Characterisation of a Novel Acetyl Xylan Esterase (BaAXE) Screened from the Gut Microbiota of the Common Black Slug (Arion ater)

Madubuike, Henry; Ferry, Natalie

Abstract

Acetyl xylan esterases (AXEs) are enzymes capable of hydrolysing the acetyl bonds in acetylated xylan, allowing for enhanced activity of backbone-depolymerizing enzymes. Bioprospecting novel AXE is essential in designing enzyme cocktails with desired characteristics targeting the complete breakdown of lignocellulose. In this article, we report the characterisation of a novel AXE identified as Gene_id_40363 in the metagenomic library analysed from the gut microbiota of the common black slug. The conserved domain description was identified with an NCBI BLASTp search using the translated nucleotide sequence as a query. The activity of the recombinant enzyme was tested on various synthetic substrates and acetylated substrates. The protein sequence matched the conserved domain described as putative hydrolase and aligned closely to an uncharacterized esterase from Buttiauxella agrestis, hence the designation as BaAXE. BaAXE showed low sequence similarity among characterized CE family proteins with an available 3D structure. BaAXE was active on 4-nitrophenyl acetate, reporting a specific activity of 78.12 U/mg and a Km value of 0.43 mM. The enzyme showed optimal activity at 40 °C and pH 8 and showed high thermal stability, retaining over 40% activity after 2 h of incubation from 40 °C to 100 °C. BaAXE hydrolysed acetyl bonds, releasing acetic acid from acetylated xylan and β-D-glucose pentaacetate. BaAXE has great potential for biotechnological applications harnessing its unique characteristics. In addition, this proves the possibility of bioprospecting novel enzymes from understudied environments.

Citation

Madubuike, H., & Ferry, N. (2022). Characterisation of a Novel Acetyl Xylan Esterase (BaAXE) Screened from the Gut Microbiota of the Common Black Slug (Arion ater). Molecules, 27(9), 2999. https://doi.org/10.3390/molecules27092999

• Journal Article Type: Article
• Acceptance Date: May 3, 2022
• Online Publication Date: May 7, 2022
• Publication Date: May 7, 2022
• Deposit Date: Jul 1, 2022
• Publicly Available Date: Jul 1, 2022
• Journal: Molecules
• Publisher: MDPI
• Volume: 27
• Issue: 9
• Pages: 2999
• DOI: https://doi.org/10.3390/molecules27092999
• Publisher URL: https://doi.org/10.3390/molecules27092999
• Related Public URLs: https://www.mdpi.com/journal/molecules