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71.
Cheryl E. Beard Mary A. Hefford Robert J. Forster Sushama Sontakke Ron M. Teather Keith Gregg 《Current microbiology》1995,30(2):105-109
A 9.5-kb shuttle vector capable of replication and selection in both Escherichia coli and Butyrivibrio fibrisolvens was constructed. Plasmid pUC118 provided replication functions and ampicillin resistance selection in E. coli. In B. fibrisolvens, replication was controlled by the native plasmid pRJF1 from strain OB156, and selectability was provided by a 3.5-kb fragment of plasmid pAM1 containing the erythromycin resistance gene. Optimum conditions for transformation were 15 kV/cm, 2 h recovery, and plating in an agar overlay on medium containing 10 g erythromycin/ml. Maximum efficiency was 1.1×105 transformants per g plasmid DNA (average 3×104), and restriction mechanisms reduced efficiency by a factor of 2×102. Nonselective growth for 200 generations gave no measurable loss of plasmid. 相似文献
72.
The pH dependence of the activity, aggregation, and secondary structure of Artocarpus hirsuta lectin was studied using intrinsic and extrinsic fluorescence, light scattering, and circular dichroism. The lectin is more stable in the neutral and acidic than in the alkaline pH range, which is also reflected in the binding constants of the lectin to methyl alpha-galactopyranoside (me alpha-gal). The aggregation of the protein due to heat denaturation is prevented at both extremes of pH. The binding of hydrophobic dye to the lectin takes place at pH 1-2, which increases with increasing temperature. The exposure of hydrophobic patches at pH 1 is reversible. The secondary structure of the lectin is intact in the pH range of 1-8 and is distorted above pH 9. Aggregation of the protein due to heat denaturation is also prevented in the presence of guanidine hydrochloride (GdnHCl). 相似文献
73.
Swati T. Gurme Shripad N. Surwase Sushama A. Patil Shekhar B. Jadhav Jyoti P. Jadhav 《Indian journal of microbiology》2013,53(2):194-198
l-DOPA (3,4-dihydroxyphenyl-l-alanine) is the most widely used drug for treatment of Parkinson’s disease. In this study Yarrowia lipolytica-NCIM 3472 biomass was used for transformation of l-tyrosine to l-DOPA. The process parameters were optimized using response surface methodology (RSM). The optimum values of the tested variables for the production of l-DOPA were: pH 7.31, temperature 42.9 °C, 2.31 g l?1 cell mass and 1.488 g l?1 l-tyrosine. The highest yield obtained with these optimum parameters along with recycling of the cells was 4.091 g l?1. This optimization of process parameters using RSM resulted in 4.609-fold increase in the l-DOPA production. The statistical analysis showed that the model was significant. Also coefficient of determination (R2) was 0.9758, indicating a good agreement between the experimental and predicted values of l-DOPA production. The highest tyrosinase activity observed was 7,028 U mg?1 tyrosine. l-DOPA production was confirmed by HPTLC and HPLC analysis. Thus, RSM approach effectively enhanced the potential of Y. lipolytica-NCIM 3472 as an alternative source to produce l-DOPA. 相似文献
74.
75.
Sushama M Gaikwad Madhura M Gurjar M Islam Khan 《European journal of biochemistry》2002,269(5):1413-1417
Unfolding, inactivation and dissociation of the lectin from Artocarpus hirsuta seeds were studied by chemical (guanidine hydrochloride, GdnHCl) and thermal denaturation. Conformational transitions were monitored by intrinsic fluorescence and circular dichroism. The gradual red shift in the emission maxima of the native protein from 335 to 356 nm, change in the ellipticity at 218 nm and simultaneous decrease in the sugar binding activity were observed with increasing concentration of GdnHCl in the pH range between 4.0 and 9.0. The unfolding and inactivation by GdnHCl were partially reversible. Gel filtration of the lectin in presence of 1-6 m GdnHCl showed that the protein dissociates reversibly into partially unfolded dimer and then irreversibly into unfolded inactive monomer. Thermal denaturation was irreversible. The lectin loses activity rapidly above 45 degrees C. The exposure of hydrophobic patches, distorted secondary structure and formation of insoluble aggregates of the thermally inactivated protein probably leads to the irreversible denaturation. 相似文献
76.
The effect of carnitine administration on levels of lipid peroxide and activities of superoxide dismutase and catalase was
studied in rats administered isoproterenol to induce myocardial infarction. Levels of fatty acid were lower in rats pretreated
with carnitine at the peak period and given isoproterenol than the levels in isoproterenoltreated control rats. Lipid peroxides
were decreased in the heart at peak infarction in carnitine-treated rats compared to the levels in isoproterenol-treated controls.
Activities of superoxide dismutase and catalase showed no change in carnitine-treated animals given isoproterenol compared
to those in normal control rats, while they decreased in animals treated with isoproterenol alone. 相似文献