Isolation and characterization of a salt-tolerant γ-glutamyl transpeptidase from xerophilic Aspergillus sydowii

从嗜干曲霉(Aspergillus sydowii)中分离和鉴定一种耐盐γ-谷氨酰转肽酶

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Abstract

Xerophilic Aspergillus molds isolated from halo-alkaliphilic and dry environments are attractive genetic resources for obtaining salt- and osmo-adaptive enzymes. A. sydowii MA0196 secreted the largest amount of γ-glutamyl transpeptidase (GGT) during solid-state fermentation at a low initial water activity (a (w) = 0.85). Gel filtration analysis revealed that the molecular mass of the purified native enzyme (MA0196 GGT) was 120 kDa. SDS-PAGE analysis showed that MA0196 GGT consists of two subunits with molecular masses of 56.4 and 33 kDa, indicating production from a proenzyme via autoproteolysis. Deglycosylation of the subunits by N-glycosidase F yielded 40.9 and 19.6 kDa species. MA0196 GGT retained transpeptidase and hydrolysis activities and their catalytic efficiency (k (cat)/K (m)) under high salt and low water activity. The enzyme displayed broad substrate specificity toward γ-glutamyl acceptors such as amino acids and the imidazole dipeptides, carnosine and anserine. Carnosine and L-glutamine were converted into γ-glutamyl-β-alanyl-L-histidine by MA0196 GGT with a 32.9% yield in the presence of 2% (v/v) dimethyl sulfoxide. Phylogenetic analysis indicated that MA0196 GGT forms a distinct lineage from A. oryzae and A. sojae GGTs. These excellent properties indicate that MA0196 GGT can be used in salted fermentation and for producing bioactive peptides. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-022-03259-3.

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