Functional metagenomics of oil-impacted mangrove sediments reveals high abundance of hydrolases of biotechnological interest

Júlia Ronzella Ottoni1, Lucélia Cabral1, Sanderson Tarciso Pereira de Sousa1, Gileno Vieira Lacerda Júnior1, Daniela Ferreira Domingos1, Fábio Lino Soares Junior2, Mylenne Calciolari Pinheiro da Silva2, Joelma Marcon2, Armando Cavalcante Franco Dias2, Itamar Soares de Melo3, Anete Pereira de Souza4, Fernando Dini Andreote2, Valéria Maia de Oliveira1
1Division of Microbial Resources, University of Campinas, Paulínia, Brazil
2Department of Soil Sciences, “Luiz de Queiroz” College of Agriculture, University of São Paulo, Piracicaba, Brazil
3Laboratory of Environmental Microbiology, EMBRAPA Environment, Jaguariúna, Brazil
4Center of Molecular Biology and Genetic Engineering, University of Campinas, Campinas, Brazil

Tóm tắt

Mangroves are located in coastal wetlands and are susceptible to the consequences of oil spills, what may threaten the diversity of microorganisms responsible for the nutrient cycling and the consequent ecosystem functioning. Previous reports show that high concentration of oil favors the incidence of epoxide hydrolases and haloalkane dehalogenases in mangroves. This finding has guided the goals of this study in an attempt to broaden the analysis to other hydrolases and thereby verify whether oil contamination interferes with the prevalence of particular hydrolases and their assigned microorganisms. For this, an in-depth survey of the taxonomic and functional microbial diversity recovered in a fosmid library (Library_Oil Mgv) constructed from oil-impacted Brazilian mangrove sediment was carried out. Fosmid DNA of the whole library was extracted and submitted to Illumina HiSeq sequencing. The resulting Library Oil_Mgv dataset was further compared with those obtained by direct sequencing of environmental DNA from Brazilian mangroves (from distinct regions and affected by distinct sources of contamination), focusing on hydrolases with potential use in biotechnological processes. The most abundant hydrolases found were proteases, esterases and amylases, with similar occurrence profile in all datasets. The main microbial groups harboring such hydrolase-encoding genes were distinct in each mangrove, and in the fosmid library these enzymes were mainly assigned to Chloroflexaceae (for amylases), Planctomycetaceae (for esterases) and Bradyrhizobiaceae (for proteases). Assembly and analysis of Library_Oil Mgv reads revealed three potentially novel enzymes, one epoxide hydrolase, one xylanase and one amylase, to be further investigated via heterologous expression assays.

Tài liệu tham khảo

Adrio JL, Demain AL (2014) Microbial enzymes: tools for biotechnological processes. Biomolecules 4:117–139. doi:10.3390/biom4010117 Andreote FD, Jiménez DJ, Chaves D et al (2012) The microbiome of Brazilian mangrove sediments as revealed by metagenomics. PLoS ONE. doi:10.1371/journal.pone.0038600 Cabral L, Júnior GVL, de Sousa STP et al (2016) Anthropogenic impact on mangrove sediments triggers differential responses in the heavy metals and antibiotic resistomes of microbial communities. Environ Pollut 216:460–469. doi:10.1016/j.envpol.2016.05.078 Cañas C, Suzuki Y, Marchisone C et al (2014) Interaction of branch migration translocases with the holliday junction-resolving enzyme and their implications in holliday junction resolution. J Biol Chem 289:17634–17646. doi:10.1074/jbc.M114.552794 Dalmaso GZL, Ferreira D, Vermelho AB (2015) Marine extremophiles a source of hydrolases for biotechnological applications. Mar Drugs 13:1925–1965 de Vasconcellos SP, Angolini CFF, García INS et al (2010) Reprint of: screening for hydrocarbon biodegraders in a metagenomic clone library derived from Brazilian petroleum reservoirs. Org Geochem 41:1067–1073. doi:10.1016/j.orggeochem.2010.08.003 Dos Santos HF, Cury JC, do Carmo FL et al (2011) Mangrove bacterial diversity and the impact of oil contamination revealed by pyrosequencing: bacterial proxies for oil pollution. PLoS ONE 6:1–8. doi:10.1371/journal.pone.0016943 Duarte CM, Losada IJ, Hendriks IE et al (2013) The role of coastal plant communities for climate change mitigation and adaptation. Nat Clim Change 3:961–968. doi:10.1038/nclimate1970 Fuerst JA, Sagulenko E (2011) Beyond the bacterium: planctomycetes challenge our concepts of microbial structure and function. Nat Rev Microbiol 9:403–413. doi:10.1038/nrmicro2578 Ghizelini AM, Mendonça-Hagler LCS, Macrae A (2012) Microbial diversity in Brazilian mangrove sediments—a mini review. Braz J Microbiol 43:1242–1254. doi:10.1590/S1517-83822012000400002 Glass EM, Meyer F (2011) The metagenomics RAST server: a public resource for the automatic phylogenetic and functional analysis of metagenomes. In: Handbook of molecular microbial ecology I: metagenomics and complementary approaches. Wiley, Hoboken, NJ, p 325–331. doi:10.1002/9781118010518.ch37 Gomes NC, Borges LR, Paranhos R et al (2008) Exploring the diversity of bacterial communities in sediments of urban mangrove forests. FEMS Microbiol Ecol 66:96–109. doi:10.1111/j.1574-6941.2008.00519.x Gomes NCM, Flocco CG, Costa R et al (2010) Mangrove microniches determine the structural and functional diversity of enriched petroleum hydrocarbon-degrading consortia. FEMS Microbiol Ecol 74:276–290. doi:10.1111/j.1574-6941.2010.00962.x Hall T (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98 Holguin G, Vazquez P, Bashan Y (2001) The role of sediment microorganisms in the productivity, conservation, and rehabilitation of mangrove ecosystems: an overview. Biol Fertil Soils 33:265–278. doi:10.1007/s003740000319 Huang X, Madan A (1999) CAP3: a DNA sequence assembly program. Genome Res 9:868–877. doi:10.1101/gr.9.9.868 HuiJie L, CaiYun Y, Yun T et al (2011) Using population dynamics analysis by DGGE to design the bacterial consortium isolated from mangrove sediments for biodegradation of PAHs. Int Biodeterior Biodegrad 65:269–275. doi:10.1016/j.ibiod.2010.11.010 Jain R, Chan MK (2007) Support for a potential role of E. coli oligopeptidase A in protein degradation. Biochem Biophys Res Commun 359:486–490. doi:10.1016/j.bbrc.2007.05.142 Jiménez DJ, Dini-Andreote F, Ottoni JR et al (2015) Compositional profile of α/β-hydrolase fold proteins in mangrove soil metagenomes: Prevalence of epoxide hydrolases and haloalkane dehalogenases in oil-contaminated sites. Microb Biotechnol 8:604–613. doi:10.1111/1751-7915.12157 Kaushik S, Sowdhamini R (2014) Distribution, classification, domain architectures and evolution of prolyl oligopeptidases in prokaryotic lineages. BMC Genomics 15:985. doi:10.1186/1471-2164-15-985 Kilbane JJ (2006) Microbial biocatalyst developments to upgrade fossil fuels. Curr Opin Biotechnol 17:305–314 Kim YJ, Choi GS, Kim SB et al (2006) Screening and characterization of a novel esterase from a metagenomic library. Protein Expr Purif 45:315–323. doi:10.1016/j.pep.2005.06.008 Lage OM, Bondoso J (2012) Bringing Planctomycetes into pure culture. Front Microbiol 3:1–6. doi:10.3389/fmicb.2012.00405 Li S, Yang X, Yang S et al (2012) Technology prospecting on enzymes: application, marketing and engineering. Comput Struct Biotechnol J 2:e201209017. doi:10.5936/csbj.201209017 López-López O, Cerdán ME, González Siso MI (2014) New extremophilic lipases and esterases from metagenomics. Curr Protein Pept Sci 15:445–455. doi:10.2174/1389203715666140228153801 Maie N, Pisani O, Jaffe R (2008) Mangrove tannins in aquatic ecosystems: their fate and possible influence on dissolved organic carbon and nitrogen cycling. Limnol Oceanogr 53:160–171 Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York Matos RG, Bárria C, Moreira RN et al (2014) The importance of proteins of the RNase II/RNB-family in pathogenic bacteria. Front Cell Infect Microbiol 4:68. doi:10.3389/fcimb.2014.00068 Matsui M, Fowler JH, Walling LL (2006) Leucine aminopeptidases: diversity in structure and function. Biol Chem 387:1535–1544 Meyer F, Paarmann D, D’Souza M et al (2008) The metagenomics RAST server–a public resource for the automatic phylogenetic and functional analysis of metagenomes. BMC bioinformatics 9:386. doi:10.1186/1471-2105-9-386 Naumoff DG, Ivanova AA, Dedysh SN (2014) Phylogeny of β-xylanases from Planctomycetes. Mol Biol 48:439–447. doi:10.1134/S0026893314030145 Overbeek R, Begley T, Butler RM et al (2005) The subsystems approach to genome annotation and its use in the project to annotate 1000 genomes. Nucleic Acids Res 33:5691–5702. doi:10.1093/nar/gki866 Rainey FA, Weiss N, Stackebrandt E (1995) Phylogenetic analysis of the genera Cellulomonas, Promicromonospora, and Jonesia and proposal to exclude the genus Jonesia from the family Cellulomonadaceae. Int J Syst Bacteriol 45:649–652 Rao MB, Tanksale AM, Ghatge MS, Deshpande VV (1998) Molecular and biotechnological aspects of microbial proteases. Microbiol Mol Biol Rev 62:597–635 Reef R, Feller IC, Lovelock CE (2010) Nutrition of mangroves. Tree Physiol 30:1148–1160. doi:10.1093/treephys/tpq048 Rice P, Longden I, Bleasby A (2000) EMBOSS: the European molecular biology open software suite. Trends Genet 16:276–277. doi:10.1016/j.cocis.2008.07.002 Sadeghi A, Soltani BM, Jouzani GS et al (2014) Taxonomic study of a salt tolerant Streptomyces sp. strain C-2012 and the effect of salt and ectoine on lon expression level. Microbiol Res 169:232–238. doi:10.1016/j.micres.2013.06.010 Sahoo K, Dhal NK (2009) Potential microbial diversity in mangrove ecosystems: a review. Indian J Mar Sci 38:249–256. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425 Sathya R, Ushadevi T (2014) Industrially important enzymes producing streptomyces species from mangrove sediments. Int J Pharm Pharm Sci 6:233–237 Schneider E, Freundlieb S, Tapio S, Boos W (1992) Molecular characterization of the malT-dependent periplasmic alpha-amylase of Escherichia coli encoded by malS. J Biol Chem 267:5148–5154 Spalding MD, Ruffo S, Lacambra C et al (2014) The role of ecosystems in coastal protection: adapting to climate change and coastal hazards. Ocean Coast Manag 90:50–57. doi:10.1016/j.ocecoaman.2013.09.007 Taketani RG, Yoshiura CA, Dias ACF et al (2010) Diversity and identification of methanogenic archaea and sulphate-reducing bacteria in sediments from a pristine tropical mangrove. Antonie van Leeuwenhoek 97:401–411. doi:10.1007/s10482-010-9422-8 Tamura K, Peterson D, Peterson N et al (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739. doi:10.1093/molbev/msr121 Thatoi H, Behera BC, Mishra RR (2013) Ecological role and biotechnological potential of mangrove fungi: a review. Mycology 4:54–71. doi:10.1080/21501203.2013.785448 Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680. doi:10.1093/nar/22.22.4673 Thompson CE, Beys-da-Silva WO, Santi L et al (2013) A potential source for cellulolytic enzyme discovery and environmental aspects revealed through metagenomics of Brazilian mangroves. AMB Express 3:65. doi:10.1186/2191-0855-3-65 Tiwari SP, Srivastava R, Singh CS et al (2015) Amylases: an overview with special reference to alpha amylase. J Glob Biosci 4:1886–1901 Tsai YL, Olson BH (1991) Rapid method for direct extraction of DNA from soil and sediments. Appl Environ Microbiol 57:1070–1074 Varon-Lopez M, Dias ACF, Fasanella CC et al (2014) Sulphur-oxidizing and sulphate-reducing communities in Brazilian mangrove sediments. Environ Microbiol 16:845–855. doi:10.1111/1462-2920.12237 Willumsen P, Karlson U, Stackebrandt E, Kroppenstedt RM (2001) Mycobacterium frederiksbergense sp. nov., a novel polycyclic aromatic hydrocarbon-degrading Mycobacterium species. Int J Syst Evol Microbiol 51:1715–1722. doi:10.1099/00207713-51-5-1715 Yu R, Gan P, MacKay AA et al (2010) Presence, distribution, and diversity of iron-oxidizing bacteria at a landfill leachate-impacted groundwater surface water interface. FEMS Microbiol Ecol 71:260–271. doi:10.1111/j.1574-6941.2009.00797.x Zmasek CM, Godzik A (2014) Phylogenomic analysis of glycogen branching and debranching enzymatic duo. BMC Evol Biol 14:183. doi:10.1186/s12862-014-0183-2