Bioinformatics Analysis of Diadenylate Cyclase Regulation on Cyclic Diadenosine Monophosphate Biosynthesis in Exopolysaccharide Production by <i>Leuconostoc mesenteroides</i> DRP105

Lactic acid bacteria exopolysaccharides (EPS) have a variety of excellent biological functions and are widely used in the food and pharmaceutical industries. The complex metabolic system of lactic acid bacteria and the mechanism of EPS biosynthesis have not been fully analyzed, which limits the wide...

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Main Authors: Wenna Yu, Liansheng Yu, Tengxin Li, Ziwen Wang, Renpeng Du, Wenxiang Ping
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Fermentation
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Online Access:https://www.mdpi.com/2311-5637/11/4/196
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author Wenna Yu
Liansheng Yu
Tengxin Li
Ziwen Wang
Renpeng Du
Wenxiang Ping
author_facet Wenna Yu
Liansheng Yu
Tengxin Li
Ziwen Wang
Renpeng Du
Wenxiang Ping
author_sort Wenna Yu
collection DOAJ
description Lactic acid bacteria exopolysaccharides (EPS) have a variety of excellent biological functions and are widely used in the food and pharmaceutical industries. The complex metabolic system of lactic acid bacteria and the mechanism of EPS biosynthesis have not been fully analyzed, which limits the wider application of EPS. EPS synthesis is regulated by cyclic diadenosine monophosphate (c-di-AMP), but the exact mechanism remains unclear. <i>Dac</i> and <i>pde</i> are c-di-AMP anabolic genes, <i>gtfA</i>, <i>gtfB</i> and <i>gtfC</i> are EPS synthesis gene clusters, among which <i>gtfC</i> was the key gene for EPS synthesis in <i>Leuconostoc mesenteroides</i> DRP105. In order to explore whether diadenylate cyclase (DAC) can catalyze the synthesis of c-di-AMP from ATP, the sequence of DAC was analyzed by bioinformatics based on the whole genome sequence. DAC was a CdaA type diadenylate cyclase containing the classical domain DisA_N and DGA and RHR motifs. The secondary structure was mainly composed of α-helices, and AlphaFold2 was used to model the 3D structure of the protein and evaluate the rationality of the DAC protein structure model. A total of 8 salt bridges, 21 hydrogen bonds and 221 non-bonded interactions were found between DAC and GtfC. Molecular docking simulations revealed ATP1 and ATP2 fully occupied the binding pocket of DAC and interacted directly with the binding site residues of DAC. The molecular dynamics simulations showed that the binding of DAC to ATP molecules was relatively stable. Gene and enzyme correlation analysis found that <i>dac</i> and <i>gtfC</i> gene expression were significantly positively correlated with DAC enzyme activity, c-di-AMP content and EPS production, and had no significant correlation with PDE enzyme activity responsible for c-di-AMP degradation. Bioinformatics analysis of the regulatory role of DAC in the synthesis of EPS by lactic acid bacteria was helpful to fully reveal the biosynthetic mechanism of EPS and provide theoretical basis for large-scale industrial production of EPS.
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spelling doaj-art-747e651ea4a04a2993dd8e09027fcbea2025-08-20T02:28:28ZengMDPI AGFermentation2311-56372025-04-0111419610.3390/fermentation11040196Bioinformatics Analysis of Diadenylate Cyclase Regulation on Cyclic Diadenosine Monophosphate Biosynthesis in Exopolysaccharide Production by <i>Leuconostoc mesenteroides</i> DRP105Wenna Yu0Liansheng Yu1Tengxin Li2Ziwen Wang3Renpeng Du4Wenxiang Ping5Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education & Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region & Key Laboratory of Microbiology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin 150080, ChinaEngineering Research Center of Agricultural Microbiology Technology, Ministry of Education & Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region & Key Laboratory of Microbiology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin 150080, ChinaEngineering Research Center of Agricultural Microbiology Technology, Ministry of Education & Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region & Key Laboratory of Microbiology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin 150080, ChinaEngineering Research Center of Agricultural Microbiology Technology, Ministry of Education & Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region & Key Laboratory of Microbiology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin 150080, ChinaEngineering Research Center of Agricultural Microbiology Technology, Ministry of Education & Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region & Key Laboratory of Microbiology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin 150080, ChinaEngineering Research Center of Agricultural Microbiology Technology, Ministry of Education & Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region & Key Laboratory of Microbiology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin 150080, ChinaLactic acid bacteria exopolysaccharides (EPS) have a variety of excellent biological functions and are widely used in the food and pharmaceutical industries. The complex metabolic system of lactic acid bacteria and the mechanism of EPS biosynthesis have not been fully analyzed, which limits the wider application of EPS. EPS synthesis is regulated by cyclic diadenosine monophosphate (c-di-AMP), but the exact mechanism remains unclear. <i>Dac</i> and <i>pde</i> are c-di-AMP anabolic genes, <i>gtfA</i>, <i>gtfB</i> and <i>gtfC</i> are EPS synthesis gene clusters, among which <i>gtfC</i> was the key gene for EPS synthesis in <i>Leuconostoc mesenteroides</i> DRP105. In order to explore whether diadenylate cyclase (DAC) can catalyze the synthesis of c-di-AMP from ATP, the sequence of DAC was analyzed by bioinformatics based on the whole genome sequence. DAC was a CdaA type diadenylate cyclase containing the classical domain DisA_N and DGA and RHR motifs. The secondary structure was mainly composed of α-helices, and AlphaFold2 was used to model the 3D structure of the protein and evaluate the rationality of the DAC protein structure model. A total of 8 salt bridges, 21 hydrogen bonds and 221 non-bonded interactions were found between DAC and GtfC. Molecular docking simulations revealed ATP1 and ATP2 fully occupied the binding pocket of DAC and interacted directly with the binding site residues of DAC. The molecular dynamics simulations showed that the binding of DAC to ATP molecules was relatively stable. Gene and enzyme correlation analysis found that <i>dac</i> and <i>gtfC</i> gene expression were significantly positively correlated with DAC enzyme activity, c-di-AMP content and EPS production, and had no significant correlation with PDE enzyme activity responsible for c-di-AMP degradation. Bioinformatics analysis of the regulatory role of DAC in the synthesis of EPS by lactic acid bacteria was helpful to fully reveal the biosynthetic mechanism of EPS and provide theoretical basis for large-scale industrial production of EPS.https://www.mdpi.com/2311-5637/11/4/196cyclic diadenosine monophosphatediadenylate cyclaseglucansucrasebioinformaticsexopolysaccharides
spellingShingle Wenna Yu
Liansheng Yu
Tengxin Li
Ziwen Wang
Renpeng Du
Wenxiang Ping
Bioinformatics Analysis of Diadenylate Cyclase Regulation on Cyclic Diadenosine Monophosphate Biosynthesis in Exopolysaccharide Production by <i>Leuconostoc mesenteroides</i> DRP105
Fermentation
cyclic diadenosine monophosphate
diadenylate cyclase
glucansucrase
bioinformatics
exopolysaccharides
title Bioinformatics Analysis of Diadenylate Cyclase Regulation on Cyclic Diadenosine Monophosphate Biosynthesis in Exopolysaccharide Production by <i>Leuconostoc mesenteroides</i> DRP105
title_full Bioinformatics Analysis of Diadenylate Cyclase Regulation on Cyclic Diadenosine Monophosphate Biosynthesis in Exopolysaccharide Production by <i>Leuconostoc mesenteroides</i> DRP105
title_fullStr Bioinformatics Analysis of Diadenylate Cyclase Regulation on Cyclic Diadenosine Monophosphate Biosynthesis in Exopolysaccharide Production by <i>Leuconostoc mesenteroides</i> DRP105
title_full_unstemmed Bioinformatics Analysis of Diadenylate Cyclase Regulation on Cyclic Diadenosine Monophosphate Biosynthesis in Exopolysaccharide Production by <i>Leuconostoc mesenteroides</i> DRP105
title_short Bioinformatics Analysis of Diadenylate Cyclase Regulation on Cyclic Diadenosine Monophosphate Biosynthesis in Exopolysaccharide Production by <i>Leuconostoc mesenteroides</i> DRP105
title_sort bioinformatics analysis of diadenylate cyclase regulation on cyclic diadenosine monophosphate biosynthesis in exopolysaccharide production by i leuconostoc mesenteroides i drp105
topic cyclic diadenosine monophosphate
diadenylate cyclase
glucansucrase
bioinformatics
exopolysaccharides
url https://www.mdpi.com/2311-5637/11/4/196
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