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Biotechnology and Bioengineering

Biotechnology and Bioengineering

IF: 3.5
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Deciphering the Catalytic Proficiency and Mechanism of the N‐Acetylglucosamine Deacetylase From Pantoea dispersa

Published:26 November 2024 DOI: 10.1002/bit.28894 PMID: 39592864
Wentao Yang, Xiao Chen, Li Pang, Hong Tian, Liang Yang, Bo Xia

Abstract

Glucosamine (GlcN) is a widely utilized amino monosaccharide. It is traditionally synthesized from N-acetylglucosamine (GlcNAc) via chemical processes that pose environmental threats. In pursuit of a greener alternative, our investigation explored biocatalysis with a Pantoea dispersa derived deacetylase (Pd-nagA), showcasing its efficacy as a catalyst in GlcN production. As a result, this work provides a comprehensive characterization of Pd-nagA, scrutinizes its enzymatic behavior, and delves into the deacetylation mechanism in detail. Heterologous expression methods were utilized for the production and isolation of Pd-nagA, followed by a kinetic evaluation highlighting its enzymatic activity. The complex interactions between the enzyme and its substrate were investigated by integrating classical molecular dynamics, quantum mechanics/molecular mechanics simulations, funnel metadynamics, and on-the-fly probability enhanced sampling techniques, thereby elucidating the precise deacetylation pathway. Rigorous computational analysis results demonstrated that Pd-nagA exhibited promising specificity and efficiency for GlcNAc with a high turnover rate. The catalytic residues central to the reaction were identified, and the underlying quantum reaction mechanism was detailed. Our findings suggest an approach to GlcN production using eco-friendly biocatalysis, positioning Pd-nagA at the forefront of industrial application not only because of its remarkable catalytic capabilities but also due to its potential for enzyme optimization.

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