RT Journal Article SR Electronic(1) A1 Foster, Simon J.YR 1991 T1 Cloning, expression, sequence analysis and biochemical characterization of an autolytic amidase of Bacillus subtilis 168 trpC2 JF Microbiology, VO 137 IS 8 SP 1987 OP 1998 DO https://doi.org/10.1099/00221287-137-8-1987 PB Microbiology Society, SN 1465-2080, AB By use of a functional assay for activity, an autolysin structural gene was cloned on a 3 kb EcoRI DNA fragment from a λgt11 expression library of Bacillus subtilis 168 trpC2 genomic DNA. Sequencing of the fragment showed five open reading frames, the central one of which encoded the lytic enzyme as found by subclone activity mapping and its homology to a recently sequenced autolysin gene. The protein had a deduced sequence of 272 amino acids and a molecular mass of 29957 Da. When expressed in Escherichia coli DH5α, the protein was processed to a 21 kDa form, as estimated by renaturing SDS-PAGE. The autolysin was an N-acetylmuramyl-L-alanine amidase and its activity was MgCI2-dependent (20 mM optimum) and LiC1-sensitive. The enzyme could bind to and hydrolyse a wide range of peptidoglycan substrates isolated from Gram-positive bacteria; the binding was also MgC12-dependent. Initial mapping experiments located the autolysin gene near aroD on the B. subtilis 168 chromosome., UL https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-137-8-1987