Constitutive soluble methane monooxygenase mutants of methanotrophic bacteria such as Methylosinus trichosporium A.T.C.C. 55314

Selection of mutant methanotrophic bacteria capable of efficiently degrading halogenated hydrocarbons typically found in numerous wastewater sources is described. The mutants are distinguishable from parental strains in having a unique resistance to the presence of copper while exhibiting unusually high degradation rates toward trichlorethylene. Methylosinus trichosporium A.T.C.C. 55314 strains are particularly good sources of the described mutants which may be obtained using a new method of selection and screening. The disclosed microorganisms may be immobilized on various matrices and are particularly adaptable for use in bioreactors. Further, the methanotrophic bacteria have antibiotic resistance to streptomycin or rifampicin B.Board of Regents, University of Texas Syste

Methsnotrophic Biodegradation of Trichloroethylene in a Hollow Fiber Membrane Bioreactor

Biodegradation of Trichloroethlyene (TCE) in a Hollow Fiber Membrane Bioreactor Was Investigated using a Mutant of the Methanotrophic Bacteria, Methylosinus Trichosporium 0B3b. Contaminated Water Flowed through the Lumen (I.e., Fiber Interior), and the Bacteria Circulated through the Shell Side of the Membrane Module and an External Growth Reactor. in Mass Transfer Studies with a Radial Cross-Flow Membrane Module, 78.3-99.9% of the TCE Was Removed from the Lumen at Hydraulic Residence Times of 3-15 Min in the Lumen and the Shell. in Biodegradation Experiments, 80-95% of the TCE Was Removed from the Lumen at Hydraulic Residence Times of 5-9 Min in the Lumen. the TCE Transferred to the Shell Was Rapidly Biodegraded, with Rate Constants Ranging from 0.16 to 0.9 L (Mg of TSS)-1 Day-1. Radiochemical Data Showed that over 75% of the Transferred TCE Was Biodegraded in the Shell, with the Byproducts Being Approximately Equally Divided between Carbon Dioxide and Nonvolatiles. This Study Shows that a Hollow Fiber Membrane Bioreactor System Coupled with the Mutant Strain PP358 of M. Trichosporium 0B3b is a Very Promising Technology for Chlorinated Solvent Biodegradation. © 1995, American Chemical Society. All Rights Reserved

Dissolution Kinetics of High Explosives Particles in a Saturated Sandy Soil

Solid phase high explosive (HE) residues from munitions detonation may be a persistent source of soil and groundwater contamination at military training ranges. Saturated soil column tests were conducted to observe the dissolution behavior of individual components (RDX, HMX, and TNT) from two HE formulations (Comp B and C4). HE particles dissolved readily, with higher velocities yielding higher dissolution rates, higher mass transfer coefficients, and lower effluent concentrations. Effluent concentrations were below solubility limits for all components at superficial velocities of 10–50 cm day-1. Under continuous flow at 50 cm day-1, RDX dissolution rates from Comp B and C4 were 34.6 and 97.6 μg h-1 cm-2 (based on initial RDX surface area), respectively, significantly lower than previously reported dissolution rates. Cycling between flow and no-flow conditions had a small effect on the dissolution rates and effluent concentrations; however, TNT dissolution from Comp B was enhanced under intermittent-flow conditions. A model that includes advection, dispersion, and film transfer resistance was developed to estimate the steady-state effluent concentrations

Cometabolism of Trihalomethanes by Nitrosomonas europaea

The ammonia-oxidizing bacterium Nitrosomonas europaea (ATCC 19718) was shown to degrade low concentrations (50 to 800 μg/liter) of the four trihalomethanes (trichloromethane [TCM], or chloroform; bromodichloromethane [BDCM]; dibromochloromethane [DBCM]; and tribromomethane [TBM], or bromoform) commonly found in treated drinking water. Individual trihalomethane (THM) rate constants ([Formula: see text]) increased with increasing THM bromine substitution, with TBM > DBCM > BDCM > TCM (0.23, 0.20, 0.15, and 0.10 liters/mg/day, respectively). Degradation kinetics were best described by a reductant model that accounted for two limiting reactants, THMs and ammonia-nitrogen (NH(3)-N). A decrease in the temperature resulted in a decrease in both ammonia and THM degradation rates with ammonia rates affected to a greater extent than THM degradation rates. Similarly to the THM degradation rates, product toxicity, measured by transformation capacity (T(c)), increased with increasing THM bromine substitution. Because both the rate constants and product toxicities increase with increasing THM bromine substitution, a water's THM speciation will be an important consideration for process implementation during drinking water treatment. Even though a given water sample may be kinetically favored based on THM speciation, the resulting THM product toxicity may not allow stable treatment process performance

Degradation Of Trichloroethylene By Methanol-Grown Cultures Of Methylosinus Trichosporium Ob3B Pp358

A soluble methane monooxygenase-constitutive mutant strain of Methylosinus trichosporium OB3b, strain PP358, was grown with methanol as the carbon source, and the kinetics of trichloroethylene (TCE) degradation were determined. PP358 exhibited high TCE degradation rates under both oxygen- and carbon-limiting conditions. The optimal pseudo first-order rate constant for TCE was comparable to the values measured for cells grown,vith methane. We found that growth under oxygen-limiting conditions results in increased accumulation of polyhydroxybutyrate, which in turn correlates,vith higher transformation capacities for TCE. It was also shown that methanol inhibits TCE degradation only at high concentrations. Thus, methanol-grown cultures of PP358 represent an efficient system for the biodegradation of chlorinated hydrocarbons.Gulf Coast Hazardous Substance Research CenterTexas Advanced Technology Development Program Technology Development ProgramCivil, Architectural, and Environmental EngineeringChemical Engineerin

Ammonia-Oxidizing Bacteria in Biofilters Removing Trihalomethanes Are Related to Nitrosomonas oligotropha ▿

Ammonia-oxidizing bacteria (AOB) in nitrifying biofilters degrading four regulated trihalomethanes—trichloromethane, bromodichloromethane, dibromochloromethane, and tribromomethane—were related to Nitrosomonas oligotropha. N. oligotropha is associated with chloraminated drinking water systems, and its presence in the biofilters might indicate that trihalomethane tolerance is another reason that this bacterium is dominant in chloraminated systems

Methylosinus trichosporium OB3b Mutants Having Constitutive Expression of Soluble Methane Monooxygenase in the Presence of High Levels of Copper

The methanotrophic bacterium Methylosinus trichosporium OB3b is unusually active in degrading recalcitrant haloalkanes such as trichloroethylene (TCE). The first and rate-limiting step in the degradation of TCE is catalyzed by a soluble methane monooxygenase (sMMO). This enzyme is not expressed when the cells are grown in the presence of copper at concentrations typically found in polluted groundwater. Under these conditions, M. trichosporium OB3b expresses a particulate form of the enzyme (pMMO), which has a narrow substrate specificity and does not degrade TCE at any significant rate. We have isolated M. trichosporium OB3b mutants that are deficient in pMMO and express sMMO constitutively in the presence of elevated concentrations of copper. One mutant (PP358) exhibited a TCE degradation rate which was almost twice as high as that of the wild-type strain grown under optimal conditions (without copper). All of the mutants lost the ability to express pMMO activity and to form stacked intracellular membranes characteristic of wild-type cells expressing pMMO