Development of a genetic system for Marinobacter adhaerens HP15 involved in marine aggregate formation by interacting with diatom cells

Sonnenschein, Eva C.; Gärdes, Astrid; Seebah, Shalin; Torres-Monroy, Ingrid; Grossart, Hans‐Peter; Ullrich, Matthias S.

doi:10.1016/j.mimet.2011.08.008

Cited by 25 publications

(32 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To produce single gene deletions, we selected an approach utilizing double homologous recombination and conjugation strategies. A similar approach to what we have taken was recently reported for an alternative strain of Marinobacter (11). We found that pUC19-derived plasmids did not replicate in M. aquaeolei VT8 and could thus serve as suicide vectors for genome integration studies if they contained the proper mobilization element (14), and we found the kanamycin cassette from pBBR1MCS-2 (14) to be an ideal selection marker with M. aquaeolei VT8.…”

Section: Resultssupporting

confidence: 63%

“…The M. aquaeolei VT8 conjugation procedure was derived from methods for the conjugation of Psychrobacter arcticus 273-4 and Marinobacter adhaerens (10,11). Briefly, cultures of the donor cells, E. coli WM3064, containing the specific plasmid and recipient cells, M. aquaeolei VT8, were grown separately on LB plates, then mixed at a ratio of 1:3 donor to recipient cells, spotted onto an LB plate containing DAP, and then incubated at 30°C for approximately 24 h. Cells were collected from two spots, washed with LB broth, resuspended in 100 l LB, and spread onto LB plates devoid of DAP but containing kanamycin for selection.…”

Section: Strains and Reagentsmentioning

confidence: 99%

See 1 more Smart Citation

Fatty Alcohols for Wax Esters in Marinobacter aquaeolei VT8: Two Optional Routes in the Wax Biosynthesis Pathway

Lenneman

Ohlert

Palani

et al. 2013

Appl Environ Microbiol

View full text Add to dashboard Cite

The biosynthesis of wax esters in bacteria is accomplished by a unique pathway that combines a fatty alcohol and a fatty acyl coenzyme A substrate. Previous in vitro enzymatic studies indicated that two different enzymes could be involved in the synthesis of the required fatty alcohol in Marinobacter aquaeolei VT8. In this study, we demonstrate through a series of gene deletions and transcriptional analysis that either enzyme is capable of fulfilling the role of providing the fatty alcohol required for wax ester biosynthesis in vivo, but evolution has clearly selected one of these, a previously characterized fatty aldehyde reductase, as the preferred enzyme to perform this reaction under typical wax ester-accumulating conditions. These results complement previous in vitro studies and provide the first glimpse into the role of each enzyme in vivo in the native organism.T he global cycle of oil is of interest from the standpoints of both energy and the environment, as efforts by humankind to obtain this valuable resource can result in substantial releases of crude oil through incidents such as the Deepwater Horizon oil spill of 2010 in the Gulf of Mexico. We also note that crude oil from natural deposits is routinely released into aqueous environments, such as the oceans, by natural processes where geological reserves meet surface waters. These environments have allowed natural populations of organisms, such as marine bacteria, to evolve to utilize these supplies, rich in reduced carbon, for use as a biological fuel source. A primary focus related to oil-degrading marine bacteria is the oxidation of these oils to meet energy requirements of the living cell. Interestingly, for certain marine bacteria found to utilize and degrade oils, these bacteria are also capable of producing natural lipids that have economic values similar to those obtained from harvesting sperm whales prior to the late 20th century, even when the bacteria are grown on simple organic acids or carbohydrates. We selected the marine bacterium Marinobacter aquaeolei VT8, which was isolated from an oil well off the coast of Vietnam (1), as a model bacterial species to study metabolic processes in an oil-metabolizing and neutral lipid-accumulating bacteria. In addition to growing on long-chain hydrocarbons, M. aquaeolei VT8 also produces a natural hydrocarbon, the wax ester, when grown on simple citric acid cycle intermediates, such as succinate or citrate, as the sole carbon source (1-3), indicating that all of the precursors required for the biosynthesis of wax ester are indigenous to this strain.Biosynthesis of wax esters is accomplished by the combination of several different enzymes. The wax ester synthase/acyl-coenzyme A (CoA):diacylglycerol acyltransferase (WS/DGAT) enzyme catalyzes the reaction of a fatty acyl-CoA substrate with a fatty alcohol (Fig. 1). While the fatty acyl-CoA utilized by the WS/ DGAT is proposed to come directly from the fatty acyl-CoA pool, the fatty alcohol is believed to be produced through the action of several reducta...

show abstract

Section: Resultssupporting

confidence: 63%

Section: Strains and Reagentsmentioning

confidence: 99%

Fatty Alcohols for Wax Esters in Marinobacter aquaeolei VT8: Two Optional Routes in the Wax Biosynthesis Pathway

Lenneman

Ohlert

Palani

et al. 2013

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…To understand the actual mechanisms by which diatom cells and bacteria interact and influence marine aggregate formation, a bilateral model system was established (22,23,31,57). The bacterial strain M. adhaerens HP15 was demonstrated to attach to the diatom Thalassiosira weissflogii and to induce formation of transparent exopolymeric particles (TEPs) and aggregates in cocultures.…”

mentioning

confidence: 99%

Chemotaxis of Marinobacter adhaerens and Its Impact on Attachment to the Diatom Thalassiosira weissflogii

Sonnenschein

Syit

Grossart

et al. 2012

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

ABSTRACTAlga-bacterium interactions are crucial for aggregate formation and carbon cycling in aquatic systems. To understand the initiation of these interactions, we investigated bacterial chemotaxis within a bilateral model system.Marinobacter adhaerensHP15 has been demonstrated to attach to the diatomThalassiosira weissflogiiand induce transparent exopolymeric particle and aggregate formation.M. adhaerenspossesses one polar flagellum and is highly motile. Bacterial cells were attracted to diatom cells, as demonstrated by addition of diatom cell homogenate or diatom culture supernatant to soft agar, suggesting that chemotaxis might be important for the interaction ofM. adhaerenswith diatoms. Three distinct chemotaxis-associated gene clusters were identified in the genome sequence ofM. adhaerens, with the clusters showing significant sequence similarities to those ofPseudomonas aeruginosaPAO1. Mutations in the genescheA,cheB,chpA, andchpB, which encode histidine kinases and methylesterases and which are putatively involved in either flagellum-associated chemotaxis or pilus-mediated twitching motility, were generated and mutants with the mutations were phenotypically analyzed. ΔcheAand ΔcheBmutants were found to be swimming deficient, and all four mutants were impaired in biofilm formation on abiotic surfaces. Comparison of the HP15 wild type and its chemotaxis mutants in cocultures with the diatom revealed that the fraction of bacteria attaching to the diatom decreased significantly for mutants in comparison to that for the wild type. Our results highlight the importance ofM. adhaerenschemotaxis in initiation of its interaction with the diatom. In-depth knowledge of these basic processes in interspecies interactions is pivotal to obtain a systematic understanding of organic matter flux and nutrient cycling in marine ecosystems.

show abstract

“…HP15 and Thalassiosira weissflogii M. adhaerens HP15 (Grossart et al 2004, Kaeppel et al 2012) was cultured in 500 ml Erlenmeyer flasks in liquid marine broth (MB) medium (Sonnenschein et al 2011) at 18°C under constant shaking at 250 rpm. Axenic T. weissflogii diatom cultures (CCMP 1336; received from the Provasoli-Guillard N ational Center for Culture of Marine Phytoplankton) were grown in 2000 ml Erlenmeyer flasks in f/2 medium prepared from filtered and autoclaved N orth Sea water (N SW; 15°C, 12:12 h light:dark period at 150 µm photons m −2 s −1…”

Section: Culture Conditions For Marinobacter Adhaerensmentioning

confidence: 99%

“…A proteomics approach was applied to a model interaction system comprising the diatom T. weissflogii and the marine bacterium Marinobacter adhaerens HP15. This model system had been established in the past in order to investigate bacterial colonization of diatoms and formation of marine aggregates (Sonnenschein et al 2011). It has since successfully served in a set of studies dissecting this interaction , Sonnenschein et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Proteomics analysis of the response of the marine bacterium Marinobacter adhaerens HP15 to the diatom Thalassiosira weissflogii

Stahl¹,

Ullrich²

2016

Aquat. Microb. Ecol.

View full text Add to dashboard Cite

Development of a genetic system for Marinobacter adhaerens HP15 involved in marine aggregate formation by interacting with diatom cells

Cited by 25 publications

References 59 publications

Fatty Alcohols for Wax Esters in Marinobacter aquaeolei VT8: Two Optional Routes in the Wax Biosynthesis Pathway

Fatty Alcohols for Wax Esters in Marinobacter aquaeolei VT8: Two Optional Routes in the Wax Biosynthesis Pathway

Chemotaxis of Marinobacter adhaerens and Its Impact on Attachment to the Diatom Thalassiosira weissflogii

Proteomics analysis of the response of the marine bacterium Marinobacter adhaerens HP15 to the diatom Thalassiosira weissflogii

Contact Info

Product

Resources

About