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Influence of Randomly Inserted Feruloyl Esterase A on β-Glucosidase Activity in Trichoderma reesei
- Source :
- Applied biochemistry and biotechnology. 183(1)
- Publication Year :
- 2016
-
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.
- Subjects :
- 0301 basic medicine
030106 microbiology
Bioengineering
Cellulase
Biology
Applied Microbiology and Biotechnology
Biochemistry
Fungal Proteins
03 medical and health sciences
chemistry.chemical_compound
Feruloyl esterase
Sodium dodecyl sulfate
Molecular Biology
Polyacrylamide gel electrophoresis
Trichoderma reesei
Trichoderma
Strain (chemistry)
beta-Glucosidase
General Medicine
biology.organism_classification
chemistry
biology.protein
Fermentation
Expression cassette
Carboxylic Ester Hydrolases
Biotechnology
Subjects
Details
- ISSN :
- 15590291
- Volume :
- 183
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Applied biochemistry and biotechnology
- Accession number :
- edsair.doi.dedup.....7ed084ded45d61332eb23bfb27dc67ad