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.
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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).
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Co-first author, Ming-Yang Zhou and Xiu-Lan Chen had equal contribution to this work.
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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)
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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
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DOI: https://doi.org/10.1007/s00248-009-9506-z