A Keto Reductase Involved in Steroid Degradation in Mycolicibacterium neoaurum

Phytosterols can be used by microorganisms as carbon and energy sources and completely degraded into CO₂ and H₂O. The catabolic pathway of phytosterols was well characterized in many microorganisms. Blocking the steroid core ring degradation by deletions of fadE30 and fadD3 genes, two important steroid intermediates, 3aα-H-4α-(3’-Propionic acid)-5α-hydroxy-7aβ-methylhexahydro-1-indanone-δ-lactone (sitolactone, or HIL) and 3aα-H-4α-(3’-propionic acid)-7aβ-methylhexahydro-1,5-indanedione (HIP) can be accumulated. They are currently used to synthesize nor-steroid drugs with an α-methyl group or without the methyl group at the C₁₀-position, such as estrone and norethindrone. In this study, a key gene involved in the bioconversion of HIP to HIL was identified in Mycolicibacterium neoaurum. Through heterologous expression, gene hipR was found to be involved in the reduction of the C₅ keto group of HIP to a hydroxy group, leading to spontaneously lactonization into HIL in vitro. Through gene complementation and knockout, HipR functions were verified and two HIP degradation pathways in vivo were elucidated. The finding of this research facilitated the understanding of the metabolic pathway of sterols, and was directly applied to engineering robust production strains by overexpression or knockout of related genes.     

Reactive radical intermediates formed from illuminated nifedipine

Nifedipine, (1,4-dihydro-2,6,dimethyl-4-(2-nitrophenyl)-3,5-pyridinedicarboxylic acid dimethyl ester) a calcium channel blocker widely used in treatment of hypertension, is strongly photolabile. This may represent a problem in patients taking nifedipine and in handling of nifedipine samples. Reactive radical intermediates were determined and characterized in the process of nifedipine illumination using EPR spectroscopy. On illumination of nifedipine by daylight or by a mercury lamp, a nitroxide radical, R^I_IL-NIFNO^.X was observed (in the first step), in various solvents like benzene, cyclohexane, methanol, acetonitrile, dimethylsulphoxide, or aqueous suspensions of liposomes. R^I_IL-NIF represents the nifedipine skeleton centered with phenyl group, and X is an EPR silent substituent. The generation of R^I_IL-NIFNO^.X is coupled with the formation of nitroso compound, R^I_IL-NIFNO, as characterized by UV-visible spectroscopy. In a further step, R^I_IL-NIFNO abstracts hydrogen from nifedipine skeleton under the formation of R^I_IL-NIFNO^.H radical. In addition to this, in system containing R^I_IL-NIFNO and unsaturated lipids, nitroxide radicals R_IL-INF^INO^.R_LIPIDS are formed probably via a pseudo Diels-Alder mechanism (R_LIPIDSrepresents lipidic skeleton). The unusually easy photochemical activation of nifedipine is probably stimulated by photosensitization of its nitro group interacting with suitably positioned hydrogen or carboxylic mehtyl ester group from the pyridynl ring.     

Enzymatic and hydrolytic degradation of benzylaryl ethers related to hardwood lignins

A number of 4-hydroxybenzylphenyl ethers and their acetates were synthesized as models for hardwood lignin and used as substrates in acid hydrolysis and enzymatic oxidation reactions. Under hydrolytic conditions, the acetates underwent ether cleavage at a slower rate than the free phenols. Evidence for carbonium ion intermediates is presented. Cleavage of the ether substrates by peroxidase—peroxide oxidation was much faster than by acid hydrolysis for all substrates except the acetates which did not react. Subsequent oxidation of the component parts of the ether substrates was selective: the syringyl moieties were oxidized in preference to the guaiacyl moieties. Electron spin resonance studies of the oxidation reaction showed that removal of the phenolic hydrogen atom was the first step, followed by quinone—methide formation. A mechanism is proposed to account for the oxidative degradation of the lignin models.     

Intermolecular Singlet Fission in Unsymmetrical Derivatives of Pentacene in Solution

Intermolecular singlet fission (SF) is an electronically coupled process between two chromophores, where distance dependences are decisive in terms of rates and yields. In the current work, a family of pentacene derivatives featuring different functional groups have been designed, synthesized, and probed with respect to intermolecular SF in the low, medium, and high concentration regimes rather than in the solid state. By means of advanced photophysical techniques, global analysis modeling, and ab initio calculations, a model for intermolecular SF is postulated. The model is based on an early key intermediate, which involves the diffusional encounter between one pentacene in its singlet excited‐state with another one in its ground state and which features excimer characteristics. This is followed by a transformation into a coupled triplet excited‐state. The role of the functional group appended to pentacene is analyzed with respect to steric shielding of the pentacene core as a means to prevent photophysical degradation, as well as control diffusional encounter and, subsequently, SF. The findings demonstrate the potential of new molecular materials for SF, especially in solution studies, as well as the challenges of implementing them in energy conversion schemes due to the appearance of photodegradation processes that compete with SF.     

Growth regulatory and growth inhibitory effects of Thevetia neriifolia stem extracts on Helicoverpa armigera (Lepidoptera: Noctuidae)

Abstract

Helicoverpa armigera, a serious global destructive pest of agricultural crops, is on continuous rise despite several control measures undertaken. The detrimental effects of these measures have created a dire need to explore alternate eco-safe strategies. The present study investigates the growth-regulatory and growth-arrest potential of hexane and methanol extracts of Thevetia neriifolia stems against H. armigera. Investigations revealed that larval feeding and rearing on the extract-containing diet did not result in appreciable larval mortality but delayed the larval growth and development. Both the extracts demonstrated dose-dependent effects exhibiting negative correlation between the weights gained by developmental stages and the extract concentrations. Feeding with extracts also resulted in formation of few larval–pupal and pupal–adult intermediates and significantly reduced percent adult emergence of H. armigera. Current study, however, revealed higher growth inhibitory potential of methanol extracts as compared with hexane extracts. The study attempts to provide an eco-friendly approach for H. armigera management.     

Recovery of trichloroethylene removal efficiency through short-term toluene feeding in a biofilter enriched withPseudomonas putida F1

Trichloroethylene (TCE) is an environmental contaminant provoking genetic mutation and damages to liver and central nerve system even at low concentrations. A practical scheme is reported using toluene as a primary substrate to revitalize the biofilter column for an extended period of TCE degradation. The rate of trichloroethylene (TCE) degradation byPseudomonas putida F1 at 25°C decreased exponentially with time, without toluene feeding to a biofilter column (11 cm I.D.×95 cm height). The rate of decrease was 2.5 times faster at a TCE concentration of 970 μg/L compared to a TCE concentration of 110 μg/L. The TCE itself was not toxic to the cells, but the metabolic intermediates of the TCE degradation were apparently responsible for the decrease in the TCE degradation rate. A short-term (2 h) supply of toluene (2,200 μg/L) at an empty bed residence time (EBRT) of 6.4 min recovered the relative column activity by 43% when the TCE removal efficiency at the time of toluene feeding was 58%. The recovery of the TCE removal efficiency increased at higher incoming toluene concentrations and longer toluene supply durations according to the Monod type of kinetic expression. A longer duration (1.4∼2.4 times) of toluene supply increased the recovery of the TCE removal efficieny by 20% for the same toluene load.     

The metabolic integrity of Fasciola hepatica during in vitro maintenance

Levels of metabolic intermediates and end products in F. hepatica after 24 and 48 h in Hédon-Fleig salt solution with added glucose were compared with levels obtained immediately on removal from the host. Glycogen levels dropped initially, probably due to the expulsion of eggs; thereafter they remained constant. Internal glucose concentrations increased as the parasites equilibrated with the medium. Other changes in internal pool sizes were consistent with regulation to the in vitro conditions. ATP levels increased; ATP/ADP ratios were maintained. Comparisons of mass action ratios and equilibrium constants suggest that hexokinase, pyruvate kinase and phosphofructokinase are regulatory. Output of excretory products approached linearity; from the calculated regressions the proportions of lactate, acetate and propionate were 1: 2: 4. The implications for metabolic regulation in F. hepatica are briefly discussed, and it is concluded that, for at least 48 h in vitro, energy metabolism is not adversely affected.     

High-valent intermediates of heme proteins and model compounds

Recent computational and experimental probes of high-valent intermediates in heme proteins and model compounds reveal a rich spectrum of chemical behavior that is dependent on the nature of the proximal ligand, metal center, distal- and proximal-binding site environment, porphyrin macrocycle architecture, and consequent electronic structure. The results of such studies reveal an underlying complexity, which is simply understood once one is cognizant of the 'chameleon'-like behavior of such intermediates is determined by the high-valent intermediate environment.     

Microbial energetics and stoichiometry for biodegradation of aromatic compounds involving oxygenation reactions

Oxygenation reactions significantly alter the energy and electron flows and, consequently, the overall stoichiometry for the microbial utilization of aromatic compounds. Oxygenation reactions do not yield a net release of electrons, but require an input of electrons to reduce oxygen molecules. The biodegradation pathway of phenanthrene as a model compound was analyzed to determine the impact of oxygenation reactions on overall stoichiometry using the half-reaction method. For individual oxygenation reactions, the half-reaction method for analyzing the electron and energy flows must be modified, because the reactions do not release electrons for synthesis or energy generation. Coupling the oxygenation reaction to subsequent reaction steps provides a net electron release for the coupled reactions. Modeling results indicate that oxygenation reactions increase the oxygen requirement and reduce the cell yield, compared to the conventional mineralization represented by hydroxylation reactions in place of oxygenations. The computed yields considering oxygenation reactions conform better to empirical yields reported in the literature than do yields computed by the hydroxylation single-step methods. The coupled-reaction model also is consistent with information about the ways in which micro-organisms that degrade aromatics accumulate intermediates, regulate degradation genes, and organize enzyme clusters.