Protoplast formation and regeneration in <Emphasis Type="Italic">Lactobacillus delbrueckii</Emphasis>

Method for production and regeneration of Lactobacillus delbrueckii protoplasts are described. The protoplasts were obtained by treatment with a mixture of lysozyme and mutanolysin in protoplast buffer at pH 6.5 with different osmotic stabilizers. The protoplasts were regenerated on deMan, Rogosa and Sharpe (MRS) with various osmotic stabilizers. Maximum protoplast formation was obtained in protoplast buffer with sucrose as an osmotic stabilizer using a combination of lysozyme (1 mg/ml) and mutanolysin (10 μg/ml). Maximum protoplast regeneration was obtained on MRS medium with sucrose (0.5 M) as an osmotic stabilizer. The regeneration medium was also applicable to other species of lactobacilli as well. This is, to our knowledge, the first report on protoplast formation and efficient regeneration in case of L. delbrueckii.     

An inquiry into the source of stereospecificity of lactate dehydrogenase using substrate analogues and molecular modeling.

Lactate dehydrogenase catalyzes the stereospecific hydride transfer to and from the re face of the nicotinamide coenzyme. The demonstrated probability of transfer to the si face of less than 2 x 10(-8) indicates that the free energy of any diastereotopic transition state leading to a si transfer must be over 10 kcal/mol greater than the free energy for transfer to or from the re face. The general notion of closed, desolvated active sites suggests the a priori hypothesis that steric hindrance prevents the nicotinamide ring from assuming a conformation that would lead to transfer of the pro-S hydrogen. In this paper we report that the probability of transfer of the pro-S proton is less than 9 x 10(-7) with 3-pyridinealdehyde adenine dinucleotide as coenzyme and less than 4 x 10(-7) during the lactate dehydrogenase catalyzed disproportionation of glyoxylate. Examination of the crystal structure of lactate dehydrogenase further suggests that steric exclusion does not enforce the extreme stereospecificity of the reaction. An electrostatic interaction with the macrodipole associated with the alpha 2F helix is suggested as a potential molecular source of the stereospecificity.     

The effect of antibiotics on the electrical polarisability of aqueous suspensions of bacteria.

The orientation of washed Escherichia coli bacteria in suspension induced by applied a.c. electric fields has been monitored by observing the accompanying changes in the intensity of light scattered by the suspension. The data enable the anisotropy of electrical polarisability deltaalpha to be determined. Changes in deltaalpha due to the addition of various antibiotics to the suspension have been measured as a function of both the antibiotic concentration and the temperature of the suspension. The results are taken to indicate that there is an accumulation of the antibiotic molecules at the bacterial surface.     

In vivo and in vitro sugar transport in frog intestine

In the frog intestine, both in vitro and in vivo, experiments were carried out in order to increase knowledge of the mechanism of sugar exit across the basolateral membrane of the enterocyte. The frog intestine was chosen because it lacks crypt cells and, consequently, any external fluid circuit mechanism during sugar transport can be avoided. Therefore, the sugar concentration in the absorbate collected on the serosal side is likely to be similar to that present underneath the basolateral membrane of the enterocyte. Under this condition, cell and absorbate sugar concentrations are similar; yet there is a concomitant net transintestinal sugar transport. Moreover, in in vivo experiments a net transintestinal sugar transport takes place even against a concentration difference. These results suggest that sugar exit across the basolateral membrane is not simply due to a chemically facilitated diffusion.     

Results of the study of preventive effectiveness and side effects of anti-influenza lactoglobulin in controlled epidemiologic trial]

Prophylactic activity and side action of antiinfluenza lactoglobulin were studied under conditions of controlled epidemiological trial in 6361 persons during the epidemiological rise of influenza incidence. The data obtained pointed to the absence of any protective effect of antiinfluenza lactoglobulin. Marked allergenic properties of lactoglobulin and development of sensitization in the persons under observation were found during the study of the side-effect of the preparation. Thus, the absence of prophylactic efficacy and the sensitizing action of antiinfluenza lactoglobulin pointed to the inexpediency of its use for prophylactic and therapeutic purpose.     

Crystallins in the differentiation and regeneration processes of the crystalline lens in amphibia

The published and authors' data have been summarized on (1) the spectrum and properties of crystallins in different amphibian species, (2) localization and synthesis of crystallins in different cellular compartments of the adult amphibian lens, (3) dynamics of crystallin formation during embryogenesis and (4) lens regeneration from tissues of the larval and adult amphibian eyes. The necessity of more detailed studies of crystallin synthesis during embryogenesis and lens regeneration using molecular biological and biochemical methods is stressed. The significance of this approach is illustrated by the pioneering data of Soviet scientists on crystallin polypeptides and corresponding mRNAs in development of Rana temporaria obtained with the use of DNA-RNA hybridization and immunoelectroblotting.