Identification of erythrobactin, a hydroxamate-type siderophore produced by Saccharopolyspora erythraea.

AIMS: To investigate the production of siderophores by Saccharopolyspora erythraea SGT2 and how this production is affected by the inoculum. METHODS AND RESULTS: When grown in a low-iron, chemically defined medium (CDM), the soil dwelling actinomycete S. erythraea secretes a substance that is reactive in the nonspecific chrome azurol S (CAS) assay. Importantly, the production of CAS-reactive substance is highly reduced upon the addition of 0.925 micromol l(-1) iron to the cultures and has a peak of production in the late-log to early stationary growth phase. In addition, the culture supernatants tested were negative in the Arnow and Rioux assays but positive in the Csáky procedure. Interestingly, we also found evidence that the production of this CAS-reactive substance in CDM was highly reduced, when inoculated with cells that had been previously grown to late-stationary phase. Conversely, inocula derived from late-log to early stationary cultures presented high levels of CAS activity. CONCLUSIONS: These results indicate that S. erythraea produces a hydroxamate-type siderophore that we have generically designated as erythrobactin. Additionally, the inocula growth stage plays a key role in siderophore production in S. erythraea. SIGNIFICANCE AND IMPACT OF THE STUDY: It is the first evidence for siderophore synthesis in S. erythraea and one of the first examples of non-polyketide secondary metabolite production by this organism.

A quantitative approach to characterizing cell lysis caused by mechanical agitation of Streptomyces clavuligerus.

Streptomyces clavuligerus is a commercially important actinomycete that is used to produce clavulanic acid, a beta-lactamase inhibitor. Observations during 10 batch cultivations with S. clavuligerus on defined media have led to the finding that the organism is very sensitive to shear when grown in batch cultures with increasing stirrer speed. The stirrer speed was increased to keep the dissolved oxygen level above 50% air saturation. A quantitative approach based on the calculation of elemental balances and a simple mathematical model is proposed to characterize the biomass lysis. Finally, a linear relation between biomass yield and observed specific growth rate is determined. Results show that cell lysis occurs at a high degradation rate, e.g., mu(max) = 0.16 h(-1) and k(d) = 0.07 h(-1), when the gassed power input increases above 1.1, 1.7, or 2.0 kW/m(3), respectively, depending on the medium composition. The overall biomass yield on substrate is dramatically reduced in all experiments (>30%).

Hyphal growth and fragmentation of Penicillium chrysogenum in submerged cultures.

A previously derived population model describing the average properties of hyphal elements in submerged cultures of filamentous fungi was revised, and a term for the influence fo spore germination on the average total hyphal length was added. The model was derived from a general balance for the distribution function for the hyphal elements. Based on experimental data and the derived model, simple kinetic expressions for spore germination, tip extension, branching, and hyphal break-up were set up. It is concluded that spore germination can be quantified by three parameters: (1) the time at which spore germination is initiated, (2) the time at which spore germination terminates, and (3) the fraction of viable spores in a spore suspension. The frequency of spore germination can be described with the B-distribution. For growth kinetics it is concluded that the branching frequency is closely correlated with the total hyphal length and that the average tip-extension rate can be described with saturation kinetics with respect to the hyphal length. Finally, the rate of fragmentation is linearly related to the energy input to the bioreactor, and related to the effective hyphal length.

Pellet formation and fragmentation in submerged cultures of Penicillium chrysogenum and its relation to penicillin production.

The spores of Penicillium chrysogenum are of the noncoagulating type, and after spore germination a culture of disperse mycelia is obtained. In this study, it is shown that when the hyphal elements increase in size, they may agglomerate, and depending on the operating conditions, these agglomerates may develop into pellets with a dense core. The influence of initial spore concentration and agitation rate on agglomeration, leading to pellet formation, was studied. For a low concentration of spores in the inoculum, only a few hyphal elements agglomerate and pellets with a small diameter are obtained. At higher spore concentrations, many hyphal elements agglomerate and develop into large diameter pellets. Finally, at a very high spore concentration in the inoculum, the final hyphal element size is small and agglomerates therefore are not formed. With a high agitation rate, the agglomeration of hyphal elements is reduced. In a repeated fed-batch cultivation, where there was a shift from pellet morphology to disperse mycelia, it was found that there is no relation between macroscopic morphology and penicillin production by P. chrysogenum. The morphology was quantified throughout the repeated fed-batch cultivation, and both the pellet diameter and the concentration of pellets were affected by the agitation rate.