Effect of temperature and yeast extract on microbial respiration of sediments from a shallow coastal subsurface and vadose zone.

As a part of our study on microbial heterogeneity in subsurface environments, we have examined the microbial respiration of sediment samples obtained from a coastal site near Oyster, VA. The sediments at the site are unconsolidated, fine to coarse beach sand and gravel. A Columbus Instruments Micro-Oxymax Respirometer was used to measure the rate of carbon dioxide (CO2) production during the respiration of the sediment samples. The rate of respiration of the sediment samples ranged from 0.035-0.6 microL CO2/h/g of the sediment. The sediment samples showing maximum (0.6 microL CO2/h/g) and minimum (0.035 microL CO2/h/g) production of CO2 were selected to study the effect of micronutrient-yeast extract (0.5 and 1.0 micrograms/g of the sediment) and water (0.5 and 1.0 mL) on the rate of CO2 production. The rate of CO2 production increased with the addition of water, but increased approx 2 orders of magnitude (from 0.26 to an average of 23.5 microL CO2/h/g) when 1.0 g/g yeast extract was added to the sediment samples. In these coastal sediments, temperature, depth, and addition of water influenced microbial activity, but the addition of 1.0 microgram/g yeast extract as a micronutrient rapidly increased the rate of CO2 production 2 orders of magnitude.

Cell-free extract(s) of Pseudomonas putida catalyzes the conversion of cyanides, cyanates, thiocyanates, formamide, and cyanide-containing mine waters into ammonia.

Our isolate, Pseudomonas putida, is known to be capable of utilizing cyanides as the sole source of carbon (C) and nitrogen (N) both in the form of free cells and cells immobilized in calcium alginate. In the present study, the cell-free extract(s) were prepared from the cells of P. putida grown in the presence of sodium cyanide. The ability of enzyme(s) to convert cyanides, cyanates, thiocyanates, formamide and cyanide-containing mine waters into ammonia (NH3) was studied at pH 7.5 and pH 9.5. The kinetic analysis of cyanide and formamide conversion into NH3 at pH 7.5 and pH 9.5 by the cell-free extract(s) of P. putida was also studied. The Km and Vmax values for cyanide/formamide were found to be 4.3/8 mM and 142/227 mumol NH3 released mg protein-1 min-1 respectively at pH 7.5 and 5/16.67 mM and 181/434 mumol NH3 released mg protein-1 h-1 respectively at pH 9.5. The study thus concludes that the cell-free extract(s) of P. putida is able to metabolize not only cyanides, cyanates, thiocyanates, and formamide but also cyanide-containing mine waters to NH3.

Biodegradation of cyanides, cyanates and thiocyanates to ammonia and carbon dioxide by immobilized cells of Pseudomonas putida.

Pseudomonas putida utilizes cyanide as the sole source of carbon and nitrogen. Agar, alginate, and carrageenan were screened as the encapsulating matrices for P. putida. Alginate-immobilized cells of P. putida degraded sodium cyanide (NaCN) more efficiently than non-immobilized cells or cells immobilized in agar or carrageenan. The end products of biodegradation of cyanide were identified as ammonia (NH3) and carbon dioxide (CO2). These products changed the medium pH. In bioreactors, the rate of cyanide degradation increased with an increase in the rate of aeration. Maximum utilization of cyanide was observed at 200 ml min-1 of aeration. Immobilized cells of P. putida degraded cyanides, cyanates and thiocyanates to NH3 and CO2. Use of Na[14C]-CN showed that 70% of carbon of Na[14C]-CN was converted into 14CO2 and only 10% was associated with the cell biomass. The substrate-dependent kinetics indicated that the Km and Vmax values of P. putida for the substrate, NaCN were 14 mM and 29 nmol of oxygen consumed mg protein-1 min-1 respectively.