02185nas a2200265   4500008004100000022001400041245009100055210006900146260007300215300001000288490000700298520124000305653002201545653003601567653001801603653002301621100002401644700002501668700001901693700002101712700002501733700002001758700002401778856011701802       2012                        eng d  a0141-813000aPurification and characterization of YxeI, a penicillin acylase from Bacillus subtilis0 aPurification and characterization of YxeI a penicillin acylase f  aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcJAN  a25-300 v503 a<p>The paper reports the purification and characterization of the first penicillin acylase from Bacillus subtilis. YxeI, the protein annotated as hypothetical, coded by the gene yxeI in the open reading frame between iol and hut operons in B. subtilis was cloned and expressed in Eshcherichia coli, purified and characterized. The purified protein showed measurable penicillin acylase activity with penicillin V. The enzyme was a homotetramer of 148 kDa. The apparent Km of the enzyme for penicillin V and the synthetic substrate 2-nitro-5-(phenoxyacetamido)-benzoic acid was 40 mM and 0.63 mM, respectively, and the association constants were 8.93 x 10(2) M-1 and 2.51 x 10(5) M-1, respectively. It was inhibited by cephalosporins and conjugated bile salts, substrates of the closely related bile acid hydrolases. It had good sequence homology with other penicillin V acylases and conjugated bile acid hydrolases, members of the Ntn hydrolase family. The N-terminal nucleophile was a cysteine which is revealed by a simple removal of N-formyl-methionine. The activity of the protein was affected by high temperature, acidic pH and the presence of the denaturant guanidine hydrochloride. (C) 2011 Elsevier B.V. All rights reserved.</p>10aBacillus subtilis10aConjugated bile acid hydrolases10aNtn hydrolase10aPenicillin acylase1 aRathinaswamy, Priya1 aGaikwad, Sushama, M.1 aSuresh, C., G.1 aPrabhune, Asmita1 aBrannigan, James, A.1 aDodson, Guy, G.1 aPundle, Archana, V.  uhttps://library.ncl.res.in/content/purification-and-characterization-yxei-penicillin-acylase-bacillus-subtilis-0