Numerical taxonomy and ecology of petroleum-degrading bacteria.

A total of 99 strains of petroleum-degrading bacteria isolated from Chesapeake Bay water and sediment were identified by using numerical taxonomy procedures. The isolates, together with 33 reference cultures, were examined for 48 biochemical, cultural, morphological, and physiological characters. The data were analyzed by computer, using both the simple matching and the Jaccard coefficients. Clustering was achieved by the unweighted average linkage method. From the sorted similarity matrix and dendrogram, 14 phenetic groups, comprising 85 of the petroleum-degrading bacteria, were defined at the 80 to 85% similarity level. These groups were identified as actinomycetes (mycelial forms, four clusters), coryneforms, Enterobacteriaceae, Klebsiella aerogenes, Micrococcus spp. (two clusters), Nocardia species (two clusters), Pseudomonas spp. (two clusters), and Sphaerotilus natans. It is concluded that the degradation of petroleum is accomplished by a diverse range of bacterial taxa, some of which were isolated only at given sampling stations and, more specifically, from sediment collected at a given station.     

Genetic engineering of coryneform bacteria

The coryneforms are a diverse group of bacteria which includes animal and plant pathogens as well as non-pathogenic bacteria. Although they are of significant economic and health importance, their genetics is poorly understood. The development of genetic engineering techniques for coryneforms and initial gene cloning studies are discussed.     

Morphology and physiology of coryneform bacteria

Four groups of coryneform bacteria, viz. soil arthrobacters, orange cheese coryneforms, orange sea-fish coryneforms and non-orange cheese coryneforms, were studied as regards morphological and physiological features. The soil arthrobacters can be divided into a simplex and a globiformis group on the basis of their ability of utilizing a number of carbon sources. The group of the orange cheese coryneforms was found to be rather homogeneous, in contrast to the groups of the orange sea-fish coryneforms and the non-orange cheese coryneforms, some strains of which deviated from the others of their group as to the majority of the characteristics tested. Mainly the physiological characteristics of each of the groups justify the division of the coryneforms into the four chief groups mentioned.     

Numerical taxonomy of phenanthrene-degrading bacteria isolated from the Chesapeake Bay

Phenanthrene-degrading bacteria were isolated from Chesapeake Bay samples by the use of a solid medium which had been overlaid with an ethanol solution of phenanthrene before inoculation. Eighteen representative strains of phenanthrene-degrading bacteria with 21 type and reference bacteria were examined for 123 characteristics representing physiological, biochemical, and nutritional properties. Relationships between strains were computed with several similarity coefficients. The phenogram constructed by unweighted-pair-group arithmetic average linkage and use of the simple Jaccard (SJ) coefficient was used to identify seven phena. Phenanthrene-degrading bacteria were identified as Vibrio parahaemolyticus and Vibrio fluvialis by their clustering with type and reference strains. Several phenanthrene-degrading bacteria resembled Enterobacteriaceae family members, although some Vibrio-like phenanthrene degraders could not be identified.     

Plasmids in coryneform bacteria of human origin

A study of plasmids in coryneform bacteria isolated from human sources is reported. Seventy of 269 strains possessed a total of 89 plasmids. These were shown to be of varying sizes and in some cases of varying structures by endonuclease restriction digest. In six of 20 strains antibiotic resistance was cured with loss of the plasmid. The diversity of plasmids is emphasized.     

Development cycles of coryneform and Nocardia-like bacteria

The growth cycles and the types of cell separation were studied in a microchamber with the collection strains of Brevibacterium ammoniagenes ATTC 6871, B. helvolum ATCC 19239, B. linens CCM 47 and ATCC 9174, B. maris VKM B-464 and B. stationis ATCC 14403, as well as with the strains of the genus Rhodococcus isolated from soils, viz. R. maris sp. nov. IMB 283 and R. luteus sp. nov. IMB 385. According to the increasing complexity of cellular morphological transformation in the life cycle, the organisms may be arranged in a series: R. maris -- B. ammoniagenes -- B. stationis -- B. linens -- B. helvolum -- R. luteus. The first three organisms are characterized by the snapping type of separation of short rod-like daughter cells. The cells of B. linens separate by both the snapping and bending types. The coccoid cells of B. helvolum ATCC 19239 produce many buds which are transformed into rod-like cells in the course of growth. In the log phase of growth, both true and false branching of the cells is observed; the latter is the result of a peculiar growth of the ends in the separated cells of B. helvolum. The cells of R. luteus form a rudimentary, rapidly fragmenting mycelium whose rod-like elements divide then by binary fission; the daughter cells separate the bending and snapping types.     

Numerical taxonomy of heavy metal-tolerant bacteria isolated from an estuary.

A total of 230 strains of metal-tolerant bacteria from water and sediment samples collected in Chesapeake Bay were isolated on a medium containing cobalt, lead, mercury, or molybdenum. In addition, a set of 71 cultures were simultaneously isolated on glucose tryptone yeast extract agar medium without metals. Twenty-three reference strains were also included in the numerical taxonomy study of these bacteria, bringing the grand total of strains examined to 324. All strains were examined for 112 biochemical, cultural, morphological, and physiological characters. The taxonomic data obtained were analyzed by computer and the simple matching (SSM) and Jaccard (SJ) coefficients were calculated. Clustering achieved by unweighted average linkage is presented and, from sorted similarity matrices and dendrograms, 294 strains, i.e., 97% of the total, were recovered in 12 phenetic groups defined at the 75 to 80% similarity level. Among the strains there were nine phena presumptively identified as Bacillus, Erwinia, Mycobacterium, Pseudomonas, and coryneforms. From the results of the taxonomic study, it is concluded that metal tolerance in estuarine water and sediment bacteria occurs among a restricted range of taxa distributed throughout the estuarine environment.     

Plasmids in coryneform bacteria of human origin

A study of plasmids in coryneform bacteria isolated from human sources is reported. Seventy of 269 strains possessed a total of 89 plasmids. These were shown to be of varying sizes and in some cases of varying structures by endonuclease restriction digest. In six of 20 strains antibiotic resistance was cured with loss of the plasmid. The diversity of plasmids is emphasized.     

Numerical taxonomy of phenanthrene-degrading bacteria isolated from the Chesapeake Bay.

Phenanthrene-degrading bacteria were isolated from Chesapeake Bay samples by the use of a solid medium which had been overlaid with an ethanol solution of phenanthrene before inoculation. Eighteen representative strains of phenanthrene-degrading bacteria with 21 type and reference bacteria were examined for 123 characteristics representing physiological, biochemical, and nutritional properties. Relationships between strains were computed with several similarity coefficients. The phenogram constructed by unweighted-pair-group arithmetic average linkage and use of the simple Jaccard (SJ) coefficient was used to identify seven phena. Phenanthrene-degrading bacteria were identified as Vibrio parahaemolyticus and Vibrio fluvialis by their clustering with type and reference strains. Several phenanthrene-degrading bacteria resembled Enterobacteriaceae family members, although some Vibrio-like phenanthrene degraders could not be identified.     

Numerical taxonomy of heavy metal-tolerant nonhalophilic bacteria isolated from hypersaline environments.

A total of 232 metal-tolerant bacterial strains were isolated from water and sediment samples collected in different hypersaline environments located in Cádiz, Huelva and Morón de la Frontera (Spain). They were isolated on a medium containing mercury, chromium, cadmium, copper or zinc. These halotolerant isolates were analyzed by numerical taxonomy techniques by using the simple matching (SSM) and Jaccard (SJ) coefficients; clustering was achieved using the unweighted pair group method with averages (UPGMA) algorithm. At the 81% and 83% similarity level, different numbers of phenons were obtained for Gram-negative and Gram-positive halotolerant microorganisms. Most of the 48 Gram-negative metal-tolerant strains studied were grouped into nine phenons, representing the genera Pseudoalteromonas, Alteromonas, Xanthomonas, Pseudomonas, Alcaligenes and Enterobacteria. The 72 Gram-positive metal-tolerant strains grouped into eight phenons, with only 15 strains left unassigned. Most of the isolates were assigned to the genus Bacillus (seven phenons), and one phenon comprised microorganisms with phenotypic characteristics similar to those of the genus Celullomonas.     

Identification of plasmid partition function in coryneform bacteria

We have identified and characterized a partition function that is required for stable maintenance of plasmids in the coryneform bacteria Brevibacterium flavum MJ233 and Corynebacterium glutamicum ATCC 31831. This function is localized to a HindIII-NspV fragment (673 bp) adjacent to the replication region of the plasmid, named pBY503, from Brevibacterium stationis IFO 12144. The function was independent of copy number control and was not associated directly with plasmid replication functions. This fragment was able to stabilize the unstable plasmids in cis but not in trans.     

Industrial production of amino acids by coryneform bacteria

In the 1950s Corynebacterium glutamicum was found to be a very efficient producer of L-glutamic acid. Since this time biotechnological processes with bacteria of the species Corynebacterium developed to be among the most important in terms of tonnage and economical value. L-Glutamic acid and L-lysine are bulk products nowadays. L-Valine, L-isoleucine, L-threonine, L-aspartic acid and L-alanine are among other amino acids produced by Corynebacteria. Applications range from feed to food and pharmaceutical products. The growing market for amino acids produced with Corynebacteria led to significant improvements in bioprocess and downstream technology as well as in molecular biology. During the last decade big efforts were made to increase the productivity and to decrease the production costs. This review gives an overview of the world market for amino acids produced by Corynebacteria. Significant improvements in bioprocess technology, i.e. repeated fed batch or continuous production are summarised. Bioprocess technology itself was improved furthermore by application of more sophisticated feeding and automatisation strategies. Even though several amino acids developed towards commodities in the last decade, side aspects of the production process like sterility or detection of contaminants still have increasing relevance. Finally one focus of this review is on recent developments in downstream technology.