Studies on the Formation of Bacitracin by Bacillus licheniformis: Effect of Inorganic Phosphate

Haavik, H. I.

doi:10.1099/00221287-84-1-226

Cited by 27 publications

(10 citation statements)

References 9 publications

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“…Addition of 0.2% KH 2 PO 4 in the production medium increased the bacitracin production upto 51.5±1.47 IU ml -1 by mutant strain (Figure 2 B). The results obtained are in accordance with the results obtained by Haavik (16), Demain (10). Maximum bacitracin production (55.8±2.06 IU ml -1 ) by mutant strain was achieved at temperature 37°C.…”

Section: Discussionsupporting

confidence: 92%

Systematic mutagenesis method for enhanced production of bacitracin by Bacillus licheniformis mutant strain UV-MN-HN-6

Aftab¹,

Haq²,

Baig³

2012

Braz. J. Microbiol.

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The purpose of the current study was intended to obtain the enhanced production of bacitracin by Bacillus licheniformis through random mutagenesis and optimization of various parameters. Several isolates of Bacillus licheniformis were isolated from local habitat and isolate designated as GP-35 produced maximum bacitracin production (14±0.72 IU ml-1). Bacitracin production of Bacillus licheniformis GP-35 was increased to 23±0.69 IU ml-1 after treatment with ultraviolet (UV) radiations. Similarly, treatment of vegetative cells of GP-35 with chemicals like N-methyl N’-nitro N-nitroso guanidine (MNNG) and Nitrous acid (HNO2) increased the bacitracin production to a level of 31±1.35 IU ml-1 and 27±0.89 IU ml-1respectively. Treatment of isolate GP-35 with combined effect of UV and chemical treatment yield significantly higher titers of bacitracin with maximum bacitracin production of 41.6±0.92 IU ml-1. Production of bacitracin was further enhanced (59.1±1.35 IU ml-1) by optimization of different parameters like phosphate sources, organic acids as well as temperature and pH. An increase of 4.22 fold in the production of bacitracin after mutagenesis and optimization of various parameters was achieved in comparison to wild type. Mutant strain was highly stable and produced consistent yield of bacitracin even after 15 generations. On the basis of kinetic variables, notably Yp/s (IU/g substrate), Yp/x (IU/g cells), Yx/s(g/g), Yp/s, mutant strain B. licheniformis UV-MN-HN-6 was found to be a hyperproducer of bacitracin.

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Section: Discussionsupporting

confidence: 92%

Systematic mutagenesis method for enhanced production of bacitracin by Bacillus licheniformis mutant strain UV-MN-HN-6

Aftab¹,

Haq²,

Baig³

2012

Braz. J. Microbiol.

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“…The increased production observed at the higher phosphate concentration was contrary to previous studies reported on antibiotic synthesis (14); however, a similar phosphate effect was reported by Haavik (8) regarding bacitracin biosynthesis. It is difficult to distinguish between a phosphate effect per se and a pHReffect since the same level of stimulation and specific yield was seen with 50 mM HEPES plus 5.7 mM phosphate in the medium.…”

Section: Resultscontrasting

confidence: 56%

Effect of phosphate and amino acids on echinomycin biosynthesis by Streptomyces echinatus

Formica¹,

Waring²

1983

Antimicrob Agents Chemother

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Streptomyces echinatus produces only echinomycin (quinomycin A), in contrast to other streptomycetes, which produce families of quinoxaline antibiotics differing in the amino acid composition of the oligopeptide (quinomycins A, B, Bo, C, D, and E) or the structure of the sulfur-containing cross bridge (triostins A, B, and C). Attempts were made to establish conditions for directed biosynthesis with S. echinatus. The lability of the peptide lactone to alkaline pH was obviated by using high levels of phosphate or HEPES [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid] buffer in the production medium. Maintaining the pH below 7.5 resulted in an apparent stimulation of production. Amino acids which serve as structural components or as precursors of echinomycin were employed singly or in combination with nitrate in a chemically defined medium. Based on specific yield (micrograms of echinomycin per milligram of mycelia [dry weight]), D-and L-serine, D-alanine, L-valine, and L-phenylalanine produced equivalent yields of antibiotic which were approximately twofold greater than yields obtained with nitrate alone. In contrast, L-alanine, P-alanine, and L-threonine produced a threeto fourfold stimulation of production. Although the other amino acids diminished antibiotic production, L-isoleucine, which ostensibly was inhibitory to production, supported the accumulation of a quinoxaline antibiotic in which the crossbridge sulfur lacked a methyl group.The family of quinoxaline antibiotics embraces two structurally related series referred to as the triostins and the quinomycins. Both series are similar in composition, consisting of two quinoxaline-2-carboxylic acid moieties attached to a cyclic octapeptide which contains a sulfur cross-linkage. They are distinguished in that the triostins contain a disulfide cross bridge, whereas the quinomycins contain a thioacetal cross bridge (3). The circularity of the oligopeptide is maintained by lactone bonds between the hydroxyl group of D-serine and the carboxyl group of the N-methyl amino acid (Fig. 1).Echinomycin (quinomycin A) differs from other natural quinomycin congeners in that its oligopeptide contains 2 mol each of N-methyl-Lvaline, D-serine, L-alanine, and L-cysteine (the latter as N-methyl-L-cysteine and N-methyl-Smethyl cysteine) (11). The major difference among the other natural congeners resides in the N-methyl amino acid residue which can occur as mono-or dimethylated alloisoleucines (11,15,18) with or without N-methyl valine in the other half-molecule.

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“…Acti- nomycin synthesis appears to be far less sensitive to the inorganic phosphate concentration in the medium than the corresponding fermentations of prodigiosin, monamycin, pyocyanine, or vancomycin (57). By contrast, Haavik (21) reported that optimal synthesis of bacitracin was observed at levels of inorganic phosphate in excess of 100 mM, which represents a 20-fold greater amount than required for maximal actinomycin formation.…”

Section: Resultsmentioning

confidence: 99%

Development of a Chemically Defined Medium for the Synthesis of Actinomycin D by Streptomyces parvulus

Williams

Katz

1977

Antimicrob Agents Chemother

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A chemically defined medium, consisting of D-fructose, L-glutamic acid, L-histidine, K2HPO4, MgSO4 * 7H2O, ZnSO4 * 7H2O, CaCl2 * 2H2O, FeSO4 * 7H20, CoC12 -6H2O, and deionized water, was developed for synthesis of high yields (500 to 600 ,ug/ml) of actinomycin D by Streptomyces parvulus. Under these nutritional conditions, growth and actinomycin formation did not follow a typical trophophase-idiophase pattern. The amino acids appeared to have a sparing action on the utilization ofD-fructose which was slowly and incompletely metabolized during mycelium development and antibiotic production. A significant repression of actinomycin synthesis by S. parvulus was observed when Dglucose (0.01 to 0.25%) was added to the culture medium. The repression was not due to a decline in the pH of the medium during glucose catabolism.Because of our continued interest in the mechanism of directed biosynthesis (30), we have turned our attention to the metabolic activities of Streptomyces parvulus which synthesizes virtually the single antibiotic component, actinomycin D (40). Although the medium described previously (16, 32) proved satisfactory for the production of actinomycin mixtures by Streptomyces antibioticus, the synthesis of actinomycin D by S. parvulus, under the same conditions, was rather poor and extremely variable. Studies were, therefore, initiated to establish the nutritional factors that would support growth and consistently high yields of the antibiotic by this organism. This report describes a series of experiments that have culminated in the development of a chemically defmed medium in which production of high titers of actinomycin D was achieved.MATERIALS AND METHODS Organism and conditions of cultivation. S. parvulus (ATCC 12434) was maintained on slants of glucose-yeast extract-malt extract agar medium (26). Incubation was carried out for 3 to 5 days at 30°C until gray spores had developed; slants were then kept at 40C for further use.For preparation of vegetative inoculum, spores were scraped gently from the surface of a slant(s) with 2.5 ml of NZ-Amine medium (31), and 2-ml quantities of the spore suspension were inoculated into a 250-ml Erlenmeyer flask containing 100-ml amounts of NZ-Amine medium. After incubation on a gyratory shaking incubator (New Brunswick Scientific Co., Inc., New Brunswick, N.J.) for 48 h at 3000, the mycelium was harvested by centrifugation (30 min at 9;000 rev/min), washed twice with and then suspended in 100 ml of saline as described previously (31, 34). Aliquots (3 ml) ofthe suspension served as inoculum for each 250-ml Erlenmeyer flask containing 100 ml of chemically defined medium.Basal medium for nutritional studies. The medium (D-galactose, 10 g; 1-glucose, 1.0 g; L-glutamic acid * HCI, 2.5 g; MgSO4 * 7H20,25 mg; ZnSO4 *7H20, 25 mg; CaCl2 *2H20, 25 mg; FeSO4 7H20, 25 mg; deionized water, 1,000 ml; pH 7.1) routinely utilized for actinomycin production with S. antibioticus and Streptomyces chrysomallus (16, 33, 34, 60) was employed initially for the studies with S. parvulus. Ca...

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Studies on the Formation of Bacitracin by Bacillus licheniformis: Effect of Inorganic Phosphate

Cited by 27 publications

References 9 publications

Systematic mutagenesis method for enhanced production of bacitracin by Bacillus licheniformis mutant strain UV-MN-HN-6

Systematic mutagenesis method for enhanced production of bacitracin by Bacillus licheniformis mutant strain UV-MN-HN-6

Effect of phosphate and amino acids on echinomycin biosynthesis by Streptomyces echinatus

Development of a Chemically Defined Medium for the Synthesis of Actinomycin D by Streptomyces parvulus

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