Characterization of a Novel Fructosyltransferase from
            <i>Lactobacillus reuteri</i>
            That Synthesizes High-Molecular-Weight Inulin and Inulin Oligosaccharides

Hijum, Sacha A. F. T. van; Geel-Schutten, G.H. van; Rahaoui, H.; Maarel, van der Marc; Dijkhuizen, Lubbert

doi:10.1128/aem.68.9.4390-4398.2002

Cited by 157 publications

(170 citation statements)

References 45 publications

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“…1). The LPKAG motif was also reported to occur in L. johnsonii NCC 533 inulosucrase InuJ (Anwar et al, 2008) and is similar to the consensus sequence LPXTG, which is well-conserved in Gram-positive cell-wall-associated proteins (Fischetti et al, 1990;van Hijum et al, 2002van Hijum et al, , 2004Waldherr et al, 2008). The C terminus of InuGB contains a 17-fold repeat of the motif PXX preceded by a C-terminal variable region of 43 aa.…”

Section: (I) Signal Peptides and N-terminal Variable Regionsmentioning

confidence: 99%

“…Isolation of ftf genes from L. gasseri DSM 20604 and 20077 was initiated by PCR using the degenerate primers 5FTF and 6FTFi, which are based on conserved amino acid sequences of FTFs of Grampositive origin (van Hijum et al, 2002). Amplicons of 228 and 235 bp were amplified this way from the genomic DNA of L. gasseri DSM 20604 and L. 20077, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Activities of these enzymes were also tested on raffinose (500 mM) using the same pH and temperature conditions, but with 5000 U l 21 enzyme concentration and 18 h incubation time. Recombinant L. reuteri 121 inulosucrase (InuD699His) (van Hijum et al, 2002) was used as a positive control.…”

Section: Methodsmentioning

confidence: 99%

“…Protein expression in overnight cultures (600 ml each, grown at 37 uC in LB medium) of E. coli BL21 (DE3 star) harbouring pETInuGA-R, pETInuGA-RM, pETInuGB-R and pETLevG was induced with 0.1 mM IPTG for 1 h at 200 r.p.m. The cells were harvested by centrifugation at 3500 g for 15 min and the FTF proteins were purified to homogeneity by Ni-NTA and anion-exchange chromatography as described by van Hijum et al (2002).…”

Section: Methodsmentioning

confidence: 99%

“…Currently, only six inulosucrase genes/enzymes have been characterized from Gram-positive bacteria, namely in Streptococcus mutans (Rosell & Birkhed, 1974), Leuconostoc citreum CW28 (Olivares-Illana et al, 2002), Lactobacillus reuteri 121 (van Hijum et al, 2002), Bacillus sp. (Wada et al, 2003), L. reuteri TMW 1.106 (Schwab et al, 2007) and Lactobacillus johnsonii NCC 533 (Anwar et al, 2008), while numerous levansucrases are known (http://www.cazy.org/fam/GH68.html) from both Grampositive and Gram-negative bacteria.…”

Section: Introductionmentioning

confidence: 99%

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Inulin and levan synthesis by probiotic Lactobacillus gasseri strains: characterization of three novel fructansucrase enzymes and their fructan products

et al. 2010

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Fructansucrase enzymes polymerize the fructose moiety of sucrose into levan or inulin fructans, with b(2-6) and b(2-1) linkages, respectively. Here, we report an evaluation of fructan synthesis in three Lactobacillus gasseri strains, identification of the fructansucrase-encoding genes and characterization of the recombinant proteins and fructan (oligosaccharide) products. High-performance anion-exchange chromatography and nuclear magnetic resonance analysis of the fructo-oligosaccharides (FOS) and polymers produced by the L. gasseri strains and the recombinant enzymes revealed that, in situ, L. gasseri strains DSM 20604 and 20077 synthesize inulin (and oligosaccharides) and levan products, respectively. L. gasseri DSM 20604 is only the second Lactobacillus strain shown to produce inulin polymer and FOS in situ, and is unique in its distribution of FOS synthesized, ranging from DP2 to DP13. The probiotic bacterium L. gasseri DSM 20243 did not produce any fructan, although we identified a fructansucrase-encoding gene in its genome sequence. Further studies showed that this L. gasseri DSM 20243 gene was prematurely terminated by a stop codon. Exchanging the stop codon for a glutamine codon resulted in a recombinant enzyme producing inulin and FOS. The three recombinant fructansucrase enzymes characterized from three different L. gasseri strains have very similar primary protein structures, yet synthesize different fructan products. An interesting feature of the L. gasseri strains is that they were unable to ferment raffinose, whereas their respective recombinant enzymes converted raffinose into fructan and FOS.

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Section: (I) Signal Peptides and N-terminal Variable Regionsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Inulin and levan synthesis by probiotic Lactobacillus gasseri strains: characterization of three novel fructansucrase enzymes and their fructan products

et al. 2010

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Enzymatic Oligosaccharide Synthesis

Buchholz

Seibel

2010

Encyclopedia of Industrial Biotechnology

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Oligosaccharides (OS) have found manyfold interest in the field of food, pharmaceuticals, and cosmetics due to different specific properties. Food, sweeteners, and food ingredients represent important sectors where OS are used in major amounts. Commercial products comprise isomaltulose, and derivatives, isomalto‐oligosaccharides (IMOS), cyclodextrins (CD), fructosyl‐oligosaccharides (FOS), galactosyl‐oligosaccharides (Gal‐OS), and other specialties. They have been commercialized in Japan, Europe, and worldwide as sweeteners, prebiotics, etc. Nearly all major industrial processes involving carbohydrates are, or include biotechnological transformations. This is due to the complex nature of carbohydrates where stereo‐ and regioselectivity are highly complex and difficult to control. Enzymes and microorganisms in general work highly selective and efficient. Furthermore, they work in water solution and provide for high yield in general. Important examples of commercialized products will be presented in this review. Increasing attention has been devoted to the sophisticated roles of OS and glycosylated compounds, at cell‐ or membrane surfaces, and their function, for example, in infection and cancer proliferation. The challenge for synthesis is obvious. Recent trends and promising developments for industrial use will be mentioned.

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Production of Heterologous Storage Polysaccharides in Potato Plants

Huang¹,

Vincken²,

Visser³

et al. 2018

Annual Plant Reviews Online

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Starch is the most important storage polysaccharide in higher plants. This polysaccharide is used in many industrial applications as it is abundant, renewable and biodegradable and it can be modified into a wide range of products used in food, animal feed, pharmaceuticals and industry. With the understanding of the starch biosynthesis pathway and the isolation of many of the genes involved in this process, the potential for producing novel starches with improved functionality through genetic modification of plants has become more clear. Several different strategies have been used to achieve the rational design of renewable polymers to meet specific requirements. One of these strategies is the regulation of starch biosynthesis and granule assembly in higher plants. A second strategy involves the expression of heterologous genes encoding biosynthetic or modifying enzymes from other organisms to produce novel products with novel functional properties. In this chapter, these recent developments in starch modification in potato plants, such as alteration of starch composition and production of novel storage polysaccharides, are reviewed.

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Characterization of a Novel Fructosyltransferase from Lactobacillus reuteri That Synthesizes High-Molecular-Weight Inulin and Inulin Oligosaccharides

Cited by 157 publications

References 45 publications

Inulin and levan synthesis by probiotic Lactobacillus gasseri strains: characterization of three novel fructansucrase enzymes and their fructan products

Inulin and levan synthesis by probiotic Lactobacillus gasseri strains: characterization of three novel fructansucrase enzymes and their fructan products

Enzymatic Oligosaccharide Synthesis

Production of Heterologous Storage Polysaccharides in Potato Plants

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