Bacillus subtilis extraction

Fig. 3.1 CE-MS analysis of Bacillus subtilis extracts. Ion electropherograms are shown for selected intermediates of central carbon metabolism, energy eqnivalents, and redox cofactors. Separation was achieved in the negative mode by CZE in coated capillaries with reversed inner-wall polarity. (Reprinted with permission from [127].)...

Trachypogon plumosus I O-extract, roots inhib d. E. coli, Bacillus subtilis Staph, aureus, Strep, haemolyticus 118... 

Black, G. E. Snyder, A. Heroux, K. Chemotaxonomic differentiation between the Bacillus cereus group and Bacillus subtilis by phospholipid extracts analyzed with electospray tandem mass spectrometry. J. Microbiol. Meth. 1997, 28, 187-190. 

Figure 11.3 Positive ion FIESMS spectra of crude cell extracts from Escherichia coli HB101 (A), Bacillus sphaericus DSM 28 (B), and Bacillus licheniformis NTCC 10341 (C). (D) A pseudo-3D plot of the first three discriminant functions (DF1-3) obtained from positive ion whole-cell DIESMS spectra of seven Bacillus subtilis strains (a-g) (E) the corresponding abridged dendrogram obtained from the same information as in D. (Adopted from Vaidyanathan et al.57)...

Mutagenic activity. Petroleum ether extract of the aerial parts, in the ration of Drosophila at concentrations of 0.5, 1, and 5% of the diet, was active . Petroleum ether extract of the dried leaf, administered by gastric intubation to male mice at a dose of 50 mg/kg, was active h Water and methanol extracts of the seed, on agar plate at a concentration of 100 mg/mL, were inactive on Bacillus subtilis H-17 (Rec+) and Salmonella typhimurium TAIOO and TA98. Metabolic activation had no effect on the results h... 

Antibacterial activity. Methylene chloride extract of the dried aerial parts of the plants, on agar plate at a concentration of 1 g/mL, was active on Bacillus subtilis L Methanol extract of the shade-dried plant, on agar plate at a concentration of 0.6 mg/mL, was inactive on Staphylococcus aureus. A concentration of 10 mg/mL was inactive on Escherichia coli and Pseudomonas aureugi-nosab L... 

Anti-amoebic activity. Ethanol (80%) extract of the dried rhizome was inactive on Entamoeba histolytica, minimum inhibitory concentration (MIC) greater than 1 mg/ mL The extract, administered intragas-trically to male hamsters at a dose of 800 mg/kg, was active vs experimentally induced hepatic amebiasis . A dose of 250 mg/kg, administered intragastrically to rats on days 1-5, produced weak activity and a dose of 500 mg/kg was active " ". Anti-atherosclerotic activity. Ethanol (50%) extract of the dried rhizome, administered intragastrically to male rabbits at a dose of 500 mg/kg, reduced atherogenic index from 4.7 to 1.2 on the aorta . Antibacterial activity. Decoction of the dried entire plant, on agar plate, was inactive on Proteus vulgaris, Staphylococcus aureus, and Staphylococcus epidermidis MIC 125 mg/mL. Bacillus subtilis, Bordetella... 

Yersinia enterolitica °. Ethanol (90%) extract of the dried rhizome, on agar plate at a concentration of 500 mg/disc, produced weak activity on Bacillus subtilis, Escherichia coli. Streptococcus aureus, and Streptococcus faecalis L Hot water extract of the dried rhizome, on agar plate at a concentration of 50 mg/disc, was inactive on Salmonella typhimurium TAIOO and TA98" . 

The four-plate test was initially based on the German Hemmstoff-test with an additional plate of Sarcina lulea at pH 8.0, designed for the detection of lower levels of macrolides, and a fourth plate of Escherichia coli at pH 7.2 for the detection of sulfonamides (74,75). The modified version adopted by the European Community for screening carcasses is based on three plates with Bacillus subtilis BGA at pH values of 6.0, 8.0, and 7.2 with added trimethoprim, respectively, and a fourth plate with Micrococcus luteus NCTC 8340 at pH 8.0 (74). This test as described elsewhere (76) is intended to detect residues of -lactams, tetracyclines, aminoglycosides, sulfonamides, and macrolides in muscle tissue of slaughtered animals, without any prior extraction or cleanup. 

Later studies concerned the detection of both sulfonamides and sulfonamide potentiators in fish tissues. Mixtures containing ethyl acetate, sodium sulfate, and sodium hydroxide (80), or ethyl acetate and sodium hydroxide (115), have been all used for extracting residues of sulfadiazine and trimethoprim from trout and salmon tissues. In the former approach, para-aminobenzoic acid was employed to neutralize sulfadiazine prior to the microbiological assay of trimethoprim, whereas in the latter approach Bacillus subtilis (ATCC) was directly used for sulfadiazine detection. The limit of detection for sulfadiazine using Bacillus subtilis was found to be 0.04 ppm. The test indicator organism used in both approaches to detect trimethoprim down to 0.1 ppm was Bacillus pumilus (CN60 Welcome Research Laboratories, London) (115,116). 

Nishimura and Maruo (118) extracted and RNase from cells of Bacillus subtilis strain H, which is quite different from the extracellular RNases of the same strain. It is remarkable that the digestion products of RNA by the enzyme are exclusively four nucleoside 2, 3 -cyclic phosphates. 

Most biologically active natural peptides are linear, but bacitracin is a leading member of the so-called cyclic peptide type of antibiotics. The commercial material, extracted from Bacillus subtilis, is a mixture of several compounds in which bacitracin A predominates. It exerts its action by inhibiting peptidoglycan synthesis and membrane function. Bacitracin has been a useful antibiotic since the 1960s, but its systemic use results in a number of toxic side effects, including nephrotoxicity. One cannot be sure which components of the mixture are responsible for the toxicity, and separation of natural constituents is complex and difficult. For this reason, an efficient synthesis of bacitracin A would be useful. 

Real-time PCR was also performed by on-chip thermal cycling for the detection of Hantavirus, HIV, orthopoxviruses (266-281 bp), Borrelia burgdorferi, human (3-actin (294 bp), and the human complement C6 gene (73 bp) [944]. In another report, real-time PCR was performed for DNA extracted from Bacillus subtilis in a plastic microfluidic cassette. The spore samples (lOVmL) were sampled, filtered, and sonicated in the presence of 6-pm glass beads for spore disruption to release the DNA [945],... 

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