Selection of microbial biocatalysts for the reduction of cyclic and heterocyclic ketones

The reduction of carbonyl compounds plays an important role in the synthesis of complex chiral molecules. In particular, enantiopure substituted cyclic and heterocyclic compounds are useful intermediates for the synthesis of several antiviral, antitumor, and antibiotic agents, and recently, they have also been used as organocatalysts for C-C addition. Alcohol dehydrogenases (ADH) are enzymes involved in the transformation of prochiral ketones to chiral hydroxyl compounds. While significant scientific effort has been paid to the use of aliphatic and exocyclic ketones as ADH substrates, reports on (hetero)cyclic carbonyl compounds as substrates of these enzymes are scarce. In the present study, 109 bacteria and 36 fungi were screened, resulting in 10 organisms belonging to both kingdoms capable of transforming cyclic and heterocyclic ketones into the corresponding alcohols. Among them, Erwinia chrysanthemi could quantitatively reduce cyclododecanone and Geotrichum candidum could stereoselectively reduce N-Boc-3-piperidone and N-Boc-3-pyrrolidinone to their corresponding (S)-alcohols; however, the anti-Prelog isomer was obtained when acetophenone was the substrate.     

Toxicity assessment of compounds in soil using a simple respirometric technique

A laboratory procedure using a simple respirometric technique was evaluated to determine the microbial toxicity in soil of three toxicant compounds: two pesticides, chlorpyrifos and glyphosate; and diesel oil. The microbial toxicity was tested using the specific oxygen uptake rate (SOUR) method, evaluating the soil samples for both the reduction in maximum SOUR (SOUR_max) and the cumulative oxygen demands after 20 h (OD₂₀). Consumption rate curves were produced for the lowest concentrations assessed: diesel (2460 ppm), chlorpyrifos (62.5 ppm), and glyphosate (250 ppm) (limiting amounts considered as local soil contamination). In comparison with the control, these showed drastic reductions in SOUR_max, demonstrating the high sensitivity of this SOUR method. The SOUR_max provides a better indication of the microbial toxicity of these contaminants compared to the OD₂₀ because of the different effects of these toxic compounds on microbial communities in the soil. Increasing concentrations of these toxic compounds resulted in different responses, evaluated as percentage inhibition by these different xenobiotic compounds. For these reasons, the microbial toxicity of xenobiotic compounds can be better recognized with SOUR_max as compared to other respirometric methodologies.     

Generation of genetic diversity by DNA rearrangements in resting bacteria

Abstract Transpositional DNA rearrangements importantly contribute to the genomic plasticity of bacteria, their viruses and plasmids. Interestingly, enzymatically mediated transposition is not limited to propagating bacteria, but it also occurs in prolonged periods of rest. As revealed with RFLP analysis, genetic polymorphism increases steadily upon storage of bacteria in the resting phase. These results are discussed here in the wider context of accumulating knowledge on molecular mechanisms contributing to overall spontaneous mutagenesis, which is the result of a multitude of specific, often enzyme-mediated processes of variation generation.     

Effects of gas periodic stimulation on key enzyme activity in gas double-dynamic solid state fermentation (GDD-SSF)

The heat and mass transfer have been proved to be the important factors in air pressure pulsation for cellulase production. However, as process of enzyme secretion, the cellulase formation has not been studied in the view of microorganism metabolism and metabolic key enzyme activity under air pressure pulsation condition. Two fermentation methods in ATPase activity, cellulase productivity, weight lose rate and membrane permeability were systematically compared. Results indicated that gas double-dynamic solid state fermentation had no obviously effect on cell membrane permeability. However, the relation between ATPase activity and weight loss rate was linearly dependent with r = 0.9784. Meanwhile, the results also implied that gas periodic stimulation had apparently strengthened microbial metabolism through increasing ATPase activity during gas double-dynamic solid state fermentation, resulting in motivating the production of cellulase by Trichoderma reesei YG3. Therefore, the increase of ATPase activity would be another crucial factor to strengthen fermentation process for cellulase production under gas double-dynamic solid state fermentation.     

Studies on litter characterization using 13C NMR and assessment of microbial activity in natural forest and plantation crops’ (teak and rubber) soil ecosystems of Kerala, India

The leaf litter is the major source of soil organic matter in natural and many plantation crop ecosystems. Quantity and quality of organic matter in a soil ecosystem is of utmost importance in regulating the soil health. Hence assessment of quality of organic matter input, viz., litter is important and is attempted in this study. The leaf litter of rubber (Hevea brasiliensis), pueraria (Pueraria phaseoloides), mucuna (Mucuna bracteata), teak (Tectona grandis) and forest (mixed species) were analyzed using solid state ¹³C nuclear magnetic resonance (NMR) to study the relative abundance of different carbon compounds present. The spectra revealed that litter of all species studied contain relatively larger amounts of polysaccharides compared to other C containing compounds. Also it could be observed that the alkyl-C to O-alkyl-C ratio of rubber litter was much higher compared to that of others. Aromatics and carbonyl compounds were also present in all litter species. The resource quality based on alkyl-C to O-alkyl-C ratio of the litter samples studied can be arranged in the order pueraria > teak > mucuna > forest > rubber. The respiration rate, substrate induced respiration rate and biomass-C (Cmic) of the litter samples were estimated. It could be observed that litter associated microbial activity decreased as alkyl-C to O-alkyl-C ratio increased. Resource quality derived from the NMR spectra and the litter biological properties were complementary. Soil samples (0–15 cm) from the five soil ecosystems (rubber, pueraria, mucuna, teak and forest) were analyzed for respiration rate, substrate induced respiration rate, Cmic, total-C and total-N. The forest soil had higher respiration rate, total-C and total-N compared to cultivated soil systems. Pueraria, mucuna and teak soils were comparable for their biological properties while rubber soil recorded comparatively lower microbial activity.     

Analysis of a microbial electrochemical cell using the proton condition in biofilm (PCBIOFILM) model

Common to all microbial electrochemical cells (MXCs) are the anode-respiring bacteria (ARB), which transfer electrons to an anode and release protons that must transport out of the biofilm. Here, we develop a novel modeling platform, Proton Condition in BIOFILM (PCBIOFILM), with a structure geared towards mechanistically explaining: (1) how the ARB half reaction produces enough acid to inhibit the ARB by low pH; (2) how the diffusion of alkalinity carriers (phosphates and carbonates) control the pH gradients in the biofilm anode; (3) how increasing alkalinity attenuates pH gradients and increases current; and (4) why carbonates enable higher current density than phosphates. Analysis of literature data using PCBIOFILM supports the hypothesis that alkalinity limits the maximum current density for MXCs. An alkalinity criterion for eliminating low-pH limitation - 12 mg CaCO₃/mg BOD - implies that a practical MXC can achieve a maximum current density with an effluent quality comparable to anaerobic digestion.     

Production of Bacillus thuringiensis Berliner var. kurstaki Grown in Alternative Media

Agro - industrial residues and by - products available in southeastern Brazil were used as ingredients for low - cost culture media for liquid fermentation of Bacillus thuringiensis var. kurstaki. Highest spore yield was obtained with a medium containing cheese whey , soya bean milk and molasses (WSM) . Crystals and spores were produced in all media and potency of the final product was highest for nutrient broth + yeast extract medium (NBY) . There was no correlation between the number of spores in the fermented media and the potency of the preparations . Considering all three factors , the potencies , costs and yields of the final products , lowest relative cost was obtained with BMM medium ( Bombyx mori pupae + molasses) . NBY and WSM had intermediate relative cost approximately nine times higher than BMM . The cost analysis suggests that BMM medium should be preferred for local production of B. thuringiensis var . kurstaki in comparison to other media tested . The results also demonstrate the importance of considering yields , cost and potency of the B. thuringiensis preparations in selecting the production medium .     

Competition for nitrate between denitrifying Pseudomonas stutzeri and nitrate ammonifying enterobacteria

Abstract Competition for nitrate between nitrate ammonifying enterobacteria and a denitrifying pseudomonad was studied in electron acceptor-limited chenostats. In pure cultures, using different carbon and energy sources, the C/N-ratio needed for denitrification is far lower than that required for nitrate ammonification. In mixed cultures of Citrobacter freundii and Pseudomonas stutzeri , competing for nitrate with l -lactate as electron donor, the nitrate ammonifying organism dominated at dilution rates of D ≤ 0.14 h⁻¹. Competition for both nitrate and l -lactate at a dilution rate of D = 0.05 h⁻¹ always resulted in the coexistence of both species. Using glucose as additional carbon source, the final ratio of nitrate ammonifying and denitrifying organism depended on the C/N-ratio as well as on the dilution rate. The results of the study are discussed with respect to field data.     

The Protozoan-Metazoan Trophic Link In Pelagic Ecosystems

ABSTRACT The evidence for a qualitatively and quantitatively important trophic link between planktonic Protozoa and higher order metazoan consumers is reviewed. the available data are obtained primarily, but not exclusively, from laboratory studies of calanoid copepod consumers and tintinnid ciliate prey from marine estuarine and nearshore environments. the data indicates that the protozoan-metazoan link is of similar magnitude and importance in the pelagic ecosystems of freshwaters. It is proposed that planktonic Protozoa constitute a high quality, nitrogen-rich food in the diets of their metazoan consumers. Implications of die trophic link to the consumers, prey, and ecosystem are discussed.