Characterization of Levansucrase from<i>Rahnella aquatilis</i>JCM-1683

Ohtsuka, Koutaro; Hino, Shiro; Fukushima, Takayuki; Ozawa, O.; Kanematsu, Tadashi; Uchida, Takatsugu

doi:10.1271/bbb.56.1373

Cited by 69 publications

(41 citation statements)

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“…Previously, Ohtsuka et al [19] and Hettwer et al [20] have shown that the activity and stability of the levansucrases from the Gram-negative bacteria Rahnella aquatilis and Pseudomonas syringae are not affected by the presence of calcium or EDTA.…”

Section: Sequence Alignments Of Members Of Family Gh68mentioning

confidence: 99%

Mutational analysis of the role of calcium ions in the Lactobacillus reuteri strain 121 fructosyltransferase (levansucrase and inulosucrase) enzymes

et al. 2005

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Section: Sequence Alignments Of Members Of Family Gh68mentioning

confidence: 99%

Mutational analysis of the role of calcium ions in the Lactobacillus reuteri strain 121 fructosyltransferase (levansucrase and inulosucrase) enzymes

et al. 2005

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“…phaseolicola pointed to an involvement of lipopolysaccharides or membrane particles in stabilizing the enzyme in solution (25). Substrate, inhibitor, and acceptor specifities differ widely in levansucrases from different species (5,6,15,21,23,29,30,34,41,46). In B. subtilis levansucrase, an aspartyl residue was detected as the substrate-binding amino acid (4).…”

Section: ϩmentioning

confidence: 99%

“…Levansucrases have been identified in a number of gramnegative species such as Pantoea agglomerans (ϭErwinia herbicola) (5,24), Erwinia amylovora (16), Zymomonas mobilis (30), Acetobacter suboxydans (29), Gluconobacter oxydans (8), and a Rhanella aquatilis isolate (34) as well as in gram-positive bacteria such as Bacillus subtilis (6,15,47), Bacillus polymyxa (21), Bacillus natto (46), Bacillus amyloliquefaciens (31), Streptococcus mutans (39), and Actinomyces viscosus (35). The enzyme (sucrose:2,6-␤-D-fructan:6-D-fructosyltransferase; EC 2.4.1.10) conducts three characteristic reactions: (i) synthesis of levan from sucrose by transfructosylation while releasing glucose, (ii) hydrolysis of levan to monosaccharides of fructose, and (iii) exchange of [ 14 C]glucose in the reaction fructose-2,1-glucose ϩ [ The product levan consists of D-fructofuranosyl residues linked predominantly ␤- (2,6), but with extensive branching through ␤-(2,1) linkages (19).…”

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Purification and characterization of an extracellular levansucrase from Pseudomonas syringae pv. phaseolicola

1995

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Levansucrase (EC 2.4.1.10), an exoenzyme of Pseudomonas syringae pv. phaseolicola, was purified to homogeneity from the cell supernatant by chromatography on TMAE-Fraktogel and butyl-Fraktogel. The enzyme has molecular masses of 45 kDa under denaturing conditions and 68 kDa during gel filtration of the native form. In isoelectric focusing, active bands appeared at pH 3.55 and 3.6. Maximum sucrose cleaving activities were measured at pH 5.8 to 6.6 and 60؇C. The enzyme was highly tolerant to denaturing agents, proteases, and repeated freezing and thawing. The molecular weight of the produced levan depended on temperature, salinity, and sucrose concentration. The enzyme had levan-degrading activity and did not accept raffinose as a substrate. Comparison of the N-terminal amino acid sequence with the predicted amino acid sequence of levansucrases from Erwinia amylovora and Zymomonas mobilis showed 88 and 69% similarity, respectively, in amino acids 5 to 20. No similarity could be detected to levansucrases of gram-positive bacteria in the first 20 amino acids. By comparison of all levansucrases which have been sequenced to date, the enzyme seems to be conserved in the gram-negative bacteria. The rheological behavior of the product levan prompted a new assessment of the enzyme's role in pathogenesis. Depending on formation conditions, levan solutions exclude other polymer solutions. This behavior supports the presumption that the levansucrase is important in the early phase of infection by creating a separating layer between bacteria and plant cell wall to prevent the pathogen from recognition.

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“…Interestingly, the formation of levan can be quantitatively replaced by the formation of homo-and hetero--(2-6)-FOSs in the presence of various acceptors. 3,4) Most of the research reported to date has been carried out on a few LSs from Zymomonas mobilis 5) and Bacillus subtilis 4,6) and, to a lesser extent, on LSs from Lactobacillus reuteri, 7) Gluconacetobacter diazotrophicus 8) and Bacillus megaterium.…”

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Purification and Characterization of Levansucrases fromBacillus amyloliquefaciensin Intra- and Extracellular Forms Useful for the Synthesis of Levan and Fructooligosaccharides

Tian¹,

Inthanavong²,

Karboune³

2011

Bioscience, Biotechnology, and Biochemistry

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The intra-and extracellular levansucrase (LS) activities produced by Bacillus amyloliquefaciens were promoted by supplementing the sucrose medium with yeast and peptone as nitrogen sources. These activities were purified by polyethylene glycol (PEG) fractionation for the first time. PEGs of low molecular weight selectively fractionated the intracellular LS activity rather than the extracellular LS activity. Contrary to other LSs, B. amyloliquefaciens LSs exhibited high levan-forming activity over a wide range of sucrose concentrations. The optimum temperatures for the intra-(25-30 C) and extracellular (40 C) LS transfructosylation activities were lower than those for the hydrolytic activities (45-50 C; 50 C). In addition, the catalytic efficiency for the transfructosylation activity of intracellular LS was higher than that of extracellular LS. These differences between intra-and extracellular LSs reveal the occurrence of certain conformational changes to LS upon protein secretion and/or purification. This study is the first to highlight that B. amyloliquefaciens LSs synthesized a variety of FOSs from various saccharides, with lactose and maltose being the best fructosyl acceptors.

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Characterization of Levansucrase fromRahnella aquatilisJCM-1683

Cited by 69 publications

References 1 publication

Mutational analysis of the role of calcium ions in the Lactobacillus reuteri strain 121 fructosyltransferase (levansucrase and inulosucrase) enzymes

Mutational analysis of the role of calcium ions in the Lactobacillus reuteri strain 121 fructosyltransferase (levansucrase and inulosucrase) enzymes

Purification and characterization of an extracellular levansucrase from Pseudomonas syringae pv. phaseolicola

Purification and Characterization of Levansucrases fromBacillus amyloliquefaciensin Intra- and Extracellular Forms Useful for the Synthesis of Levan and Fructooligosaccharides

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