Abstract
The deep-sea water of the South Pacific Gyre (SPG, 20°S–45°S) is a cold and ultra-oligotrophic environment that is the source of cold-adapted enzymes. However, the characteristic features of psychrophilic enzymes derived from culturable microbes in the SPG remained largely unknown. In this study, the degradation properties of 174 cultures from the deep water of the SPG were used to determine the diversity of cold-adapted enzymes. Thus, the abilities to degrade polysaccharides, proteins, lipids, and DNA at 4, 16, and 28 °C were investigated. Most of the isolates showed one or more extracellular enzyme activities, including amylase, chitinase, cellulase, lipase, lecithinase, caseinase, gelatinase, and DNase at 4, 16, and 28 °C. Moreover, nearly 85.6 % of the isolates produced cold-adapted enzymes at 4 °C. The psychrophilic enzyme-producing isolates distributed primarily in Alteromonas and Pseudoalteromonas genera of the Gammaproteobacteria. Pseudoalteromonas degraded 9 types of macromolecules but not cellulose, Alteromonas secreted 8 enzymes except for cellulase and chitinase. Interestingly, the enzymatic activities of Gammaproteobacteria isolates at 4 °C were higher than those observed at 16 or 28 °C. In addition, we cloned and expressed a gene encoding an α-amylase (Amy2235) from Luteimonas abyssi XH031T, and examined the properties of the recombinant protein. These cold-active enzymes may have huge potential for academic research and industrial applications. In addition, the capacity of the isolates to degrade various types of organic matter may indicate their unique ecological roles in the elemental biogeochemical cycling of the deep biosphere.
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References
Amoozegar MA, Malekzadeh F, Malik KA (2003) Production of amylase by newly isolated moderate halophile, Halobacillus sp. strain MA-2. J Mol Biol 52:353–359
Chen XL, Xie BB, Bian F, Zhao GY, Zhao HL, He HL, Cheng Z, Zhang YZ (2009) Ecological function of myroilysin, a novel bacterial M12 metalloprotease with elastinolytic activity and a synergistic role in collagen hydrolysis, in biodegradation of deep-sea high-molecular weight organic nitrogen. Appl Environ Microbiol 75:1838–1844
Cottrel M, Kirchman DL (2000) Natural assemblages of marine proteobacteria and members of the Cytophaga-Flavobacter cluster consuming low-and high-molecular-weight dissolved organic matter. Appl Environ Microbiol 66:1692–1697
D’Hondt S, Inagaki F, Alvarez Zarikian CA, The Expedition 329 Scientists (2011) South Pacific Gyre subseafloor life. Proc IODP, 329: Tokyo (Integrated Ocean Drilling Program Management International, Inc). doi:10.2204/iodp.proc.329
D’Hondt S, Rutherford S, Spivack AJ (2002) Metabolic activity of subsurface life in deep-sea sediments. Science 295:2067–2070. doi:10.1126/science.1064878
D’Hondt S, Jørgensen BB, Miller DJ, Batzke A, Padilla CN, Acosta JL (2004) Distributions of microbial activities in deep subseafloor sediments. Science 306:2216–2221. doi:10.1126/science.1101155
D’Hondt S, Spivack AJ, Pockalny R, Ferdelman TG, Fischer JP, Kallmeyer J, Abrams LJ, Smith DC, Graham D, Hasiuk F, Schrum H, Stancin AM (2009) Subseafloor sedimentary life in the South Pacific Gyre. Proc Nati Acad Sci USA 106:11651–11656
D’Hondt S, Inagaki F, Alvarez Zarikian CA (2010) South Pacific Gyre microbiology [C] IODP Sci Prosp 329
Fang JS, Zhang L, Li JT, Kato C, Tamburini C, Zhang YZ, Dang HY, Wang GY, Wang FP (2015) The POM-DOM piezophilic microorganism continuum (PDPMC)—the role of piezophilic microorganisms in the global ocean carbon cycle. Sci China 58:106–115
Feller G, Gerday C (2003) Psychrophilic enzymes: hot topics in cold adaptation. Nat Rev Microbiol 1:200–208
Feller G, Lonhienne T, Deroanne C, Libioulle C, Van BJ, Gerday C (1992) Purification, characterization, and nucleotide sequence of the thermolabile alpha-amylase from the antarctic psychrotroph Alteromonas haloplanctis A23. J Biol Chem 267:5217–5221
Feller G, Payan F, Theys F, Qian M, Haser R, Gerday C (1994) Stability and structural analysis of alpha-amylase from the antarctic psychrophile Alteromonas haloplanctis A23. Eur J Biochem 222:441–447
Gerday C, Aittaleb M, Bentahir M, Chessa JP, Claverie P, Collins T, D’Amico S, Dumont J, Garsoux G, Georlette D, Hoyoux A, Lonhienne T, Meuwis MA, Feller G (2000) Cold-adapted enzymes: from fundamentals to biotechnology. Trends Biotechnol 18:103–107
Groudieva T, Kambourova M, Yusef H, Royter M, Grote R, Trinks H, Antranikian G (2004) Diversity and cold-active hydrolytic enzymes of culturable bacteria associated with Arctic sea ice, Spitzbergen. Extremophiles 8:475–488
Hedges JI (1992) Global biogeochemical cycles: progress and problems. Mar Chem 39:67–93
Hopkinson CS, Vallion JJ, Nolin A (2002) Decomposition of dissolved organic matter from the continental margin. Deep Sea Res Part II 49:4461–4478
Hoppe HG (1983) Significance of exoenzymatic activities in the ecology of brackish water: measurements by means of methylumbelliferyl-substrates. Mar Ecol Progr Ser 11:99–308
Joseph B, Ramteke PW, Thomas G (2008) Cold active microbial esterases: some hot issues and recent developments. Biotechnol Adv 26:457–470
Kirchman D (2002) The ecology of Cytophaga-Flavobacteria in aquatic environments. FEMS Microbio Ecol 39:91–100
Li Z, Qin Y, Fan XY, Shi XC, Zhang XH (2014) Diversity of culturable bacteria in the bottom seawater of the South Pacific Gyre. Period Ocean Univ China 44:052–059
Liu J, Zhang Z, Dang H, Lu J, Cui Z (2011) Isolation and characterization of a cold-active amylase from marine Wangia sp. C52. Afr J Microbiol Res 5:1156–1162
Lu MS, Fang YW, Li HZ, Liu HF, Wang SJ (2010) Isolation of a novel cold-adapted amylase-producing bacterium and study of its enzyme production conditions. Ann Microbiol 60:557–563
Margesin R, Fauster V, Fonteyne PA (2005) Characterization of cold-active pectate lyases from psychrophilic Mrakia frigida. Lett Appl Microbiol 40:453–459
Marshall CJ (1997) Cold-adapted enzymes. Trends Biotechnol 15:359–364
Mojallali L, Shahbani ZH, Rajaei S, Akbari NK, Haghbeen K (2013) A novel approximately 34-kDa alpha-amylase from psychrotroph Exiguobacterium sp. SH3: Production, purification, and characterization. Biotechnol Appl Biochem 61:118–125
Ogawa H, Fukuda R, Koike I (1999) Vertical distributions of dissolved organic carbon and nitrogen in the Southern Ocean. Deep-Sea Res Part I: Oceanogr Res Pap 46:1809–1826
Pfannkuche O (1992) Organic carbon flux through the benthic community in the temperate abyssal northeast Atlantic. In: Deep-sea food chains and the global carbon cycle, vol 360. Kluwer, The Netherlands, pp 183–198
Pinchuk GE, Ammons C, Culley DE, Li SMW, McLean JS, Romine MF, Nealson KH, Fredrickson JK, Beliaev AS (2008) Utilization of DNA as a sole source of phosphorus, carbon, and energy by Shewanella spp.: ecological and physiological implications for dissimilatory metal reduction. Appl Environ Microbiol 74:1198–1208
Qin YJ, Huang ZQ, Liu ZD (2014a) A novel cold-active and salt-tolerant a-amylase from marine bacterium Zunongwangia profunda: molecular cloning, heterologous expression and biochemical characterization. Extremophiles 18:271–281
Qin Y, Huang Z, Liu Z (2014b) A novel cold-active and salt-tolerant alpha-amylase from marine bacterium Zunongwangia profunda: molecular cloning, heterologous expression and biochemical characterization. Extremophiles 18(2):271–281
Souza CP, Almeida BC, Colwell RR, Rivera ING (2011) The importance of chitin in the marine environment. Mar Biotechnol 13:823–830
Suzuki MT, Rappé MS, Haimberger ZW, Winfield H, Adair N, Ströbel J, Giovannoni SJ (1997) Bacterial diversity among small-subunit rRNA gene clones and cellular isolates from the same seawater sample. Appl Environ Microbiol 63:983–989
Talbot V, Micheline B (1997) Bacterial proteolytic activity in sediments of the Subantarctic Indian Ocean Sector. Deep Sea Res Part II 44:1069–1084
Vetter YA, Deming JW (1999) Growth rates of marine bacterial isolates on particulate organic substrates solubilized by freely released extracellular enzymes. Microb Ecol 37:86–94
Zaccone R, Caruso G, Cal C (2002) Heterotrophic bacteria in the northern Adriatic Sea: seasonal changes and ectoenzyme profile. Mar Environ Res 54:1–19
Zhang XH (2007) Marine microbiology. Ocean University of China, Qingdao
Zhang JW, Zeng RY (2008) Purification and characterization of a cold-adapted α-amylase produced by Nocardiopsis sp. 7326 isolated from Prydz Bay, Antarctic. Mar Biotechnol 10:75–82
Zhang L, Wang XL, Yu M, Qiao YL, Zhang XH (2015) Genomic analysis of Luteimonas abyssi XH031T: insights into its adaption to the subseafloor environment of South Pacific Gyre and ecological role in biogeochemical cycle. BMC Genom 16:1092–1105
Zhou MY, Chen XL, Zhao HL, Dang HY, Luan XW, Zhang XY, He HL, Zhou BC, Zhang YZ (2009) Diversity of both the cultivable protease-producing bacteria and their extracellular proteases in the sediments of the South China Sea. Microb Ecol 58:582–590
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This work was supported by the National Natural Science Foundation of China (No. 41276141), and the National High Technology Research and Development Program of China (863 Programs, No. 2013AA092103).
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Communicated by H. Atomi.
L. Zhang and Y. Wang contributed equally to this work.
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Zhang, L., Wang, Y., Liang, J. et al. Degradation properties of various macromolecules of cultivable psychrophilic bacteria from the deep-sea water of the South Pacific Gyre. Extremophiles 20, 663–671 (2016). https://doi.org/10.1007/s00792-016-0856-4
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DOI: https://doi.org/10.1007/s00792-016-0856-4