Resistance of Streptomyces cinnamonensis to butyrate and isobutyrate: production and properties of a new anti-isobutyrate (AIB) factor

Abstract

Butyiate and isobutyrate (after isomerization to n-butyrate) are specific precursors for the biosynthesis of monensin A in Streptomyces cinnamonensis. High concentrations of both butyrate and isobutyrate (greater than 20 and 10 mM, respectively) were toxic to S. cinnamonensis plated on solid medium. Spontaneous mutants resistant to these substances were isolated. These new strains produced monensins at even higher concentrations of butyrate or isobutyrate, with an increased yield of monensin A. S. cinnamonensis produced an anti-isobutyrate (AIB) factor, which was originally found to be excreted by some isobutyrate-resistant strains growing on solid medium containing isobutyrate. On plates, the AIB factor efficiently counteracted toxic concentrations not only of isobutyrate, but also of acetate, propionate, butyrate, 2-methylbutyrate, valerate and isovalerate against S. cinnamonensis as well as other Streptomyces species. Although the AIB factor enabled normal growth, sporulation and monensin production on plates, it did not have positive effects on submerged cultures of S. cinnamonensis with isobutyrate. The partial purification of the AIB factor was achieved. The role of the AIB factor during spore germination on solid medium containing isobutyrate or its homologues is discussed.

Excretion of butyraldehyde, isobutyraldehyde and valeraldehyde by Streptomyces cinnamonensis

“…The seed cultures were grown for 3 days and then used in the amount of 5% (v/v) for inoculation. Submerged cultivations of the strain S. cinnamonensis C-100-5 on a complex soybean flour medium were performed as described previously [8]. The cultures were incubated at 34°C on a rotary shaker at 2.7 Hz for 80 h.…”

Na+-Coupled Respiration and Reshaping of Extracellular Polysaccharide Layer Counteract Monensin-Induced Cation Permeability in Prevotella bryantii B14

“…Monensin and its derivates were originally isolated from the soil bacteria Streptomyces cinnamonensis , which synthesizes the compound via the precursor butyrate and iso-butyrate [ 9 , 10 , 11 ]. A self-intoxication by monensin is prevented due to the anti-isobutyrate factor (AIB), which allows S. cinnamonensis to resist not only monensin, but also toxic concentrations of short-chain fatty acids (SCFAs) [ 12 ].…”

Ethanol Production in Actinomycetes after Expression of Synthetic adhB and pdc

“…Isolation of plasmid and chromosomal DNA, conjugation and other gene cloning techniques for actinomycetes were performed using protocols that have been described by Kieser et al [14].…”

“…Tü 6071 [12], S. fradiae Tü 2717 [13], S. cinnamonensis [14] S. collinus [15], S. coelicolor CH999 [16], S. lividans 1326 [17], S. coelicolor M1154 [18] and S. coelicolor A3(2) [17] were grown at 28°C in 100 ml of liquid medium. HA medium (yeast extract, 4 g/l; malt extract, 10g/l; D-glucose, 4 g/l; CaCl 2 ,1 M), TSB medium (tryptic soy broth, 30 g/l), NL111 medium (Lab lemco meat extract, 20 g/l; malt extract, 100g/l; CaCO 3 10g/l), NL 19 medium (D-mannitol, 20 g/l; soy flour,20g/l) and E1 medium (starch 20 g/l; D-glucose, 20 g/l; yeast extract, 2.5 g/l; pharmamedia cottonseed flour, 3 g/l; CaCO 3, 1g/l; NaCl, 2 g/l; KHPO 4 , 1 g/l; MgSO 4 , 1 g/l) containing appropriate antibiotics for selection had been tested for best production levels, which could be obtained using NL 111 medium.…”