Characterization of a benzoate permease mutant of Pseudomonas putida

A spontaneous mutant of Pseudomonas putida (PRS 2017) has been isolated which is incapable of growth on benzoate, does not induce the enzymes of the catechol branch of the -ketoadipate pathway when grown in the presence of benzoate, cannot accumulate radioactively labeled benzoate, yet grows well with mandelate as sole source of carbon and energy. This strain apparently lacks a benzoate permease, which in the wild type shows a K _mof about 0.1 mM for benzoate, is inducible, and is not under the control of the regulatory system which governs the induction of the enzymes of the catechol branch of the -ketoadapate pathway. The lesion in PRS 2017 is apparently single site and maps near other genes governing benzoate dissimilation.Dedicated to R. Y. Stanier on the occasion of his 60th birthday     

The cellulase system of a Cytophaga species.

Cellulases (EC 3.2.1.4) of a Cytophaga species WTHC 2421 (ATCC 29474) were found in the soluble portion of the cell (the periplasm and the cytoplasm) and on the membrane. Cell-free cellulases were not found. Most of the carboxymethylcellulase activity associated with reduction of viscosity was membrane bound, whereas most of the carboxymethylcellulose (CMC) saccharifying activity was soluble. The CMC-saccharifying activity was increased 534 X by purification procedures which included ammonium sulfate precipitation and molecular exclusion chromatography with Sephadex G-75 and Biogel p-100. Periplasmic carboxymethycellulase had a molecular weight of 6250 and cytoplasmic carboxymethylcellulase had a molecular weight of 8650. Analytical ultracentrifugation of the periplasmic carboxymethylcellulase (CMCase) indicated that it had a low molecular density. The chromatographic fraction containing periplasmic CMCase also contained enzyme activity against crystalline cellulose. The activity against crystalline cellulose was 238 X higher than the activity shown by the whole cell. The reaction of the enzyme with either CMC or dewaxed cotton produced only glucose. The enzyme was slightly inhibited by the presence of 0.01% (w/v) glucose, lactose, or cellobiose, but it was not affected by sucrose, and exhibited increased activity in the presence of xylose and fructose.     

Antigens of Neisseria gonorrhoeae: Characterization by Gel Filtration, Complement Fixation, and Agar-Gel Diffusion of Antigens of Gonococcal Protoplasm

With the use of the agar-gel-diffusion and complement-fixation techniques, it was shown that protoplasm from different gonococcal isolates reacted with sera from some humans with a history of gonorrhea but did not react with "normal" human sera. The reactive antigen(s) could be partially separated from the other antigens by passing the gonococcal protoplasm through Sephadex G-200. The antigen(s) reacting in the gel-diffusion and complement-fixation tests appeared in the same fraction. On the basis of Sephadex gel filtration, the molecular weight of this antigen(s) is probably greater than 200,000.     

Preparation of Cell Walls and Protoplasm of Neisseria with the Ribi Cell Fractionator

The varied pressures required for disruption of Neisseria gonorrhoeae and other species of Neisseria when the Sorvall-Ribi refrigerated cell fractionator is used in the preparation of cell walls and cellular protoplasm are reported. Optimal disruption pressure for the gonococcus was considerably less than that required for other members of the genus Neisseria. Pressures varied from 8,000 psi for N. gonorrhoeae F62, colony type 4, to 22,000 psi for the nonpathogenic Neisseria-N. sicca, N. flava, and N. catarrhalis. Representative electron photomicrographs are shown.     

A synthetic tripeptide which increases survival of normal liver cells, and stimulates growth in hepatoma cells

A synthetic peptide with a structure analogous to a growth promoting human serum tripeptide was found to possess activities which, at nanomolar concentrations, increased the survival of normal hepatocytes from regenerating rat liver and which enhanced growth of a line of cultured hepatoma cells. The synthetic tripeptide (glycyl-histidyl-lysine) also stimulated the incorporation of labeled uridine and thymidine into trichloroacetic acid precipitable material.     

Three-dimensional structure of the elastase of Pseudomonas aeruginosa at 1.5-A resolution

Pseudomonas aeruginosa elastase (PAE) is a zinc metalloprotease with 301 amino acids. We have crystallized and solved the three-dimensional structure of PAE, using data to 1.5-A resolution, and have refined the native molecular structure to R = 0.188. The overall tertiary structure of the PAE molecule is similar to that of thermolysin, with which it shares 28% amino acid sequence identity. Nearly all of the active site residues that might potentially interact with substrates are identical in the two proteins. However, the active site cleft is significantly more "open" in PAE than in thermolysin.