Influence of vitamins and osmolites on growth and bacteriocin production by Lactobacillus salivarius CRL 1328 in a chemically defined medium.
The aim of this study was to analyze the influence of vitamins, glycerol, and salts on the growth and bacteriocin production by Lactobacillus salivarius CRL 1328, a human vagina isolate, by using a chemically defined medium to determine the optimal conditions for salivaricin production. The single omission of <span class="smallcap">d</span>-biotin, thiamine, p-aminobenzoic acid, folic acid, or cyanocobalamin did not affect the bacterial growth, whereas the removal of nicotinic acid, riboflavin, and pyridoxal produced a decrease of about 30% in the growth rate. Maximum salivaricin activity was observed after the addition of 5 or 10 g/L of NaCl. On the basis of the nutritional requirements and the levels of salivaricin production, a new optimized and simplified defined medium (SDM-NaCl) for L. salivarius CRL 1328 bacteriocin production was formulated. The kinetics of salivaricin production in SDM-NaCl and in the complex media LAPTg revealed that bacteriocin production was growth linked. A combination of tricine - sodium dodecyl sulfate polyacrylamide gel electrophoresis (Tricine-SDS-PAGE), Lumitein protein gel staining, and a bioassay for antibacterial activity indicated that the molecular mass of salivaricin CRL 1328 is about 4.5 kDa. The partially purified bacteriocin, obtained from SDM-NaCl after concentration, allowed for the design of a relatively simple method for the recovery of a biologically active protein.
Vera Pingitore
Esteban
E
Centro de Referencia para Lactobacilos (CERELA-CONICET), Chacabuco 145 (T4000ILC), San Miguel de Tucuman 4000, Argentina.
Hebert
Elvira María
EM
Sesma
Fernando
F
Nader-Macías
María Elena
ME
D001430:Bacteriocins / Q000096:biosynthesis* / Q000494:pharmacology; D003470:Culture Media; D013293:Enterococcus faecalis / Q000187:drug effects / Q000254:growth & development; D005260:Female; D005990:Glycerol / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D007778:Lactobacillus / Q000187:drug effects / Q000254:growth & development* / Q000302:isolation & purification / Q000378:metabolism*; D008826:Microbial Sensitivity Tests; D009997:Osmotic Pressure; D012965:Sodium Chloride / Q000378:metabolism / Q000494:pharmacology*; D014621:Vagina / Q000382:microbiology; D014815:Vitamins / Q000378:metabolism / Q000494:pharmacology*
D001430:Bacteriocins; D003470:Culture Media; D014815:Vitamins; D012965:Sodium Chloride; D005990:Glycerol