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Heterologous production of a feruloyl esterase fromPleurotus sapidussynthesizing feruloyl-saccharide esters
- Source :
- Biotechnology and Applied Biochemistry. 63:852-862
- Publication Year :
- 2015
- Publisher :
- Wiley, 2015.
-
Abstract
- The feruloyl esterase (FAE) gene EST1 from the basidiomycete Pleurotus sapidus was heterologously expressed in Escherichia coli and Pichia pastoris. Catalytically active recombinant Est1 was secreted using P. pastoris as a host. For expression in P. pastoris, the expression vector pPIC9K was applied. The EST1 gene was cloned with an N-terminal α-mating factor pre-pro sequence and expressed under the control of a methanol inducible alcohol oxidase 1 promotor. Est1 was purified to homogeneity using ion exchange and hydrophobic interaction chromatography. The recombinant Est1 showed optima at pH 5.0 and 50 °C, and released ferulic acid from saccharide esters and from the natural substrate destarched wheat bran. Substrate specificity profile and descriptor-based analysis demonstrated unique properties, showing that Est1 did not fit into the current FAE classification model. Transferuloylation synthesis of feruloyl-saccharide esters was proven for mono- and disaccharides.
- Subjects :
- 0301 basic medicine
Biomedical Engineering
Bioengineering
medicine.disease_cause
Applied Microbiology and Biotechnology
Pichia pastoris
law.invention
Ferulic acid
03 medical and health sciences
chemistry.chemical_compound
law
Feruloyl esterase
Drug Discovery
medicine
Escherichia coli
Expression vector
biology
Process Chemistry and Technology
Hydrophilic interaction chromatography
General Medicine
biology.organism_classification
Alcohol oxidase
030104 developmental biology
chemistry
Biochemistry
Recombinant DNA
Molecular Medicine
Biotechnology
Subjects
Details
- ISSN :
- 08854513
- Volume :
- 63
- Database :
- OpenAIRE
- Journal :
- Biotechnology and Applied Biochemistry
- Accession number :
- edsair.doi...........799650cbe9136536ab02887a769a098b