Properties and localisation of mycobacillin-synthesising enzyme system in Bacillus subtilis B3.
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Biomedical subjects
Publications and source records attributed to S K Bose.
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Mycobacillin lacks amino groups but contains two free alpha-carboxyl groups, indicating the presence of two side-chain peptide linkages. The five aspartic acid residues of mycobacillin are all in alpha-peptide linkage whereas the two glutamic acid residues are in gamma-linkage. Mycobacillin does not react with hydroxylamine to give hydroxamate, indicating the absence of anhydride, lactone and ester linkages. This is also confirmed by i.r. spectroscopy and titration of the molecule. Of the 15 peptides obtained from partial hydrolysates of mycobacillin, 12 contain aspartic acid. Results obtained by treatment of hydrolysates of aspartic acid-containing peptides with d-amino acid oxidase and l-glutamate decarboxylase (containing l-aspartate decarboxylase activity) indicate that residue 5 is l-aspartic acid and residues 2, 8, 11 and 13 are d-aspartic acid. The d- or l-peptide sequence and nature of peptide linkages in mycobacillin are proposed on the basis of these findings and the amino acid sequence reported earlier.
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Degradation of bacterial deoxyribonucleic acid (DNA) after infection with T4 bacteriophage was studied in an endonuclease I-deficient host. The kinetics of degradation were similar to those seen in other hosts with a normal level of this enzyme. Irradiation of extracellular phage with ultraviolet (UV) destroyed the capacity of the infecting virus to induce extensive breakdown of host DNA, which was, however, converted to high-molecular-weight material. Addition of chloramphenicol to T4-infected cells provided data which can be interpreted to indicate the involvement of at least two endodeoxyribonucleases and one exodeoxyribonuclease having a high degree of specificity. A model is proposed showing the sequential action of two endodeoxyribonucleases followed by an exodeoxyribonuclease in the degradation of host DNA. The appearance of these hydrolytic enzymes requires protein synthesis. Infections leading to partial degradation only (UV-irradiated phages, gene 46 mutants) effectively inhibited the synthesis of bacterial messenger ribonucleic acid and of beta-galactosidase.
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The individual colonies of a partially degenerated culture of Aspergillus versicolor vary widely in their ability to produce an antibiotic active against Trichophyton rubrum. Mutagenic treatment with nitrous acid or ultraviolet light gave potent colonies. Nitrous acid gave better results in terms of stability and yield. This treatment may therefore prove useful in the regeneration of partially degenerated cultures.
Versicolin is a new antifungal agent isolated from the culture broth of a new strain of Aspergillus versicolor. The antibiotic is specifically active against pathogenic fungi, particularly Trichophyton rubrum which causes 90% of the skin infections occurring in Calcutta and eastern India.
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The kinetics of degradation of bacterial deoxyribonucleic acid (DNA) after infection of Escherichia coli with T4D, ultraviolet-irradiated T4D, and two amber mutants, N122 and N94, was studied by zone sedimentation through linear glycerol gradients. Within 5 min after infection with any of the bacteriophages, breakdown of host genome was evident. The first product was a high-molecular-weight material (50S to 70S) and further degradation appeared to occur in discrete steps. Rapid and extensive breakdown of bacterial DNA was seen after infection with am N122 and T4D. Infection with ultraviolet-irradiated phage or with am N94 resulted in an accumulation of high-molecular-weight material. These results suggest that the observed degradation of host DNA begins early and requires sequential action of several phage-induced endo- as well as exodeoxyribonucleases.
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