title: Insight into glucosidase II from the red marine microalga Porphyridium sp. (Rhodophyta).
abstract: N-glycosylation of proteins is one of the most important post-translational modifications that occur in various organisms, and is of utmost importance for protein function, stability, secretion, and loca-lization. Although the N-linked glycosylation pathway of proteins has been extensively characterized in mammals and plants, not much information is available regarding the N-glycosylation pathway in algae. We studied the α 1,3-glucosidase glucosidase II (GANAB) glycoenzyme in a red marine microalga Porphyridium sp. (Rhodophyta) using bioinformatic and biochemical approaches. The GANAB-gene was found to be highly conserved evolutionarily (compo-sed of all the common features of α and β subunits) and to exhibit similar motifs consistent with that of homolog eukaryotes GANAB genes. Phylogenetic analysis revealed its wide distribution across an evolutionarily vast range of organisms; while the α subunit is highly conserved and its phylogenic tree is similar to the taxon evolutionary tree, the β subunit is less conserved and its pattern somewhat differs from the taxon tree. In addition, the activity of the red microalgal GANAB enzyme was studied, including functional and biochemical characterization using a bioassay, indicating that the enzyme is similar to other eukaryotes ortholog GANAB enzymes. A correlation between polysaccharide production and GANAB activity, indicating its involvement in polysaccharide biosynthesis, is also demonstrated. This study represents a valuable contribution toward understanding the N-glycosylation and polysaccharide biosynthesis pathways in red microalgae.
authors.affiliation: Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer-Sheva, 8410501, Israel.|Department of Chemical Engineering, Sami Shamoon College of Engineering, Beer-Sheva, 8410001, Israel.
authors.forename: Oshrat
authors.initials: O
authors.lastname: Levy-Ontman
authors.affiliation: Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer-Sheva, 8410501, Israel.
authors.forename: Merav
authors.initials: M
authors.lastname: Fisher
authors.affiliation: Department of Chemical Engineering, Sami Shamoon College of Engineering, Beer-Sheva, 8410001, Israel.
authors.forename: Yoram
authors.initials: Y
authors.lastname: Shotland
authors.affiliation: Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer-Sheva, 8410501, Israel.|Protalix Biotherapeutics, Carmiel, 2161401, Israel.
authors.forename: Yoram
authors.initials: Y
authors.lastname: Tekoah
authors.affiliation: Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer-Sheva, 8410501, Israel.
authors.forename: Shoshana
authors.initials: S
authors.lastname: Malis Arad
keywords: Porphyridium sp.; glucosidase II; glycosylation; polysaccharide; red microalgae