Oxidation of 5-hydroxymethylfurfural with a novel aryl alcohol oxidase from Mycobacterium sp. MS1601
(2022) In Microbial Biotechnology 15(8). p.2176-2190- Abstract
Bio-based 5-hydroxymethylfurfural (HMF) serves as an important platform for several chemicals, among which 2,5-furan dicarboxylic acid (FDCA) has attracted considerable interest as a monomer for the production of polyethylene furanoate (PEF), a potential alternative for fossil-based polyethylene terephthalate (PET). This study is based on the HMF oxidizing activity shown by Mycobacterium sp. MS 1601 cells and investigation of the enzyme catalysing the oxidation. The Mycobacterium whole cells oxidized the HMF to FDCA (60% yield) and hydroxymethyl furan carboxylic acid (HMFCA). A gene encoding a novel bacterial aryl alcohol oxidase, hereinafter MycspAAO, was identified in the genome and was cloned and expressed in... (More)
Bio-based 5-hydroxymethylfurfural (HMF) serves as an important platform for several chemicals, among which 2,5-furan dicarboxylic acid (FDCA) has attracted considerable interest as a monomer for the production of polyethylene furanoate (PEF), a potential alternative for fossil-based polyethylene terephthalate (PET). This study is based on the HMF oxidizing activity shown by Mycobacterium sp. MS 1601 cells and investigation of the enzyme catalysing the oxidation. The Mycobacterium whole cells oxidized the HMF to FDCA (60% yield) and hydroxymethyl furan carboxylic acid (HMFCA). A gene encoding a novel bacterial aryl alcohol oxidase, hereinafter MycspAAO, was identified in the genome and was cloned and expressed in Escherichia coli Bl21 (DE3). The purified MycspAAO displayed activity against several alcohols and aldehydes; 3,5 dimethoxy benzyl alcohol (veratryl alcohol) was the best substrate among those tested followed by HMF. 5-Hydroxymethylfurfural was converted to 5-formyl-2-furoic acid (FFCA) via diformyl furan (DFF) with optimal activity at pH 8 and 30–40°C. FDCA formation was observed during long reaction time with low HMF concentration. Mutagenesis of several amino acids shaping the active site and evaluation of the variants showed Y444F to have around 3-fold higher kcat/Km and ~1.7-fold lower Km with HMF.
(Less)
- author
- Sayed, Mahmoud LU ; Gaber, Yasser LU ; Junghus, Fredrik LU ; Martín, Eric Valdés ; Pyo, Sang-Hyun LU and Hatti-Kaul, Rajni LU
- organization
- publishing date
- 2022
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Microbial Biotechnology
- volume
- 15
- issue
- 8
- pages
- 2176 - 2190
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- pmid:35349220
- scopus:85127247036
- ISSN
- 1751-7907
- DOI
- 10.1111/1751-7915.14052
- project
- STEPS – Sustainable Plastics and Transition Pathways, Phase 2
- language
- English
- LU publication?
- yes
- additional info
- Publisher Copyright: © 2022 The Authors. Microbial Biotechnology published by Society for Applied Microbiology and John Wiley & Sons Ltd.
- id
- 3fd50f88-885e-4597-a651-3ab51a4a269c
- date added to LUP
- 2022-04-20 17:32:56
- date last changed
- 2024-11-05 00:36:19
@article{3fd50f88-885e-4597-a651-3ab51a4a269c, abstract = {{<p>Bio-based 5-hydroxymethylfurfural (HMF) serves as an important platform for several chemicals, among which 2,5-furan dicarboxylic acid (FDCA) has attracted considerable interest as a monomer for the production of polyethylene furanoate (PEF), a potential alternative for fossil-based polyethylene terephthalate (PET). This study is based on the HMF oxidizing activity shown by <i>Mycobacterium </i>sp. MS 1601 cells and investigation of the enzyme catalysing the oxidation. The <i>Mycobacterium </i>whole cells oxidized the HMF to FDCA (60% yield) and hydroxymethyl furan carboxylic acid (HMFCA). A gene encoding a novel bacterial aryl alcohol oxidase, hereinafter MycspAAO, was identified in the genome and was cloned and expressed in <i>Escherichia coli</i> Bl21 (DE3). The purified MycspAAO displayed activity against several alcohols and aldehydes; 3,5 dimethoxy benzyl alcohol (veratryl alcohol) was the best substrate among those tested followed by HMF. 5-Hydroxymethylfurfural was converted to 5-formyl-2-furoic acid (FFCA) via diformyl furan (DFF) with optimal activity at pH 8 and 30–40°C. FDCA formation was observed during long reaction time with low HMF concentration. Mutagenesis of several amino acids shaping the active site and evaluation of the variants showed Y444F to have around 3-fold higher k<sub>cat</sub>/K<sub>m</sub> and ~1.7-fold lower K<sub>m</sub> with HMF.</p>}}, author = {{Sayed, Mahmoud and Gaber, Yasser and Junghus, Fredrik and Martín, Eric Valdés and Pyo, Sang-Hyun and Hatti-Kaul, Rajni}}, issn = {{1751-7907}}, language = {{eng}}, number = {{8}}, pages = {{2176--2190}}, publisher = {{John Wiley & Sons Inc.}}, series = {{Microbial Biotechnology}}, title = {{Oxidation of 5-hydroxymethylfurfural with a novel aryl alcohol oxidase from <i>Mycobacterium </i>sp. MS1601}}, url = {{http://dx.doi.org/10.1111/1751-7915.14052}}, doi = {{10.1111/1751-7915.14052}}, volume = {{15}}, year = {{2022}}, }