World Journal of Microbiology and Biotechnology

, Volume 27, Issue 4, pp 841–850

Molecular cloning and expression of an extracellular α-amylase gene from an Antarctic deep sea psychrotolerant Pseudomonas stutzeri strain 7193

Authors

  • Jinwei Zhang
    • Key Laboratory of Marine Biogenetic ResourcesState Oceanic Administration
    • Third Institute of Oceanography, State Oceanic Administration
    • Dove Marine Laboratory, School of Marine Science and TechnologyNewcastle University Upon Tyne
    • Key Laboratory of Marine Biogenetic ResourcesState Oceanic Administration
    • Third Institute of Oceanography, State Oceanic Administration
Original Paper

DOI: 10.1007/s11274-010-0526-0

Cite this article as:
Zhang, J. & Zeng, R. World J Microbiol Biotechnol (2011) 27: 841. doi:10.1007/s11274-010-0526-0

Abstract

Psychrotolerant Pseudomonas stutzeri strain 7193 capable of producing an extracellular α-amylase was isolated from deep sea sediments of Prydz Bay, Antarctic. The 59678-Da protein (AmyP) was encoded by 1665-bp gene (amyP). The deduced amino acid sequence was identified with four regions, which are conserved in amylolytic enzymes and form a catalytic domain, and was predicted to be maltotetraose forming extracellular amylase by using the I-TASSER online server. Purification of AmyP amylases from both the recombinant of Escherichia coli Top 10 F′ and strain 7193 was conducted. Biochemical characterization revealed that the optimal amylase activity was observed at pH 9.0 and temperature 40°C. The enzymes were unstable at temperatures above 30°C, and only retain half of their highest activity after incubation at 60°C for 5 min. Thin-layer chromatography analysis of the products of the amylolytic reaction showed the presence of maltotetraose, maltotriose, maltose and glucose in the starch hydrolysate.

Keywords

Deep sea sedimentExtracellular α-amylaseSequencing and expressionCharacterization

Copyright information

© Springer Science+Business Media B.V. 2010