Synthesis of Isomaltooligosaccharides by Saccharomyces cerevisiae Cells Expressing Aspergillus niger α-Glucosidase

ACS Omega
Mary Casa-VillegasJulio Polaina

Abstract

The α-glucosidase encoded by the aglA gene of Aspergillus niger is a secreted enzyme belonging to family 31 of glycoside hydrolases. This enzyme has a retaining mechanism of action and displays transglycosylating activity that makes it amenable to be used for the synthesis of isomaltooligosaccharides (IMOs). We have expressed the aglA gene in Saccharomyces cerevisiae under control of a galactose-inducible promoter. Recombinant yeast cells expressing the aglA gene produced extracellular α-glucosidase activity about half of which appeared cell bound whereas the other half was released into the culture medium. With maltose as the substrate, panose is the main transglycosylation product after 8 h of incubation, whereas isomaltose is predominant after 24 h. Isomaltose also becomes predominant at shorter times if a mixture of maltose and glucose is used instead of maltose. To facilitate IMO production, we have designed a procedure by which yeast cells can be used directly as the catalytic agent. For this purpose, we expressed in S. cerevisiae gene constructs in which the aglA gene is fused to glycosylphosphatidylinositol anchor sequences, from the yeast SED1 gene, that determine the covalent binding of the hybrid protein to the cell ...Continue Reading

References

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Citations

Jan 8, 2020·International Journal of Molecular Sciences·Triinu VisnapuuTiina Alamäe
May 8, 2021·Applied Microbiology and Biotechnology·Cecelia A Garcia, Jeffrey G Gardner
Oct 27, 2021·Critical Reviews in Food Science and Nutrition·Ayyappan Palaniappan, Mohammad Naushad Emmambux

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Datasets Mentioned

BETA
D45356.1

Methods Mentioned

BETA
transglycosylation
electrophoresis
glycosylation
transglucosylation
ion exchange chromatography

Software Mentioned

PyMOL Molecular Graphics System

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