As a whole, a mild HP increase to 5MPa was lethal for A. borkumensis SK2 while doubling such a HP re-established some cell replication and improved cell integrity and activity, indicating that some pressure-resistance mechanism was triggered. Conversely, respiration capacity was generally enhanced by increased HP (Fig. nov., a novel hydrocarbonoclastic marine bacterium isolated from Antarctic coastal sea water. Appl. Under 10MPa, SK2 upregulated the synthetic pathway of the osmolyte ectoine, whose concentration increased from 0.45 to 4.71 fmoles cell1. Wang, W., Wang, L. & Shao, Z. Diversity and abundance of oil-degrading bacteria and alkane hydroxylase (alkB) genes in the subtropical seawater of Xiamen Island. Abe, F., Kato, C. & Horikoshi, K. Pressure-regulated metabolism in microorganisms. Article While the bioremediation potential remained as low as that observed at 5MPa (Fig. Int. Starting material (1g) of total RNA was depleted of rRNAs using Ribo-Zero magnetic bead based capture-probe system (Illumina, Hayward, USA). 1998; Alcanivorax dieselolei Liu and Shao 2005; Alcanivorax gelatiniphagus Kwon et al. 5, 10021008 (2003). The cells were then washed three times with fresh ASM medium to remove the residual n-alkane-13C2, air-dried and subjected to carbon (13C) isotopic analysis. Microbiol. Microbiol. ID ACCA_ALCBS Reviewed; 323 AA. 1(5), 130 (2005). 108, 12071216 (2010). Article Manilla-Prez E, Lange AB, Hetzler S, Steinbchel A. Appl Microbiol Biotechnol. 1). Some species, such as Alcanivorax borkumensis, produce surfactants that solubilize the oil, while other bacteria degrade the oil into carbon dioxide. PNAS 109(50), 2022920234 (2012). Curr Opin Biotechnol. An aliquot of the bacterial suspensions (200l) was then added to the bottom of the chamber using a micropipette. Lai, Q., Li, W. & Shao, Z. Unlike most other cells, it cannot consume more common substances such as sugars or amino acids as a source of energy. PubMed Central Environ Microbiol. Sign up for the Nature Briefing newsletter what matters in science, free to your inbox daily. Microbiol 63, 4559 (2009). Application of a HP equivalent to 5MPa was lethal, as it essentially inactivated SK2: final cell number was lower than what initially inoculated, decreasing to almost undetectable values (5847*106 cells mL1) with more than 90% of such surviving cells being damaged (Fig. Water Res. 4, 173182 (2006). The highest values of solubilized dodecane were observed under 5MPa (Fig. Chemosph. For in vitro experiments, pET-almA was transfected into E. coli BL21 cells (Novagen), and almA was overexpressed and purified as described below. 1A,B, respectively), while little difference in pH was observed under 5 and 10MPa as compared to sterile controls (Fig. nov., isolated from deep sea water of Indian Ocean. Aminoacyl-tRNA binding to ribosomes determines a conformational change in the latter that leads to an increase in volume59. For example, the genome of Alcanivorax borkumensis includes genes for alkane metabolism, biosurfactant production and biofilm formation, uptake of a variety of inorganic nutrients, and efflux. Furthermore, the majority of the genes expressing subunits of the Na+-translocating NADH-quinone reductase were either upregulated or unaffected (5/6, Table 2), supporting the hypothesis that the microbial respiration chain under HP may follow different pathways with respect to that occurring at ambient pressure. The mini-Tn5 insertion sites in the selected mini-Tn5 mutants were identified via inverse PCR. Isolation and characterization of a novel biosurfactant produced by hydrocarbon-degrading bacterium Alcanivorax dieselolei B-5. Propane respiration jump-starts microbial response to a deep oil spill. Somero, G. N. Adaptations to high hydrostatic pressure. Aseptically transfer this aliquot back into the broth tube. 48(2), 339348 (1998). 14(9), 24052416 (2012). To confirm the results of the initial screening and further analyse these mutants, they were replicated in OmniTray plates containing ASM supplemented with 0.5% (wt/vol) sodium acetate or with C16 or C32 alkanes as the sole carbon source. nov., a novel marine bacterium that obligately utilizes hydrocarbons. Ni, B., Huang, Z., Fan, Z., Jiang, C. Y. How are microbes used to clean up the environment? Natl Acad. Early stationary phase cells were collected by centrifugation at 4000rpm for 10 min at 4C (Sorval RC5c PLUS, Beckman, Suarle, Belgium) and resuspended in fresh ONR7a medium at an initial optical density (OD610) of 0.1000.005, corresponding to 14030106 cells mL1. Alcanivorax borkumensis (A. borkumensis) is a Gram-negative, rod-shaped hydrocarbonoclastic ("oil-degrading") bacterium. S1). Environ Microbiol. Microbiol. Casselman, A. Compatible solutes in luminescent bacteria of the genera Vibrio, Photobacterium and Photorhabdus (Xenorhabdus): occurrence of ectoine, betaine and glutamate. The carbon (13C) isotopic values are presented as versus the stated standard. CAS Energetic hurdles due to extreme, stressing conditions are known to impact ATP intracellular balance at high pH53 and salinity54, with acid55 or HP stress (about 40MPa56). However, according to sequence abundance they considered Alcanivorax contribution to oil degradation rather negligible20. High quality libraries were quantified using the Qubit Fluorometer (Life Technologies, Carlsbad, CA, USA), concentration normalized and samples pooled according to number of reads. CAS & Nagata, S. Efficient production of ectoine using ectoine-excreting strain. Gas-phase was analyzed with a Compact GC (Global Analyser Solutions, Breda, The Netherlands), equipped with a Molsieve 5A pre-column and two channels. To test the growth of the mutants on various alkane substrates, the wild-type B5 and mutant strains were grown in 100ml of ASM containing the appropriate antibiotics in 250ml Erlenmeyer flasks at 28C on a rotary shaker (200r.p.m.). Mutants exhibiting no growth or abnormal growth on C32 alkane were re-arrayed from the original library into new 96-well microtitre plates containing 200l of modified LB medium supplemented with nalidixic acid and streptomycin. Environ. Valentine, D. L. et al. Google Scholar. Help Topics FAQ. Signal transduction in bacterial chemotaxis. Yakimov, M. M., Timmis, K. N. & Golyshin, P. N. Obligate oil-degrading marine bacteria. Mooney, A., O'Leary, N. D. & Dobson, A. D. Cloning and functional characterization of the styE gene, involved in styrene transport in Pseudomonas putida CA-3. FEMS Microbiol. Use several drops of the suspension to inoculate a #2698 agar slant and/or plate. Article Saline Sys. Int. Global and overview maps 01100 Metabolic pathways 01110 Biosynthesis of secondary metabolites 01120 Microbial metabolism in diverse environments 01200 Carbon metabolism Hydrocarbon-degrading bacteria and the bacterial community response in Gulf of Mexico beach sands impacted by the Deepwater Horizon oil spill. Carbon (13C) isotopic analyses were performed using a Carlo Erba 1108 Elemental Analyzer (EA) connected via a Conflo II interface to a Finnigan MAT Delta Plus XL isotope ratio mass spectrometer56. Nat. Sanders, J. W., Venema, G. & Kok, J. and transmitted securely. Wang, W. & Shao, Z. nov., a new, hydrocarbon-degrading and surfactant-producing marine bacterium. Struct. Doumenq, P. et al. The https:// ensures that you are connecting to the Unlike other cells, it does not consume common substances like sugars or amino acids as an energy source. S.B. Blum, J. et al. PMC In the latter, the large majority of the genes was either upregulated or remained unaffected under HP (18/21, Table 2). The exact protective mechanisms exerted by ectoine are not clear as well as the triggering effect leading to its enhanced production. The .gov means its official. To evaluate gene expression, the number of mRNA copies present in 1,000 A. dieselolei B-5 or mutant cells was determined using quantitative real-time PCR. An impaired metabolic response to hydrostatic pressure explains Alcanivorax borkumensis recorded distribution in the deep marine water column. The role of HP on A. borkumensis physiological and molecular response has never been investigated, leaving a knowledge gap about its role, deep-sea distribution and activity. Microbiol. nov., a novel marine bacterium that obligately utilizes hydrocarbons. 185, 32323237 (2003). DSM 11573 ) Add to Cart Open Pricelist. Microbiol. A. borkumensis is a rod-shaped bacterium without flagella that obtains its energy primarily from consuming alkanes (a type of hydrocarbon).It is aerobic, meaning it uses oxygen to gain energy, and it is halophilic, meaning it tends to live in environments that contain salt, such as salty ocean water.It is also Gram-negative, which essentially means it has a relatively thin cell wall. Notification list. 46(3), 907919 (2012). Thus, A. borkumensis lacks proper adaptation to HP but activates resistance mechanisms. Extrem 6(6), 507514 (2002). Microbiol. Transposon mutagenesis was used for functional genome analysis of Alcanivorax SK2 to reveal the genetic basis of other environmentally relevant phenotypes, such as biofilm formation, adaptation to UV exposure, and to growth at either low temperature or high salinity. Notwithstanding the very different cell number (Fig. Environ. CAS Such a prominent role in petroleum-affected environments is due to some critical features possessed by Alcanivorax, including the ability to efficiently use branched-chain alkanes6 and the capacity to enhance the bioavailability of hydrophobic compounds in water. Natl Acad. Alcanivorax borkumensis is a marine bacterium that uses exclusively petroleum oil hydrocarbons as sources of carbon and energy (and is therefore designated "hydrocarbonoclastic"). Variations in pH were mirrored by CO2 production, as the latter significantly decreased under mild HP to low, comparable values at 5 and 10MPa (P>0.05, Fig. Blackhart, B. D. & Zusman, D. R. Frizzy genes of Myxococcus xanthus are involved in control of frequency of reversal of gliding motility. Google Scholar. Bio. 2016 Sep 15;110(1):378-382. doi: 10.1016/j.marpolbul.2016.06.038. Kish, A. et al. Application of 10MPa HP resulted into a reduced expression of the majority of the genes (56%, 1242/2202), while only a minor fraction was upregulated (16%, 354/2202) and the rest remained unaffected (28%, 606/2202). Lai, Q. et al. Disclaimer, National Library of Medicine Asterisk indicates that values obtained under 10MPa were significantly higher (P<0.05) than those observed under 0.1MPa. Metagenome, metatranscriptome and single-cell sequencing reveal microbial response to Deepwater Horizon oil spill. For the MCP protein ligand-binding analysis, an injection series (2l for the first injection and 10l for subsequent injections) was conducted using a 250-l syringe filled with the test solutions (25 injections at 120-min intervals or 16 injections at 100-min intervals). 21(4), 12061212 (2005). Its negligible presence in oil-contaminated deep waters (as observed during the Deepwater Horizon accident) raises the hypothesis that it may lack adaptive mechanisms to hydrostatic pressure (HP). Metallomics 3, 775787 (2011). RNA was isolated from pelleted cells using the Rneasy Mini Kit (Qiagen, Antwerp, Belgium) following manufacturers instructions. One unit was defined as the amount of enzyme required to hydroxylate 1mol of alkane per min. Biotechnol prog. Bars indicate 95% confidence intervals. Epub 2014 Sep 24. While the use of booms with skimmers and dispersants may account for recovering or dissolving the largest fraction of the spilled oil1, the last, fine hydrocarbons removal step relies on bacterial degradation. AlmR, a gene regulatory protein of LC-alkane metabolism In a mutant library screen, the mutant strain MRB5 exhibited better growth than the wild-type strain when a C 32 alkane was used as the.
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