The relationship between antibiotic production and culture growth rate in Saccharopolyspora erythraea and Streptomyces hygroscopicus was manipulated by changing the growth‐limiting substrate. Carbon‐ and nitrogen‐limited cultures were studied and antibiotic synthesis was obtained in both cases in Saccharopolyspora erythraea cultures and in nitrogen‐limited Streptomyces hygroscopicus cultures. In all cultures where antibiotic was detected, onset of antibiotic production coincided with the minimal protein synthesis rate. Further investigation in Saccharopolyspora erythraea cultures indicated that this corresponded to minimum ratio of charged to uncharged tRNA, i.e. when uncharged tRNA accumulated. This latter phenomenon was investigated in the presence of a protein synthesis inhibitor.
Microbial biofilms can be valuable in many biotechnological applications, including bioremediation. We have previously constructed a recombinant strain of Caulobacter crescentus JS4022/p723-6H by inserting a hexahistidine peptide to a permissive site of the host surface layer (S-layer) protein RsaA. This engineered strain is highly effective in removal of cadmium from water as free cells. In this communication, we examined the biofilms formed by self-immobilized JS4022/p723-6H and evaluated their ability to retrieve cadmium from contaminated water samples. According to light and electron microscopic observations, JS4022/p723-6H cells developed a uniform monolayer biofilm on borosilicate surfaces through their intrinsic appendage, a stalk with an adhesive holdfast. The density of the biofilms reached a maximum after 48 h of incubation and was not affected by exposure to at least 1 ppm cadmium. When 0.4 ppm Cd(II) was added to the growth medium, immobilized JS4022/p723-6H removed 76.9% of the total metal, whereas the control strain only removed 13.5%. When a water sample collected from Lake Erie was spiked with various amounts of CdCl2, immobilized JS4022/p723-6H was able to sequester 44 approximately 51% of the total metal, compared to 37 approximately 42% accumulated by the control strain. By combining two powerful techniques, cell surface display and self-immobilization, we achieved complete separation of dissolved heavy metals from contaminated water in a single step. This study laid down the foundation to cost-effectively construct large-scale bioreactors with high efficiency and specificity to retrieve environmental contaminants from water.
Against Enterobacter aerogenes 13048, Serratia marcescens 13880, Klebsiella pneumoniae 10031, Pseudomonas aeruginosa 10145, Escherichia coli 9723, Lactobacillus casei 7469, Lactobacillus plantarum 8014, Leuconostoc dextranicum 8086, and Streptococcus faecalis 8043, the mean minimal inhibitory concentrations of three cyclic hydroxamic acids, 3-amino-3,4-dihydro-1-hydroxycarbostyril, the 6-chloro analog, and the 7-chloro analog, were 0.6, 0.6, and 0.2 ug/ml, and those of the corresponding lactams, 3-amino-3,4-dihydrocarbostyril, the 6-chloro analog, and the 7-chloro analog, were 60, 60, and 6 ug/ml, respectively. Under the same assay conditions the mean minimal inhibitory concentrations of chloramphenicol and kanamycin were both 2 pug/ml. In addition, the cyclic hydroxamic acids but not the lactams inhibited the growth of Candida albicans at miniimal inhibitory concentrations ranging from 20 to 200 pg/ml, at pH 7, as compared with that of amphotericin B, at 2 pg/ml.
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