Abstract
Bacillus brevis strain Nagano and its gramicidin S-negative mutant, BI-7, were compared with respect to germination of their spores produced in several media. Germination initiation occurred in the presence of nutrient broth orL-alanine but not with inosine, glucose, glycerol or fructose; the process was activated by heat. Parental and mutant spores behaved similarly in these experiments. During outgrowth, parental spores remained in this phase of germination much longer than did mutant spores, but only when the parental spores had been harvested from a sporulation medium where significant gramicidin S synthesis had occurred. When parental spores were extracted or treated with an enzyme that hydrolyzes gramicidin S, rapid outgrowth occurred. Adding exogenous gramicidin S or the extract from parental spores to mutant spores lengthened the outgrowth in a dose-dependent manner. The uptake of labeledL-alanine by parental spores was delayed compared to mutant spores in the presence or absence of chloramphenicol. These data suggest a mechanism of action for gramicidin S whereby it interferes in membrane function, such as transport or energy metabolism, in outgrowing spores.
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Abbreviations
- GS:
-
Gramicidin S
- CFU:
-
colony-forming units
References
Asatani M, Kurahashi K (1977) Carbohydrate metabolism inBacillus brevis ATCC 9999. J Biochem 81:813–822
Danders W, Marahiel MA (1981) Control of RNA synthesis by gramicidin S during germination and outgrowth ofBacillus brevis spores. FEMS Microbiol Lett 10:277–283
Demain AL, Piret JM (1979) Relationship between antibiotic biosynthesis and sporulation. In: Luckner M, Schreiber K (eds) Regulation of secondary product and plant hormone metabolism, vol 55. Pergamon Press, New York, pp 183–188
Egorov NS, Vyipiyach AN, Zharikova GG, Maksimov VN (1975) Effect of various factors on spore germination in S and P-variants ofBacillus brevis. Mikrobiologiya 44:237–240
Frangou-Lazaridis M, Lazaridis I, Nandi S, Seddon B (1980) Developmental significance of gramicidin S produced during sporulaiion ofBacillus brevis (Nagano). Studies of RNA synthesis using gramicidin S-negative mutants. Soc Gen Microbiol Quarterly 7:114
Haavik HI, Thomassen S (1973) A bacitracin-negative mutant ofB. licheniformis which is able to sporulate. J Gen Microbiol 76:451–454
Hsieh LK, Vary JC (1975) Germination and peptidoglycan solubilization inBacillus megaterium spores. J Bacteriol 123:463–470
Hyatt MZ, Levinson HS (1964) Effect of sugars and other carbon compounds on germination and postgerminative development ofBacillus megaterium spores. J Bacteriol 88:1403–1415
Iwaki M, Shimura K, Kanda M, Kaji E, Saito Y (1972) Some mutants ofBacillus brevis deficient in gramicidin S formation. Biochem Biophys Res Commun 48:113–118
Kaprel'yants AS, Nikiforov VV, Miroshnikov AI, Snezhkova LG, Eremin VA, Ostrovskii DN (1977) Bacterial membranes and mechanism of action of gramicidin S. Biokhimiya 42:329–337
Katz E, Demain AL (1977) The peptide antibiotics ofBacillus: chemistry, biogenesis and possible function. Bacteriol Rev 41:449–474
Keynan A (1978) Spore structure and its relations to resistance, dormancy and germination. In: Chambliss G, Vary JC (eds) Spores VII. American Society for Microbiology, Washington, D.C., pp 45–53
Lonshon CA, Setlow P (1982)Bacillus megaterium spore protease: Purification, radioimmunoassay and analysis of antigen level and localization during growth, sporulation and spore germination. J Bacteriol 150:303–311
Marahiel M, Danders W, Krause M, Kleinkauf H (1979) Biological role of gramicidin S in spore functions. Studies on gramicidin S-negative mutants ofBacillus brevis ATCC 9999. Eur J Biochem 99:49–55
Matteo CC, Glade M, Tanaka A, Piret J, Demain AL (1975) Microbiological studies on the formation of gramicidin S synthetases. Biotechnol Bioeng 17:129–142
Mitronova TN, Zharikova GG (1965) A method for obtaining spores from the four forms ofBacillus brevis vai. G.B. Mikrobiologiya 34:835–839
Mukherjee PK, Paulus H (1977) Biological function of gramicidin. Studies on gramicidin-negative mutants. Proc Natl Acad Sci USA 74:780–784
Nandi S, Seddon B (1978) Evidence for gramicidin S functioning as a bacterial hormone specifically regulating spore outgrowth inBacillus brevis strain Nagano. Biochem Soc Trans 6:409–411
Pache W, Chapman D, Hillaby R (1972) Interaction of antibiotics with membranes: Polymyxin B and gramicidin S. Biochim Biophys Acta 255:358–364
Pandey NK, Gollakota KG (1977) Induction of microcycle sporulation inBacillus brevis spp. AG4 by amethopterin. Arch Microbiol 114:189–191
Piret JM, Demain AL (1981) Role of gramicidin S in the producer organism,Bacillus brevis. In: Levinson HS, Sonenshein AL, Tipper DJ (eds) Sporulation and germination. American Society for Microbiology, Washington, D.C., pp 243–245
Piret JM, Demain AL (1982) Sporulation and spore properties ofBacillus brevis and its gramicidin S-negative mutant. Submitted
Ray B, Bose SK (1971) Polypeptide antibiotics-negative sporeformer mutants ofBacillus subtilis. J Gen Appl Microbiol 17:491–498
Rossignol DP, Vary JC (1979) Biochemistry ofL-proline-triggered germination ofBacillus megaterium spores. J Bacteriol 138:431–441
Sadoff HL (1972) Sporulation antibiotics ofBacillus species. In: Halvorson HO, Hansen R, Campbell LL (eds) Spores V. American Society for Microbiology, Washington, D.C., pp 157–166
Sarkar N, Paulus H (1972) Function of peptide antibiotics in sporulation. Nature (New Biology) 239:228–230
Schaeffer P (1969) Sporulation and the production of antibiotics, exoenzymes and exotoxins. Bacterial Rev 33:48–71
Seddon B, Nandi S (1978) Biochemical aspects of germination and outgrowth ofBacillus brevis Nagano and control by gramicidin S. Biochem Soc Trans 6:412–413
Setlow P (1975) Purification and properties of some unique low molecular weight basic proteins degraded during germination ofBacillus magaterium. J Biol Chem 250:8168–8173
Seto-Young DL, Ellar DJ (1979) Membrane changes during germination ofBacillus megaterium KM spores. Microbios 26:7–15
Tochikubo K, Hayakawa Y, Kojima K (1981) Mutual relationship between antibiotics and resting spores ofBacillus subtilis: binding of cyclic polypeptide and aminoglycoside antibiotics to spores and their inhibitory effect on outgrowth and vegetative growth. Microbiol Immunol 25:113–126
Vinter V, Slepecky RA (1965) Direct transition of outgrowing bacterial spores to new sporangia without intermediate cell division. J Bacteriol 90:803–807
Yukioka M, Saito Y, Otani S (1966) Enzymatic hydrolysis of gramicidin S. J Biochem 60:295–302
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Piret, J.M., Demain, A.L. Germination initiation and outgrowth of spores ofBacillus brevis strain Nagano and its gramicidin S-negative mutant. Arch. Microbiol. 133, 38–43 (1982). https://doi.org/10.1007/BF00943767
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DOI: https://doi.org/10.1007/BF00943767