Identification of a New Marine Bacterial Strain SD8 and Optimization of Its Culture Conditions for Producing Alkaline Protease

PLoS One. 2015 Dec 30;10(12):e0146067. doi: 10.1371/journal.pone.0146067. eCollection 2015.

Abstract

While much attention has been given to marine microorganisms for production of enzymes, which in general are relatively more stable and active compared to those from plants and animals, studies on alkaline protease production from marine microorganisms have been very limited. In the present study, the alkaline protease producing marine bacterial strain SD8 isolated from sea muds in the Geziwo Qinhuangdao sea area of China was characterized and its optimal culture conditions were investigated. Strain SD8 was initially classified to belong to genus Pseudomonas by morphological, physiological and biochemical characterizations, and then through 16S rDNA sequence it was identified to be likely Pseudomonas hibiscicola. In addition, the culture mediums, carbon sources and culture conditions of strain SD8 were optimized for maximum production of alkaline protease. Optimum enzyme production (236U/mL when cultured bacteria being at 0.75 mg dry weight/mL fermentation broth) was obtained when the isolate at a 3% inoculum size was grown in LB medium at 20 mL medium/100mL Erlenmeyer flask for 48h culture at 30°C with an initial of pH 7.5. This was the first report of strain Pseudomonas hibiscicola secreting alkaline protease, and the data for its optimal cultural conditions for alkaline protease production has laid a foundation for future exploration for the potential use of SD8 strain for alkaline protease production.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aquatic Organisms / enzymology
  • Aquatic Organisms / growth & development
  • Aquatic Organisms / metabolism
  • Bacterial Proteins / biosynthesis*
  • Culture Media
  • Electrophoresis, Agar Gel
  • Endopeptidases / biosynthesis*
  • Fermentation
  • Phylogeny
  • Pseudomonas / enzymology
  • Pseudomonas / growth & development
  • Pseudomonas / metabolism*
  • RNA, Ribosomal, 16S / genetics

Substances

  • Bacterial Proteins
  • Culture Media
  • RNA, Ribosomal, 16S
  • Endopeptidases
  • alkaline protease

Grants and funding

This study is financially supported by Natural Science Foundation of Hebei Province (D2014203102 to HC) and Ministry of Education Specialized Research Fund for the Doctoral Program of Higher Education (20131333120010 to HC). LW is supported by Australian National Health and Medical Research Council (NHMRC) Postgraduate Research Scholarship, and CJX is supported by NHMRC Senior Research Fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.