Abstract: The present invention relates to an isolated nucleic acid molecule which encodes one or more of the enzymes which catalyze one or more steps in the desulfurization of thiophene, or a homologue or active fragment thereof. The invention also includes a recombinant microorganism containing one or more such heterologous nucleic acid molecules. The invention also provides a method for desulfurizing a fossil fuel containing thiophene and/or one or more substituted thiophenes. The method comprises contacting the fossil fuel with an organism containing a recombinant nucleic acid molecule which encodes an enzyme which catalyzes the desulfurization of thiophene.

Abstract: The invention relates to a novel microorganism, designated Sphingomonas sp. strain AD109, which is capable of selectively desulfurizing dibenzothiophene. The invention also includes isolated proteins and nucleic acid sequences obtained from this microorganism. In another embodiment, the invention provides a method of using this microorganism or enzyme preparations derived therefrom in the biocatalytic desulfurization of a fossil fuel containing organic sulfur compounds.

Abstract: The present invention relates to a method for the desulfurization of a fossil fuel containing one or more organosulfur compounds. In one embodiment, the method comprises the steps of (1) contacting the fossil fuel with a biocatalyst capable of converting the organosulfur compound to an oxyorganosulfur compound which is separable from the fossil fuel; and (2) separating the oxyorganosulfur compound from the fossil fuel. The oxyorganosulfur compound can then be isolated, discarded or further processed, for example, via desulfurization by a biocatalyzed process or an abiotic process, such as hydrodesulfurization.

Abstract: The invention relates to the discovery that the rate of reaction of the desulfurization of fossil fuels is enhanced by the addition of a flavoprotein to the biocatalyst. The invention is drawn to a method for enhancing the rate of desulfurizing a fossil fuel containing organic sulfur compounds, comprising the steps of:a) contacting the fossil fuel with an aqueous phase containing a biocatalyst capable of cleaving carbon-sulfur bonds and a rate-enhancing amount of a flavoprotein, thereby forming a fossil fuel and aqueous phase mixture;b) maintaining the mixture of step (a) under conditions sufficient for cleavage of the carbon-sulfur bonds of the organic sulfur molecules by the biocatalyst, thereby resulting in a fossil fuel having a reduced organic sulfur content; andc) separating the fossil fuel having a reduced organic sulfur content from the resulting aqueous phase.

Abstract: The present invention relates to compounds having utility as surfactants which are derived from intermediates produced in petroleum biodesulfurization processes. The compounds of the invention include acyloxybiphenylsulfinates, acyloxybiphenylsulfonates, alkyl sulfinatobiphenyl ethers and alkyl sulfonatobiphenyl ethers. The invention also provides methods of producing these compounds.

Abstract: The invention relates to the unexpected discovery that the removal of the sulfinate group from an organosulfinate compound can be improved in the presence of an effective amount of a Lewis acid. The invention includes a method of removing the sulfinic acid functional group from an organosulfinic acid compound, comprising reacting the organosulfinic acid compound with an effective amount of a Lewis acid.

Abstract: The present invention provides novel recombinant pseudomonads which contain a heterologous nucleic acid molecule comprising a nucleotide sequence encoding one or more desulfurization enzymes which are components of a biodesulfurization catalyst. The invention also provides a method of desulfurizing a carbonaceous material, such as a fossil fuel, which comprises organosulfur compounds. The method includes the steps of The method comprises the steps of (1) contacting the fossil fuel with an aqueous phase containing a recombinant biocatalyst which is capable of cleaving carbon-sulfur bonds and, optionally, a flavoprotein, thereby forming a fossil fuel and aqueous phase mixture; (2) maintaining the mixture under conditions sufficient for cleavage of the carbon-sulfur bonds of the organosulfur molecules by the biocatalyst, thereby resulting in a fossil fuel having a reduced organic sulfur content; and (3) separating the fossil fuel having a reduced organic sulfur content from the resulting aqueous phase.

Abstract: The invention provides a novel flavin reductase, which has been isolated from Rhodococcus strain sp. IGTS8 and purified to homogeneity. The flavin reductase is of use in the biocatalytic desulfurization of a fossil fuel containing organic sulfur compounds. The method comprises the steps of (1) contacting the fossil fuel with an aqueous phase containing a biocatalyst capable of cleaving carbon-sulfur bonds and a rate-enhancing amount of the flavin reductase of the present invention, thereby forming a fossil fuel and aqueous phase mixture; (2) maintaining the mixture under conditions sufficient for cleavage of the carbon-sulfur bonds of the organic sulfur molecules by the biocatalyst, thereby resulting in a fossil fuel having a reduced organic sulfur content; and (3) separating the fossil fuel having a reduced organic sulfur content from the resulting aqueous phase.

Abstract: This invention relates to recombinant DNA molecule containing a gene or genes which encode a biocatalyst capable of desulfurizing a fossil fuel which contains organic sulfur molecules. For example, the present invention encompasses a recombinant DNA molecule containing a gene or genes of a strain of Rhodococcus rhodochrous.

Abstract: The invention relates to the discovery that the rate of reaction of the desulfurization of fossil fuels is enhanced by the addition of an oxidoreductase to the biocatalyst. The invention is drawn to a method for enhancing the rate of desulfurizing a fossil fuel containing organic sulfur compounds, comprising the steps of:a) contacting the fossil fuel with an aqueous phase containing a biocatalyst capable of cleaving carbon-sulfur bonds and a rate-enhancing amount of an oxidoreductase, thereby forming a fossil fuel and aqueous phase mixture;b) maintaining the mixture of step (a) under conditions sufficient for cleavage of the carbon-sulfur bonds of the organic sulfur molecules by the biocatalyst, thereby resulting in a fossil fuel having a reduced organic sulfur content; andc) separating the fossil fuel having a reduced organic sulfur content from the resulting aqueous phase.

Abstract: The invention relates to the discovery that the rate of reaction of the desulfurization of fossil fuels is enhanced by the addition of an oxidoreductase to the biocatalyst. The invention is drawn to a method for enhancing the rate of desulfurizing a fossil fuel containing organic sulfur compounds, comprising the steps of:a) contacting the fossil fuel with an aqueous phase containing a biocatalyst capable of cleaving carbon-sulfur bonds and a rate-enhancing amount of an oxidoreductase, thereby forming a fossil fuel and aqueous phase mixture;b) maintaining the mixture of step (a) under conditions sufficient for cleavage of the carbon-sulfur bonds of the organic sulfur molecules by the biocatalyst, thereby resulting in a fossil fuel having a reduced organic sulfur content; andc) separating the fossil fuel having a reduced organic sulfur content from the resulting aqueous phase.

Abstract: The invention provides a novel flavin reductase, which has been isolated from Rhodococcus strain sp. IGTS8 and purified to homogeneity. The flavin reductase is of use in the biocatalytic desulfurization of a fossil fuel containing organic sulfur compounds. The method comprises the steps of (1) contacting the fossil fuel with an aqueous phase containing a biocatalyst capable of cleaving carbon-sulfur bonds and a rate-enhancing amount of the flavin reductase of the present invention, thereby forming a fossil fuel and aqueous phase mixture; (2) maintaining the mixture under conditions sufficient for cleavage of the carbon-sulfur bonds of the organic sulfur molecules by the biocatalyst, thereby resulting in a fossil fuel having a reduced organic sulfur content; and (3) separating the fossil fuel having a reduced organic sulfur content from the resulting aqueous phase.

Abstract: A method and an apparatus for separating a water/organic/solid emulsion, wherein the solid comprises particles having a length of about 50 .mu.m or less, are disclosed. The method comprises the steps of (1) directing the water/organic/solid emulsion from an emulsion source through a first hydrocyclone; (2) inverting the phase of the overflow emulsion from the first hydrocyclone; (3) directing the overflow emulsion through one or more subsequent hydrocyclones arranged in series; and (4) collecting the aqueous phase and the organic phase. The solid can be a biocatalyst, such as an intact bacterial, fungal or yeast cell. The organic phase is a liquid material substantially comprising carbon, which is substantially immiscible with water, such as petroleum.

Abstract: The invention relates to the discovery that the rate of reaction of the desulfurization of fossil fuels is enhanced by the addition of a flavoprotein to the biocatalyst. The invention is drawn to a method for enhancing the rate of desulfurizing a fossil fuel containing organic sulfur compounds, comprising the steps of:a) contacting the fossil fuel with an aqueous phase containing a biocatalyst capable of cleaving carbon-sulfur bonds and a rate-enhancing amount of a flavoprotein, thereby forming a fossil fuel and aqueous phase mixture;b) maintaining the mixture of step (a) under conditions sufficient for cleavage of the carbon-sulfur bonds of the organic sulfur molecules by the biocatalyst, thereby resulting in a fossil fuel having a reduced organic sulfur content; andc) separating the fossil fuel having a reduced organic sulfur content from the resulting aqueous phase.

Abstract: The invention relates to a method of removing metals from a fossil fuel comprising the steps of contacting the fossil fuel with a biocatalyst selected from the group consisting of an enzyme which degrades porphyrin molecules under conditions suitable for the removal of the metals from the fossil fuel; and separating the metals from the fossil fuel. Preferred embodiments of the biocatalyst include heme oxygenase and cytochrome C reductase, such as cytochrome C reductase from Bacillus megaterium, Catharanthus roseuse, Escherichia coli, animal cells, plant cells or yeast cells. The cytochrome C reductase can be contacted with the fossil fuel in an aqueous medium as a substantially cell-free preparation or cell preparation. In one embodiment of the invention, the metals are recovered from the resulting metal containing stream.

Abstract: The invention relates to a method of removing metals from a fossil fuel comprising the steps of contacting the fossil fuel with an oxygenase which degrades porphyrin molecules under conditions suitable for the removal of the metals from the fossil fuel; and separating the metals from the fossil fuel. Preferred embodiments of the oxygenase include heme oxygenase and cytochrome C reductase, such as cytochrome C reductase from Bacillus megaterium, Catharanthus roseuse, Escherichia coli, animal cells, plant cells or yeast cells. The cytochrome C reductase can be contacted with the fossil fuel in an aqueous medium as a substantially cell-free preparation or cell preparation. In one embodiment of the invention, the metals are recovered from the resulting metal containing stream.

Abstract: This invention relates to a recombinant DNA molecule containing a gene or genes which encode a biocatalyst capable of desulfurizing a fossil fuel which contains organic sulfur molecules. For example, the present invention encompasses a recombinant DNA molecule containing a gene or genes of a strain of Rhodococcus rhodochrous.

Abstract: A method for reducing the viscosity of viscous petroleum liquids, such as heavy crude oil and bitumen, is disclosed. The method is appropriate for use with viscous petroleum liquids that contain sulfur-bearing heterocycles, the physicochemical properties of the heterocycles contributing significantly to the viscosity of the liquid. The method comprises contacting the viscous petroleum liquid with a biocatalyst that converts sulfur-bearing heterocycles into molecules that lack physicochemical properties condusive to viscosity. The biocatalyst works in a sulfur-specific manner, such that the sulfur-bearing heterocycle is altered at the sulfur heteroatom thereof. Through biocatalysis, carbon-sulfur bonds are cleaved and/or polar substituents such as hydroxyl groups are joined to the sulfur heteroatom, the hydrocarbon framework of the sulfur-bearing heterocycle, or both. Preferred biocatalysts for viscosity reduction include preparations of Rhodococcus rhodochrous ATCC No.

Abstract: The invention relates to a method of separating a multiple phase liquid medium comprising a first liquid phase and a second liquid phase wherein the medium is contacted with a first filter, said filter having been wetted by a wetting agent miscible with the first liquid phase but immiscible with the second liquid phase; whereby the first liquid phase passes through the filter, thus obtaining a filtrate substantially free of the second liquid phase. In a preferred embodiment, separation of a multiple phase liquid medium comprising liquid fossil fuel, water and biocatalyst employing one or two filters is disclosed. One filter will preferentially collect either the liquid fossil fuel or aqueous phase as the filtrate. The retentate will then flow to the second filter which will collect the component not removed before, i.e. the aqueous phase or liquid fossil fuel, as the filtrate. The remaining retentate, containing the biocatalyst, can then, preferably, be recycled.

Abstract: A method for the deep desulfurization of a liquid fossil fuel containing organic sulfur comprising aromatic sulfur-bearing heterocycles is described, wherein the fossil fuel is (a) subjected to hydrodesulfurization or microbial desulfurization, (b) contacted with a biocatalyst in an aqueous medium in an amount and under conditions sufficient for the conversion of aromatic sulfur-bearing heterocycles to inorganic sulfur, wherein the biocatalyst comprises bacteria or a substantially cell-free preparation thereof having the capability of the parent microorganism for catalyzing the removal of sulfur from aromatic sulfur-bearing heterocycles, thereby preparing a deeply desulfurized fossil fuel; and (c) separated from the aqueous medium.