Synthesis and characterisation of galactosyl glycerol by β-galactosidase catalysed reverse hydrolysis of galactose and glycerol

Wei, Wei; Qi, Danping; Zhao, Haizhen; Lu, Zhaoxin; Lv, Fengting; Bie, Xiaomei

doi:10.1016/j.foodchem.2013.05.145

Cited by 23 publications

(27 citation statements)

References 31 publications

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“…Using a combination of the COSY and TOCSY spectra it was possible to track the scalar coupling but only from H-1 through to H-4 (typical of a galactose residue); the corresponding carbons were obtained from the HSQC spectrum. The position of these C-1 & H-1 resonances perfectly match those published for 1-O--galactopyranosylglycerol (Wei et al 2013).…”

Section: Discussionsupporting

confidence: 82%

Extraction of the same novel homoglycan mixture from two different strains of Bifidobacterium animalis and three strains of Bifidobacterium breve

Alhudhud

Sadiq

Ngo

et al. 2018

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Three strains of Bifidobacterium breve (JCM 7017, JCM 7019 and JCM 2258) and two strains of Bifidobacterium animalis subsp. lactis (AD011 and A1dOxR) were grown in broth cultures or on plates, and a standard exopolysaccharide extraction method was used in an attempt to recover exocellular polysaccharides. When the extracted materials were analysed by NMR it was clear that mixtures of polysaccharides were being isolated including exopolysaccharides (EPS) cell wall polysaccharides and intracellular polysaccharides. Treatment of the cell biomass from the B. breve strains, or the B. animalis subsp. lactis AD011 strain, with aqueous sodium hydroxide provided a very similar mixture of polysaccharides but without the EPS. The different polysaccharides were partially fractionated by selective precipitation from an aqueous solution upon the addition of increasing percentages of ethanol. The polysaccharides extracted from B. breve JCM 7017 grown in HBM media supplemented with glucose (or isotopically labelled D-glucose-1-C) were characterised using 1D and 2D-NMR spectroscopy. Addition of one volume of ethanol generated a medium molecular weight glycogen (Mw=1×10 Da, yield 200 mg/l). The addition of two volumes of ethanol precipitated an intimate mixture of a low molecular weight β-(1→6)-glucan and a low molecular weight β-(1→6)-galactofuranan which could not be separated (combined yield 46 mg/l). When labelled D-glucose-1-C was used as a carbon supplement, the label was incorporated into >95% of the anomeric carbons of each polysaccharide confirming they were being synthesised in situ. Similar H NMR profiles were obtained for polysaccharides recovered from the cells of B. animalis subsp. lactis AD011and A1dOxR (in combination with an EPS), B. breve JCM 7017, B. breve JCM 7019, B. breve JCM 2258 and from an EPS (-ve) mutant of B. breve 7017 (a non-EPS producer).

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Section: Discussionsupporting

confidence: 82%

Extraction of the same novel homoglycan mixture from two different strains of Bifidobacterium animalis and three strains of Bifidobacterium breve

Alhudhud

Sadiq

Ngo

et al. 2018

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“…Therefore, reverse hydrolysis has been considered as an economically feasible approach for industrial applications (Julio and Andrew 2007). Several glycosidases including α-rhamnosidase (Lu et al 2015;Martearena et al 2008), β-glucosidase (Vic et al 1997), β-galactosidase (Majumder et al 2008;Wei et al 2013), α-mannosidase (Maitin et al 2004;Singh et al 2000), and endo-α-N-acetylgalactosaminidase (Ashida et al 2010) have been used for glycoside synthesis through reverse hydrolysis reaction. It has been experimentally proved that the transglycosylation efficiency could be improved by mutagenesis of the glycosidases, and the sitedirected mutagenesis based on rational protein design is regarded as the common strategy to improve the catalytic properties in glycosidases on the basis of the solved homological 3D structures (Choi et al 2008;Hansson et al 2001;Hinz et al 2006;Lundemo et al 2013;Pollet et al 2010;Sun et al 2014;Zeuner et al 2014).…”

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confidence: 99%

Site-directed mutagenesis of α-l-rhamnosidase from Alternaria sp. L1 to enhance synthesis yield of reverse hydrolysis based on rational design

Liu

Yin³

et al. 2016

Appl Microbiol Biotechnol

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The α-L-rhamnosidase catalyzes the hydrolytic release of rhamnose from polysaccharides and glycosides and is widely used due to its applications in a variety of industrial processes. Our previous work reported that a wild-type α-L-rhamnosidase (RhaL1) from Alternaria sp. L1 could synthesize rhamnose-containing chemicals (RCCs) though reverse hydrolysis reaction with inexpensive rhamnose as glycosyl donor. To enhance the yield of reverse hydrolysis reaction and to determine the amino acid residues essential for the catalytic activity of RhaL1, site-directed mutagenesis of 11 residues was performed in this study. Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp, Asp, Asp, Glu, Arg, His, and Trp) and may form the hydrophobic pocket in stabilizing donor (Trp, Tyr, Tyr, and Trp) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. From the three positive variants, mutant W261Y accelerated the reverse hydrolysis with a prominent increase (43.7 %) in relative yield compared to the wild-type enzyme. Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp. Mutant W261Y also exhibited a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme.

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“…It should be noted that, according to literature, in transglycosylation reactions galactose preferentially binds to the primary hydroxyl group of the acceptor. Examples of products meeting that relation include: β-d-galactopyranosyl-(1→1)-d-fructose (using β-ga lac tosidase from Kluyveromyces lactis) (29), 3-O-β-d-ga lac to pyranosyl-glycerol (Kluyveromyces lactis; 25) or salicin galactoside (Aspergillus oryzae; 26). It may be assumed that in the studied case that combination will also be prevailing, and so the dominant fraction of the product is most probably 6-O-β-galactopyranosyl-d-gluconic acid.…”

Section: Determination Of the Transgalactosylation Productmentioning

confidence: 99%

“…No data regarding the use of that method for the synthesis of bionic acids are available in the literature. The transglycosylation activity of that enzyme has been used mostly for production of galactooligosaccharides or lactulose (22), but also for production of galactosyl derivatives of xylose (23), sorbitol (24), glycerol (25), and other substances of potential biological activity (e.g. salicin galactoside (26)).…”

Section: Introductionmentioning

confidence: 99%

Synthesis of Galactosyl Derivative of Gluconic Acid with Transglycosylation Activity of β-galactosidase

Wojciechowska

Klewicki

Sójka

et al. 2017

Food Technol. Biotechnol.

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SummaryBionic acids are bioactive compounds demonstrating numerous interesting properties. They are widely produced by chemical or enzymatic oxidation of disaccharides. This paper focuses on the galactosyl derivative of gluconic acid as a result of a new method of bionic acid synthesis which utilises the transglycosylation properties of β-galactosidase and introduces lactose as a substrate. Products obtained in such a process are characterised by different structures (and, potentially, properties) than those resulting from traditional oxidation of disaccharides. The aim of this study is to determine the effect of selected parameters (concentration and ratio of substrates, dose of the enzyme, time, pH, presence of salts) on the course of the reaction carried out with the enzymatic preparation Lactozym, containing β-galactosidase from Kluyveromyces lactis. Research has shown that increased dry matter content in the baseline solution (up to 50 %, by mass per volume) and an addition of NaCl contribute to higher yield. On the other hand, reduced content of the derivative is a result of increased pH from 7.0 to 9.0 and an addition of magnesium and manganese salts. Moreover, exceeding the β-galactosidase dose over approx. 35 000 U per 100 g of lactose also leads to reduced yield of the process. The most favourable molar ratio of sodium gluconate to lactose is 2.225:0.675. Depending on the conditions of the synthesis, the product concentration ranged between 17.3 and 118.3 g/L of the reaction mixture, which corresponded to the mass fraction of 6.64-23.7 % of dry matter. The data obtained as a result of the present study may be useful for designing an industrial process.

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Synthesis and characterisation of galactosyl glycerol by β-galactosidase catalysed reverse hydrolysis of galactose and glycerol

Cited by 23 publications

References 31 publications

Extraction of the same novel homoglycan mixture from two different strains of Bifidobacterium animalis and three strains of Bifidobacterium breve

Extraction of the same novel homoglycan mixture from two different strains of Bifidobacterium animalis and three strains of Bifidobacterium breve

Site-directed mutagenesis of α-l-rhamnosidase from Alternaria sp. L1 to enhance synthesis yield of reverse hydrolysis based on rational design

Synthesis of Galactosyl Derivative of Gluconic Acid with Transglycosylation Activity of β-galactosidase

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