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Effect of fungal mycelia on the HPLC–UV and UV–vis spectrophotometric assessment of mycelium-bound epoxide hydrolase using glycidyl phenyl ether

Paper ID Volume ID Publish Year Pages File Format Full-Text
32914 44950 2016 11 PDF Available
Title
Effect of fungal mycelia on the HPLC–UV and UV–vis spectrophotometric assessment of mycelium-bound epoxide hydrolase using glycidyl phenyl ether
Abstract

•Matrix effects in mycelium-bound epoxide hydrolase-catalysed reactions were evaluated for the first time.•Mycelia retain some of the final product produced.•Calibration standards spiked before extraction were used to compensate for matrix retention effects.•The validated calibration method was successfully applied to real samples.

The use of mycelia as biocatalysts has technical and economic advantages. However, there are several difficulties in obtaining accurate results in mycelium-catalysed reactions. Firstly, sample extraction, indispensable because of the presence of mycelia, can bring into the extract components with a similar structure to that of the analyte of interest; secondly, mycelia can influence the recovery of the analyte. We prepared calibration standards of 3-phenoxy-1,2-propanediol (PPD) in the pure solvent and in the presence of mycelia (spiked before or after extraction) from five fungi (Aspergillus niger, Aspergillus tubingensis, Penicillium aurantiogriseum, Penicillium sp. and Aspergillus terreus). The quantification of PPD was carried out by HPLC–UV and UV–vis spectrophotometry. The manuscript shows that the last method is as accurate as the HPLC method. However, the colorimetric method led to a higher data throughput, which allowed the study of more samples in a shorter time. Matrix effects were evaluated visually from the plotted calibration data and statistically by simultaneously comparing the intercept and slope of calibration curves performed with solvent, post-extraction spiked standards and pre-extraction spiked standards. Significant differences were found between the post- and pre-extraction spiked matrix-matched functions. Pre-extraction spiked matrix-matched functions based on A. tubingensis mycelia, selected as the reference, were validated and used to compensate for low recoveries. These validated functions were successfully applied to the quantification of PPD achieved during the hydrolysis of glycidyl phenyl ether by mycelium-bound epoxide hydrolases and equivalent hydrolysis yields were determined by HPLC–UV and UV–vis spectrophotometry. This study may serve as starting point to implement matrix effects evaluation when mycelium-bound epoxide hydrolases are studied.

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Effect of fungal mycelia on the HPLC–UV and UV–vis spectrophotometric assessment of mycelium-bound epoxide hydrolase using glycidyl phenyl ether
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Publisher
Database: Elsevier - ScienceDirect
Journal: New Biotechnology - Volume 33, Issue 4, 25 June 2016, Pages 449–459
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us