title: Molecular and biochemical characterization of alpha-glucosidase and alpha-mannosidase and their clustered genes from the thermoacidophilic archaeon Picrophilus torridus.
abstract: The genes encoding a putative alpha-glucosidase (aglA) and an alpha-mannosidase (manA) appear to be physically clustered in the genome of the extreme acidophile Picrophilus torridus, a situation not found previously in any other organism possessing aglA or manA homologs. While archaeal alpha-glucosidases have been described, no alpha-mannosidase enzymes from the archaeal kingdom have been reported previously. Transcription start site mapping and Northern blot analysis revealed that despite their colinear orientation and the small intergenic space, the genes are independently transcribed, both producing leaderless mRNA. aglA and manA were cloned and overexpressed in Escherichia coli, and the purified recombinant enzymes were characterized with respect to their physicochemical and biochemical properties. AglA displayed strict substrate specificity and hydrolyzed maltose, as well as longer alpha-1,4-linked maltooligosaccharides. ManA, on the other hand, hydrolyzed all possible linkage types of alpha-glycosidically linked mannose disaccharides and was able to hydrolyze alpha3,alpha6-mannopentaose, which represents the core structure of many triantennary N-linked carbohydrates in glycoproteins. The probable physiological role of the two enzymes in the utilization of exogenous glycoproteins and/or in the turnover of the organism's own glycoproteins is discussed.
authors.affiliation: Institut für Mikrobiologie und Genetik, Georg-August-Universität Göttingen, Grisebachstr. 8, D-37077 Göttingen, Germany.
authors.forename: Angel
authors.initials: A
authors.lastname: Angelov
authors.forename: Mateusz
authors.initials: M
authors.lastname: Putyrski
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Liebl
mesh_terms: D019843:Archaeal Proteins / Q000235:genetics / Q000378:metabolism; D015152:Blotting, Northern; D003001:Cloning, Molecular; D004187:Disaccharides / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D019766:Genes, Archaeal; D006023:Glycoproteins / Q000378:metabolism; D008320:Maltose / Q000378:metabolism; D005810:Multigene Family; D009844:Oligosaccharides / Q000378:metabolism; D012329:RNA, Bacterial / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D013379:Substrate Specificity; D019603:Thermoplasmales / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D024363:Transcription Initiation Site; D014158:Transcription, Genetic; D000520:alpha-Glucosidases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D043323:alpha-Mannosidase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D019843:Archaeal Proteins; D004187:Disaccharides; D006023:Glycoproteins; D009844:Oligosaccharides; D012329:RNA, Bacterial; D012333:RNA, Messenger; D011994:Recombinant Proteins; C021705:maltooligosaccharides; D008320:Maltose; C048068:mannopentaose; D000520:alpha-Glucosidases; D043323:alpha-Mannosidase