Marina Strain Research Articles

An EntD-like phosphopantetheinyl transferase gene from Photobacterium profundum SS9 complements pfa genes of Moritella marina strain MP-1 involved in biosynthesis of docosahexaenoic acid

The EntD-like phosphopantetheinyl transferase (PPTase) gene, cloned from the eicosapentaenoic acid-producing bacterium Photobacterium profundum strain SS9, has an ORF of 690 bp encoding a 230-amino acid protein. When this PPTase gene was expressed in Escherichia coli with pfaA, pfaB, pfaC and pfaD derived from Moritella marina MP-1, which were four of five essential genes for biosynthesis of docosahexaenoic acid (DHA), the DHA production of the recombinant was 2% (w/w) of total fatty acids. This is the first report showing that the EntD-like PPTase is involved in producing n-3 polyunsaturated fatty acids.

Pigment composition and adaptation in free-living and symbiotic strains of Acaryochloris marina

Acaryochloris marina strains have been isolated from several varied locations and habitats worldwide demonstrating a diverse and dynamic ecology. In this study, the whole cell photophysiologies of strain MBIC11017, originally isolated from a colonial ascidian, and the free-living epilithic strain CCMEE5410 are analyzed by absorbance and fluorescence spectroscopy, laser scanning confocal microscopy, sodium dodecyl sulfate polyacrylamide gel electrophoresis and subsequent protein analysis. We demonstrate pigment adaptation in MBIC11017 and CCMEE5410 under different light regimes. We show that the higher the incident growth light intensity for both strains, the greater the decrease in their chlorophyll d content. However, the strain MBIC11017 loses its phycobiliproteins relative to its chlorophyll d content when grown at light intensities of 40 microE m(-2) s(-1) without shaking and 100 microE m(-2) s(-1) with shaking. We also conclude that phycobiliproteins are absent in the free-living strain CCMEE5410.

Recombinant production of docosahexaenoic acid in a polyketide biosynthesis mode in Escherichia coli

The docosahexaenoic acid (DHA) biosynthesis gene cluster (pDHA3) from the DHA-producing Moritella marina strain MP-1 includes the genes pfaA, pfaB, pfaC, and pfaD, which are similar to the genes of polyketide biosynthesis. When this cluster was co-expressed in Escherichia coli with M. marina MP-1 pfaE, which encodes phosphopantetheinyl transferase, DHA was biosynthesized. The maximum production of DHA (5% of total fatty acids) was observed at 15 degrees C. This is the first report of the recombinant production of DHA in a polyketide biosynthesis mode.

A phosphopantetheinyl transferase gene essential for biosynthesis of n − 3 polyunsaturated fatty acids from Moritella marina strain MP-1

A phosphopantetheinyl transferase (PPTase) gene ( pfaE), cloned from the docosahexaenoic acid (DHA)-producing bacterium Moritella marina strain MP-1, has an open reading frame of 861 bp encoding a 287-amino acid protein. When the pfaE gene was expressed with pfaA– D, which are four out of five essential genes for biosynthesis of eicosapentaenoic acid (EPA) derived from Shewanella pneumatophori SCRC-2738 in Escherichia coli, the recombinant produced 12% EPA of total fatty acids. This suggests that pfaE encodes a PPTase required for producing n − 3 polyunsaturated fatty acids, which is probably involved in the synthesis of DHA in M. marina strain MP-1.

There are High Levels of Functional and Genetic Diversity in Oxyrrhis marina

Oxyrrhis marina, a widely distributed marine protist, is used to model heterotrophic flagellate responses in microbial food webs. Although clonal variability occurs in protists, assessments of intraspecific diversity are rare; such assessments are critical, particularly where species are used as models in ecological studies. To address the extent of intraspecific variation within O. marina, we assessed diversity among 11 strains using 5.8S rDNA and ITS sequences. The 5.8S rDNA and ITS regions revealed high divergence between strains: 63.1% between the most diverse. To compare O. marina diversity relative to other alveolates, 18S rDNA sequences for five strains were analysed with sequences from representatives of the major alveolate groups. 18S rDNA also revealed high divergence in O. marina. Additionally, consistent with phylogenies based on protein coding genes, maximum likelihood analysis indicated that O. marina was monophyletic and ancestral to the dinoflagellates. To assess ecophysiological differences, growth rates of seven O. marina strains were measured at 10 salinities (10-55 per thousand). Two salinity responses occurred: one group achieved highest growth rates at high salinities; the other grew best at low salinities. There was no clear correlation between molecular, ecophysiological, or geographical differences. However, salinity tolerance was associated with habitat type: intertidal strains grew best at high salinities; open-water strains grew best at low salinities. These data indicate the need to examine many strains of a species in both phylogenetic and ecological studies, especially where key-species are used to model ecological processes.

Enhancement of polyunsaturated fatty acid production by cerulenin treatment in polyunsaturated fatty acid-producing bacteria

When docosahexaenoic acid (DHA)-producing Moritella marina strain MP-1 was cultured in the medium containing 0.5 microg cerulenin ml-1, an inhibitor for fatty acid biosynthesis, the cells grew normally, but the content of DHA in the total fatty acids increased from 5.9-19.4%. The DHA yield of M. marina strain MP-1 cells also increased from 4 to 13.7 mg l-1 by cerulenin treatment. The same effect of cerulenin was observed in eicosapentaenoic acid (EPA)-producing Shewanella marinintestina strain IK-1 grown in the medium containing 7.5 microg cerulenin ml-1, and the cerulenin treatment increased the EPA yield from 1.6 to 8 mg l-1. The use of cerulenin is, therefore, advantageous to increase the content of intracellular polyunsaturated fatty acids (PUFA) in particular PUFA-containing phospholipids in bacterial cells.

Presence of Ecophysiologically Diverse Populations within Cobetia marina Strains Isolated from Marine Invertebrate, Algae and the Environments

Over the last decade, taxonomic surveys have recovered sixteen strains of Halomonas-like marine heterotrophic bacteria from different ecological habitats. The sixteen strains were isolated from three N.W. Pacific Ocean habitats: seawater, the mussel Crenomytilus grayanus and the degraded thallus of brown alga Fucus evanescens. These strains were subjected to a taxonomic investigation of their phenotypic/physiological, genetic, and phylogenetic features. Analysis indicates these bacteria belong to Cobetia marina. The study found all strains tolerated CdCl2 concentrations up to 875 mM. Taxonomically, the sixteen strains belong to the same species, nevertheless, their physiological features revealed distinguishing characteristics. For instance, strain KMM 296, recovered from the mussel Crenomytilus grayanus, was distinct from other C. marina strains by its ability to produce highly active alkaline phosphatase. The majority of C. marina strains that were isolated from degraded alga thallus appeared to have a particular metabolic specialisation by utilizing a range of easily assimilable monosaccharides. Notably, despite a high level of genetic similarity (80% of DNA relatedness), the phenotypic features of the strains isolated from degraded alga thallus differed with the type strain C. marina LMG 2217T. These differences suggest an ecologically adapted population of C. marina at a subspecies level.

Temperature and nutrient availability control growth rate and fatty acid composition of facultatively psychrophilic Cobetia marina strain L-2.

A facultative psychrophilic bacterium, strain L-2, that grows at 0 and 5 degrees C as minimum growth temperatures in complex and defined media, respectively, was isolated. On the basis of taxonomic studies, strain L-2 was identified as Cobetia marina. The adaptability of strain L-2 to cold temperature was higher than that of the type strain and of other reported strains of the same species. When the bacterium was grown at 5-15 degrees C in a defined medium, it produced a high amount of trans-unsaturated fatty acids. By contrast, in a complex medium in the same temperature range it produced a low amount of trans-unsaturated fatty acids. In the complex medium at 5 degrees C, the bacterium exhibited a three-fold higher growth rate than that obtained in the defined medium. Following a temperature shift from 11 to 5 degrees C, strain L-2 grew better in complex than in defined medium. Furthermore, when the growth temperature was shifted from 0 to 5 degrees C both the growth rate and the yield of strain L-2 growing in complex medium was markedly enhanced. These phenomena suggest that an upshift of the growth temperature had a positive effect on metabolism. The effects of adding complex medium components to the defined medium on bacterial growth rate and fatty acid composition at 5 degrees C were also studied. The addition of yeast extract followed by peptone was effective in promoting rapid growth, while glutamate addition was less effective, resulting in a cis-unsaturated fatty acid ratio similar to that of cells grown in the complex medium. These results suggest that the rapid growth of strain L-2 at low temperatures requires a high content of various amino acids rather than the presence of a high ratio of cis-unsaturated fatty acids in the cell membrane.

Characterization of the gene encoding the beta-lactamase of the psychrophilic marine bacterium Moritella marina strain MP-1.

The beta-lactamase gene (mbla) of the psychrophilic marine bacterium Moritella marina strain MP-1 was identified in a previously isolated genomic DNA fragment and it was expressed in Escherichia coli cells. The mbla gene encoded a protein consisting of 287 amino acid residues. Its predicted amino acid sequence showed approximately 50% identity with that of a number of class A beta-lactamases, especially with that of CARB/PSE type of beta-lactamases (carbenicillinases). E. coli transformed with the plasmid containing mbla grew on an ampicillin-containing plate at 37 degrees C but not at 42 degrees C, suggesting that the beta-lactamase of this bacterium is heat-labile.

Biosynthesis of fatty acids in the docosahexaenoic acid-producing bacterium Moritella marina strain MP-1

We have isolated the fatty acid biosynthetic (fab) gene cluster taking part in the synthesis of middle-chain fatty acids and a genomic segment which was homologous with the eicosapentaenoic acid-biosynthetic gene cluster from the docosahexaenoic acid (DHA)-producing bacterium Moritella marina strain MP-1. This segment was presumed to include the DHA-biosynthetic gene cluster of M. marina strain MP-1. When M. marina strain MP-1 was cultured in medium containing cerulenin, a fatty acid synthesis inhibitor, decreases in levels of middle-chain fatty acids and remarkable increases in levels of DHA were observed. These results suggest that the synthesis of middle-chain fatty acids works independently of the synthesis of DHA.

Biosynthesis of fatty acids in the docosahexaenoic acid-producing bacterium Moritella marina strain MP-I

We have isolated the fatty acid biosynthetic (fab) gene cluster taking part in the synthesis of middle-chain fatty acids and a genomic segment which was homologous with the eicosapentaenoic acid-biosynthetic gene cluster from the docosahexaenoic acid (DHA)-producing bacterium Moritella marina strain MP-1. This segment was presumed to include the DHA-biosynthetic gene cluster of M. marina strain MP-1. When M. marina strain MP-1 was cultured in medium containing cerulenin, a fatty acid synthesis inhibitor, decreases in levels of middle-chain fatty acids and remarkable increases in levels of DHA were observed. These results suggest that the synthesis of middle-chain fatty acids works independently of the synthesis of DHA.

Variable expression of gliding and swimming motility in Deleya marina

Surface-associated motility has been observed in the Deleya marina type strain ATCC 25374 (strain 219). Slime tracks and a complex growth pattern, characteristic of gliding motility, developed on semisolid marine-agar motility plates. Cell movement observed by light microscopy consisted of rapid glides and flips by single cells and groups of cells. Following the development of the gliding cell growth pattern, a subpopulation of swimming cells appeared. The variation in motility was random and reversible in subculture. Electron microscopic comparisons of cells of the two motility types showed that gliding cells had no obvious motility organelles, whereas swimming cells had polar flagella. Variable expression of gliding and swimming motility was also observed in D. marina strain 140 (ATCC 27129) and in two other species of the Deleya genus. Key words: gliding, morphological variation, Deleya, biofouling.