Surface display of novel transglycosylating α-glucosidase from Aspergillus neoniger on Pichia pastoris for synthesis of isomaltooligosaccharides

Trisha, Tissopi and Sandeep, Kumar and Ashok, Sadhu and Sarma, Mutturi (2022) Surface display of novel transglycosylating α-glucosidase from Aspergillus neoniger on Pichia pastoris for synthesis of isomaltooligosaccharides. Journal of Biochemical Engineering, 181. pp. 1-11.

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Abstract

In the present study, α-glucosidase gene of Aspergillus neoniger, agdA, was fused with glycosylphosphatidylinositol (GPI)-anchored glycoprotein gene of Pichia pastoris, GCW61 at its C-terminus for functional expression in Pichia pastoris X-33. The expression of this chimeric gene was induced using methanol, and the protein was displayed on the surface of X-33 strain with cell pellet activity of 4.65 U/gDCW. The surface display of recombinant α-glucosidase was confirmed using immunofluorescence microscopy and flow cytometry. The strain displaying α-glucosidase protein was also cultivated in bioreactor using fed-batch cultivation to obtain a maximum activity of 0.158 U/gDCW and with cell density of 45.35 gDCW/L. These cells were later used as whole-cell biocatalysts (20 g wet cell weight/L) for production of isomaltooligosaccharides (IMOs) using 250 g/L of maltose as substrate. The IMO mixture contains isomaltose, panose, maltotetraose, isomaltotriose and three other higher DP oligosaccharides with a net yield of 0.4 g/g maltose. Amongst these, the panose yield was observed to be highest (0.25 g panose/ g maltose). Here we establish that the surface display of novel transglycosylating α-glucosidase on Pichia pastoris using GCW61p as an anchor protein can be used as whole-cell biocatalysts for synthesis of isomaltooligosaccharides.

Item Type:	Article
Uncontrolled Keywords:	Yeast surface display, α-Glucosidase, Fed-batch Transglycosylation, Isomaltooligosaccharides
Subjects:	600 Technology > 08 Food technology > 29 Microbiological food > 02 Fungi 600 Technology > 08 Food technology > 29 Microbiological food > 04 Yeast
Divisions:	Food Microbiology
Depositing User:	Somashekar K S
Date Deposited:	05 Mar 2025 09:48
Last Modified:	05 Mar 2025 09:48
URI:	http://ir.cftri.res.in/id/eprint/19169

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