Kinetic modeling of Stickland reactions-coupled methanogenesis for a methanogenic culture.

Studying amino acid catabolism-coupled methanogenesis is the important standpoints to decipher the metabolic behavior of a methanogenic culture. L-Glycine and L-alanine are acted as sole carbon and nitrogen sources for acidogenic bacteria. One amino acid is oxidized and another one is reduced for acetate production via pyruvate by oxidative deamination process in the Stickland reactions. Herein, we have developed a kinetic model for the Stickland reactions-coupled methanogenesis (SRCM) and simulated objectively to maximize the rate of methane production. We collected the metabolic information from enzyme kinetic parameters for amino acid catabolism of Clostridium acetobutylicum ATCC 824 and methanogenesis of Methanosarcina acetivorans C2A. The SRCM model of this study consisted of 18 reactions and 61 metabolites with enzyme kinetic parameters derived experimental data. The internal or external metabolic flux rate of this system found to control the acidogenesis and methanogenesis in a methanogenic culture. Using the SRCM model, flux distributions were calculated for each reaction and metabolite in order to maximize the methane production rate from the glycine-alanine pair. Results of this study, we demonstrated the metabolic behavior, metabolite pairing while mutually interact, and advantages of syntrophic metabolism of amino acid-directed methane production in a methanogenic starter culture.

Biodegradation of insecticide monocrotophos by Bacillus subtilis KPA-1, isolated from agriculture soils.

Twenty bacterial strains, which are capable of degrading monocrotophos, were isolated from five soil samples collected from agriculture soils in India. The ability of the strains to mineralize monocrotophos was investigated under different culture conditions. A potential strain degrading monocrotophos was selected and named KPA-1. The strain was identified as a Bacillus subtilis on the basis of the results of its cellular morphology, physiological and chemotaxonomic characteristics, and phylogenetic conclusion of 16S ribosomal DNA (rDNA) gene sequences. Organophosphate hydrolase (opdA gene) involved in the initial biodegradation of monocrotophos in KPA-1 was quantitatively expressed, which was a constitutively expressed cytosolic enzyme. RT-qPCR data revealed that KPA-1 harboring opdA gene in an early stage was significantly downregulated from opdA gene in a degradation stage (1.5 fold more) with a p value of 0.0375 (p < 0.05). We have optimized culture conditions for the efficient degradation (94.2 %) of monocrotophos under aerobic conditions. Growth and degradation kinetic studies proved that KPA-1 was able to grow in minimal salt medium containing 1000 ppm monocrotophos as the only carbon source. Hence, KPA-1 culture has a great potential utility for the bioremediation of agriculture soils contaminated with organophosphorus pesticides, particularly monocrotophos.