Skip to main content

Advertisement

Log in

Diversity of Both the Cultivable Protease-Producing Bacteria and Their Extracellular Proteases in the Sediments of the South China Sea

  • Microbiology of Aquatic Systems
  • Published:
Microbial Ecology Aims and scope Submit manuscript

Abstract

Protease-producing bacteria are known to play an important role in degrading sedimentary particular organic nitrogen, and yet, their diversity and extracellular proteases remain largely unknown. In this paper, the diversity of the cultivable protease-producing bacteria and their extracellular proteases in the sediments of the South China Sea was investigated. The richness of the cultivable protease-producing bacteria reached 106 cells/g in all sediment samples. Analysis of the 16S rRNA gene sequences revealed that the predominant cultivated protease-producing bacteria are Gammaproteobacteria affiliated with the genera Pseudoalteromonas, Alteromonas, Marinobacter, Idiomarina, Halomonas, Vibrio, Shewanella, Pseudomonas, and Rheinheimera, with Alteromonas (34.6%) and Pseudoalteromonas (28.2%) as the predominant groups. Inhibitor analysis showed that nearly all the extracellular proteases from the bacteria are serine proteases or metalloproteases. Moreover, these proteases have different hydrolytic ability to different proteins, reflecting they may belong to different kinds of serine proteases or metalloproteases. To our knowledge, this study represents the first report of the diversity of bacterial proteases in deep-sea sediments.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3

Similar content being viewed by others

References

  1. Annette SE, Megan LP, Libby AS, Sarah Q, Philip CB (2004) Bacterial diversity and ecosystem function of filamentous microbial mats from aphotic (cave) sulfidic springs dominated by chemolithoautotrophic “Epsilonproteobacteria”. FEMS Microbiol Ecol 51:31–53

    Article  Google Scholar 

  2. Bianchi A, Calafat A, Wit RD, Garcin J, Tholosan O, Cacho I, Canals M, Fabrés J, Grout H, Masqué P, Sanchez-Cabeza J-A, Sempéré R (2003) Microbial activity at the deep water sediment boundary layer in two highly productive systems in the Western Mediterranean: the Almeria–Oran front and the Malaga upwelling. Oceanol Acta 25:315–324

    Article  Google Scholar 

  3. Brunnegard J, Grandel S, Stahl H, Tengberg A, Hall POJ (2004) Nitrogen cycling in deep-sea sediments of the Porcupine Abyssal Plain, NE Atlantic. Prog Oceanogr 63:159–181

    Article  Google Scholar 

  4. Buesseler KO, Trull TW, Steinberg DK, Silver MW, Siegel DA, Saitoh S-I, Lamborg CH, Lam PJ, Karl DM, Jiao NZ, Honda MC, Elskens M, Dehairs F, Browng SL, Boyd PW, Bishop JKB, Bidigare RR (2008) VERTIGO (VERtical Transport In the Global Ocean): a study of particle sources and flux attenuation in the North Pacific. Deep-Sea Res II 55:1522–1539

    Article  Google Scholar 

  5. Chen DF, Huang YY, Yuan XL, Cathles LMIII (2005) Seep carbonates and preserved methane oxidizing archaea and sulfate reducing bacteria fossils suggest recent gas venting on the seafloor in the Northeastern South China Sea. Mar Petrol Geol 22:613–621

    Article  Google Scholar 

  6. Chen X-L, Zhang Y-Z, Gao P-J, Luan X-W (2003) Two different proteases produced by a deep-sea psychrotrophic strain Pseudoalteromonas sp. SM9913. Mar Biol 143:989–993

    Article  CAS  Google Scholar 

  7. Chen X-L, Xie B-B, Lu J-T, He H-L, Zhang Y-Z (2007) A novel type of subtilase from the psychrophilic bacterium Pseudoalteromonas sp. SM9913: catalytic and structural properties of deseasin MCP-01. Microbiol-SGM 153:2116–2125

    Article  CAS  Google Scholar 

  8. Chen Z, Yan W, Chen MH, Wang SH, Lu J, Zhang F, Xiang R, Xiao SB, Yan B, Gu SC (2006) Discovery of seep carbonate nodules as new evidence for gas venting on the northern continental slope of South China Sea. Chin Sci Bull 51:1228–1237

    Article  CAS  Google Scholar 

  9. D’Hondt S, Jørgensen BB, Miller DJ, Batzke A, Blake R, Cragg BA, Cypionka H, Dickens GR, Ferdelman T, Hinrichs K-U, Holm NG, Mitterer R, Spivack A, Wang G, Bekins B, Engelen B, Ford K, Gettemy G, Rutherford SD, Sass H, Skilbeck CG, Ailleo IW, Guerin G, House CH, Inagaki F, Meister P, Naehr T, Niitsuma S, Parkes RJ, Schippers A, Smith DC, Teske A, Wiegel J, Padilla CN, Acosta JLS (2004) Distribution of microbial activities in deep subseafloor sediments. Science 306:2216–2221

    Article  PubMed  Google Scholar 

  10. Han XQ, Suess E, Huang YY, Wu NY, Bohrmann G, Su X, Eisenhauer A, Rehder G, Fang YX (2008) Jiulong methane reef: Microbial mediation of seep carbonates in the South China Sea. Mar Geol 249:243–256

    Article  Google Scholar 

  11. Huston AL, Deming JW (2002) Relationships between microbial extracellular enzymatic activity and suspended and sinking particulate organic matter: seasonal transformations in the North Water. Deep-Sea Res II 49:5211–5225

    Article  CAS  Google Scholar 

  12. Hunter EM, Mills HJ, Kostka JE (2006) Microbial community diversity associated with carbon and nitrogen cycling in permeable shelf sediments. Appl Environ Microbiol 72:5689–5701

    Article  PubMed  CAS  Google Scholar 

  13. Inagaki F, Suzuki M, Takai K, Oida H, Sakamoto T, Aoki K, Nealson K, Horikoshi K (2003) Microbial communities associated with geological horizons in coastal subseafloor sediments from the Sea of Okhotsk. Appl Environ Microbiol 69:7224–7235

    Article  PubMed  CAS  Google Scholar 

  14. Kobayashi T, Koide O, Mori K, Shimamura S, Matsuura T, Miura T, Takaki Y, Morono Y, Nunoura T, Imachi H, Inagaki F, Takai K, Horikoshi K (2008) Phylogenetic and enzymatic diversity of deep subseafloor aerobic microorganisms in organics- and methane-rich sediments off Shimokita Peninsula. Extremophiles 12:519–527

    Article  PubMed  CAS  Google Scholar 

  15. Kumar S, Tamura K, Nei M (2004) MEGA3: integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment. Brief Bioinform 5:150–163

    Article  PubMed  CAS  Google Scholar 

  16. Kurata A, Miyazaki M, Kobayashi T, Nogi Y, Horikoshi K (2007) Alkalimonas collagenimarina sp. nov., a psychrotolerant, obligate alkaliphile isolated from deep-sea sediment. Int J Syst Evol Microbiol 57:1549–1553

    Article  PubMed  CAS  Google Scholar 

  17. Olivera NL, Sequeiros C, Nievas ML (2007) Diversity and enzyme properties of protease-producing bacteria isolated from sub-Antarctic sediments of Isla de Los Estados, Argentina. Extremophiles 11:517–526

    Article  PubMed  CAS  Google Scholar 

  18. Polymenakou PN, Bertilsson S, Tselepides A, Stephanou EG (2005) Bacterial community composition in different sediments from the eastern Mediterranean Sea: a comparison of four 16S ribosomal DNA clone libraries. Microbioal Ecol 50:447–462

    Article  CAS  Google Scholar 

  19. Süß J, Engelen B, Cypionka H, Sass H (2004) Quantitative analysis of bacterial communities from Mediterranean sapropels based on cultivation-dependent methods. FEMS Microbiol Ecol 51:109–121

    Article  PubMed  Google Scholar 

  20. Talbot V, Bianchi M (1997) Bacterial proteolytic activity in sediments of the Subantarctic Indian Ocean Sector. Deep-Sea Res II 44:1069–1084

    Article  CAS  Google Scholar 

  21. Xiong H, Song L, Xu Y, Tsoi M-Y, Dobretsov S, Qian P-Y (2007) Characterization of proteolytic bacteria from the Aleutian deep-sea and their proteases. J Ind Microbiol Biotechnol 34:63–71

    Article  PubMed  CAS  Google Scholar 

  22. Xu H, Wu M, Wang X, Yang J, Wang C (2008) Bacterial diversity in deep-sea sediment from northeastern Pacific Ocean. Acta Ecol Sinica 28:479–485

    Article  CAS  Google Scholar 

  23. Zeng R, Zhang R, Zhao J, Lin N (2003) Cold-active serine alkaline protease from the psychrophilic bacterium Pseudomonas strain DY-A: enzyme purification and characterization. Extremophiles 7(4):335–337

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The sediment samples were collected during the 2007 South China Sea Open Cruise by R/V Shiyan 3, South China Sea Institute of Oceanology, CAS. The work was supported by Hi-Tech Research and Development program of China (2006AA09Z414, 2007AA091903, 2007AA021306), National Natural Science Foundation of China (30770040, 40776032, 40576069), Program for New Century Excellent Talents in University (NCET-06-0578), Foundation for Young Excellent Scientists in Shandong Province (2006BS02002), and COMRA Program (DYXM-115-02-2-6, DYXM-115-02-2-20).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yu-Zhong Zhang.

Additional information

Co-first author, Ming-Yang Zhou and Xiu-Lan Chen had equal contribution to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Fig. S1

The protease-producing bacteria from the sediments of the South China Sea growing on the screening plates. One-gram sediment (wet weight) of every sample was serially tenfold diluted to 10−6 dilution with sterile artificial sea water. Aliquots of 100 μl diluted deep-sea sediment samples (10−2–10−6 dilution) were spread on screening plates and incubated at 15°C for a proper time to form detectable colonies with clear hydrolytic zone. The results of 10−4 diluted samples were shown in Fig. S1. (DOC 8425 kb)

Fig. S2

The neighbor-joining phylogenetic tree of the strains in branch 1 in Fig. 3 based on the 16S rRNA gene sequences. (DOC 38 kb)

Fig. S3

The neighbor-joining phylogenetic tree of the strains in branch 2 in Fig. 3 based on the 16S rRNA gene sequences. (DOC 26 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhou, MY., Chen, XL., Zhao, HL. et al. 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 (2009). https://doi.org/10.1007/s00248-009-9506-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00248-009-9506-z

Keywords

Navigation