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M A Pisano

Publications and source records attributed to M A Pisano.

At least 37 records · Page 2Linked to original sources

Purification and properties of polyol dehydrogenase from Cephalosporium chrysogenus.

A polyol dehydrogenase of broad specificity was purified 178-fold from extracts of the filamentous fungus Cephalosporium chrysogenum. The enzyme was found to act as an oxido-reductase in two substrate-coenzyme systems: D-sorbitol (or xylitol)-nicotinamide-adenine dinucleotide (NAD) and D-mannitol-nicotinamide adenine dinucleotide phosphate (NADP). The dehydrogenase was composed of five isozymes, which, as a mixture, exhibited these properties: Km to D-sorbitol and D-mannitol, 7.15 to 10(-2) M; PH optimum, 9 to 10; molecular weight, 300,000 subunit weight, 29,000; PI, 5.8 to 7.5. The NADP-linked activity was labile to treatment with heat or ethylenediaminetetraacetic acid. Mixed substrate assays support the hypothesis that both NAD-, and NADP-linked activities are associated with isozymes of a single dehydrogenase.

Acremonium↗

Synthesis and antimicrobial activity of N-substituted N'-cyano-S-(triorganostannyl)isothioureas.

Six N-substituted N'-cyano-S-(trimethylstannyl)isothioureas were synthesized by the reaction of (trimethylstannyl)cyanamide with various organic isothiocyanates. The IR spectrum of each compound was obtained over the 4000-30-cm(-1) range, and some bands were assigned. The six new compounds and five previously synthesized N-substituted N'-cyano-S-(triphenylstannyl)isothioureas were tested for and were found to exhibit antifungal activity. N-Phenyl-N'-cyano-S-(triphenylstannyl)isothiourea was also investigated for antibacterial activity and was observed to be especially inhibitory toward Gram-positive species. The antimicrobial activity of two compounds was compared to that of the oxygen analogs of these compounds.

Bacteria↗

Production of cephalosporin C by Paecilomyces persicinus P-10.

After the growth of Paecilomyces persicinus P-10 in a glucose-peptone medium, filtrates were collected and analyzed for antibiotic antivity. Activities against Salmonella gallinarum ATCC 3030 and Alcaligenes faecalis ATCC 8750 (penicillin N-resistant strain) were obtained. Part of the former activity was readily inactivated by penicillinase. The fraction active against A. faecalis was isolated by passage through Amberlite XAD-2 and Amberlite IRA-68. The powder eventually obtained was subjected to paper chromatography followed by bioautography, and the activity obtained corresponded to that of a sample of cephalosporin C. Thin-layer chromatography was also employed to verify the presence of cephalosporin C in the P-10 powder. The active solids were further purified by means of paper chromatography in a solvent system consisting of n-butanol-acetic acid-water (60:15:25, vol/vol). The material obtained from this procedure yielded an infrared absorption spectrum identical to that of cephalosporin C. Similarly, the ultraviolet absorption of the purified preparation coincided with that of cephalosporin C. Exposure of the purified solids to cephalosporinase resulted in rapid inactivation of the antibiotic. In addition to penicillin N and cephalosporin C, filtrates of P. persicinus P-10 also contained deacetylcephalosporin C, deacetoxycephalosporin C, and cephalosporin P.

Cephalosporins↗

Characteristics of a bacteriocin derived from Streptococcus faecalis var. zymogenes antagonistic to Diplococcus peumoniae.

A bacteriocin-producing strain of Streptococcus faecalis var. zymogenes (E-1) was isolated from clinical material (conjunctiva). The active substance differed from bacteriocins described by other investigators primarily in its spectrum of antibacterial activity, especially by its marked inhibition of Diplococcus pneumoniae. The E-1 bacteriocin also inhibited nonhemolytic strains of enterococci as well as one-third of the Viridans group of streptococcal strains investigated. The degree of inhibition, however, as indicated by the size of the zones against the latter organisms, was significantly reduced. No activity was detected against any of the strains belonging to the following groups of bacteria: hemolytic enterococci, beta-hemolytic streptococci, nonhemolytic streptococci, staphylococci, and various gram-negative species. Similarly, three strains each of Bacillus cereus and Listeria monocytogenes and one strain of Erysipelothrix insidiosa were not inhibited. The bacteriocin was able to diffuse through bacterial membranes as well as cellulose dialyzer tubing. It was inactivated by heating to 80 C for 20 min but resisted inactivation by either trypsin or chloroform.

Agar↗

Proteinase production by a species of Cephalosporium.

An unidentified Cephalosporium species produced an extracellular proteinase when grown in a variety of fermentation media under submerged culture conditions. Maximal enzyme yields were obtained in a medium containing 2% corn meal, 1% soybean meal, and 0.5% CaCO(3) in tap water. Optimal proteinase production in this medium occurred within a 72- to 96-hr growth period. High enzyme yields were also attained with media in which cottonseed meal, Fermatein, Pharmamedia, or soybean-alpha-protein was substituted for the soybean meal. The substitution of these ingredients for the corn meal resulted in significantly decreased proteinase yields. The addition of minerals or vitamins to the corn meal-soybean meal fermentation medium failed to enhance proteinase production. The enzyme was most active in an alkaline environment; maximal caseinolysis occurred at pH 7.5, whereas pH 8.5 was optimal for either hemoglobin or beta-lactoglobulin hydrolysis. Enzymatic activity was also noted with either bovine albumin fraction V or soybean-alpha-protein substrates, whereas ovalbumin was not susceptible to enzymatic attack. The enzyme was stable within the pH range of 3.0 to 9.5 at 25 C for 2 hr, and at 5 C for 24 hr. The proteinase was stable upon heating for 10 min at 35 to 45 C, but it was totally inactivated at 70 C. The proteinase was unaffected by soybean inhibitor, partially inactivated by lima bean inhibitor, and completely inactivated by ovomucoid inhibitor.

Acremonium↗

Steroid transformations by species of Cephalosporium and other fungi.

A total of 58 cultures, tentatively identified as species of the genus Cephalosporium, were screened in flask fermentations for their ability to effect conversions of progesterone (Delta(4)-pregnene-3,20-dione) and Reichstein's Substance S (Delta(4)-pregnene-17alpha,21-diol-3,20-dione). A large number of transformations were observed by means of a series of five paper chromatography systems rated for analysis of steroid compounds ranging in polarity from progesterone to polyhydroxylated steroids. Five different transformation products were selected for isolation and identification. For purposes of recovery, conversions were conducted under submerged conditions in either 4- or 200-liter fermentors in which the broth was agitated and aerated. The steroid substrate was dissolved in acetone and added aseptically to the growing culture in a final concentration of 0.025%. After the conversions were effected, the whole broth was extracted with chloroform, and the transformation products were recovered, either by direct crystallization from solvents or through the use of silica gel columns. It was determined that C. ciferrii 21C converted progesterone to Delta(4)-androstene-3,17-dione. Kendall's Compound F (Delta(4)-pregnene-11beta,17alpha,21-triol-3,20-dione) was converted to its 20beta-ol analogue by Geotrichum sp. 51C (during these studies, a number of cultures were taxonomically reclassified). Cephalosporium sp. 27C formed the Delta(1)-analogue of Reichstein's Substance S, and Cephalosporium sclerotigenum 31C and Verticillium aphidum both converted Substance S to the 6beta-hydroxy derivative. Paecilomyces persicinus 22C converted Substance S to a product believed to be a dihydroxylated derivative.

Ascomycota↗

Synergistic effects in sonochemical sterilization.

The synergistic effects observed during the sterilization of Bacillus subtilis var. niger ATCC 9372 by the combined action of ethylene or propylene oxide with high-intensity airborne sound waves (34.8 kc/sec) were investigated. It has been shown that the number of surviving spores deposited on paper strips decreases exponentially with the sound intensity at sample level. Reductions of the order of one-third of the time required for standard propylene oxide sterilization have been observed by using the combined action of sound waves with gaseous sterilization. At the present time, maximal synergistic effects seem to be achieved for the following experimental conditions: propylene oxide concentration, 500 to 1,000 mg/liter; acoustic intensity, 161 to 162 db; contact time, 80 min; temperature, 60 C; and relative humidity, 40%. The basic mechanism involved in sonochemical sterilization seems to be more of a physical (accelerated gas diffusion) than a chemical nature.

Bacillus subtilis↗

Production of alanine by Fusarium moniliforme.

Fusarium moniliforme grown in a chemically defined medium in submerged culture accumulated amino acids extracellularly. Alanine and glutamic acid were present in greatest amounts, with traces of glycine, lysine, threonine, and valine detectable. Increasing the glucose and urea concentrations of the medium increased yields of alanine. Further increases in alanine production occurred with elevated levels of mineral salts in the medium, whereas the addition of a vitamin mixture proved to be inhibitory. Chemical changes resulting from the growth of F. moniliforme in the final fermentation medium disclosed maximal alanine production, mycelial weight, and glucose consumption after 72 hr of incubation at 28.5 C. Total soluble nitrogen, by contrast, was minimal at the same time period. The pH remained in the alkaline range throughout the fermentation.

Alanine↗

Sterilizing effects of high-intensity airborne sonic and ultrasonic waves.

The lethal effects of high-intensity airborne sonic (9.9 kc/sec) and ultrasonic waves (30.4 kc/sec) on spores of Bacillus subtilis var. niger ATCC 9372 were determined. The spores, which were deposited on filter-paper strips, were exposed to sound waves for periods varying from 1 to 8 hr, at a temperature of 40 C and a relative humidity of 40%. Significant reductions in the viable counts of spores exposed to airborne sonic or ultrasonic irradiations were obtained. The antibacterial activity of airborne sound waves varied with the sound intensity level, the period of irradiation, and the distance of the sample from the sound source. At similar intensity levels, the amplitude of motion of the sound waves appeared to be a factor in acoustic sterilization.

Bacillus subtilis↗