Autotrophic Growth of Thiobacillus acidophilus in the Presence of a Surface-Active Agent, Tween 80

Cellular protein, pH, dissolved oxygen concentration, and static surface tension were measured during growth of Thiobacillus acidophilus on elemental sulfur in the absence and presence of up to 5,000 mg of Tween 80 per liter. The decrease in pH and the increase in sulfate production were observed to be less accurate measurements of growth when compared with the increase in cellular protein. The doubling time of the bacterium decreased approximately 50% with the addition of 500 mg of Tween 80 per liter. The bacteria did not appear to synthesize any wetting agents as demonstrated by the constant surface tension of the medium during growth. Morphological alterations in the presence of Tween 80 were also observed.     

Growth of Iron-Oxidizing Thiobacilli in the Presence of Chalcopyrite and Galena

Iron-oxidizing thiobacilli were adapted to grow on a chalcopyrite and a galena ore concentrate. When grown on the chalcopyrite concentrate, the bacteria exhibited a doubling time of 38.4 ± 2.9 h, with a final cellular protein concentration of 185 μg/ml and solubilization of 10.3 g of copper per liter. When grown on the galena ore concentrate, the generation time was 39.6 ± 2.7 h, with a final cellular protein concentration of 120 μg/ml. Galena was converted to lead salts soluble in 1 M ammonium acetate to a concentration of 20.2 g of lead per liter. X-ray diffraction and refractive-image analysis indicated that the smaller-sized particles were favored in this process. Galena was converted to anglesite, and soluble copper was liberated from chalcopyrite with the concurrent formation of jarosite.     

Pseudomonas oleovorans as a tool in bioconversions of hydrocarbons: growth,morphology and conversion characteristics in different two-phase systems

The bioconversion of hydrocarbons by Pseudomonas oleovorans has been studied in two-phase systems. In these systems, the hydrocarbon substrate is present in sufficient amounts to form the bulk apolar phase. High cell densities (up to 20 mg dry mass per ml water phase) are reached when the apolar phase consists of n-octane, 1-octene or 1-decene. There is considerable cell damage after incubation for 50–70 h. Loss of cell viability and membrane damage as observed by freeze-fracture electron microscopy correlate with a loss of hydrocarbon oxidation, measured as the conversion of 1-octene to 1,2-epoxyoctane. The final yield of oxidized hydrocarbon in the apolar substrate phase can be increased substantially by replacing the damaged cells with freshly grown cells. Yields up to 150 mg 1,2-epoxyoctane per ml 1-octene and up to 20–25 mg 1,2-epoxyoctane per ml culture were obtained with four cycles of the cell renewal procedure. Several other substrates in addition to octene were tested in the optimized two-phase system. Of these, 1-decene was converted into (R)-1,2-epoxydecane with an optical purity of 60%, while allylbenzene was converted into chiral 1,2-epoxy-3-phenylpropane. Some of the future applications of the conversion products are discussed.     

Reperfusion-induced arrhythmias are not prevented by uric acid in the isolated rat heart

Oxygen-derived free radicals have been implicated in ventricular arrhythmogenesis during coronary reperfusion following an acute ischemic event. We have investigated the possibility that uric acid, a potentially important physiological antioxidant (inhibits lipid peroxidation and scavenges various radical species during oxidation to allantoin), or oxonic acid (inhibitor of uricase enzyme), are able to prevent reperfusion-induced ventricular dysrhythmias in isolated buffer-perfused rat hearts. Rat hearts (n = 12/group) underwent 15 minutes occlusion; arrhythmias were monitored during ischemia and for 10 minutes of reperfusion. There was no difference in the incidence of ventricular fibrillation or ventricular tachycardia in either uric acid or oxonic acid treated hearts compared to untreated controls. Mean duration of ventricular fibrillation appeared to be reduced in hearts treated with 10(-3) and 10(-4) M oxonic acid compared to controls but these data did not achieve a level of statistical significance. These results demonstrate that uric acid and oxonic acid failed to prevent reperfusion-mediated ventricular dysrhythmias in this experimental preparation. Although oxygen-derived free radicals may contribute to the initiation of either ischemia- or reperfusion-induced arrhythmogenesis, our findings provide little support for this hypothesis.     

Improved enantioselective conversion of styrene epoxides and meso-epoxides through epoxide hydrolases with a mutated nucleophile-flanking residue

In epoxide hydrolase from Agrobacterium radiobacter (EchA), phenylalanine 108 flanks the nucleophilic aspartate and forms part of the substrate-binding pocket. The influence of mutations at this position on the activity and enantioselectivity of the enzyme was investigated. Screening for improved enantioselectivity towards para-nitrophenyl glycidyl ether (pNPGE) using spectrophotometric progress curve analysis yielded five different mutants with 3- to 7-fold improved enantioselectivity. The increase in enantioselectivity was in most cases the result of an enhanced catalytic efficiency toward the preferred enantiomer. Several mutations at position F108 resulted in a higher activity toward cis-disubstituted meso-epoxides, which were converted to a single product enantiomer. Mutant F108C converted cis-2,3-epoxybutane to (2R,3R)-2,3-butanediol of >99% ee with a 7-fold improved activity, and mutant F108A hydrolyzed cyclohexene oxide to (1R,2R)-1,2-cyclohexanediol of >99% ee with a more than 150-fold higher activity than wild-type enzyme. It is concluded that single amino acid substitutions in the active site of epoxide hydrolase can result in enzyme variants with catalytic properties that are suitable for preparative scale production of (S)-epoxides and chiral vicinal diols in high yield and with excellent ee.     

Tori lines mitigate seabird bycatch in a pelagic longline fishery

Reviews in Fish Biology and Fisheries - Albatross bycatch has been increasing over the past decade in the US tuna longline fishery of the central North Pacific. A controlled field...     

Influence of Cardiac Decentralization on Cardioprotection

The role of cardiac nerves on development of myocardial tissue injury after acute coronary occlusion remains controversial. We investigated whether acute cardiac decentralization (surgical) modulates coronary flow reserve and myocardial protection in preconditioned dogs subject to ischemia-reperfusion. Experiments were conducted on four groups of anesthetised, open-chest dogs (n = 32): 1- controls (CTR, intact cardiac nerves), 2- ischemic preconditioning (PC; 4 cycles of 5-min IR), 3- cardiac decentralization (CD) and 4- CD+PC; all dogs underwent 60-min coronary occlusion and 180-min reperfusion. Coronary blood flow and reactive hyperemic responses were assessed using a blood volume flow probe. Infarct size (tetrazolium staining) was related to anatomic area at risk and coronary collateral blood flow (microspheres) in the anatomic area at risk. Post-ischemic reactive hyperemia and repayment-to-debt ratio responses were significantly reduced for all experimental groups; however, arterial perfusion pressure was not affected. Infarct size was reduced in CD dogs (18.6±4.3; p = 0.001, data are mean±1SD) compared to 25.2±5.5% in CTR dogs and was less in PC dogs as expected (13.5±3.2 vs. 25.2±5.5%; p = 0.001); after acute CD, PC protection was conserved (11.6±3.4 vs. 18.6±4.3%; p = 0.02). In conclusion, our findings provide strong evidence that myocardial protection against ischemic injury can be preserved independent of extrinsic cardiac nerve inputs.     

Steady-state and pre-steady-state kinetic analysis of halopropane conversion by a rhodococcus haloalkane dehalogenase

Haloalkane dehalogenase from Rhodococcus rhodochrous NCIMB 13064 (DhaA) catalyzes the hydrolysis of carbon-halogen bonds in a wide range of haloalkanes. We examined the steady-state and pre-steady-state kinetics of halopropane conversion by DhaA to illuminate mechanistic details of the dehalogenation pathway. Steady-state kinetic analysis of DhaA with a range of halopropanes showed that bromopropanes had higher k(cat) and lower K(M) values than the chlorinated analogues. The kinetic mechanism of dehalogenation was further studied using rapid-quench-flow analysis of 1,3-dibromopropane conversion. This provided a direct measurement of the chemical steps in the reaction mechanism, i.e., cleavage of the carbon-halogen bond and hydrolysis of the covalent alkyl-enzyme intermediate. The results lead to a minimal mechanism consisting of four main steps. The occurrence of a pre-steady-state burst, both for bromide and 3-bromo-1-propanol, suggests that product release is rate-limiting under steady-state conditions. Combining pre-steady-state burst and single-turnover experiments indicated that the rate of carbon-bromine bond cleavage was indeed more than 100-fold higher than the steady-state k(cat). Product release occurred with a rate constant of 3.9 s(-1), a value close to the experimental k(cat) of 2.7 s(-1). Comparing the kinetic mechanism of DhaA with that of the corresponding enzyme from Xanthobacter autotrophicus GJ10 (DhlA) shows that the overall mechanisms are similar. However, whereas in DhlA the rate of halide release represents the slowest step in the catalytic cycle, our results suggest that in DhaA the release of 3-bromo-1-propanol is the slowest step during 1,3-dibromopropane conversion.     

The structure, catalytic mechanism and regulation of adenylyl cyclase

The recent structure determinations of the mammalian effector enzyme adenylyl cyclase reveal the structure of its catalytic core, provide new insights into its catalytic mechanism and suggest how diverse signaling molecules regulate its activity.