Laccase activity measurement by FTIR spectral fingerprinting

Research output: Contribution to journalJournal article – Annual report year: 2019Researchpeer-review

Standard

Laccase activity measurement by FTIR spectral fingerprinting. / Perna, Valentina; Baum, Andreas; Ernst, Heidi A.; Agger, Jane W.; Meyer, Anne S.

In: Enzyme and Microbial Technology, Vol. 122, 2019, p. 64-73.

Research output: Contribution to journalJournal article – Annual report year: 2019Researchpeer-review

Harvard

APA

CBE

MLA

Vancouver

Author

Bibtex

@article{addaf61c55aa4b7d8bcbd0b9f826e931,
title = "Laccase activity measurement by FTIR spectral fingerprinting",
abstract = "Laccases (EC 1.10.3.2) are enzymes known for their ability to catalyze the oxidation of phenolic compounds using molecular oxygen as the final electron acceptor. Laccase activity is commonly determined by monitoring spectrophotometric changes (absorbance) of the product or substrate during the enzymatic reaction. Fourier Transform Infrared Spectroscopy (FTIR) is a fast and versatile technique where spectral evolution profiling, i.e. assessment of the spectral changes of both substrate and products during enzymatic conversion in real time, can be used to assess enzymatic activity when combined with multivariate data analysis. We employed FTIR to monitor enzymatic oxidation of monolignols (sinapyl, coniferyl and p-coumaryl alcohol), sinapic acid, and sinapic aldehyde by four different laccases: three fungal laccases from Trametes versicolor, Trametes villosa and Ganoderma lucidum, respectively, and one bacterial laccase from Meiothermus ruber. By coupling the FTIR measurements with Parallel Factor Analysis (PARAFAC) we established a quantitative assay for assessing laccase activity. By combining PARAFAC modelling with Principal Component Analysis we show the usefulness of this technology as a multivariate tool able to compare and distinguish different laccase reaction patterns. We also demonstrate how the FTIR approach can be used to create a reference system for laccase activity comparison based on a relatively low number of measurements. Such a reference system has potential to function as a high-throughput method for comparing reaction pattern similarities and differences between laccases and hereby identify new and interesting enzyme candidates in large sampling pools.",
keywords = "Laccase, Enzyme activity assay, Spectral evolution profiles, FTIR, PARAFAC, High-throughput",
author = "Valentina Perna and Andreas Baum and Ernst, {Heidi A.} and Agger, {Jane W.} and Meyer, {Anne S.}",
year = "2019",
doi = "10.1016/j.enzmictec.2018.12.009",
language = "English",
volume = "122",
pages = "64--73",
journal = "Enzyme and Microbial Technology",
issn = "0141-0229",
publisher = "Elsevier Inc",

}

RIS

TY - JOUR

T1 - Laccase activity measurement by FTIR spectral fingerprinting

AU - Perna, Valentina

AU - Baum, Andreas

AU - Ernst, Heidi A.

AU - Agger, Jane W.

AU - Meyer, Anne S.

PY - 2019

Y1 - 2019

N2 - Laccases (EC 1.10.3.2) are enzymes known for their ability to catalyze the oxidation of phenolic compounds using molecular oxygen as the final electron acceptor. Laccase activity is commonly determined by monitoring spectrophotometric changes (absorbance) of the product or substrate during the enzymatic reaction. Fourier Transform Infrared Spectroscopy (FTIR) is a fast and versatile technique where spectral evolution profiling, i.e. assessment of the spectral changes of both substrate and products during enzymatic conversion in real time, can be used to assess enzymatic activity when combined with multivariate data analysis. We employed FTIR to monitor enzymatic oxidation of monolignols (sinapyl, coniferyl and p-coumaryl alcohol), sinapic acid, and sinapic aldehyde by four different laccases: three fungal laccases from Trametes versicolor, Trametes villosa and Ganoderma lucidum, respectively, and one bacterial laccase from Meiothermus ruber. By coupling the FTIR measurements with Parallel Factor Analysis (PARAFAC) we established a quantitative assay for assessing laccase activity. By combining PARAFAC modelling with Principal Component Analysis we show the usefulness of this technology as a multivariate tool able to compare and distinguish different laccase reaction patterns. We also demonstrate how the FTIR approach can be used to create a reference system for laccase activity comparison based on a relatively low number of measurements. Such a reference system has potential to function as a high-throughput method for comparing reaction pattern similarities and differences between laccases and hereby identify new and interesting enzyme candidates in large sampling pools.

AB - Laccases (EC 1.10.3.2) are enzymes known for their ability to catalyze the oxidation of phenolic compounds using molecular oxygen as the final electron acceptor. Laccase activity is commonly determined by monitoring spectrophotometric changes (absorbance) of the product or substrate during the enzymatic reaction. Fourier Transform Infrared Spectroscopy (FTIR) is a fast and versatile technique where spectral evolution profiling, i.e. assessment of the spectral changes of both substrate and products during enzymatic conversion in real time, can be used to assess enzymatic activity when combined with multivariate data analysis. We employed FTIR to monitor enzymatic oxidation of monolignols (sinapyl, coniferyl and p-coumaryl alcohol), sinapic acid, and sinapic aldehyde by four different laccases: three fungal laccases from Trametes versicolor, Trametes villosa and Ganoderma lucidum, respectively, and one bacterial laccase from Meiothermus ruber. By coupling the FTIR measurements with Parallel Factor Analysis (PARAFAC) we established a quantitative assay for assessing laccase activity. By combining PARAFAC modelling with Principal Component Analysis we show the usefulness of this technology as a multivariate tool able to compare and distinguish different laccase reaction patterns. We also demonstrate how the FTIR approach can be used to create a reference system for laccase activity comparison based on a relatively low number of measurements. Such a reference system has potential to function as a high-throughput method for comparing reaction pattern similarities and differences between laccases and hereby identify new and interesting enzyme candidates in large sampling pools.

KW - Laccase

KW - Enzyme activity assay

KW - Spectral evolution profiles

KW - FTIR

KW - PARAFAC

KW - High-throughput

U2 - 10.1016/j.enzmictec.2018.12.009

DO - 10.1016/j.enzmictec.2018.12.009

M3 - Journal article

VL - 122

SP - 64

EP - 73

JO - Enzyme and Microbial Technology

JF - Enzyme and Microbial Technology

SN - 0141-0229

ER -