New Article Published in Applied Microbiology and Biotechnology (IF 3.9)

Happy to share our recent collaborative paper of BioSIM (Sérgio F. Sousa) with the group of Prof. Lígia Rodrigues at Univ Minho, published in the Applied Microbiology and Biotechnology journal (IF 3.9).

In this study, a compost metagenomic library was screened using function-driven methods and automated halo area extraction via MATLAB, leading to the discovery of a promising xylanase-producing clone, LP4. The xylanase from this clone, named Xyl4, belongs to glycosyl hydrolase family 10 (GH10). When expressed in Escherichia coli BL21(DE3), Xyl4’s enzyme activity doubled compared to the LP4 clone. Structural prediction and docking studies of Xyl4 with xylohexaose revealed the importance of amino acids Glu133 and Glu238 for catalysis. Additionally, it was suggested that replacing Phe316 with a bulkier Trp could enhance Xyl4 activity. These findings offer insights into improving Xyl4’s catalytic performance for industrial applications.

Heterologous expression and structure prediction of a xylanase identified from a compost metagenomic library

Joana Sousa, Cátia Santos-Pereira, Joana S. Gomes, Ângela M. A. Costa, Andréia O. Santos, Ricardo Franco-Duarte, João M. M. Linhares, Sérgio F. Sousa, Sara C. Silvério Vieira, Lígia R. Rodrigues

Applied Microbiology and Biotechnology, 108, 329 (2024) | DOI: 10.1007/s00253-024-13169-4

Abstract:

Xylanases are key biocatalysts in the degradation of the β‐1,4‐glycosidic linkages in the xylan backbone of hemicellulose. These enzymes are potentially applied in a wide range of bioprocessing industries under harsh conditions. Metagenomics has emerged as powerful tools for the bioprospection and discovery of interesting bioactive molecules from extreme ecosystems with unique features, such as high temperatures. In this study, an innovative combination of function-driven screening of a compost metagenomic library and automatic extraction of halo areas with in-house MATLAB functions resulted in the identification of a promising clone with xylanase activity (LP4). The LP4 clone proved to be an effective xylanase producer under submerged fermentation conditions. Sequence and phylogenetic analyses revealed that the xylanase, Xyl4, corresponded to an endo-1,4-β-xylanase belonging to glycosyl hydrolase family 10 (GH10). When xyl4 was expressed in Escherichia coli BL21(DE3), the enzyme activity increased about 2-fold compared to the LP4 clone. To get insight on the interaction of the enzyme with the substrate and establish possible strategies to improve its activity, the structure of Xyl4 was predicted, refined, and docked with xylohexaose. Our data unveiled, for the first time, the relevance of the amino acids Glu133 and Glu238 for catalysis, and a close inspection of the catalytic site suggested that the replacement of Phe316 by a bulkier Trp may improve Xyl4 activity. Our current findings contribute to enhancing the catalytic performance of Xyl4 towards industrial applications..

Leave a comment