Authors:J. Raj, N. Singh, S. Prasad, A. Seth, and T. Bhalla
The free and agar immobilized cells of
NHB-2 having nitrilase (EC 188.8.131.52) activity were used to catalyse the transformation of benzonitrile to benzoic acid. The whole cells of
NHB-2 were immobilized in agar which exhibited maximum conversion of benzonitrile to benzoic acid in 0.1 M potassium phosphate buffer pH 7.5 (free cells) 8.0 (immobilized cells), temperature 40 °C, cells 2 mg dcm ml
reaction mixture and benzonitrile (4% v/v) in 4 h (free cells). The effect of temperature on the stability of nitrilase was studied and cells retained 100% activity at 30 °C and lost 50% activity at 40 °C. In a fed batch mode of reaction 108 and 84 gl
benzoic acid was produced using free and agar entrapped cells (2 g dcm). The agar immobilized cells were recycled up to three times and 80, 62, 20 gl
benzoic acid was again produced respectively in each of three cycles and a total 244 g benzoic acid was produced by recycling the same mass of immobilized biocatalyst.
Authors:S. Omanwar, J. R. Rao, G. Butchaiah, S. H. Basagoudanavar, and R. K. Singh
The mechanically transmitted haemoflagellate,Trypanosoma evansicauses 'surra', a wasting disease of domestic animals and is highly endemic in distribution in Southeast Asia. The detection ofT. evansiis important for improving the epizootiological and animal health status of the region. The specificity and sensitivity of polymerase chain reaction (PCR) using oligonucleotide primers constructed fromT. evansirepetitive DNA sequences were studied in the present investigation. Using the assay, it was possible to amplify template DNA ofT. evansiderived from buffaloes, camels and horses to a threshold sensitivity level of 0.5 pg and to detect DNA from as few as five organisms in 10 (l crude blood samples. Following experimental infection of calves with 5 × 105T. evansi, positive signals could be observed as early as 12 h post-infection. DNAs from two common haemoflagellates of cattle,Babesia bigeminaandTheileria annulatawere not amplified with the primers.
Authors:S. H. Basagoudanavar, J. R. Rao, Swati Omanwar, A. K. Tiwari, R. K. Singh, R. S. Kataria, and G. Butchaiah
A highly reproducible, dominant, monomorphic fragment of 473 base pair (bp) amplified from the genome of Trypanosoma evansi by arbitrary primer — polymerase chain reaction (AP-PCR) was labelled with digoxigenin and investigated for its potential as DNA probe. Dot-blot hybridisation of total genomic DNA with the probe proved useful in detecting bubaline, cameline and equine strains of T. evansi down to 10 pg of parasite template DNA. No cross-hybridisation was seen with Babesia bigemina, Theileria annulata and the bubaline host DNA. This probe may facilitate laboratory identification of T. evansi in developing countries, without the inherent risk associated with radioisotopes.