Characterization of an α-glucosidase enzyme conserved in Gardnerella spp. isolated from the human vaginal microbiome

Bhandari, P.; Tingley, J. P.; Abbott, D. W.; Hill, J. E.

doi:10.1101/2020.05.11.086124

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bioRxiv · 10.1101/2020.05.11.086124

Characterization of an α-glucosidase enzyme conserved in Gardnerella spp. isolated from the human vaginal microbiome

Bhandari, P.·Tingley, J. P.·Abbott, D. W.·Hill, J. E.

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Abstract

Gardnerella spp. in the vaginal microbiome are associated with bacterial vaginosis, a dysbiosis in which a lactobacilli dominant microbial community is replaced with mixed aerobic and anaerobic bacteria including Gardnerella species. The co-occurrence of multiple Gardnerella species in the vaginal environment is common, but different species are dominant in different women. Competition for nutrients, particularly glycogen present in the vaginal environment, could play an important role in determining the microbial community structure. Digestion of glycogen into products that can be taken up and further processed by bacteria requires the combined activities of several enzymes collectively known as amylases, which belong to glycoside hydrolase family 13 (GH13) within the CAZy classification system. GH13 is a large and diverse family of proteins, making prediction of their activities challenging. SACCHARIS annotation of the GH13 family in Gardnerella resulted in identification of protein domains belonging to eight subfamilies. Phylogenetic analysis of predicted amylase sequences from 26 Gardnerella genomes demonstrated that a putative -glucosidase-encoding sequence, CG400_06090, was conserved in all species in the genus. The predicted -glucosidase enzyme was expressed, purified and functionally characterized. The enzyme was active on a variety of maltooligosaccharides over a broad pH range (4.0 - 8) with maximum activity at pH 7. The Km, kcat and kcat/Km values for the substrate 4-nitrophenyl -D-glucopyranoside were 8.3 M, 0.96 min-1 and 0.11 M-1min-1 respectively. Glucose was released from maltose, maltotriose, maltotetraose and maltopentaose, but no products were detected on thin layer chromatography when the enzyme was incubated with glycogen. Our findings show that Gardnerella spp. produce an -glucosidase enzyme that may contribute to the complex and multistep process of glycogen metabolism by releasing glucose from maltooligosaccharides.

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Bhandari, P., Tingley, J. P., Abbott, D. W., Hill, J. E.. 2020-05-11. Characterization of an α-glucosidase enzyme conserved in Gardnerella spp. isolated from the human vaginal microbiome. https://doi.org/10.1101/2020.05.11.086124

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