Proteomic Analysis Reveals Enzymes for β-D-Glucan Formation and Degradation in <i>Levilactobacillus brevis</i> TMW 1.2112
Bacterial exopolysaccharide (EPS) formation is crucial for biofilm formation, for protection against environmental factors, or as storage compounds. EPSs produced by lactic acid bacteria (LAB) are appropriate for applications in food fermentation or the pharmaceutical industry, yet the dynamics of f...
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MDPI AG
2022-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/6/3393 |
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| author | Julia A. Bockwoldt Chen Meng Christina Ludwig Michael Kupetz Matthias A. Ehrmann |
| author_facet | Julia A. Bockwoldt Chen Meng Christina Ludwig Michael Kupetz Matthias A. Ehrmann |
| author_sort | Julia A. Bockwoldt |
| collection | DOAJ |
| description | Bacterial exopolysaccharide (EPS) formation is crucial for biofilm formation, for protection against environmental factors, or as storage compounds. EPSs produced by lactic acid bacteria (LAB) are appropriate for applications in food fermentation or the pharmaceutical industry, yet the dynamics of formation and degradation thereof are poorly described. This study focuses on carbohydrate active enzymes, including glycosyl transferases (GT) and glycoside hydrolases (GH), and their roles in the formation and potential degradation of <i>O2</i>-substituted (1,3)-β-D-glucan of <i>Levilactobacillus</i> (<i>L</i>.) <i>brevis</i> TMW 1.2112. The fermentation broth of <i>L</i>. <i>brevis</i> TMW 1.2112 was analyzed for changes in viscosity, β-glucan, and D-glucose concentrations during the exponential, stationary, and early death phases. While the viscosity reached its maximum during the stationary phase and subsequently decreased, the β-glucan concentration only increased to a plateau. Results were correlated with secretome and proteome data to identify involved enzymes and pathways. The suggested pathway for β-glucan biosynthesis involved a β-1,3 glucan synthase (GT2) and enzymes from maltose phosphorylase (MP) operons. The decreased viscosity appeared to be associated with cell lysis as the β-glucan concentration did not decrease, most likely due to missing extracellular carbohydrate active enzymes. In addition, an operon was discovered containing known moonlighting genes, all of which were detected in both proteome and secretome samples. |
| first_indexed | 2024-03-09T19:40:50Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T19:40:50Z |
| publishDate | 2022-03-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-d379226c3dd8488ea07129aa2e254f692023-11-24T01:38:54ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-03-01236339310.3390/ijms23063393Proteomic Analysis Reveals Enzymes for β-D-Glucan Formation and Degradation in <i>Levilactobacillus brevis</i> TMW 1.2112Julia A. Bockwoldt0Chen Meng1Christina Ludwig2Michael Kupetz3Matthias A. Ehrmann4Lehrstuhl für Mikrobiologie, Technische Universität München, 85354 Freising, GermanyBayerisches Zentrum für Biomolekulare Massenspektrometrie (BayBioMS), Technische Universität München, 85354 Freising, GermanyBayerisches Zentrum für Biomolekulare Massenspektrometrie (BayBioMS), Technische Universität München, 85354 Freising, GermanyLehrstuhl für Brau- und Getränketechnologie, Technische Universität München, 85354 Freising, GermanyLehrstuhl für Mikrobiologie, Technische Universität München, 85354 Freising, GermanyBacterial exopolysaccharide (EPS) formation is crucial for biofilm formation, for protection against environmental factors, or as storage compounds. EPSs produced by lactic acid bacteria (LAB) are appropriate for applications in food fermentation or the pharmaceutical industry, yet the dynamics of formation and degradation thereof are poorly described. This study focuses on carbohydrate active enzymes, including glycosyl transferases (GT) and glycoside hydrolases (GH), and their roles in the formation and potential degradation of <i>O2</i>-substituted (1,3)-β-D-glucan of <i>Levilactobacillus</i> (<i>L</i>.) <i>brevis</i> TMW 1.2112. The fermentation broth of <i>L</i>. <i>brevis</i> TMW 1.2112 was analyzed for changes in viscosity, β-glucan, and D-glucose concentrations during the exponential, stationary, and early death phases. While the viscosity reached its maximum during the stationary phase and subsequently decreased, the β-glucan concentration only increased to a plateau. Results were correlated with secretome and proteome data to identify involved enzymes and pathways. The suggested pathway for β-glucan biosynthesis involved a β-1,3 glucan synthase (GT2) and enzymes from maltose phosphorylase (MP) operons. The decreased viscosity appeared to be associated with cell lysis as the β-glucan concentration did not decrease, most likely due to missing extracellular carbohydrate active enzymes. In addition, an operon was discovered containing known moonlighting genes, all of which were detected in both proteome and secretome samples.https://www.mdpi.com/1422-0067/23/6/3393<i>Levilactobacillus brevis</i> TMW 1.2112β-glucanexopolysaccharideglycosyltransferaseglycosyl hydrolasemoonlighting proteins |
| spellingShingle | Julia A. Bockwoldt Chen Meng Christina Ludwig Michael Kupetz Matthias A. Ehrmann Proteomic Analysis Reveals Enzymes for β-D-Glucan Formation and Degradation in <i>Levilactobacillus brevis</i> TMW 1.2112 International Journal of Molecular Sciences <i>Levilactobacillus brevis</i> TMW 1.2112 β-glucan exopolysaccharide glycosyltransferase glycosyl hydrolase moonlighting proteins |
| title | Proteomic Analysis Reveals Enzymes for β-D-Glucan Formation and Degradation in <i>Levilactobacillus brevis</i> TMW 1.2112 |
| title_full | Proteomic Analysis Reveals Enzymes for β-D-Glucan Formation and Degradation in <i>Levilactobacillus brevis</i> TMW 1.2112 |
| title_fullStr | Proteomic Analysis Reveals Enzymes for β-D-Glucan Formation and Degradation in <i>Levilactobacillus brevis</i> TMW 1.2112 |
| title_full_unstemmed | Proteomic Analysis Reveals Enzymes for β-D-Glucan Formation and Degradation in <i>Levilactobacillus brevis</i> TMW 1.2112 |
| title_short | Proteomic Analysis Reveals Enzymes for β-D-Glucan Formation and Degradation in <i>Levilactobacillus brevis</i> TMW 1.2112 |
| title_sort | proteomic analysis reveals enzymes for β d glucan formation and degradation in i levilactobacillus brevis i tmw 1 2112 |
| topic | <i>Levilactobacillus brevis</i> TMW 1.2112 β-glucan exopolysaccharide glycosyltransferase glycosyl hydrolase moonlighting proteins |
| url | https://www.mdpi.com/1422-0067/23/6/3393 |
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