Dataset Information

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

ABSTRACT: 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 ?-(1?1), ?-(1?3) and ?-(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.

SUBMITTER: Garcia-Gonzalez M

PROVIDER: S-EPMC6801145 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature

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

Garcia-Gonzalez Martin M Plou Francisco J FJ Cervantes Fadia V FV Remacha Miguel M Poveda Ana A Jiménez-Barbero Jesús J Fernandez-Lobato Maria M

Microbial biotechnology 20191001 6

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<sup>-1</sup> , 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 ...[more]

PMID: 31576667

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Project description:BackgroundHeterotrophic microbial oils are potentially a more sustainable alternative to vegetable or fossil oils for food and fuel applications. However, as almost all work in the area is conducted on the laboratory scale, such studies carry limited industrial relevance and do not give a clear indication of what is required to produce an actual industrial process. Metschnikowia pulcherrima is a non-pathogenic industrially promising oleaginous yeast which exhibits numerous advantages for cost-effective lipid production, including a wide substrate uptake, antimicrobial activity and fermentation inhibitor tolerance. In this study, M. pulcherrima was fermented in stirred tank reactors of up to 350 L with 250-L working volume in both batch and semi-continuous operation to highlight the potential industrial relevance. Due to being food-grade, suitable for handling at scale and to demonstrate the oligosaccharide uptake capacity of M. pulcherrima, enzyme-hydrolysed starch in the form of glucose syrup was selected as fermentation feedstock.ResultsIn batch fermentations on the 2-L scale, a lipid concentration of 14.6 g L-1 and productivity of 0.11 g L-1 h-1 were achieved, which was confirmed at 50 L (15.8 g L-1; 0.10 g L-1 h-1). The maximum lipid production rate was 0.33 g L-1 h-1 (daily average), but the substrate uptake rate decreased with oligosaccharide chain length. To produce 1 kg of dry yeast biomass containing up to 43% (w/w) lipids, 5.2 kg of the glucose syrup was required, with a lipid yield of up to 0.21 g g-1 consumed saccharides. In semi-continuous operation, for the first time, an oleaginous yeast was cultured for over 2 months with a relatively stable lipid production rate (around 0.08 g L-1 h-1) and fatty acid profile (degree of fatty acid saturation around 27.6% w/w), and without contamination. On the 250-L scale, comparable results were observed, culminating in the generation of nearly 10 kg lipids with a lipid productivity of 0.10 g L-1 h-1.ConclusionsThe results establish the importance of M. pulcherrima for industrial biotechnology and its suitability to commercially produce a food-grade oil. Further improvements in the productivity are required to make M. pulcherrima lipid production industrial reality, particularly when longer-chain saccharides are involved.

Dataset Information

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

Publications

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

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