title: Expression of low endotoxin 3-O-sulfotransferase in Bacillus subtilis and Bacillus megaterium.
abstract: A key enzyme for the biosynthesis and bioengineering of heparin, 3-O-sulfotransferase-1 (3-OST-1), was expressed and purified in Gram-positive Bacillus subtilis and Bacillus megaterium. Western blotting, protein sequence analysis, and enzyme activity measurement confirmed the expression. The enzymatic activity of 3-OST-1 expressed in Bacillus species were found to be similar to those found when expressed in Escherichia coli. The endotoxin level in 3-OST-1 from B. subtilis and B. megaterium were 10(4)-10(5)-fold lower than that of the E. coli-expressed 3-OST-1, which makes the Bacillus expression system of particular interest for the generation of pharmaceutical grade raw heparin from nonanimal sources.
authors.affiliation: College of Life Science and Technology, Beijing University of Chemical Technology, Box 75, Beijing, 100029, China.
authors.forename: Wenya
authors.initials: W
authors.lastname: Wang
authors.forename: Jacob A
authors.initials: JA
authors.lastname: Englaender
authors.forename: Peng
authors.initials: P
authors.lastname: Xu
authors.forename: Krunal K
authors.initials: KK
authors.lastname: Mehta
authors.forename: Jiraporn
authors.initials: J
authors.lastname: Suwan
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Dordick
authors.forename: Fuming
authors.initials: F
authors.lastname: Zhang
authors.forename: Qipeng
authors.initials: Q
authors.lastname: Yuan
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Linhardt
authors.forename: Mattheos
authors.initials: M
authors.lastname: Koffas
mesh_terms: D001410:Bacillus megaterium / Q000201:enzymology*; D001412:Bacillus subtilis / Q000201:enzymology*; D001426:Bacterial Proteins / Q000378:metabolism; D004731:Endotoxins / Q000378:metabolism; D015238:Sulfotransferases / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D004731:Endotoxins; D015238:Sulfotransferases