Details
| Original language | English |
|---|---|
| Pages (from-to) | 852-862 |
| Number of pages | 11 |
| Journal | Biotechnology and Applied Biochemistry |
| Volume | 63 |
| Issue number | 6 |
| Publication status | Published - 14 Aug 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.
Keywords
- basidiomycete, esterification, feruloyl esterase, functional expression, Pichia pastoris, Pleurotus sapidus
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Chemical Engineering(all)
- Bioengineering
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Medicine
- Engineering(all)
- Biomedical Engineering
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
- Pharmacology, Toxicology and Pharmaceutics(all)
- Drug Discovery
- Chemical Engineering(all)
- Process Chemistry and Technology
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In: Biotechnology and Applied Biochemistry, Vol. 63, No. 6, 14.08.2015, p. 852-862.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Heterologous production of a feruloyl esterase from Pleurotus sapidus synthesizing feruloyl-saccharide esters
AU - Kelle, Sebastian
AU - Nieter, Annabel
AU - Krings, Ulrich
AU - Zelena, Katerina
AU - Linke, Diana
AU - Berger, Ralf G.
PY - 2015/8/14
Y1 - 2015/8/14
N2 - 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.
AB - 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.
KW - basidiomycete
KW - esterification
KW - feruloyl esterase
KW - functional expression, Pichia pastoris
KW - Pleurotus sapidus
UR - http://www.scopus.com/inward/record.url?scp=84950106048&partnerID=8YFLogxK
U2 - 10.1002/bab.1430
DO - 10.1002/bab.1430
M3 - Article
C2 - 26272349
AN - SCOPUS:84950106048
VL - 63
SP - 852
EP - 862
JO - Biotechnology and Applied Biochemistry
JF - Biotechnology and Applied Biochemistry
SN - 0885-4513
IS - 6
ER -