pH was measured using a pH meter (Herisau, Metrohm, Switzerland). Ohwada, T. & Sagisaka, S. An immediate and steep increase in ATP concentration in response to reduced turgor pressure in Escherichia coli B. Arch Biochem Biophys. However, application of HPs twice as high as this lethal one triggered some HP-resistance mechanism. Nature Communications (Nat Commun) ADS 99 cytoplasmic and membrane-associated proteins of alkane-grown cells were Trends Microbiol. strain ADP1 is induced by a broad range of n-alkanes and requires the transcriptional activator AlkR. Center of Microbial Ecology and Technology (CMET), University of Gent, Coupure Links 653, Gent, B 9000, Belgium, Department of Food, Environmental and Nutritional Sciences (DeFENS), University of Milano, Via Celoria 2, Milano, 20133, Italy, Marta Barbato,Sara Borin&Daniele Daffonchio, Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology (KAUST), 4700, Thuwal, 23955-6900, Kingdom of Saudi Arabia, You can also search for this author in To ascertain this hypothesis the transcriptomic response of cells grown under 0.1 and 10MPa was compared. 2016 Sep 15;110(1):378-382. doi: 10.1016/j.marpolbul.2016.06.038. DNA integrity and synthesis was not compromised by mild HP (Table S3), while several transcriptional regulators, elongators and termination factors were upregulated (Table S2), likely serving cell replication purposes. Appl Env Microbiol. 23(4), 483501 (1999). PubMed Following the classical capillary method55, these assays were performed by placing a few drops of a suspension in a small chamber consisting of a U-shaped glass tube between a microscope and a coverslip. To select mutants defective in lipid metabolism, ONR7a selection plates containing Cm and Ndx as selective antibiotics and also containing either Nile red (NR; 0.5 g/ml) or the . Environ. Sci.USA 108, 93909395 (2011). Commun. Generation of this cofactor takes advantage of metabolic intermediates derived from fatty acids biosynthesis52, which was enhanced under HP (Table 1). Extrem 2(3), 217222 (1998). Yakimov, M. M., Timmis, K. N. & Golyshin, P. N. Obligate oil-degrading marine bacteria. 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. Nuc Acid Res. Our experiments on strain SK2 indicate that it is a piezosensitive microbe and that mild HP remarkably affects A. borkumensis growth and physiology, explaining its low abundance in the sea water column. Microbiol. Nuttapol Noirungsee, Steffen Hackbusch, Rudolf Mller, Lakshmi Tripathi, Victor U. Irorere, Ibrahim M. Banat, Scientific Reports Ziervogel, K. et al. For solid alkanes (C24, C28, C32 and C36), the ligand solutions were dissolved in hexane. Nat. 80, 523533 (2012). A., Soetaert, W. K. & Vandamme, E. J. Optimization of ectoine synthesis through fed-batch fermentation of Brevibacterium epidermis . Zhang, L. H., Lang, Y. J. Appl. & Heipieper, H. J. A. De Lorenzo, V., Herrero, M., Jakubzik, U. 94, 437448 (2012). Alcanivorax borkumensis is an ubiquitous model organism for hydrocarbonoclastic bacteria, which dominates polluted surface waters. Alcanivorax borkumensis, a marine bacterium highly specialized in degrading linear and branched alkanes, plays a key ecological role in the removal of marine oil spills.It contains several alternative enzyme systems for terminal hydroxylation of alkanes, including three P450 cytochromes (P450-1, P450-2 and P450-3). 2013 Jul;79(14):4282-93. doi: 10.1128/AEM.00694-13. Microbiol. Use the Previous and Next buttons to navigate three slides at a time, or the slide dot buttons at the end to jump three slides at a time. S1), suggesting that access to dodecane was not a limiting factor. . Kostka, J. E. et al. Provided by the Springer Nature SharedIt content-sharing initiative. CAS nov., a novel marine bacterium that obligately utilizes hydrocarbons. 175(22), 71707177 (1993). TopHat is a fast splice junction mapper for RNA-Seq reads, which aligns sequencing reads to the reference genome using the sequence aligner Bowtie2. The FadL family: unusual transporters for unusual substrates. The specific activity is expressed as Umg1 of enzyme. & Funhoff, E. G. Alkane hydroxylases involved in microbial alkane degradation. Predicting the complex structure and functional motions of the outer membrane transporter and signal transducer FecA. FOIA PubMed J 94, 24822491 (2008). Complete genomes available for important petroleum-related microorganisms Taxis Deep-sea bacteria enriched by oil and dispersant from the Deepwater Horizon spill. Relationship between membrane damage and cell death in pressure-treated Escherichia coli cells: differences between exponential- and stationary-phase cells and variation among strains. Following transfer of the mutants, a C32 alkane powder was added to the OmniTray plates as previously described11. However, previous studies failed to observe accumulation of ectoine under increased HP in both piezophilic36 and piezosensitive bacteria37. Optical density was measured at 610nm (OD610) with a spectrophotometer (Isis 9000, Dr Lange, Germany). van den Berg, B. 61(6), 13701374 (2011). & Rojo, F. Expression of the Pseudomonas putida OCT plasmid alkane degradation pathway is modulated by two different global control signals: evidence from continuous cultures. 1C), CO2:O2 molar ratio slightly increased (0.033) and uptake of some critical nutrients restored (Fig. The authors wish to acknowledge the contribution of the Reviewers, whose comments remarkably improved the quality of the manuscript. Alcanivorax borkumensis strain SK2 is a cosmopolitan hydrocarbonoclastic marine bacterium, with a specialized metabolism adapted to the degradation of petroleum oil hydrocarbons. On the contrary, to our knowledge the HP occurring at the depth of the DWH spill has been neglected as a possible factor to explain the low abundance of Alcanivorax detected after the oil spill. Free and Open Access to Biodiversity Data. In the meantime, to ensure continued support, we are displaying the site without styles The exact protective mechanisms exerted by ectoine are not clear as well as the triggering effect leading to its enhanced production. oil), polychlorinated biphenyls (PCBs), polyaromatic hydrocarbons (PAHs), heterocyclic compounds (such as pyridine or quinoline . Hydrocarbon-degrading bacteria and the bacterial community response in Gulf of Mexico beach sands impacted by the Deepwater Horizon oil spill. 21(4), 12061212 (2005). A transposon library consisting of approximately 20,000 colonies was obtained based on the mini-Tn5 transposon element. Microbiol 63, 4559 (2009). 11, 24772490 (2009). Notably, genes encoding for ribosomal proteins were largely upregulated (45/53, Table S5), suggesting that mild HP deeply affected this multicomponent protein complex. Yakimov, M. M. et al. The marine oil-degrading bacterium Alcanivorax borkumensis SK2 has attracted significant interest due to its hydrocarbonoclastic lifestyle, its alkane-centered metabolism, and for playing an important ecological role in cleaning up marine oil spills. The Alcanivorax genus was initially described by Yakimov and co-workers in 1998, who isolated A. borkumensis SK2 and proposed it as the type strain. Other pathways involved in the production of energy, such as glycolysis-gluconeogenesis and the pentose phosphate pathway, were generally downregulated (data not shown). Isolation and characterization of a novel biosurfactant produced by hydrocarbon-degrading bacterium Alcanivorax dieselolei B-5. conceived the project and co-wrote the paper. Correspondence to & Sayavedra-Soto, L. A. 77(22), 79627974 (2011). Pressure-induced cell membrane damaged analysis and total cell count was adapted after67 using flow cytometry: SYBR Green I and Propidium Iodide were used in combination to discriminate cells with intact and damaged cytoplasmic membranes using a protocol previously described68. However, according to sequence abundance they considered Alcanivorax contribution to oil degradation rather negligible20. Isopropyl -D-1-thiogalactopyranoside (0.4mM) was then added, and the culture was maintained at 30C for 3h. Cellular lysates of recombinant E. coli were prepared by sonification of cell suspensions in 50mM phosphate buffer (pH 8.0). Microbial activities and dissolved organic matter dynamics in oil-contaminated surface seawater from the Deepwater Horizon oil spill site. & Mackay, D. Environmentally relevant physical-chemical properties of hydrocarbons: a review of data and development of simple correlations. Synergistic PAH biodegradation by a mixed bacterial consortium: based on a multi-substrate enrichment approach Environ Microbiol. Mol. Natl Acad. Genes involved in alkanes activation45 were upregulated under HP (Table 1). In all experiments, a blank control (without chemoeffector) was included. Epub 2014 Sep 24. Do you need support? Haines, J. et al. Alcanivorax borkumensis participates in the biochemical process known as hydrocarbon degradation. Several organic and inorganic solutes accumulate intracellularly under osmotic and thermal stress to maintain turgor pressure, cell hydration or stabilize macromolecular structures, while little is known about solutes implicated in counteracting the destabilizing effects exerted by HP32. Sign up for the Nature Briefing newsletter what matters in science, free to your inbox daily. Remaining RNA (including mRNAs, lin-cRNAs and other RNA species) was subsequently purified (Agencourt RNA- Clean XP, Beckman Coulter, Brea, CA, USA) and fragmented using enzymatic fragmentation. Research Microbiol. 13, 11681178 (2011). Biophys. The chemotactic response was measured by placing the open end of a 1-l capillary tube containing the chemoeffector solution in the pool of bacterial cells in the chamber. USA 109, E1481E1488 (2012). CAS Alkanes were added in the form of either 350l of liquid alkanes (C8, C12, C16 and pristane) or 0.1% (w/v) solid alkane (C18, C24, C28, C32 and C36) per 100ml of ASM medium. Adaptation of the hydrocarbonoclastic bacterium Alcanivorax borkumensis SK2 to alkanes and toxic organic compounds: a physiological and transcriptomic approach. 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. 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. Biotechnol. Alcanivorax borkumensis is a ubiquitous marine petroleum oil-degrading bacterium with an unusual physi-ology specialized for alkane metabolism. Appl. 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. Microb Biotechnol. Metabolism; A. borkumensis and biosurfactants; Biotechnological applications; References; Alcanivorax borkumensis. 1). For the OmpS protein ligand-binding analysis, an injection series (2l for the first injection and 8l for subsequent injections) was conducted using a 250-l syringe filled with the test solutions (20 injections at 100-min intervals). Int J Sys Bacteriol. 15(3), 205214 (2004). In the present study this would result into the generation of dodecanoyl-CoA (terminal oxidation) and/or decanoyl-CoA and acetyl-CoA (subterminal oxidation). Handbook of Hydrocarbon and Lipid Microbiology (ed Timmis, K. N.).15291543 (Springer-Verlag, 2010). Notification list. It has several efficient enzymes that. 1 overestimates the effective mineralization rate and it rather represents the upper limit for dodecane degradation by the cells. A.S. conceived the paper, designed and performed the experiments, analysed the data and wrote the paper. Bioessays 28, 922 (2006). Microbiol. Sabirova JS, Becker A, Lnsdorf H, Nicaud JM, Timmis KN, Golyshin PN. Environ Microbiol. Panke, S., Meyer, A., Huber, C. M., Witholt, B. Article Our findings indicate that A. borkumensis SK2 might not be enriched under the mild HP of the DWH deep waters, suggesting that the lack of such a selection factor would restore the capacity of Alcanivorax to grow. Article Proc. Environ. 5:5755 doi: 10.1038/ncomms6755 (2014). Alcanivorax borkumensis skyrocketed, helping to get rid of the oil. Hydrostatic pressure-induced changes in cellular protein synthesis. Based on the obtained densitograms, the speed (V) of the bacterial front was calculated using the following formula: V=(XnX1)/(tnt1)[mmh1], where Xn and X1 are the positions of the bacterial front at times tn and t1, respectively. Article Oger, P. M. & Jebbar, M. The many ways of coping with pressure. The carbon (13C) isotopic values are presented as versus the stated standard. Get the most important science stories of the day, free in your inbox. 7(9), 14261441 (2005). 40, 12611272 (2001). 45(1), 116123 (1995). PO43 uptake was completely inhibited when increasing HP to 5MPa, although some activity was restored with further HP increase to 10MPa (Fig. Syutsubo, K., Kishira, H. & Harayama, S. Development of specific oligonucleotide probes for the identification and in situ detection of hydrocarbon-degrading Alcanivorax strains. strain H1 utilized 4-fluorocinnamic acid for growth under aerobic conditions. Saline Sys. cell is metabolism ().Cellular processes such as the building or breaking down of complex molecules occur through series of stepwise, interconnected . The details of these procedures are described in the Supplementary Methods. 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. Liu, J. et al. The resultant spectra were recorded in electron impact mode at 70eV. 7(11), 20912104 (2013). Int. An aliquot of the bacterial suspensions (200l) was then added to the bottom of the chamber using a micropipette. Muccio, Z. RNA stranded libraries were pre-amplified and purified (AgencourtAMPure XP). CAS Starting material (1g) of total RNA was depleted of rRNAs using Ribo-Zero magnetic bead based capture-probe system (Illumina, Hayward, USA). Oil Chem. Natl Acad. Adaptation of the hydrocarbonoclastic bacterium Alcanivorax borkumensis SK2 to alkanes and toxic organic compounds: a physiological and transcriptomic approach. The following PCR programme was utilized for these assays: 95C for 3min, followed by 30 cycles of 95C for 30s, 5052C for 45s and 72C for 2min, with a final extension at 72C for 5min. Harayama, S., Kasai, Y. 2.3 Culture conditions A. pH homeostasis and ATP synthesis: studies of two processes that necessitate inward proton translocation in extremely alkaliphilic Bacillus species . nov., a novel alkane-degrading bacterium isolated from sea water and deep-sea sediment. Hydrocarbon-degrading bacteria feed on the hydrocarbons in the oil droplet, breaking them into inorganic compounds. Camilli, R. et al. 312139, Integrated Biotechnological Solutions for Combating Marine Oil Spills) and by the Geconcentreerde Onderzoeksactie (GOA) of Ghent University (BOF15/GOA/006). Matsumura, P. & Marquis, R. E. Energetics of streptococcal growth inhibition by hydrostatic pressure. On the contrary, it was considered that downregulated genes had a 0.50 fold change. 2C). Analyst 134, 213222 (2009). 81(17), 58555866 (2015). Asterisks indicate statistical significance (P<0.05), that is: in (A), mean average value at 0.1MPa is lower than at 5 and 10MPa; (B), mean average value at 0.1MPa is higher than at 10MPa. The authors declare no competing financial interests. 33(4), 885892 (1977). But it can use other organic compounds also. Biotechnol. Dyksterhouse, S. E., Gray, J. P., Herwig, R. P., Lara, J. C. & Staley, J. T. Cycloclasticus pugetii gen. nov., sp. During cultivation on n-alkanes as sole source of carbon and energy, all strains produced both extracellular and cell-bound surface-active glucose lipids which reduced the surface tension of water from 72 to 29 mN m-1 (16). An official website of the United States government. On the contrary, some of the typical HP-responsive pool of genes (such as sigma factors, chaperonins and outer membrane proteins) were marginally affected by HP increase (Table S6). 1A) as also observed for total (Fig. The long-chain alkane metabolism network of Alcanivorax dieselolei. Lin, S., Hanson, R. E. & Cronan, J. E. Biotin synthesis begins by hijacking the fatty acid synthetic pathway. The A. dieselolei B5 genome contains 4,448 genes29. FEMS Microbiol Lett. On the contrary, protein translation was highly affected, as several translation (4/6) and elongation factors (3/3) were upregulated (Table S4). The most closely studied of these rod-shaped bacteria, Alcanivorax borkumensis, was first identified in 1998 near the Isle of Borkum in the North Sea. Evol. Microbiology 147, 16211630 (2001). Wang, W. & Shao, Z. Diversity of flavin-binding monooxygenase genes (almA) in marine bacteria capable of degradation long-chain alkanes. ONE SENTENCE SUMMARY A. borkumensis adapt their interfacial properties over time to evolve their biofilm phenotype and increase their oil consumption Provided is a composition that may include an aqueous phase, and extremophilic bacteria having signal transduction chemotaxis machinery and nanoemulsion oil droplets including metal-silica nanoparticles attached to a surface thereof. Alkane solutions were prepared by saturating the chemotaxis buffer (410.75mM NaCl, 9.39mM KCl, 21.5mM Na2HPO4, 22.41mM KH2PO4 and 0.123mM EDTA, pH 7.0 ) with excess crystals for several days and then passing them through polycarbonate filters (0.45m) to remove the remaining solid alkanes (maintained at 10C). A. borkumensis primarily uses alkanes as its source of energy/carbon, however, there are a few other organic compounds that it can utilize. S2). Article Reactors were incubated in a temperature-controlled room at 20C for 4 days reaching the stationary phase. Densitometric chemotaxis assays were performed as previously described55 with minor modifications. Sen, T. Z. et al. Head, I. M., Jones, D. M. & Rling, W. F. Marine microorganisms make a meal of oil. Tracking hydrocarbon plume transport and biodegradation at Deepwater Horizon. USA 104, 513518 (2007). To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/, Scoma, A., Barbato, M., Borin, S. et al. 65(4), 15641569 (1999). Scientists are working on ways to genetically engineer this bacterium to be more . Gas-phase was analyzed with a Compact GC (Global Analyser Solutions, Breda, The Netherlands), equipped with a Molsieve 5A pre-column and two channels. Yakimov, M. M. et al. Google Scholar. Environ. Proc. Curr Op Biotechnol. Centrifuge was pre-refrigerated at 4C and cells centrifuged at 13000rpm for 5min (Sorval RC5c PLUS, Beckman, Suarle, Belgium). Kostka, J. E. et al. Alcanivorax borkumensis is a marine bacterium that uses exclusively petroleum oil hydrocarbons as sources of carbon and energy (and is therefore designated "hydrocarbonoclastic"). PubMedGoogle Scholar. CAS 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. These substances act as a fertilizer for the bacteria and help them grow at an increased rate. Internet Explorer). DT 05-SEP-2006, sequence version 1. J. Bacteriol. First and second strand synthesis were performed and double stranded cDNA was purified (AgencourtAMPure XP). Res Microbiol. Unlike most other cells, it cannot consume more common substances such as sugars or amino acids as a source of energy. nov., an aromatic hydrocarbon-degrading bacterium from marine sediments. J Mol Microbiol Biotechnol. Natl Acad. Studies on the fate of the deep oil plume occurring after the DWH spill indicated that bacteria other than Alcanivorax were mainly enriched during hydrocarbon biodegradation16,17,18,19,20,21. Matturro B, Frascadore E, Cappello S, Genovese M, Rossetti S. Mar Pollut Bull. The control mixture contained no enzyme. FEMS Microbiol. Biol. J Bacteriol. ID ACCA_ALCBS Reviewed; 323 AA. 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. Separation was performed at 120C for 1min, and the temperature was then increased from 120C to 280C at a rate of 15Cmin1. 55, 11811186 (2005). 1). Nat. Uptake of PO43 was completely inhibited (Fig. In fact, genes related with fatty acids degradation were generally downregulated (6/9, Table 1) while those expressing enzymes linked with fatty acids biosynthesis were more highly expressed or remained unaffected (6/7, Table 1). to approximately A600=0.5. 14(9), 24052416 (2012). Microbiol 54, 141148 (2004). 2C) when comparing atmospheric and mild HP may be considered as an indication of the shift in cell metabolism under increased HP. Considering the ubiquity of Alcanivorax in polluted surface marine waters, including those in the Gulf of Mexico after the DWH oil spill23, the low frequency of Alcanivorax in the DWH oil plume in the deep water column raises the hypothesis that this organism may not effectively respond to HP. Involvement of BmoR and BmoG in n-alkane metabolism in Pseudomonas butanovora. The wild-type B5 strain and the OTB5-1, OTB5-2 and OTB5-3 mutant strains were grown in 100ml of ASM liquid medium containing the appropriate antibiotics. Extrem 6(6), 507514 (2002). Our hypothesis is that mild HP (up to 10MPa, equivalent to1 km in the marine water column, approximately the depth of the DWH oil spill) may be sufficiently stressing to exert an impact on A. borkumensis metabolism and potentially impair its remarkable oil-bioremediation capacity. and 1mm thickness; Agilent Technologies). Parales, E. & Ditty, L. Chemotaxis. Genes were grouped according to orthologous clusters using the database provided by Ortholuge DB70. Chemosph. Throne-Holst, M., Wentzel, A., Ellingsen, T. E., Kotlar, H.-K. & Zotchev, S. B. However, A. borkumensis SK2 is also known to be able to withdrawn oxidized n-alkanes from the degradation pathway and incorporate them as their corresponding fatty acids in the membrane47, a mechanism already observed in other hydrocarbon-degrading microbes (e.g., Marinobacter hydrocarbonoclasticus48 and Rhodococcus erythropolis49). 1(5), 130 (2005). sharing sensitive information, make sure youre on a federal 2015; Alcanivorax hongdengensis Wu et al. 44, 519528 (2001). As it stands, the increased HP impact on A. borkumensis cells (Fig. PubMed Microbiol. Thus, A. borkumensis lacks proper adaptation to HP but activates resistance mechanisms. and transmitted securely. 2B), resulting in a CO2:O2 molar ratio of 0.019. Statistical significance was assessed using a nonparametric test (Mann-Whitney test) which considered a two-sided distribution with 95% CI. Microbiol. Phenotypic analysis showed a restricted nutritional profile, high halotolerance, the absence of fermentative metabolism and a low G+C content. Blackhart, B. D. & Zusman, D. R. Frizzy genes of Myxococcus xanthus are involved in control of frequency of reversal of gliding motility. Further provided is a method for treating an oil reservoir, that may include introducing into the oil reservoir an extremophilic bacteria having signal . Eco 44, 373381 (2003). https://doi.org/10.1038/srep31316. All RNA polymerase genes (4/4, Table S4) were upregulated, while the tRNA regulator pseudouridine synthase was either upregulated or unaffected (3/3, Table S4). Natl Acad. and JavaScript. Pollard, E. C. & Weller, P. K. The effect of hydrostatic pressure on the synthetic processes in bacteria. Transposon mutants from the 96-well library plates were replicated in OmniTray plates (Nunc) containing solid artificial sea water medium (ASM) without a carbon source (24g NaCl, 7.0g MgSO47H2O, 1g NH4NO3, 0.7g KCl, 2.0g KH2PO4, 3.0g Na2HPO4 and 10ml of a trace element solution, pH 7.5). Would you like email updates of new search results? 1(1), 6370 (1999). Appl Env Microbiol. Int J Sys Ev Microbiol. By submitting a comment you agree to abide by our Terms and Community Guidelines. State Key Laboratory Breeding Base of Marine Genetic Resources, Third Institute of Oceanography, SOA, Xiamen, 361005, China, Key Laboratory of Marine Genetic Resources, Third Institute of Oceanography, SOA, Xiamen 361005, China, Key Laboratory of Marine Genetic Resources of Fujian Province, Xiamen 361005, China, Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources, Xiamen, 361005, China, You can also search for this author in Epub 2016 Jun 14. Cufflinks takes the alignment results from TopHat to assemble the aligned sequences into transcripts, constructing a map of the transcriptome, based on a previously reported transcriptome annotation69. Google Scholar. 6(9), 682688 (2010). Feng, L. et al. performed the experiments and analyzed the data. Sci. These suspensions were stored at 4C for no more than 24h before the chemotaxis assays were performed. Science 330(6001), 204208 (2010). Several environmental factors such as low temperature or lack of nutrients were proposed as conditional for the enrichment of Alkanivorax in the deep water oil plume22, but supporting evidence was not supplied20. 52, 901911 (2002). & Timmis, K. N. Mini-Tn5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in Gram-negative eubacteria. Curr Opin Biotechnol. Cell replication and metabolism in A. borkumensis cultures were examined under atmospheric (0.1MPa) and mild (5 or 10MPa) HP. This "hydrocarbonoclastic" bacterium degrades an exceptionally broad range of alkane hydrocarbons but few other substrates. 108, 12071216 (2010). Gutierrez, T. et al. While 5MPa essentially inactivated SK2, further increase to 10MPa triggered some resistance mechanism, as indicated by higher total and intact cell numbers. Hazen, T. C. et al. Camilli, R. et al. Provided that the averaged bacterial cell dry weight (CDW) is 1012g40,41, estimates of the highest ectoine accumulation capacity in A. borkumensis SK2 cells at 10MPa would yield 0.59gectoine g1CDW.
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