Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in <i>Lactiplantibacillus plantarum</i> and <i>Lentilactibacillus buchneri</i>
In <i>Lactococcus lactis</i> and some other lactic acid bacteria, respiratory metabolism has been reported upon supplementation with only heme, leading to enhanced biomass formation, reduced acidification, resistance to oxygen, and improved long-term storage. Genes encoding a complete re...
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MDPI AG
2021-07-01
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| author | Nisit Watthanasakphuban Ludovika Jessica Virginia Dietmar Haltrich Clemens Peterbauer |
| author_facet | Nisit Watthanasakphuban Ludovika Jessica Virginia Dietmar Haltrich Clemens Peterbauer |
| author_sort | Nisit Watthanasakphuban |
| collection | DOAJ |
| description | In <i>Lactococcus lactis</i> and some other lactic acid bacteria, respiratory metabolism has been reported upon supplementation with only heme, leading to enhanced biomass formation, reduced acidification, resistance to oxygen, and improved long-term storage. Genes encoding a complete respiratory chain with all components were found in genomes of <i>L. lactis</i> and <i>Leuconostoc mesenteroides</i>, but menaquinone biosynthesis was found to be incomplete in Lactobacillaceae (except <i>L. mesenteroides</i>). <i>Lactiplantibacillus plantarum</i> has only two genes (<i>menA</i>, <i>menG</i>) encoding enzymes in the biosynthetic pathway (out of eight), and <i>Lentilactobacillus buchneri</i> has only four (<i>menA</i>, <i>menB</i>, <i>menE</i>, and <i>menG</i>). We constructed knock-out strains of <i>L. lactis</i> defective in <i>menA</i>, <i>menB</i>, <i>menE</i>, and <i>menG</i> (encoding the last steps in the pathway) and complemented these by expression of the extant genes from <i>Lactipl. plantarum</i> and <i>Lent. buchneri</i> to verify their functionality. Three of the <i>Lactipl. plantarum</i> biosynthesis genes, <i>lpmenA1</i>, <i>lpmenG1</i>, and <i>lpmenG2</i>, as well as <i>lbmenB</i> and <i>lbmenG</i> from <i>Lent. buchneri</i>, reconstituted menaquinone production and respiratory growth in the deficient <i>L. lactis</i> strains when supplemented with heme. We then reconstituted the incomplete menaquinone biosynthesis pathway in <i>Lactipl. plantarum</i> by expressing six genes from <i>L. lactis</i> homologous to the missing genes in a synthetic operon with two inducible promoters. Higher biomass formation was observed in <i>Lactipl. plantarum</i> carrying this operon, with an OD<sub>600</sub> increase from 3.0 to 5.0 upon induction. |
| first_indexed | 2024-03-10T09:31:42Z |
| format | Article |
| id | doaj.art-aecd1afd67e04aa9b0d3f6640407146d |
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| issn | 2076-2607 |
| language | English |
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| spelling | doaj.art-aecd1afd67e04aa9b0d3f6640407146d2023-11-22T04:26:19ZengMDPI AGMicroorganisms2076-26072021-07-0197147610.3390/microorganisms9071476Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in <i>Lactiplantibacillus plantarum</i> and <i>Lentilactibacillus buchneri</i>Nisit Watthanasakphuban0Ludovika Jessica Virginia1Dietmar Haltrich2Clemens Peterbauer3Department of Food Sciences and Technology, BOKU-University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, AustriaDepartment of Food Sciences and Technology, BOKU-University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, AustriaDepartment of Food Sciences and Technology, BOKU-University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, AustriaDepartment of Food Sciences and Technology, BOKU-University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, AustriaIn <i>Lactococcus lactis</i> and some other lactic acid bacteria, respiratory metabolism has been reported upon supplementation with only heme, leading to enhanced biomass formation, reduced acidification, resistance to oxygen, and improved long-term storage. Genes encoding a complete respiratory chain with all components were found in genomes of <i>L. lactis</i> and <i>Leuconostoc mesenteroides</i>, but menaquinone biosynthesis was found to be incomplete in Lactobacillaceae (except <i>L. mesenteroides</i>). <i>Lactiplantibacillus plantarum</i> has only two genes (<i>menA</i>, <i>menG</i>) encoding enzymes in the biosynthetic pathway (out of eight), and <i>Lentilactobacillus buchneri</i> has only four (<i>menA</i>, <i>menB</i>, <i>menE</i>, and <i>menG</i>). We constructed knock-out strains of <i>L. lactis</i> defective in <i>menA</i>, <i>menB</i>, <i>menE</i>, and <i>menG</i> (encoding the last steps in the pathway) and complemented these by expression of the extant genes from <i>Lactipl. plantarum</i> and <i>Lent. buchneri</i> to verify their functionality. Three of the <i>Lactipl. plantarum</i> biosynthesis genes, <i>lpmenA1</i>, <i>lpmenG1</i>, and <i>lpmenG2</i>, as well as <i>lbmenB</i> and <i>lbmenG</i> from <i>Lent. buchneri</i>, reconstituted menaquinone production and respiratory growth in the deficient <i>L. lactis</i> strains when supplemented with heme. We then reconstituted the incomplete menaquinone biosynthesis pathway in <i>Lactipl. plantarum</i> by expressing six genes from <i>L. lactis</i> homologous to the missing genes in a synthetic operon with two inducible promoters. Higher biomass formation was observed in <i>Lactipl. plantarum</i> carrying this operon, with an OD<sub>600</sub> increase from 3.0 to 5.0 upon induction.https://www.mdpi.com/2076-2607/9/7/1476menaquinonedemethlymenaquinonerespirationlactic acid bacteria |
| spellingShingle | Nisit Watthanasakphuban Ludovika Jessica Virginia Dietmar Haltrich Clemens Peterbauer Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in <i>Lactiplantibacillus plantarum</i> and <i>Lentilactibacillus buchneri</i> Microorganisms menaquinone demethlymenaquinone respiration lactic acid bacteria |
| title | Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in <i>Lactiplantibacillus plantarum</i> and <i>Lentilactibacillus buchneri</i> |
| title_full | Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in <i>Lactiplantibacillus plantarum</i> and <i>Lentilactibacillus buchneri</i> |
| title_fullStr | Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in <i>Lactiplantibacillus plantarum</i> and <i>Lentilactibacillus buchneri</i> |
| title_full_unstemmed | Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in <i>Lactiplantibacillus plantarum</i> and <i>Lentilactibacillus buchneri</i> |
| title_short | Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in <i>Lactiplantibacillus plantarum</i> and <i>Lentilactibacillus buchneri</i> |
| title_sort | analysis and reconstitution of the menaquinone biosynthesis pathway in i lactiplantibacillus plantarum i and i lentilactibacillus buchneri i |
| topic | menaquinone demethlymenaquinone respiration lactic acid bacteria |
| url | https://www.mdpi.com/2076-2607/9/7/1476 |
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