PubMed Health⌕ Search

Biomedical subjects

H J Phaff

Publications and source records attributed to H J Phaff.

50 records · Page 3Linked to original sources

Growth of Saccharomycopsis schionning under continuous gassing.

Two strains of Saccharomycopsis guttulata, JB-1 and JB-3, isolated from stomach contents of domestic rabbits, were grown under different gas phases, and their growth rates were compared. Strain JB-1 grew exponentially at a maximal growth rate under a continuous gas phase of 15% CO(2), 2% O(2) in nitrogen. High cell yields with low cell granulation were obtained. The growth rates were almost the same between oxygen concentrations of 0.25 and 20% at 15% CO(2). Poor growth and early cell granulation occurred in the absence of oxygen at 15% CO(2). Growth increased at 2% O(2) in direct proportion to the carbon dioxide concentration up to 10 to 15% CO(2). A very high carbon dioxide content (e.g. 98%) was somewhat inhibitory. Cell granulation always occurred during the maximal stationary phase in media at pH 4, but was relatively slight at pH 5.6 or higher. Strain JB-3 responded to various gas phases in a similar manner except that it grew slowly in the absence of oxygen at 15% CO(2) (pH 4). The effect of an optimal gas phase on the growth of strain JB-1 was examined in relation to other environmental conditions. In the presence of 15% CO(2), 2% O(2), this strain grew exponentially in yeast autolysate-Proteose Peptone-glucose medium at 37 C at pH 2, 4, and 5.6 at approximately the same rate; the growth rate was somewhat lower at pH 6.2. Under similar conditions, strain JB-1 grew at 30 C and pH 4 at one-sixth its maximal growth rate. Cell granulation was greatly reduced at this temperature. With adequate CO(2) strain JB-1 also grew at a reduced rate in a yeast autolysate medium previously reported not to support growth. Results indicate that continuous gassing with an optimal gas phase increases the growth rate to the extent that the growth rate surpasses the death rate by a significant margin; as a result, granulated cells can be avoided almost entirely in the log phase.

Animals↗

Deoxyribonucleic acid base composition in yeasts.

The deoxyribonucleic acid base composition of 15 species of yeasts was determined to obtain further clues to or supporting evidence for their taxonomic position. Species examined belonged to the genera Saccharomyces, Debaryomyces, Lodderomyces, Metschnikowia, and Candida. The range of moles per cent guanine plus cytosine (GC content) for all yeasts examined extended from 34.9 to 48.3%. The sporogenous species and the asporogenous yeasts spanned the range with 36.6 to 48.3% GC and 34.9 to 48% GC, respectively. Three Saccharomyces species (S. rosei and related species) exhibited significantly higher GC contents than S. cerevisiae, whereas the fermentative species D. globosus revealed a%GC more aligned to the S. rosei group than to the nonfermentative D. hansenii. Similar GC contents were demonstrated by L. elongasporus and its proposed imperfect form C. parapsilosis. The range of GC contents of various strains of three Metschnikowia species studied was 6.1%, with the type strain of M. pulcherrima having the highest GC content (48.3%) of all of the yeasts examined.

Candida↗

Nucleotide composition of deoxyribonucleic acid of some species of Cryptococcus, Rhodotorula, and Sporobolomyces.

The buoyant density of deoxyribonucleic acid (DNA) from nine species and two varieties of Cryptococcus, three species and two varieties of Rhodotorula, and six species of Sporobolomyces was determined by CsCl density gradient equilibrium centrifugation. Several species were represented by two to four different strains. Expressed in moles per cent of guanine plus cytosine (GC content) the ranges were 49 to 65%, 52 to 70%, and 51 to 65% for Cryptococcus, Rhodotorula, and Sporobolomyces, respectively. For each genus, the GC content was distributed into two discrete groups with averages ranging from 52 to 54 and 60 to 66, respectively. An analysis of these results suggested that the determination of GC content of DNA had a taxonomic value for these yeast genera.

Centrifugation, Density Gradient↗

Exo-beta-glucanases in yeast.

1. A number of yeast species were examined for the presence of beta-glucanases. Extracts obtained by cell disruption of Saccharomyces cerevisiae, Fabospora fragilis and Hansenula anomala hydrolysed laminarin and pustulan with the production of glucose. Enzymic activities were also detected in the culture fluids of F. fragilis and H. anomala grown aerobically in buffered mineral medium with glucose as the carbon source. 2. F. fragilis and H. anomala possessed approximately sevenfold higher beta-(1-->3)-glucanase activity than S. cerevisiae. 3. Intracellular exo-beta-glucanase from baker's yeast was purified 344-fold from the dialysed cell extract. 4. Exo-beta-glucanase from F. fragilis was purified 114-fold from the dialysed culture fluid and 423-fold from the dialysed intracellular extract. The purified extracellular and intracellular enzymes had similar properties and essentially the same specific activity, 79 enzyme units/mg. of protein. 5. Extracellular exo-beta-glucanase of H. anomala was purified 600-fold. 6. The optimum pH of the enzymes from F. fragilis, S. cerevisiae and H. anomala was 5.5 in each case. Chromatographic evidence indicated that the three enzymes remove glucosyl units sequentially from laminarin as well as pustulan. 7. The ratio of activities towards laminarin and pustulan remained constant during purification of the exo-beta-glucanase obtained from the three species, suggesting a single enzyme. Additional evidence for its unienzymic nature are: (i) the two activities were destroyed at exactly the same rate on heating of the purified enzyme from F. fragilis at three different temperatures; (ii) the competitive inhibitor glucono-delta-lactone gave the same value of K(i) when tested with either substrate; (iii) quantitative application of the ;mixed-substrate' method with the purified enzyme of S. cerevisiae gave data that were in excellent agreement with those calculated on the assumption of a single enzyme. 8. The purified exo-beta-glucanases of the different species of yeast had different kinetic constants. The ratios of maximal velocities and K(m) values with laminarin and pustulan differed markedly. Comparison of V(max.) and K(m) values suggests that the rapid release of spores from asci in F. fragilis might be explained in terms of an enzyme with higher maximal velocity and higher affinity to the ascus wall than that present in baker's yeast. 9. The estimated molecular weights for exo-beta-glucanases from F. fragilis, S. cerevisiae and H. anomala were 22000, 40000 and 30000 respectively.

Chromatography↗

Successive Microbial Populations in Calimyrna Figs.

Smyrna-type (Calimyrna) figs have essentially sterile internal tissue until visited by the pollinating fig wasp, Blastophaga psenes, which introduces a specific microflora consisting of Candida guilliermondii var. carpophila and Serratia plymuthica. This flora persists and develops in numbers throughout the ripening period until maturity of the fruit. These organisms do not cause spoilage. The presence of C. guilliermondii var. carpophila appears to increase the attractiveness of the fruit to drosophilae. Drosophila (mainly D. melanogaster) carry spoilage yeasts and bacteria on their exterior body parts, and introduce these organisms during ovipositing in the fruit cavity. The spoilage yeasts consist almost entirely of apiculate yeasts (Hanseniaspora valbyensis, H. uvarum, and Kloeckera apiculata) and of Torulopsis stellata, which cause active fermentative spoilage. Spoilage bacteria (primarily Acetobacter melanogenus) are also introduced with the yeasts. Organic acids are produced by these yeasts as well as by the Acetobacter. A number of minor spoilage yeasts were also identified.

Journal Article↗

Ultrastructure of the cell wall of Schizosaccharomyces pombe following treatment with various glucanases.

The ultrastructure of isolated cell walls of Schizosaccharomyces pombe was studied by electron microscopy after treatment with the following purified enzymes: endo-beta-(1-->3)-glucanase, endo-beta-(-->6)-glucanase, and endo-alpha-(1-->3)-glucanase produced by Bacillus circulans; exo-beta-(1-->3)-glucanase and endo-beta-(1-->3)-glucanase produced by Schizosaccharomyces japonicus var. versatilis. The exo-beta-(1-->3)-glucanase had no detectable effect on the walls, but amorphous wall material was removed by action of the endo-beta-(1-->3)- and endo-beta-(1-->6)-glucanases of B. circulans to reveal a wall component consisting of densely interwoven microfibrils. The fibrils were hydrolyzed by treatment with the Schiz. japonicus endo-beta-(1-->3)-glucanase followed by B. circulans endo-alpha-(1-->3)-glucanase--suggesting that they were composed of -beta-(1-->3)-linked glucan and alpha-(1-->3)-linked glucan. The presence of a fibrillar component in untreated walls was evident after negative staining.

Actin Cytoskeleton↗