Levansucrase from Leuconostoc mesenteroides NTM048 produces a levan exopolysaccharide with immunomodulating activity

Ishida, Ryuichi; Sakaguchi, Kouta; Matsuzaki, Chiaki; Katoh, Toshihiko; Ishida, Noriko; Yamamoto, Kenji; Hisa, Keiko

doi:10.1007/s10529-015-2024-9

Cited by 18 publications

(18 citation statements)

References 13 publications

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“…3), the Michaelis constant of levansucrase to sucrose was 25.63 mM. The Km value of the enzyme in this study was similar to the Km value (24mM) of the levansucrase from Leuconostoc mesenteroides NTM048 (Ishida et al 2016).…”

Section: Enzyme Kineticssupporting

confidence: 81%

Biosynthesis and Structural Characterization of Levan by a Recombinant Levansucrase From Bacillus Subtilis ZW019

Wang

Zhao

et al. 2021

Preprint

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ObjectivesThe yield of levan extracted from microbial fermentation broth is low, so in vitro catalytic synthesis of levan by levansucrase is expected to be one of the industrial production approaches of levan. ResultsA recombinant plasmid Pet-28A-AcmA-Z constructed in the previous study was used to produce levansucrase. The recombinant levansucrase could be easily purified in one step and the purified enzyme had a single band clearly visible in SDS-PAGE. The conditions for enzymatic reactions was optimal at pH 5.2 and 40 ℃, and the activity of enzymes was stimulated by K+ and Ca2+. The yield of levan biosynthesis from 10% (w/v) sucrose with 6.45 U/g sucrose of levansucrase was 30.6 g/L. The molecular weight of the levan was about 1.56×106 Da, as measured by GPC. HPIC analysis showed that the monosaccharide composition of the levan was fructose and glucose. The results of FTIR and NMR analysis indicated that the polymer produced by the recombinant levansucrase was β-(2, 6) levan.ConclusionsThe results of this study provide a basis for large-scale production of levan by enzymatic method.

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Section: Enzyme Kineticssupporting

confidence: 81%

Biosynthesis and Structural Characterization of Levan by a Recombinant Levansucrase From Bacillus Subtilis ZW019

Wang

Zhao

et al. 2021

Preprint

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“…Surprisingly, another unidentified Leuconostoc species accounted for 42.7% of bacterial abundance. Nevertheless, the synthesis of the additional fructo-oligosaccharides found in type-1 aguamiel may be the result of fructosyltransferases from Leuconostoc and Zymomonas , similar to the levan- and inulo-sucrases reported for L. mesenteroides, L. paramesenteroides, L. citreum, L.kimchii , and L. plantarum ( 32 – 36 ), or the levansucrases from Z. mobilis . These bacteria, isolated from fresh aguamiel immediately after scraping, are also the most abundant ones in accumulated aguamiel and play a central role in pulque fermentation, where Z. mobilis produces most of the ethanol while Leuconostoc species account for most of the pre- and probiotic compounds.…”

Section: Resultsmentioning

confidence: 74%

Evolution of Fructans in Aguamiel (Agave Sap) During the Plant Production Lifetime

et al. 2020

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Aguamiel is the sap collected from agave , while pulque is the result of the natural fermentation of aguamiel . Despite its ancestral origin and numerous publications on pulque production, little is known about the evolution and concentration of sugars and fructo-oligosaccharides in aguamiel , either during its daily accumulation or through the agave production lifetime. In this study, we examined aguamiel composition in three agave plants during their productive lifetime (4 to 9 months). After each collection, the agave pine is scraped to induce aguamiel to flow into an internally created cavity ( cajete ), producing a residual bagasse ( metzal ). We found that the concentration of agave fructans and sucrose, as well as the fructan profile, change during the aguamiel production process. During the daily collection, a small amount of agave fructans released from the pine by scraping is drawn into the cajete with the first milliliters of sap where it is then diluted with the inflow of aguamiel . The main component of aguamiel is the sucrose produced in high concentration in the leaves through photosynthesis and then hydrolyzed in the cajete as aguamiel accumulates. We also describe how the fructan profile changes during the accumulation of aguamiel in the cajete . In addition to the varying amount of sucrose that is hydrolyzed in the aguamiel accumulated, we found that fructo-oligosaccharides are either diluted, consumed, or hydrolyzed, depending on the plant and its production stage, thus yielding different fructan profiles. New fructo-oligosaccharides are, in some cases, synthesized by bacteria present in aguamiel . These profiles were also observed in aguamiel collected from ten different plants in the same production region. We also found that a considerable amount of agave fructans is lost in metzal (bagasse), the agave material that is scraped and thrown away twice a day during the production process.

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“…Although there have been many reports in the literature on recombinant levansucrase expression, purification, and characterization (Ishida et al 2016;Jang et al 2001;Kim et al 1998;Li et al 2017;Liu et al 2017;Lu et al 2014), levansucrases from extremophilic microorganisms have been largely overlooked. In this work, levansucrase enzyme from H. smyrnensis AAD6 (HsLsc) was recombinantly expressed, purified, and characterized.…”

Section: Discussionmentioning

confidence: 99%

“…Several metal ions and detergents are known to have significant impacts on various levansucrases (Gao et al 2017;Ishida et al 2016;Shaheen et al 2017).…”

Section: Aq6mentioning

confidence: 99%

Levansucrase from Halomonas smyrnensis AAD6T: first halophilic GH-J clan enzyme recombinantly expressed, purified, and characterized

Kırtel

Menéndez

Versluys

et al. 2018

Appl Microbiol Biotechnol

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Fructans, homopolymers of fructose produced by fructosyltransferases (FTs), are emerging as intriguing components in halophiles since they are thought to be associated with osmotic stress tolerance and overall fitness of microorganisms and plants under high-salinity conditions. Here, we report on the full characterization of the first halophilic FT, a levansucrase from Halomonas smyrnensis AAD6 (HsLsc; EC 2.4.1.10). The encoding gene (lsc) was cloned into a vector with a 6xHis Tag at its C-terminus, then expressed in Escherichia coli. The purified recombinant enzyme (47.3 kDa) produces levan and a wide variety of fructooligosaccharides from sucrose, but only in the presence of high salt concentrations (> 1.5 M NaCl). HsLsc showed Hill kinetics and pH and temperature optima of 5.9 and 37 °C, respectively. Interestingly, HsLsc was still very active at salt concentrations close to saturation (4.5 M NaCl) and was selectively inhibited by divalent cations. The enzyme showed high potential in producing novel saccharides derived from raffinose as both fructosyl donor and acceptor and cellobiose, lactose, galactose, and ʟ-arabinose as fructosyl acceptors. With its unique biochemical characteristics, HsLsc is an important enzyme for future research and potential industrial applications in a world faced with drought and diminishing freshwater supplies.

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Levansucrase from Leuconostoc mesenteroides NTM048 produces a levan exopolysaccharide with immunomodulating activity

Abstract: Leuconostoc mesenteroides, a probiotic strain, possessed levansucrase which catalyzed the produced levan that had immunomodulating activity.

Cited by 18 publications

References 13 publications

Biosynthesis and Structural Characterization of Levan by a Recombinant Levansucrase From Bacillus Subtilis ZW019

Biosynthesis and Structural Characterization of Levan by a Recombinant Levansucrase From Bacillus Subtilis ZW019

Evolution of Fructans in Aguamiel (Agave Sap) During the Plant Production Lifetime

Levansucrase from Halomonas smyrnensis AAD6T: first halophilic GH-J clan enzyme recombinantly expressed, purified, and characterized

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