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Isolation and analysis of the protoplast membrane of Bacillus megaterium.

ABSTRACT: 1. Optimum conditions were found for the lysis of Bacillus megaterium KM by lysozyme. The age of culture, density of suspension and concentration of lysozyme affected the rate of lysis. 2. Protoplast membranes were isolated by centrifugation of lysates and were exhaustively washed. 3. Treatment with chloroform removed some lipid from the membranes, but about half of the total membrane lipid could be extracted only after partial acid hydrolysis. 4. The defatted membranes consisted of protein together with variable amounts of RNA; carbohydrate was almost absent. 5. Lipid accounted for 23% of the weight of the membrane, and included both neutral lipid and phospholipid. In both classes, branched-chain C(15) acids made up about 80% of the total fatty acid. 6. The phospholipid was a kephalin, and contained small quantities of several amino acids.

SUBMITTER: Yudkin MD

PROVIDER: S-EPMC1264938 | biostudies-other | 1966 Mar

REPOSITORIES: biostudies-other

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Project description:Bacterial metabolites exhibit a variety of biologically active compounds including antibacterial and antifungal activities. It is well known that Bacillus is considered to be a promising source of bioactive secondary metabolites. Most plant pathogens have an incredible ability to mutate and acquire resistance, causing major economic losses in the agricultural field. Therefore, it is necessary to use the natural antibacterial compounds in microbes to control plant pathogens. This study was conducted to investigate the bio-active compounds of Bacillus megaterium L2. According to the activity guidance of Agrobacterium tumefaciens T-37, Erwinia carotovora EC-1 and Ralstonia solanacearum RS-2, five monomeric compounds, including erucamide (1), behenic acid (2), palmitic acid (3), phenylacetic acid (4), and β-sitosterol (5), were fractionated and purified from the crude ethyl acetate extract of B. megaterium. To our knowledge, all compounds were isolated from the bacterium for the first time. To understand the antimicrobial activity of these compounds, and their minimum inhibitory concentrations (MICs) (range: 0.98∼500 μg/mL) were determined by the broth microdilution method. For the three tested pathogens, palmitic acid exhibited almost no antibacterial activity (>500 μg/mL), while erucamide had moderate antibacterial activity (MIC = 500 μg/mL). Behenic acid showed MICs of 250 μg/mL against T-37 and RS-2 strains with an antibacterial activity. β-sitosterol showed significant antimicrobial activity against RS-2. β-sitosterol showed remarkable antimicrobial activity against RS-2 with an MIC of 15.6 μg/mL. In addition, with the antimicrobial activity, against T-37 (62.5 μg/mL) and against EC-1 (125 μg/mL) and RS-2 (15.6 μg/mL) strains notably, phenylacetic acid may be interesting for the prevention and control of phytopathogenic bacteria. Our findings suggest that isolated compounds such as behenic acid, β-sitosterol, and phenylacetic acid may be promising candidates for natural antimicrobial agents.

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Isolation and analysis of the protoplast membrane of Bacillus megaterium.

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