01780nas a2200193   4500008004100000022001400041245017000055210006900225260006700294300001200361490000700373520095200380100002901332700002601361700002501387700002701412700001901439856012801458       2013                        eng d  a1744-309100aCloning, overexpression, crystallization and preliminary X-ray crystallographic analysis of a slow-processing mutant of penicillin G acylase from Kluyvera citrophila0 aCloning overexpression crystallization and preliminary Xray crys  a111 RIVER ST, HOBOKEN 07030-5774, NJ USAbWILEY-BLACKWELLcAUG  a925-9290 v693 a<p>{Kluyvera citrophila penicillin G acylase (KcPGA) has recently attracted increased attention relative to the well studied and commonly used Escherichia coli PGA (EcPGA) because KcPGA is more resilient to harsh conditions and is easier to immobilize for the industrial hydrolysis of natural penicillins to generate the 6-aminopenicillin (6-APA) nucleus, which is the starting material for semi-synthetic antibiotic production. Like other penicillin acylases, KcPGA is synthesized as a single-chain inactive pro-PGA, which upon autocatalytic processing becomes an active heterodimer of alpha and beta chains. Here, the cloning of the pac gene encoding KcPGA and the preparation of a slow-processing mutant precursor are reported. The purification, crystallization and preliminary X-ray analysis of crystals of this precursor protein are described. The protein crystallized in two different space groups, P1, with unit-cell parameters a = 54.0</p>1 aVarshney, Nishant, Kumar1 aRamasamy, Sureshkumar1 aBrannigan, James, A.1 aWilkinson, Anthony, J.1 aSuresh, C., G.  uhttps://library.ncl.res.in/content/cloning-overexpression-crystallization-and-preliminary-x-ray-crystallographic-analysis-0