Production of isomelezitose from sucrose by engineered glucansucrases

Côté, Gregory L.; Dunlap, Christopher A.; Vermillion, Karl E.; Skory, Christopher D.

doi:10.1515/amylase-2017-0008

Cited by 6 publications

(2 citation statements)

References 27 publications

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“…Besides, Görl et al () transformed a sucrose isomerase into an isomelezitose synthase, obtaining a protein variant by mutation that yielded 70% of isomelezitose using 100 g l −1 sucrose, but this amount was calculated based on sucrose consumed (~ 32%) instead of the total sucrose added to the reaction mixture, which means to produce about 22.4 g l −1 of this trisaccharide. More recently, Côté et al () obtained an engineered glucansucrase from the bacteria Leuconostoc mesenteroides that produced isomelezitose very efficiently using ~ 800 g l −1 sucrose. Compared to the wild type enzyme (DsrI), which only synthesized traces of isomelezitose, the mutated variant (Dsr1‐L441E) converted 57% of sucrose in this trisaccharide.…”

Section: Resultsmentioning

confidence: 99%

Efficient production of isomelezitose by a glucosyltransferase activity in Metschnikowia reukaufii cell extracts

García-González

Plou

Cervantes

et al. 2019

Microbial Biotechnology

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Summary Metschnikowia reukaufii is a widespread yeast able to grow in the plants’ floral nectaries, an environment of extreme conditions with sucrose concentrations exceeding 400 g l−1, which led us into the search for enzymatic activities involved in this sugar use/transformation. New oligosaccharides were produced by transglucosylation processes employing M. reukaufii cell extracts in overload‐sucrose reactions. These products were purified and structurally characterized by MS‐ESI and NMR techniques. The reaction mixture included new sugars showing a great variety of glycosidic bonds including α‐(1→1), α‐(1→3) and α‐(1→6) linkages. The main product synthesized was the trisaccharide isomelezitose, whose maximum concentration reached 81 g l−1, the highest amount reported for any unmodified enzyme or microbial extract. In addition, 51 g l−1 of the disaccharide trehalulose was also produced. Both sugars show potential nutraceutical and prebiotic properties. Interestingly, the sugar mixture obtained in the biosynthetic reactions also contained oligosaccharides such as esculose, a rare trisaccharide with no previous NMR structure elucidation, as well as erlose, melezitose and theanderose. All the sugars produced are naturally found in honey. These compounds are of biotechnological interest due to their potential food, cosmeceutical and pharmaceutical applications.

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Section: Resultsmentioning

confidence: 99%

Efficient production of isomelezitose by a glucosyltransferase activity in Metschnikowia reukaufii cell extracts

García-González

Plou

Cervantes

et al. 2019

Microbial Biotechnology

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“…citreum NRRL B-1355) also modified the profile and the yields of the oligosaccharides obtained from sucrose. It also allowed to generate variants producing isomelezitoze (α-D-Glu p -(1→6)-β-D-Fru f -(2↔1)-α-D-Glu p ), an osmoprotectant, in yields up to 57% [ 164 ]. In ASR, docking experiments also suggested the presence of a +2 subsite and a +2′ subsite highlighting two important amino acids: Trp675 of Motif III and Asp772, respectively.…”

Section: Structure–function Relationshipsmentioning

confidence: 99%

Bacterial α-Glucan and Branching Sucrases from GH70 Family: Discovery, Structure–Function Relationship Studies and Engineering

et al. 2021

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Glucansucrases and branching sucrases are classified in the family 70 of glycoside hydrolases. They are produced by lactic acid bacteria occupying very diverse ecological niches (soil, buccal cavity, sourdough, intestine, dairy products, etc.). Usually secreted by their producer organisms, they are involved in the synthesis of α-glucans from sucrose substrate. They contribute to cell protection while promoting adhesion and colonization of different biotopes. Dextran, an α-1,6 linked linear α-glucan, was the first microbial polysaccharide commercialized for medical applications. Advances in the discovery and characterization of these enzymes have remarkably enriched the available diversity with new catalysts. Research into their molecular mechanisms has highlighted important features governing their peculiarities thus opening up many opportunities for engineering these catalysts to provide new routes for the transformation of sucrose into value-added molecules. This article reviews these different aspects with the ambition to show how they constitute the basis for promising future developments.

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Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2017–2018

Harvey

2021

Mass Spectrometry Reviews

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This review is the tenth update of the original article published in 1999 on the application of matrix‐assisted laser desorption/ionization mass spectrometry (MALDI) mass spectrometry to the analysis of carbohydrates and glycoconjugates and brings coverage of the literature to the end of 2018. Also included are papers that describe methods appropriate to glycan and glycoprotein analysis by MALDI, such as sample preparation techniques, even though the ionization method is not MALDI. Topics covered in the first part of the review include general aspects such as theory of the MALDI process, new methods, matrices, derivatization, MALDI imaging, fragmentation and the use of arrays. The second part of the review is devoted to applications to various structural types such as oligo‐ and poly‐saccharides, glycoproteins, glycolipids, glycosides, and biopharmaceuticals. Most of the applications are presented in tabular form. The third part of the review covers medical and industrial applications of the technique, studies of enzyme reactions, and applications to chemical synthesis. The reported work shows increasing use of combined new techniques such as ion mobility and highlights the impact that MALDI imaging is having across a range of diciplines. MALDI is still an ideal technique for carbohydrate analysis and advancements in the technique and the range of applications continue steady progress.

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Production of isomelezitose from sucrose by engineered glucansucrases

Cited by 6 publications

References 27 publications

Efficient production of isomelezitose by a glucosyltransferase activity in Metschnikowia reukaufii cell extracts

Efficient production of isomelezitose by a glucosyltransferase activity in Metschnikowia reukaufii cell extracts

Bacterial α-Glucan and Branching Sucrases from GH70 Family: Discovery, Structure–Function Relationship Studies and Engineering

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2017–2018

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