Functional and structural analyses of an expansin-like protein from the antarctic yeast glaciozyma antarctica pi12 reveal strategies of nutrient scavenging in the sea ice environment

• Main Authors: Mohamad Nor, N., Hashim, N. H. F., Quay, D. H. X., Mahadi, N. C., Illias, R. M., Abu Bakar, F. D., Murad, A. M. A.
• Format: Article
• Published: Elsevier BV 2020
• Subjects: TP Chemical technology
• Online Access: http://eprints.utm.my/id/eprint/87668/; http://www.dx.doi.org/10.1016/j.ijbiomac.2019.12.099

Genome data mining of the Antarctic yeast, Glaciozyma antarctica PI12 revealed an expansin-like protein encoding sequence (GaEXLX1). The GaEXLX1 protein is 24.8 kDa with a high alkaline pI of 9.81. Homology modeling of GaEXLX1 showed complete D1 and D2 domains of a conventional expansin. The protein exhibited 36% sequence similarity to Clavibacter michiganensis EXLX1 (PDB: 4JCW). Subsequently, a recombinant GaEXLX1 protein was produced using Escherichia coli expression system. Incubation with Avicel, filter paper and cotton fiber showed that the protein can disrupt the surface of crystalline and pure cellulose, suggesting a cell wall modification activity usually exhibited by expansin-like proteins. Binding assays displayed that GaEXLX1 can bind to polymeric substrates, including those postulated to be present in the sea ice ecosystem such as crab chitin and moss lichenan. GaEXLX1 may assist in the recognition and loosening of these substrates in the sea ice prior to hydrolysis by other extracellular enzymes. Similar loosening mechanism to classical expansin-like protein has been postulated for this psychrophilic protein based on several conserved residues of GaEXLX1 involved in binding interaction identified by docking analyses.