Phylogeny of marine Bacillus isolates from the Gulf of Mexico.

The phylogeny of 11 pigmented, aerobic, spore-forming isolates from marine sources was studied. Forty-two biochemical characteristics were examined, and a 16S rDNA sequence was obtained for each isolate. In a phylogenetic tree based on 16S sequencing, four isolates (NRRL B-14850, NRRL B-14904, NRRL B-14907, and NRRL B-14908) clustered with B.

Phylogenetic position of the genus Hydrogenobacter.

The genus Hydrogenobacter consists of extremely thermophilic, obligately chemolithotrophic organisms that exhibit anaerobic anabolism but aerobic catabolism. Preliminary studies of the phylogenetic position of these organisms based on limited 16S ribosomal DNA sequence data suggested that they belong to one of the earliest branching orders of the Bacteria. In this study, the complete 16S ribosomal DNA sequences of two type strains, Hydrogenobacter thermophilus TK-6 and Calderobacterium hydrogenophilum Z-829, and another isolate, Hydrogenobacter sp.

Rapid in situ hybridization technique using 16S rRNA segments for detecting and differentiating the closely related gram-positive organisms Bacillus polymyxa and Bacillus macerans.

A rapid, sensitive, inexpensive in situ hybridization technique, using 30-mer 16S rRNA probes, can specifically differentiate two closely related Bacillus spp., B. polymyxa and B.

Comparative sequence analyses on the 16S rRNA (rDNA) of Bacillus acidocaldarius, Bacillus acidoterrestris, and Bacillus cycloheptanicus and proposal for creation of a new genus, Alicyclobacillus gen. nov.

Comparative 16S rRNA (rDNA) sequence analyses performed on the thermophilic Bacillus species Bacillus acidocaldarius, Bacillus acidoterrestris, and Bacillus cycloheptanicus revealed that these organisms are sufficiently different from the traditional Bacillus species to warrant reclassification in a new genus, Alicyclobacillus gen. nov. An analysis of 16S rRNA sequences established that these three thermoacidophiles cluster in a group that differs markedly from both the obligately thermophilic organisms Bacillus stearothermophilus and the facultatively thermophilic organism Bacillus coagulans, as well as many other common mesophilic and thermophilic Bacillus species.

How close is close: 16S rRNA sequence identity may not be sufficient to guarantee species identity.

16S rRNA (genes coding for rRNA) sequence comparisons were conducted with the following three psychrophilic strains: Bacillus globisporus W25T (T = type strain) and Bacillus psychrophilus W16AT, and W5. These strains exhibited more than 99.5% sequence identity and within experimental uncertainty could be regarded as identical.

Phylogenetic diversity in the genus Bacillus as seen by 16S rRNA sequencing studies.

Comparative sequence analysis of 16S ribosomal (r)RNAs or DNAs of Bacillus alvei, B. laterosporus, B. macerans, B.

PCR amplification of 16S rDNA from lyophilized cell cultures facilitates studies in molecular systematics.

The sequence of the major portion of a Bacillus cycloheptanicus strain SCH(T) 16S rRNA gene is reported. This sequence suggests that B. cycloheptanicus is genetically quite distinct from traditional Bacillus strains (e.

An Escherichia coli mutant conditionally altered in respiratory chain components.

Nitrosoguanidine mutagenesis was employed to isolate an Escherichia coli mutant conditionally altered in respiratory chain components. Mutant R25 was able to grow on glucose, fructose, and glycerol but failed to grow on succinate and acetate (suc-). Also, R25 exhibited leaky growth on DL-lactate, fumarate, and malate (lct*).

Characterization of an Escherichia coli mutant pleiotropically altered in membrane-bound oxidoreductase activities.

An Escherichia coli mutant pleiotropically altered in membrane-bound oxidoreductase activities was isolated following nitrosoguanidine treatment. Mutant R23 was able to grow on glucose, but was unable to grow on succinate or other oxidizable substrates as a sole energy source. Isolated membranes prepared from R23 failed to oxidize succinate and formate; while NADH was oxidized at a reduced rate by membranes.

Activation studies by phospholipids on the purified cytochrome c4:o oxidase of Azotobacter vinelandii.

A modified procedure is described that was used to solubilize and purify the TMPD-dependent cytochrome c4:o oxidase from Azotobacter vinelandii. Two functional components (Fractions I and V) were obtained after DEAE-cellulose chromatography. Fraction V contained both cytochrome c4 (3.

Comparative Cytochrome Oxidase and Superoxide Dismutase Analyses on Strains of Azotobacter vinelandii and Other Related Free-Living Nitrogen-Fixing Bacteria.

Quantitative N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD) oxidase and superoxide dismutase (SOD) analyses were performed on representative organisms of the family Azotobacteraceae. Azotobacter vinelandii, Azotobacter chroococcum, Azotobacter paspali, and Derxia gummosa exhibited high quantitative TMPD oxidase activities, and their extracts possessed very active and electrophoretically homogeneous (single gel band) Fe-type SODs. Azomonas macrocytogenes extracts had similar single Fe-type SODs, and their cells exhibited no TMPD-dependent cytochrome oxidase activity.

Isolation and purification of the cytochrome oxidase of Azotobacter vinelandii.

A membrane-bound cytochrome oxidase for Azobacter vinelandii was purified 20-fold using a detergent-solubilization procedure. Activity was monitored using as ascorbate-TMPD oxidation assay. The oxidase was 'solubilized' from a sonic-type electron-transport particle (R3 fraction) using Triton X-100 and deoxycholate.

Purification and characterization of cytochrome O from Azotobacter vinelandii.

The membrane-bound cytochrome O has been solubilized from the Azotobacter vinelandii electron transport particle and further purified by use of conventional chromatographic procedures. The spectral characteristics as well as the other properties noted for purified cytochrome O are reported herein.

Use of a quantitative oxidase test for characterizing oxidative metabolism in bacteria.

It was possible to quantitate the terminal oxidase(s) reaction using bacterial resting-cell suspensions and demonstrate the usefulness of this reaction for taxonomic purposes. Resting-cell suspensions of physiologically diverse bacteria were examined for their capabilities of oxidizing N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD) using a manometric assay. For organisms having this capability, it was possible to calculate the conventional TMPD oxidase Q(O2) value (microliters of O2 consumed per hour per milligram [dry weight]).

Tetramethyl-p-phenylenediamine oxidase reaction in Azotobacter vinelandii.

It was possible to quantitate the tetramethyl-p-phenylenediamine (TMPD) oxidase reaction in Azotobacter vinelandii strain O using turbidimetrically standarized resting cell suspensions. The Q(O2) value obtained for whole cell oxidation of ascorbate-TMPD appeared to reflect the full measure of the high respiratory oxidative capability usually exhibited by this genera of organisms. The Q(O2) value for the TMPD oxidase reaction ranged from 1,700 to 2,000 and this value was equivalent to that obtained for the oxidation of the growth substrate, e.

Characterization studies on the membrane-bound adenosine triphosphatase (ATPase) of Azotobacter vinelandii.

The adenosinetriphosphatase (ATPase) (EC 3.6.1.

Quantitation of the tetramethyl-p-phenylenediamine oxidase reaction in Neisseria species.

The tetramethyl-p-phenylenediamine oxidase reaction commonly used in the Kovacs oxidase test was quantitatively estimated for various Neisseria species employing standardized resting cell suspensions. This genus of microorganisms exhibited very high tetramethyl-p-phenylenediamine oxidase rates comparable to that of Azotobacter and Pseudomonas, and this reaction was found to be a valid measurement for the respiratory capability possessed by this group of organisms.

Preliminary characterization studies on the Neisseria catarrhalis respiratory electron transport chain.

The Neisseria catarrhalis respiratory electron transport system was examined in a sonic type particulate membrane fraction and shown to have a moderately active succinate as well as nonpyridine nucleotide-dependent dl-lactate oxidoreductase and a very active tetramethyl-p-phenylenediamine oxidase. l-Malate and l-glutamate oxidation were found to be dependent on pyridine nucleotides and exclusively associated with a soluble (or nonmembranous) fraction. The primary cytochrome components in the electron transport system appear to be c-type in nature (555 nm and 550 nm) as well as cytochrome a(1) (600 nm) and cytochrome o.