Effect of Iron and Salt on Prodigiosin Synthesis in Serratia marcescens
Serratia marcescens wild-types ATCC 264 and Nima grew but did not synthesize prodigiosin in a glycerol-alanine medium containing 10 ng of Fe per ml. Wild-type 264 required the addition of 0.2 μg of Fe per ml for maximal growth and prodigiosin synthesis; Nima required 0.5 μg of Fe per ml. Three percent, but not 0.1%, sea salts inhibited prodigiosin synthesis in a complex medium containing up to 10 μg of Fe per ml. NaCl was the inhibitory sea salt component. The inhibition was not specific for NaCl; equimolar concentrations of Na2SO4, KCl, and K2SO4 also inhibited prodigiosin synthesis. Experiments with strains 264 and Nima and with mutant WF which cannot synthesize 4-methoxy-2-2′-bipyrrole-5-carboxyaldehyde (MBC), the bipyrrole moiety of prodigiosin, and with mutant 9-3-3 which cannot synthesize the monopyrrole moiety 2-methyl-3-amylpyrrole (MAP) showed that both MBC synthesis and the reaction condensing MAP and MBC to form prodigiosin were relatively more sensitive to NaCl inhibition than the MAP-synthesizing step. The capacity of whole cells to condense MAP and MBC was present, but inactive, in cells grown in NaCl; removal of the NaCl from non-proliferating salt-grown cells restored the activity. Other evidence suggests the existence of a common precursor to the MAP- and MBC-synthesizing pathways.
Enhanced production of prodigiosin by Serratia marcescens MO-1 using ram horn peptone
This work addresses the production of prodigiosin from ram horn peptone (RHP) using MO-1, a local isolate in submerged culture. First, a novel gram-negative and rod-shaped bacterial strain, MO-1, was isolated from the body of the grasshopper (Poecilemon tauricola Ramme 1951), which was collected from pesticide-contaminated fields. Sequence analysis of 16S rDNA classified the microbe as Serratia marcescens. The substrate utilization potential (BIOLOG) and fatty acid methyl ester profile (FAME) of S. marcescens were also determined. The effect of RHP on the production of prodigiosin by S. marcescens MO-1 was investigated, and the results showed that RHP supplementation promoted the growth of MO-1 and increased the production of prodigiosin. A concentration of 0.4% (w/v) RHP resulted in the greatest yield of prodigiosin (277.74 mg/L) after 48 h when mannitol was used as the sole source of carbon. The pigment yield was also influenced by the types of carbon sources and peptones. As a result, RHP was demonstrated to be a suitable substrate for prodigiosin production. These results revealed that prodigiosin could be produced efficiently by S. marcescens using RHP.
Development of an Analytical Method for Explosive Residues in Soil
Abstract : An analytical method was developed to determine the concentrations of HMX, RDX, TNB, DNB, Tetryl, TNT and 2,4-DNT in soil. The method involves extracting a 2-g sample with 50 mL of acetonitrile using an ultrasonic bath procedure for 18 hr. A 10-mL portion of the extract is diluted with 10 mL of water, filtered through a 0.45-micrometer Millex SR filter, and analyzed by RP-HPLC using a fixed 254-nm UV detector. Separations were obtained on an LC-18 column eluted with 50:50 water-methanol. Retention times were 2.55, 3.82, 5.16, 6.25, 7.04, 8.47 and 10.15 min for HMX, RDX, TNB, DNB, Tetryl, TNT and 2,4-DNT, respectively. Confirmation of analyte identities is recommended by RP-HPLC on an LC-CN column using 50:50 water-methanol. Kinetic studies using naturally contaminated soil indicated that equilibrium was achieved within 24 hr for the majority of soils and analytes studied. The method provides linear calibration curves over a wide range of concentrations. Reporting limits were conservatively estimated at 1.6, 1.8, 1.5, 0.5, 5.5, 0.8 and 0.8 micro grams/g for HMX, RDX, TNB, DNB, Tetryl, TNT and 2,4-DNT. Analytical precision was estimated at + or - 0.44, 0.51, 0.43, 0.13, 1.24, 0.27 and 0.20 micro grams/g, respectively, in the range of homogeneous variance near the detection limits. At higher concentrations the relative standard deviation was constant at 2% for HMX, RDX, DNB and TNB, 3% for TNT and 2,4-DNT, and 6% for Tetryl.