Fungal feruloyl esterases: Functional validation of genome mining based enzyme discovery including uncharacterized subfamilies

Adiphol Dilokpimol; Miia R Mäkelä; Simona Varriale; Miaomiao Zhou; Gabriella Cerullo; Loknath Gidijala; Harri Hinkka; Joana L A Brás; Peter Jütten; Alexander Piechot; Raymond Verhaert; Kristiina S Hildén; Vincenza Faraco; Ronald P de Vries

doi:10.1016/j.nbt.2017.11.004

Fungal feruloyl esterases: Functional validation of genome mining based enzyme discovery including uncharacterized subfamilies

Adiphol Dilokpimol, Miia R Mäkelä, Simona Varriale, Miaomiao Zhou, Gabriella Cerullo, Loknath Gidijala, Harri Hinkka, Joana L A Brás, Peter Jütten, Alexander Piechot, Raymond Verhaert, Kristiina S Hildén, Vincenza Faraco, Ronald P de Vries

Molecular Plant Physiology

Fungal Physiology, Westerdijk Fungal Biodiversity Institute and Fungal Molecular Physiology, Utrecht University, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands.
Fungal Physiology, Westerdijk Fungal Biodiversity Institute & Fungal Molecular Physiology, Utrecht University, Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands; Department of Food and Environmental Sciences, Division of Microbiology and Biotechnology, P.O. Box 56, Viikinkaari 9, University of Helsinki, Helsinki, Finland, Finland.
Department of Chemical Sciences, University of Naples "Federico II", Complesso Universitario Monte S. Angelo Via Cintia, 4 IT-80126, Naples, Italy.
ProteoNic B.V., J.H. Oortweg 19-21, 2333 CH Leiden, The Netherlands.
NZYTech B.V., Estrada do Paço do Lumiar, Campus do Lumiar, Edifício E - 1° andar, 1649-038 Lisboa, Portugal.
Taros Chemicals GmbH & Co. KG, Emil-Figge-Str. 76a, 44227 Dortmund, Germany.
Division of Microbiology and Biotechnology, Department of Food and Environmental Sciences, University of Helsinki, Helsinki, Finland. Electronic address: [email protected].
Department of Chemical Sciences, University of Naples "Federico II", Complesso Universitario Monte S. Angelo Via Cintia, 4 IT-80126, Naples, Italy. Electronic address: [email protected].

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Feruloyl esterases (FAEs) are a diverse group of enzymes that specifically catalyze the hydrolysis of ester bonds between a hydroxycinnamic (e.g. ferulic) acid and plant poly- or oligosaccharides. FAEs as auxiliary enzymes significantly assist xylanolytic and pectinolytic enzymes in gaining access to their site of action during biomass saccharification for biofuel and biochemical production. A limited number of FAEs have been functionally characterized compared to over 1000 putative fungal FAEs that were recently predicted by similarity-based genome mining, which divided phylogenetically into different subfamilies (SFs). In this study, 27 putative and six characterized FAEs from both ascomycete and basidiomycete fungi were selected and heterologously expressed in Pichia pastoris and the recombinant proteins biochemically characterized to validate the previous genome mining and phylogenetical grouping and to expand the information on activity of fungal FAEs. As a result, 20 enzymes were shown to possess FAE activity, being active towards pNP-ferulate and/or methyl hydroxycinnamate substrates, and covering 11 subfamilies. Most of the new FAEs showed activities comparable to those of previously characterized fungal FAEs.

Original language	English
Pages (from-to)	9-14
Number of pages	6
Journal	New Biotechnology
Volume	41
DOIs	https://doi.org/10.1016/j.nbt.2017.11.004
Publication status	Published - 2018

Keywords

Carboxylic Ester Hydrolases/genetics
Data Mining
Fungi/enzymology
Genome, Fungal
Molecular Weight
Recombinant Proteins/biosynthesis
Reproducibility of Results
Substrate Specificity

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10.1016/j.nbt.2017.11.004

dilokpimol2Final published version, 191 KBLicence: Taverne

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Dilokpimol, A., Mäkelä, M. R., Varriale, S., Zhou, M., Cerullo, G., Gidijala, L., Hinkka, H., Brás, J. L. A., Jütten, P., Piechot, A., Verhaert, R., Hildén, K. S., Faraco, V., & de Vries, R. P. (2018). Fungal feruloyl esterases: Functional validation of genome mining based enzyme discovery including uncharacterized subfamilies. New Biotechnology, 41, 9-14. https://doi.org/10.1016/j.nbt.2017.11.004

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title = "Fungal feruloyl esterases: Functional validation of genome mining based enzyme discovery including uncharacterized subfamilies",

abstract = "Feruloyl esterases (FAEs) are a diverse group of enzymes that specifically catalyze the hydrolysis of ester bonds between a hydroxycinnamic (e.g. ferulic) acid and plant poly- or oligosaccharides. FAEs as auxiliary enzymes significantly assist xylanolytic and pectinolytic enzymes in gaining access to their site of action during biomass saccharification for biofuel and biochemical production. A limited number of FAEs have been functionally characterized compared to over 1000 putative fungal FAEs that were recently predicted by similarity-based genome mining, which divided phylogenetically into different subfamilies (SFs). In this study, 27 putative and six characterized FAEs from both ascomycete and basidiomycete fungi were selected and heterologously expressed in Pichia pastoris and the recombinant proteins biochemically characterized to validate the previous genome mining and phylogenetical grouping and to expand the information on activity of fungal FAEs. As a result, 20 enzymes were shown to possess FAE activity, being active towards pNP-ferulate and/or methyl hydroxycinnamate substrates, and covering 11 subfamilies. Most of the new FAEs showed activities comparable to those of previously characterized fungal FAEs.",

keywords = "Carboxylic Ester Hydrolases/genetics, Data Mining, Fungi/enzymology, Genome, Fungal, Molecular Weight, Recombinant Proteins/biosynthesis, Reproducibility of Results, Substrate Specificity",

author = "Adiphol Dilokpimol and M{\"a}kel{\"a}, {Miia R} and Simona Varriale and Miaomiao Zhou and Gabriella Cerullo and Loknath Gidijala and Harri Hinkka and Br{\'a}s, {Joana L A} and Peter J{\"u}tten and Alexander Piechot and Raymond Verhaert and Hild{\'e}n, {Kristiina S} and Vincenza Faraco and {de Vries}, {Ronald P}",

year = "2018",

doi = "10.1016/j.nbt.2017.11.004",

language = "English",

volume = "41",

pages = "9--14",

journal = "New Biotechnology",

issn = "1871-6784",

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Dilokpimol, A, Mäkelä, MR, Varriale, S, Zhou, M, Cerullo, G, Gidijala, L, Hinkka, H, Brás, JLA, Jütten, P, Piechot, A, Verhaert, R, Hildén, KS, Faraco, V & de Vries, RP 2018, 'Fungal feruloyl esterases: Functional validation of genome mining based enzyme discovery including uncharacterized subfamilies', New Biotechnology, vol. 41, pp. 9-14. https://doi.org/10.1016/j.nbt.2017.11.004

TY - JOUR

T1 - Fungal feruloyl esterases

T2 - Functional validation of genome mining based enzyme discovery including uncharacterized subfamilies

AU - Dilokpimol, Adiphol

AU - Mäkelä, Miia R

AU - Varriale, Simona

AU - Zhou, Miaomiao

AU - Cerullo, Gabriella

AU - Gidijala, Loknath

AU - Hinkka, Harri

AU - Brás, Joana L A

AU - Jütten, Peter

AU - Piechot, Alexander

AU - Verhaert, Raymond

AU - Hildén, Kristiina S

AU - Faraco, Vincenza

AU - de Vries, Ronald P

PY - 2018

Y1 - 2018

N2 - Feruloyl esterases (FAEs) are a diverse group of enzymes that specifically catalyze the hydrolysis of ester bonds between a hydroxycinnamic (e.g. ferulic) acid and plant poly- or oligosaccharides. FAEs as auxiliary enzymes significantly assist xylanolytic and pectinolytic enzymes in gaining access to their site of action during biomass saccharification for biofuel and biochemical production. A limited number of FAEs have been functionally characterized compared to over 1000 putative fungal FAEs that were recently predicted by similarity-based genome mining, which divided phylogenetically into different subfamilies (SFs). In this study, 27 putative and six characterized FAEs from both ascomycete and basidiomycete fungi were selected and heterologously expressed in Pichia pastoris and the recombinant proteins biochemically characterized to validate the previous genome mining and phylogenetical grouping and to expand the information on activity of fungal FAEs. As a result, 20 enzymes were shown to possess FAE activity, being active towards pNP-ferulate and/or methyl hydroxycinnamate substrates, and covering 11 subfamilies. Most of the new FAEs showed activities comparable to those of previously characterized fungal FAEs.

AB - Feruloyl esterases (FAEs) are a diverse group of enzymes that specifically catalyze the hydrolysis of ester bonds between a hydroxycinnamic (e.g. ferulic) acid and plant poly- or oligosaccharides. FAEs as auxiliary enzymes significantly assist xylanolytic and pectinolytic enzymes in gaining access to their site of action during biomass saccharification for biofuel and biochemical production. A limited number of FAEs have been functionally characterized compared to over 1000 putative fungal FAEs that were recently predicted by similarity-based genome mining, which divided phylogenetically into different subfamilies (SFs). In this study, 27 putative and six characterized FAEs from both ascomycete and basidiomycete fungi were selected and heterologously expressed in Pichia pastoris and the recombinant proteins biochemically characterized to validate the previous genome mining and phylogenetical grouping and to expand the information on activity of fungal FAEs. As a result, 20 enzymes were shown to possess FAE activity, being active towards pNP-ferulate and/or methyl hydroxycinnamate substrates, and covering 11 subfamilies. Most of the new FAEs showed activities comparable to those of previously characterized fungal FAEs.

KW - Carboxylic Ester Hydrolases/genetics

KW - Data Mining

KW - Fungi/enzymology

KW - Genome, Fungal

KW - Molecular Weight

KW - Recombinant Proteins/biosynthesis

KW - Reproducibility of Results

KW - Substrate Specificity

U2 - 10.1016/j.nbt.2017.11.004

DO - 10.1016/j.nbt.2017.11.004

M3 - Article

C2 - 29174720

SN - 1871-6784

VL - 41

SP - 9

EP - 14

JO - New Biotechnology

JF - New Biotechnology

ER -

Fungal feruloyl esterases: Functional validation of genome mining based enzyme discovery including uncharacterized subfamilies

Abstract

Keywords

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