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Regulation of Lactate Dehydrogenase pathway in Limosilactobacillus Reuteri Dsm 17938
Malmö University, Faculty of Health and Society (HS).
2021 (English)Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesisAlternative title
Reglering av Laktatdehydrogenasväg hos Limosilactobacillus Reuteri Dsm 17938 (Swedish)
Abstract [en]

Limosilactobacillus reuteri is a Gram-positive, heterofermentative and facultative anaerobic bacterium. It belongs to the Lactobacillus genus and is a part of the flora in the gastrointestinal tract. Limosilactobacillus reuteri belongs to lactic acid bacteria that has 2 metabolic routs in the Embden-Meyerhof pathway and Phosphoketolase pathway. The bacteria have an initial high Embden-Meyerhof activity but expresses a highly active Phosphoketolase throughout a fermentation process. The aim of this project was to study the effect of various organic compounds which are part of the two pathways on the flux of lactate dehydrogenase. The enzyme has an important role in the production of lactate. The investigation was initiated by bacterial fermentation in MRS broth. The bacteria were harvested at the late exponential phase, lysed and mixed with organic compounds in Triethanolamine buffer. The enzyme reaction was analysed spectrophotometrically. Michaelis-Menten principles were applied for obtaining the disassociation constant. The results show significant inhibition by adenosine diphosphate, nicotinamide adenine dinucleotide and xylulose-5-phosphate concluding that regulation of lactate dehydrogenase activity follows a different mechanism when compared to Lactococcus lactis. This primary study requires further optimisation where studies with varying concentrations of reduced nicotinamide adenine dinucleotide should be done to observe the enzyme regulation underynamic conditions. It would also benefit to find the potential activators of this enzyme.

Abstract [sv]

Limosilactobacillus reuteri är en gram-positiv, heterofermentativ och fakultativt anaerob bakterie. Den tillhör Lactobacillus genus och är en del av floran i mag-tarmkanalen. L. reuteri tillhör mjölksyrabakterier och har 2-centrala metabolavägar som följermbden-Meyerhof-vägen och fosfoketolasvägen. Limosilactobacillus reuteri har initialt en hög Embden-Meyerhof aktivitet men uttrycker en mycket aktiv fosfoketolasväg under en jäsningsprocess. Syftet med detta projekt var att undersöka effekten av olika organiska föreningar som ingår i de två vägarna på flödet av laktatdehydrogenas. Enzymet har en viktig roll i produktionen av laktat. Undersökningen påbörjades med bakteriell fermentering i MRS-buljong. Bakterierna skördades vid den sena exponentiella fasen, lyserades och blandades med organiska föreningar i trietanolamin buffert. Enzymreaktionen analyserades spektrofotometriskt. Michaelis-Menten-principer tillämpades för att erhålla disassociationskonstanten för varje reaktion. Resultaten visar signifikant inhibering av Adenosindifosfat, nikotinamidadenindinukleotid och xylulosa-5-fosfat. Slutsatsen av studien är att reglering av laktatdehydrogenas aktivitet följer en annan mekanism jämfört med Lactococcus lactis. Denna studie kräver ytterligare optimering med varierande koncentrationer av reducerad nikotinamidadenindinukleotid för att observera enzymregleringen under dynamiska förhållanden. Det skulle även gynna undersökningen att hitta potentiella aktivatorer för enzymet.

Place, publisher, year, edition, pages
2021. , p. 25
Keywords [en]
Embden-Meyerhof pathway, Enzyme kinetics, Lactate dehydrogenase, Lactic acid bacteria, Limosilactobacillus reuteri, Phosphoketolase pathway.
National Category
Microbiology
Identifiers
URN: urn:nbn:se:mau:diva-43205OAI: oai:DiVA.org:mau-43205DiVA, id: diva2:1563940
External cooperation
Lunds Tekniska Högskola
Educational program
HS Biomedicinsk laboratorievetenskap
Supervisors
Examiners
Available from: 2021-06-24 Created: 2021-06-10 Last updated: 2021-06-24Bibliographically approved

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