Abstract: Disclosed are methods using neutral red to mediate the interconversion of chemical and electrical energy. Electrically reduced neutral red has been found to promote cell growth and formation of reduced products by reversibly increasing the ratio of the reduced:oxidized forms of NAD(H) or NADP(H). Electrically reduced neutral red is able to serve as the sole source of reducing power for microbial cell growth. Neutral red is also able to promote conversion of chemical energy to electrical energy by facilitating the transfer of electrons from microbial reducing power to a fuel cell cathode.

Abstract: This invention relates in general to alkaline phosphatase and, in particular, to alkaline phosphatase from a hyperthermophilic microorganism. The invention also relates to a method of purifying a hyperthermophilic alkaline phosphatase.

Abstract: The adhB gene encoding Thermoanaerobacter ethanolicus 39E secondary-alcohol dehydrogenase (2.degree. Adh) was cloned, sequenced, and expressed in Escherichia coli. The 1056 bp gene encodes a homotetrameric recombinant enzyme consisting of 37.7 kDa subunits. The purified recombinant enzyme is optimally active above 90.degree. C. with a half-life of approximately 1.7 h at 90.degree. C. An NADP(H)-dependent enzyme, the recombinant 2.degree. Adh has 1400-fold greater catalytic efficiency in propan-2-ol versus ethanol oxidation. The enzyme was inactivated by chemical modification using dithionitrobenzoate (DTNB) and diethylpyrocarbonate, indicating that Cys and His residues are involved in catalysis. Zinc was the only metal enhancing 2.degree. Adh reactivation after DTNB modification, implicating the involvement of a strongly bound zinc in catalysis. Arrhenius plots for the oxidation of propan-2-ol by the native and recombinant 2.degree. Adhs were linear from 25.degree. C. to 90.degree. C.

Abstract: An improved process for the biomethanation of an organic substrate includes treating the substrate in a first reactor to form organic acid anions, passing an aqueous preparation containing dissolved organic anions through an ion exchanger so that the organic acid anions are adsorbed and separated from the remainder of the aqueous preparation, desorbing; the organic acid anions and passing the desorbed acids to a second reactor containing methanogenic bacteria which convert the acids to methane. In a preferred embodiment, a bicarbonate solution is produced in the second reactor and it is used to desorb the organic acid anions and regenerate the anion exchanger into the bicarbonate form. A preferred apparatus first and second reactors, and a pair of anion exchangers connected by tubing to both reactors also is disclosed.

Abstract: An improved method of preparing high maltose conversion syrups and other sweeteners from starch substrates comprises saccharifying the starch at higher temperatures than presently used with environmentally compatible thermostable .beta.-amylase and other thermostable enzymes.

Abstract: An improved process for the biomethanation of an organic substrate includes treating the substrate in a first reactor to form organic acid anions, passing an aqueous preparation containing dissolved organic anions through an anion exchanger so that the organic acid anions are adsorbed and separated from the remainder of the aqueous preparation, desorbing the organic acid anions and passing the desorbed acids to a second reactor containing methanogenic bacteria which convert the acids to methane. In a preferred embodiment, a bicarbonate solution is produced in the second reactor and it is used to desorb the organic acid anions and regenerate the anion exchanger into the bicarbonate form.

Abstract: A method is disclosed for producing catabolite repression-resistant mutant strains of C. thermosulfurogenes and C. thermohydrosulfuricum. The method comprises challenging a wild strain of the organism with nitrosoguanidine, followed by enrichment on 2-deoxyglucose and culturing an iodine stained starch-glucose containing agar. The colonies which convert starch most efficiently are catabolite repression-resistant. Pure cultures of the mutants and methods employing the mutants to prepare enzymes are also described.

Abstract: An improved process for the production of propionic acid comprises cultivating a propionic acid-producing bacterium anaerobically in a medium which contains a simple carbohydrate, which can be converted by the bacterium to pyruvate, as well as essential minerals, vitamins, and growth factors under an overpressure of hydrogen which is effective to cause a hydrogenase produced by the bacterium to suppress the oxidation of pyruvate to acetate and carbon dioxide and to promote the formation of propionic acid.

Abstract: This application represents an invention based upon the discovery of a novel thermophilic organism, its isolation from natural source and to the unique thermostable hydrolytic activity produced by this bacterium, this activity being useful for industrial purposes.

Abstract: A thermostable and thermoactive, extracellular .beta.-amylase is produced by Clostridium thermosulfurogenes. Methods of producing the .beta.-amylase and using it to convert starch to maltose are also disclosed.

Abstract: A thermostable pullulanase and a thermostable glucoamylase are produced by Clostridium thermohydrosulfuricum. Methods of producing the enzymes and using them to hydrolyze starch are also disclosed.

Abstract: Starch is converted directly to ethanol by growing a co-culture of Clostridium thermosulfurogenes and Clostridium thermohydrosulfuricum under anaerobic conditions on a starch containing substrate in a medium comprising essential vitamins, minerals and growth factors. The co-culture also produces the thermostable enzymes .beta.-amylase, glucoamylase and pullulanase.

Abstract: This application represents an invention based upon the discovery of a novel thermophilic organism, its isolation from natural source and to the unique thermostable hydrolytic activity produced by this bacterium, this activity being useful for industrial purposes.

Abstract: An improved process for the production of butanol by fermentation of carbon-containing compounds with C. acetobutylicum is disclosed. The fermentation is conducted in an aqueous medium containing a sufficient concentration of dissolved carbon monoxide to give increased yields of butanol while suppressing the formation of hydrogen and acetone. This fermentation also gives increased conversion of butyric acid to solvents.

Abstract: Vitamin B.sub.12 -active substances are produced by the fermentation of an aqueous medium consisting essentially of methanol as the main source of assimilable carbon, a source of assimilable nitrogen, cobalt ions and essential growth factors and minerals with a vitamin B.sub.12 -activity substance producing organism capable of growing under anaerobic conditions.

Abstract: A process for the propagation of microbial cells and the production of fermentation products, such as acetic and butyric acid, comprises anaerobically growing an acidogenic bacterium, such as a strain of Butyribacterium methylotrophicum ATCC 33226, in a nutrient medium containing a single carbon product, such as methanol, as the fermentation substrate or main source of assimilable carbon, accumulating the microbial cells in said medium and then separating and recovering said microbial cells and desired fermentation products from the spent media.

Abstract: A continuous spectrophotometric assay monitors the release of colored cellulose oligosaccharides during the cellulase catalyzed hydrolysis of dyed crystalline cellulose preparations. An anthraquinone derivative, Remazol Brilliant Blue R-Salt, is used to uniformly dye a variety of cellulose biopolymers without altering their respective polysaccharide conformations. The increase in dye absorbance of a cellulose filtrate is a direct measure of the total concentration of carbohydrates formed and the cellulase activity. The continuous cellulase assay has specific utility in the study of cellulase kinetics and in comparing the cellulase activities of different microorganisms.

Abstract: Production of ethanol and enzymes by fermentation of biomass with co-cultures of C. thermocellum and C. thermohydrosulfuricum.

Abstract: A partially purified NADP specific thermostable alcohol, aldehyde/ketone oxidoreductase is prepared which can react with a wide range of alcohols, ketones and aldehydes. The enzyme has a unique preference for secondary over primary alcohols. The thermostability, broad range of operating temperatures and lack of sensitivity to metal ions and complexing agents, in addition to the absolute specifity for the coenzyme, increase the utility of the enzyme in asymmetric organic synthesis and NADPH regeneration.