promoter P <subscript>pki</subscript> and T-nos terminator. Random insertion was performed via Agrobacterium tumefaciens-mediated transformation. A total of 353 mutants were obtained, and 34PTrb2 was exceptionally stable with increased FPA and BGL activity after screening for extracellular enzyme activity. Subsequently, 34PTrb2 was used as parent strain via the same method to insert the lac gene from Fomes lignosus, with promoter P <subscript>gpd</subscript> , followed by cbh1 signal peptide trss and T-nos as terminator. Several mutants successfully expressed enzyme LAC with stable activity of approximately 0.13 U/mL. The mutant 15Gsslac increased activity by 40.4% FPA compared with that of the host Rut-C30.
- Subjects :
- Carbohydrate Metabolism
Cellulose metabolism
Electrophoresis, Polyacrylamide Gel
Mitosis
Mutation genetics
Polymerase Chain Reaction
Transcription, Genetic
Transformation, Genetic
Trichoderma cytology
Filtration
Laccase metabolism
Mutagenesis, Insertional
Paper
Trichoderma enzymology
beta-Glucosidase metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1559-0305
- Volume :
- 59
- Issue :
- 8
- Database :
- MEDLINE
- Journal :
- Molecular biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 28676913
- Full Text :
- https://doi.org/10.1007/s12033-017-0018-7