Mine samples enriched for Thiobacillus ferrooxidans at 18, 12 and 6°C, after numerous transfers in medium 9K, yielded average generation times of 23, 44, and 103hr, respectively. Temperature characterization of a natural isolate, over a range of 6 to 35°C, showed that growth occurred at the temperature extremes and that the optimum temperature for growth was 25 to 30°C. A stock culture (commercially available) chosen for comparison showed a similar optimum temperature for growth, but lower growth rates at the lower temperatures examined. Growth studies of the natural isolates in the range of 2 to 35°C revealed that growth rates decreased on either side of 25 to 30°C. This effect was most pronounced in the 2 to 12°C range. These studies revealed the psychrotrophic nature of the natural T. ferrooxidans isolates, and their better growth capabilities at lower temperatures than T. ferrooxidans ATCC 33020. These observations are discussed in the light of their importance in the uranium bioleaching process, as it is currently being used.
The outer membrane of the radiation-resistant Acinetobacter sp. FO-1 was isolated by sucrose density-gradient centrifugation after disruption of bacterial cells by shaking with glass beads in a Braun Cell Homogeniser. The characteristics of the outer membrane were examined by enzymatic, chemical and electrophoretic analysis. The isolated outer membrane clearly differed from the inner membrane in the lipid-phosphorus and 2- keto-3-deoxyoctonate to protein ratios and in the enzyme activity of adenosine triphosphatase. The protein pattern of the outer membrane was determined by SDS-PAGE analysis. It showed twenty-five protein bands containing two major proteins with apparent approximate molecular weights of 39, 000 and 42, 000 daltons. The former was modified by heating. The latter was unmodified. These results were compared with the type strain of Acinetobacter calcoaceticus, ATCC 23055.
After induction, the wild type of Trametes versicolor produced two forms of laccase (A and B) in the ratio of 9 to 1. A UV-selected mutant strain of Trametes versicolor isolated on high concentrations of p-xylidine, p-cresol and phenol produced these laccases without induction in the ratio of 4 to 6. After induction with p-xylidine the production of laccase A increased significantly only in the mutant strain. Laccases A and B had catalytic activities proportional to the EPR spectroscopy-detected copper ions present. Several components were determined by electrophoretic analysis.
We investigated the biodegradation of two bipyridinium herbicides, paraquat and diquat, by Lipomyces starkeyi. This yeast was able to grow in a synthetic medium containing the herbicides as sole nitrogen sources. Growth with paraquat was more rapid than with diquat. Both herbicides were consumed from the medium within the logarithmic phase. The yeasts seemed to have low conversion activities from [methyl-14C]paraquat or [ethylene-14C]diquat to 14CO2, even when they were grown in the absence of the herbicides. Paraquat added to the culture medium augumented the degradation of both [14C]paraquat and [14C]diquat. On the other hand, diquat added to the medium did not augument degradation of the herbicides. Moreover, the addition of diquat together with paraquat abolished the augumentation caused by paraquat. The protein composition of L. starkeyi was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cells in the logarithmic phase contained more proteins than those in the stationary phase. In addition, the protein patterns of the cells in the logarithmic phase were found to vary with the nitrogen sources, i.e. paraquat, diquat and NH4Cl. The mechanisms which regulate the catabolism of the two herbicides are discussed.
Field surveys of hot springs were carried out at four Spas. Observations were made on the temperature, pH, dissolved oxygen and sulfide concentrations, current velocities, and the habitats of three (A, B, C) types of sulfur-turfs. The B-type (rods) was not present, while the A-type (sausage-shaped bacteria) grew only in the streams. In contrast, the C-type (filamentous bacteria) inhabited only the pools of hot spring water.
The DNA base composition of 74 strains in the genera Rhodosporidium, Cystofilobasidium, and Rhodotorula were determined by reversed-phase high-performance liquid chromatography (HPLC). The relative standard error of nucleoside analysis was less than 1%. The differences between the two values determined by the HPLC method and the Tm method were less than 1% in three strains determined in this study and less than 2%, with a few exceptions, compared with reference data.
Detailed taxonomic studies were made of thirty-five strains of psychrophilic yeasts with large ballistospores. These strains were isolated from dead leaves of Oryza sativa, Miscanthus sinensis, and Sasa sp. in Japan and were foundd to comprise a single, hitherto undescribed species of the genus Bullera. The species is described here as Bullera megalospora Nakase et Suzuki. Bullera megalospora resembles Bullera piricola and Sporobolomycespuniceus, but it can be distinguished from B. piricola by its inability to assimilate lactose, melibiose, and inositol, and from Sp. Puniceus in its lack of assimilation of inositol. Electrophoretic comparison of ten enzymes clearly demonstrated the differences among these three yeasts at the specific level; the similarities in their enzyme patterns were below 22%. Sporobolomyces puniceus was considered to be more closely related to B. megalospora and B. piricola than any of the other species of the genus Sporobolomyces. We propose to transfer this species to the genus Bullera as Bullera punicea (Komagata et Nakase) Nakase et Suzuki comb. nov.
Four polyglutamate (PGA)-producing Bacillus strains were isolated from "thua nao" in Thailand. Three of these did not require biotin for growth. All four produced high activities of γ-glutamyltranspeptidase (γ-GTP). Each of these strains carried a single plasmid species, and their molecular weights andd restriction patterns differ overall. The "thua nao" plasmids were found to be strongly hybridized with the "natto" plasmid, pUHl, which encodes the γ-GTP gene responsible for PGA production in B. subtilis (natto). Most of the generated fragments with HindIII from these plasmids showed higher degrees of homology with the probe, but some did not. Apparently a "natto" plasmid is distributed widely in PGA- producing Bacillus strains may develop from a common ancestral molecule. Therefore, the distribution of "natto" plasmids in PGA-producing Bacillus strains may help to distinguish B. subtilis from B. subtilis (natto).
The ubiquinone system of twenty-four strains of ballistospore-forming yeasts in the genera Sporidiobolus, Sporobolomyces, and Bullera was investigated. Most of strains are type strains. They are twenty-one known species and two unidentified species. Sporidiobolus johnsonii, Sporid. pararoseus, Sporid. ruinenii, Sporid. salmonicolor, Sporobolomyces alborubescens, Sp. foliicola, sp. gracilis, Sp. holsaticus, and Sp. Roseus had Q-10 as the major ubiquinone and Q-9 as the minor component. Bullera alba, B. armeniaca, B. aurantiaca, B. crocea, B. dendrophila, B. globospora, B. piricola, B. punicea, B. salicina, B. singularis, and B. tsugae also had Q-10 as the major ubiquinone and Q-9 as the minor component. The amounts of Q-9 are rather high (15.0-25.4%) in Sporid, pararoseus, Sporid. ruinenii, B. piricola, B. punicea, and B. salicina. The major ubiquinone of Sp. elongatus was Q-10 (H2). This yeast has Q-9 (H2) and Q-10 as the minor components. This is the 2nd finding of Q-10 (H2) in yeasts as the major ubiquinone. Three unidentified strains of two species of Bullera had Q-9 as the major ubiquinone and Q-8 and Q-10 as the minor components. Taxonomically, these strains resemble B. singularis in spite of the difference in ubiquinone systems. These yeasts may be yeast phases of certain species of the genus Itersonilia or may represent a genus yet to be established..