title: Thermostable alpha-galactosidase from Thermotoga neapolitana: cloning, sequencing and expression.
abstract: A gene encoding a thermostable alpha-galactosidase from the hyperthermophile Thermotoga neapolitana was cloned and sequenced. Sequence analysis showed that the 552-amino acid protein is similar to Escherichia coli Raf type alpha-galactosidase and belongs to Family 36 of the glycosyl hydrolases. Recombinant alpha-galactosidase expressed in E. coli has a molecular mass of ca. 61 kDa, and an optimum activity at 93 degrees C at pH 7.0. The enzyme is highly thermostable and retains 75% of activity after heating to 80 degrees C for 4 h. The potential application of the enzyme to high temperature processing of soy molasses has been demonstrated.
authors.affiliation: Food Science Department, University of Illinois, Urbana-Champaign 61801, USA.
authors.forename: M R
authors.initials: MR
authors.lastname: King
authors.forename: D A
authors.initials: DA
authors.lastname: Yernool
authors.forename: D E
authors.initials: DE
authors.lastname: Eveleigh
authors.forename: B M
authors.initials: BM
authors.lastname: Chassy
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics*; D006089:Gram-Negative Anaerobic Bacteria / Q000201:enzymology / Q000235:genetics*; D006358:Hot Temperature; D006868:Hydrolysis; D008553:Melibiose / Q000378:metabolism; D008965:Molasses; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011993:Recombinant Fusion Proteins; D017421:Sequence Analysis; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D000519:alpha-Galactosidase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D011993:Recombinant Fusion Proteins; D008553:Melibiose; D000519:alpha-Galactosidase