Gramicidin S Origin and Mode of Action

Cause, G.F.; Brazhnikova, M. G.

doi:10.1016/s0140-6736(00)88377-4

Cited by 21 publications

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“…B. brevis 47 and HPD31 have been employed successfully as host bacteria for extracellular production of foreign proteins, such as bacterial thermophilic a-amylase (38) and human epidermal growth factor (49). Additional unique characteristics of B. brevis that have been reported are gramicidin production (9) and molluscicidal activity (31).…”

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confidence: 99%

Characterization of Bacillus brevis with Descriptions of Bacillus migulanus sp. nov., Bacillus choshinensis sp. nov., Bacillus parabrevis sp. nov., and Bacillus galactophilus sp. nov.

Takagi¹,

Sato²,

Kadowaki³

et al. 1993

International Journal of Systematic Bacteriology

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Thirty-five Bacillus brevis strains obtained from culture collections, including protein-producing isolates, were taxonomically studied by using numerical analysis, DNA base composition, and DNA-DNA hybridization. Six DNA relatedness groups were represented, and these groups correlated well with clusters based on the numerical analysis. The B. brevis strains were separated into B. brevis sensu stricto, four new species, and an unidentified species of the genus Bacillus. Bacillus migulanus sp. nov., Bacillus choshinensis sp. nov., Bacillus parabrevis sp. nov., and Bacillus galactophilus sp. nov, are proposed.The heterogeneity of Bacillus brevis has been indicated by the wide range of maximum growth temperatures (10) and the results of numerical classification (13,26,27) and classification based on API test data (17). The protein producers identified as B. brevis (45) had guanine-plus-cytosine (G+C) contents ranging from 47 to 55 mol%. These observations suggest that this taxon is genetically heterogeneous. Recently, Nakamura (23) reported that the phenotypic heterogeneity of B. brevis was due to variability introduced by the presence of genetically unrelated strains. Previously, Udaka (44) and Takagi et al. (37) reported the bacterial production of large amounts of proteins in culture fluids; many of the bacteria involved appeared phenotypically to be B. brevis (37,44). These strains showed either weak or no detectable proteolytic activity in the culture supernatant (37). B. brevis 47 and HPD31 have been employed successfully as host bacteria for extracellular production of foreign proteins, such as bacterial thermophilic a-amylase (38) and human epidermal growth factor (49). Additional unique characteristics of B. brevis that have been reported are gramicidin production (9) and molluscicidal activity (31).In order to clarify the taxonomic positions of 14 B. brevis strains from culture collections and of 21 protein-producing isolates, we examined their phenotypic characteristics and processed the data obtained by using standard numerical analysis. In addition, the relationships between clusters resulting from the numerical analysis were evaluated by DNA-DNA hybridization.As a result, the above-mentioned 35 strains assigned to B. brevis were divided into six independent species, four of which are proposed below as the new species Bacillus migulanus, Bacillus choshinensis, Bacillus parabrevis, and Bacillus gala c toph ilus. MATERIALS AND METHODSBacterial strains. B. brevis strains and reference strains obtained from culture collections and other sources and the protein-producing strains described previously (37, 44) were * Corresponding author. used in this study. The sources and histories of these strains are listed in Table 1. Working stocks were cultivated on T2 agar (44) for 24 h at 30 or 37°C and were stored at a room temperature.Characterization, Unless indicated otherwise, the methods described by Gordon et al. (10) were used for characterization of organisms. Acid production from carbohydrates was determin...

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confidence: 99%

Characterization of Bacillus brevis with Descriptions of Bacillus migulanus sp. nov., Bacillus choshinensis sp. nov., Bacillus parabrevis sp. nov., and Bacillus galactophilus sp. nov.

Takagi¹,

Sato²,

Kadowaki³

et al. 1993

International Journal of Systematic Bacteriology

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“…brevis have been used for studies on GS biosynthesis: the Nagano strain in Japan, strain GB (7) in the Soviet Union, 266 VANDAMME AND DEMAIN and ATCC 9999 in the rest of the world. Most studies have been done with complex media.…”

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confidence: 99%

Nutrition of Bacillus brevis ATCC 9999, the Producer of Gramicidin S

Vandamme

Demain

1976

Antimicrob Agents Chemother

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The nutrition of the gramicidin S (GS) producer Bacillus brevis ATCC 9999 was studied with respect to growth and antibiotic production. In a complex medium containing yeast extract and peptone, only glycerol, 1)-fructose, and meso-inositol contributed to growth, whereas other hexoses, pentoses, polyols, polysaccharides, and organic acids were inactive. Glycerol and fructose were inhibitory to GS formation; inositol had no effect. In chemically defined media containing a mixture of five amino acids (glutamine, methionine, proline, arginine, and histidine) necessary for good growth, again only glycerol, ]-fructose, and meso-inositol served as effective carbon sources for growth. With respect to antibiotic formation, inositol was much poorer than glycerol or fructose. In the presence of glycerol and the five amino acids, addition of 0.1% Lphenylalanine specifically stimulated GS production. Growth kinetics in this defined medium showed two phases: early and rapid growth at the expense ofthe amino acid mixture, followed by glycerol utilization after a diauxic lag of 2 to 3 h; inositol utilization suffered a much longer lag period. GS formation was initiated after the diauxic lag. Ammonium sulfate was the best individual nitrogen source, but the combination of the five growth-stimulatory amino acids was more potent for GS production. The level of potassium phosphate influenced GS production. High levels stimulated the growth rate but progressively inhibited GS production. Methionine was the best sulfur source. A defined medium (F3/6) that supported rapid exponential growth (, = 0.53 h-') followed by progressively slower growth over a 30-to 40-h period was devised containing fructose, six amino acids, and mineral salts. GS formation was initiated as the cells left the exponential growth phase and reached high levels (1,500 mg of GS per liter; 0.19 mg of GS/mg of dry cell weight) after 40 to 50 h of incubation.The biosynthesis of the cyclic decapeptide antibiotic gramicidin S (GS) by certain strains of Bacillus brevis has been the subject of extensive research in the last decade (15,16,18). These studies have led to the discovery of a nonribosomal process for peptide biosynthesis. GS, which consists of two identical pentapeptides (DPhe-LPro-LVal-LOrn-LLeu), is synthesized by two enzymes in the presence of adenosine 5'-triphosphate and Mg2+. Synthetase II (molecular weight, 100,000) activates and racemizes L-phenylalanine, whereas synthetase I (molecular weight, 280,000) activates L-proline, L-valine, L-ornithine, and ileucine. In sequence, each amino acid is translocated from its I Contribution no.

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“…Temperature and medium composition seem to be the most important factors for cultivation. Initially, the GS producer was described as a thermotolerant strain, grown at temperatures up to 60°C (14). Later, a phenotypic dichotomy (dissociation into R and S phenotypes) was shown, and the optimal growth temperature for the GS-producing R form was determined to be 40 to 42°C, in contrast to 37°C for the unproductive S phenotype (14,55).…”

Section: Discussionmentioning

confidence: 99%

“…which has been used prominently as an antibiotic in local applications since 1944 (13,14). The peptide exerts a wide range of antimicrobial effects, including activity against grampositive and gram-negative bacteria, viruses, fungi, and singlecell pathogenic eukaryotes (5,13,19,22,23,52).…”

Section: Gramicidin S (Gs) Is a Cyclic Decapeptide ([Fpvolfpvol] Cyclo )mentioning

confidence: 99%

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The Ability ofAneurinibacillus migulanus(Bacillus brevis) To Produce the Antibiotic Gramicidin S Is Correlated with Phenotype Variation

Berditsch

Afonin

Ulrich

2007

Appl Environ Microbiol

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Phenotype instability of bacterial strains can cause significant problems in biotechnological applications, since industrially useful properties may be lost. Here we report such degenerative dissociation for Aneurinibacillus migulanus (formerly known as Bacillus brevis) an established producer of the antimicrobial peptide gramicidin S (GS). Phenotypic variations within and between various strains maintained in different culture collections are demonstrated. The type strain, ATCC 9999, consists of six colony morphology variants, R, RC, RP, RT, SC, and SP, which were isolated and characterized as pure cultures. Correlations between colony morphology, growth, GS production, spore formation, and resistance to their own antimicrobial peptide were established in this study. We found the original R form to be the best producer, followed by RC, RP, and RT, while SC and SP yielded no GS at all. Currently available ATCC 9999 T contains only 2% of the original R producer and is dominated by the newly described phenotypes RC and RP. No original R form is detected in the nominally equivalent strain DSM 2895 T (‫؍‬ATCC 9999 T ), which grows only as SC and SP phenotypes and has thus completely lost its value as a peptide producer. Two other strains from the same collection, DSM 5668 and DSM 5759, contain the unproductive SC variant and the GS-producing RC form, respectively. We describe the growth and maintenance conditions that stabilize certain colony phenotypes and reduce the degree of degenerative dissociation, thus providing a recommendation for how to revert the nonproducing smooth phenotypes to the valuable GS-producing rough ones.

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Gramicidin S Origin and Mode of Action

Cited by 21 publications

References 0 publications

Characterization of Bacillus brevis with Descriptions of Bacillus migulanus sp. nov., Bacillus choshinensis sp. nov., Bacillus parabrevis sp. nov., and Bacillus galactophilus sp. nov.

Characterization of Bacillus brevis with Descriptions of Bacillus migulanus sp. nov., Bacillus choshinensis sp. nov., Bacillus parabrevis sp. nov., and Bacillus galactophilus sp. nov.

Nutrition of Bacillus brevis ATCC 9999, the Producer of Gramicidin S

The Ability ofAneurinibacillus migulanus(Bacillus brevis) To Produce the Antibiotic Gramicidin S Is Correlated with Phenotype Variation

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