Summary A leucine aminopeptidase was purified to homogeneity fromStreptomyces rimosus culture filtrates, which are waste broth of oxytetracycline bioproduction process. Purification procedure includes ultrafiltration
and chromatography on CM-Sephadex, AH-Sepharose and FPLC Mono S column.
The enzyme is a monomer with molecular weight of 27,500 Daltons and pI of 7.3, stable in broad pH range and up to 70°C. It
is a metallo enzyme dependent on Ca2+ ions for its full activity. By its specificity it is a true aminopeptidase active on amino acid amide, arylamide, peptide
and ester bonds. The hydrolysing activity shows preference for leucine at the N-terminal position of substrates, also acts
on aromatic acids and methionine, but does not release glycine, proline, acidic amino acids orD-amino acid residues. 相似文献
Agrobacterium radiobacter, strain B6 (a strain isolated in this laboratory, which limited the occurrence of damping-off of sugar beet and influenced growth of plants in hot-house and field experiments) was found to produce an acidic exopolysaccharide in a mineral medium with various carbon sources. Hydrolyzates of the polysaccharide contained glucose, galactose, glycerol, succinic acid and pyruvic acid, whose quantitative content varied according to the carbon source used. The polysaccharide isolated from the medium containing glucose exhibited the highest physiological activity. Seeds germinated best and sugar beet roots were found to grow most rapidly in a medium containing 0.2 % (W/W) of the polysaccharide. The roots exposed for 3 d in this medium grew 2.7-fold as compared with non-treated plants. Higher sumbers of microorganisms were detected on the surface of roots treated with the polysaccharide. Growth of roots was also stimulated when immersing the seeds (30 min) in a 0.2 –0.4 % solution of this polysaccharide. After a two-fold treatment the roots were less damaged by the fungusPythium ultimum. Plants from seeds treated with the polysaccharide grew in the field soil more rapidly than the non-treated plants but worse than after bacterization of the seeds byA. radiobacter B6 and were only partially protected against the damping-off of sugar beet. 相似文献
Oudin’s principle of single immunodiffusion in agar gel was modified for quantitative determination of IgA in bronchoalveolar lavage (BAL) of normal 20–25 g mice. The reaction took place at 25 °C in 0.3 % agarose with 16.7 % pig serum against mouse IgA, and was evaluated on the basis of a relationship between the progress of the precipitin zone and the square root of time. The linear dependence of the derived constantk on the logarithmic concentration of antibody in the sample permitted to express the results as titre, corresponding to a dilution wherek = 0. Examination of seven samples of pooled blood serum of normal mice shoved taht (1) the IgA level was practically constant, (2) serum IgA possessed under given conditions similar properties as IgA from the bronchoalveolar secretion; it is therefore possible to employ pooled sera as a reliable control of the immunodiffusion system in ease of lack of reference standards with defined IgA content. Examination of 82 individual BAL samples of normal mice revealed that the mean IgA concentration in 2.5 mL samples was almost 1000 times lower than in blood serum. 相似文献
In partly purified protein complexes obtained from 22 species of theAllium genus and 6 cultivars ofAllium cepa the activity of cholinesterases was detected and measured using the method of Ellman et al. The degree of its inhibition with 10-4 M neostigmine was also tested. It was found that the activity of cholinesterase differed in individual species up to two hundred times, while the differences in the inhibitory activity of 10-4 M neostigmine occurred only in a few cases. Individual sections and cultivars could not be characterized on the basis of the differences in the activities of the cholinesterases. Of all the sections that ofPhyllodolon shows the highest average activity. In the case of the tested cultivars distinctly the lowest activity was observed in cv. Kastická. The values of the enzymatic activity measured by Ellman’s method in this plant material include the activity of specific and unspecific cholinesterases and the part uninhibitable by neostigmine. 相似文献
The seed protein patterns of 12Arachis species were compared by polyacrylamide gel electrophoresis (PAGE), similarities between patterns were measured by the Jaccard
index. Results obtained confirm the close relationships established between members of the genus on morphological grounds
and support the more recent classification schemes.A. villosa andA. correntina could well be regarded as distinct species on grounds of protein differences whileA. macedoi andA. villosulicarpa (although members of the same section, Extranervosae) show considerable differentiation of their protein patterns. Surprisingly,
the formA. ×batizogaea showed less similarity in protein pattern to those of its parental species than might have been expected. The principle value
of seed protein pattern data appears to be in distinguishing species within sections. 相似文献
Soybean (Glycine max L. Merr. cv Bragg) was grown throughout its life cycle at 330, 450, and 800 microliters CO2 per liter in outdoor controlled-environment chambers under solar irradiance. Leaf ribulose-1,5-bisphosphate carboxylase (RuBPCase) activities and ribulose-1,5-bisphosphate (RuBP) levels were measured at selected times after planting. Growth under the high CO2 levels reduced the extractable RuBPCase activity by up to 22%, but increased the daytime RuBP levels by up to 20%.
Diurnal measurements of RuBPCase (expressed in micromoles CO2 per milligram chlorophyll per hour) showed that the enzyme values were low (230) when sampled before sunrise, even when activated in vitro with saturating HCO3− and Mg2+, but increased to 590 during the day as the solar quantum irradiance (photosynthetically active radiation or PAR, in micromoles per square meter per second) rose to 600. The nonactivated RuBPCase values, which averaged 20% lower than the corresponding HCO3− and Mg2+-activated values, increased in a similar manner with increasing solar PAR. The per cent RuBPCase activation (the ratio of nonactivated to maximum-activated values) increased from 40% before dawn to 80% during the day. Leaf RuBP levels (expressed in nanomoles per milligram chlorophyll) were close to zero before sunrise but increased to a maximum of 220 as the solar PAR rose beyond 1200. In a chamber kept dark throughout the morning, leaf RuBPCase activities and RuBP levels remained at the predawn values. Upon removal of the cover at noon, the HCO3− and Mg2+-activated RuBPCase values and the RuBP levels rose to 465 and 122, respectively, after only 5 minutes of leaf exposure to solar PAR at 1500.
These results indicate that, in soybean leaves, light may exert a regulatory effect on extractable RuBPCase in addition to the well-established activation by CO2 and Mg2+.