Controlled cell lysis and protoplast formation by enhancement of inhibitors of alanine racemase by glycine.
beta-D-chloroalanine (200-800 micrograms/ml) addition to Bacillus subtilis SB 19 caused a slight inhibition of growth in MLN broth. Inhibitor plus 1% glycine initiated rapid lysis of cells (complete in 2 hours). D-alanine (500 micrograms/ml) prevented lysis. D-cycloserine behaved similarly. Inhibitors in 0.5 M sucrose and 1% glycine formed stable "primed protoplasts" which were converted to the bacillary form by addition of D-alanine. Characteristics of Dal+ cells with inhibitor are similar to Dal- mutants. Thus Dal+ cells in the presence of alanine racemase inhibitors + glycine can be used for controlled lysis and protoplast formation (suitable for PEG transformation with plasmid DNA). Inhibitors in 1% glycine exhibit a strong antibacterial action similar to potent antibiotics which inhibit cell wall formation.
D000409:Alanine / Q000378:metabolism; D000411:Alanine Racemase / Q000037:antagonists & inhibitors*; D000591:Amino Acid Isomerases / Q000037:antagonists & inhibitors*; D001433:Bacteriolysis / Q000187:drug effects*; D003523:Cycloserine / Q000494:pharmacology; D005998:Glycine / Q000494:pharmacology*; D011523:Protoplasts / Q000187:drug effects* / Q000502:physiology; D015091:beta-Alanine / Q000031:analogs & derivatives / Q000494:pharmacology
D015091:beta-Alanine; C006977:3-chloroalanine; D003523:Cycloserine; D000591:Amino Acid Isomerases; D000411:Alanine Racemase; D000409:Alanine; D005998:Glycine