Use of bacteria, proteins or peptides from Bacillus thuringiensis var kurstaki clones, Bacillus thuringiensis var israelensis clones and Bacillus sphaericus Meyer and Neide clones, to stimulate the production of phenolic compounds

摘要

Use of bacteria, proteins or peptides obtained from: Bacillus thuringiensis var kurstakiclones of Barjac and Lemille (serotype 3a and 3b), deposited at ATCC (American type culture collection) under number of 33679; Bacillus thuringiensis var israelensisclones of Barjac, deposited at ATCC under number of 35646; and Bacillus sphaericus Meyer and Neideclones, deposited at ATCC under numbers of 10208, is claimed. An independent claim is included for a process for obtaining a composition containing bacteria, proteins or peptides, comprising: culturing the bacteria using a culture medium, which is relatively simple (MS) of which composition comprises potassium dihydrogen phosphate (2.5 g/l), dipotassium dihydrogen phosphate (2.5 g/l), diammonium hydrogen phosphate (1 g/l), magnesium sulfate heptahydrate (0.2 g/l), manganese sulfate heptahydrate (7 mg/l), ferrous sulfate heptahydrate (0.01 g/l) and calcium chloride (10 mg/l), adding standard saline solution of 1% D-glucose and adjusting the pH to 7, sterilizing the medium by autoclaving at 120[deg] C for 20 minutes, inoculating a preculture of bacteria in MS medium for 15 hours at 30[deg] C under agitation, and placing a flask containing the bacteria in culture at 37[deg] C under gentle rotative agitation at 150 revolutions per minute (rpm) for 5-7 days; and (b) separating the bacteria and proteins at 4[deg] C, comprising centrifuging the contents of each flask at 4500 rpm for 30 minutes, washing the pellet in sodium chloride solution (1M) twice with distilled water containing phenylmethylsulfonyl fluoride (1 mM) and EDTA (10 mM) and then resuspended in a demineralized water and purifying the protein part of a discontinuous sucrose gradient by Thomas and Ellar method (1983) [ultracentrifugation at 25000 rpm at 48[deg] C for 16 hours], concentrating into protein, which is determined by the Bradford method after alkaline solubilization of the extract and electrophoresis denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) for detecting proteins using Thomas and Ellar (1983) method.