Cloning, Overexpression, and Characterization of Halostable, Solvent-Tolerant Novel β-Endoglucanase from a Marine Bacterium Photobacterium panuliri LBS5T (DSM 27646T) View Full Text


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Article Info

DATE

2016-02

AUTHORS

Kamal Deep, Abhijit Poddar, Subrata K. Das

ABSTRACT

A 1329 nucleotide long endoglucanase gene was amplified from marine bacterium Photobacterium panuliri strain LBS5(T).The enzyme sequence was novel as protein-based similarity search revealed that it shared maximum similarity of 99% with hypothetical protein of P. aquae and 40% with endoglucanase of P. marinum AK15. The gene was cloned, overexpressed in Escherichia coli BL21 (DE3), and purified up to homogeneity using Ni-NTA affinity chromatography. The purified enzyme, designated as Cel8, was monomeric and has a molecular mass of 53 kDa. The enzyme was halostable and exhibited optimal carboxymethyl cellulase (CMCase) activity and stability at 2 M NaCl. Optimal activity was obtained at 40 °C and at pH 4. The enzyme exhibited remarkable stability in different organic solvents (50%, v/v), and activity increased nearly 1.5-fold in presence of butanol, isopropanol, petroleum ether, benzene, acetone, and n-hexane. It was active in Ca(2+), Ba(2+), and Ni(2+) and inhibited by Co(2+), Cd(2+), Zn(2+), Cu(2+), and Hg(2+). Under normal physiological conditions, the enzyme has 25% helix, 30% sheets, and 56% irregularities, whereas salt leads to helix to sheet transition in enzyme. Three-dimensional reconstruction analysis revealed that the enzyme has (α/β)8 structure and a TIM barrel fold-like structure at the central groove of enzyme. This is the first evidenced report on halostable, organic solvent tolerant cellulase in the marine bacterial genus Photobacterium. More... »

PAGES

695-709

References to SciGraph publications

  • 2008-10. Cloning and enzymatic characterization of a xylanase gene from a soil-derived metagenomic library with an efficient approach in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2012-08. Purification and characterization of an organic solvent-tolerant alkaline cellulase from a halophilic isolate of Thalassobacillus in BIOTECHNOLOGY LETTERS
  • 2015-06. The Phyre2 web portal for protein modeling, prediction and analysis in NATURE PROTOCOLS
  • 2004-02. Purification and characterisation of alkaline cellulase produced by a novel isolate, Bacillus sphaericus JS1 in JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
  • 2006-02. Salt-activated endoglucanase of a strain of alkaliphilic Bacillus agaradhaerens in ANTONIE VAN LEEUWENHOEK
  • 2012-08. Purification and characterization of an organic-solvent-tolerant cellulase from a halotolerant isolate, Bacillus sp. L1 in JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
  • 2012-01. Effects of salts on activity of halophilic cellulase with glucomannanase activity isolated from alkaliphilic and halophilic Bacillus sp. BG-CS10 in EXTREMOPHILES
  • 2014-11. Photobacterium panuliri sp. nov., an Alkalitolerant Marine Bacterium Isolated from Eggs of Spiny Lobster, Panulirus penicillatus from Andaman Sea in CURRENT MICROBIOLOGY
  • 2015-12. Isolation, production, purification and characterization of an organic-solvent-thermostable alkalophilic cellulase from Bacillus vallismortis RG-07 in BMC BIOTECHNOLOGY
  • 2010-05. Ligand docking and binding site analysis with PyMOL and Autodock/Vina in JOURNAL OF COMPUTER-AIDED MOLECULAR DESIGN
  • 2004-11. Inhibition of ethanol-producing yeast and bacteria by degradation products produced during pre-treatment of biomass in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 1970-08. Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 in NATURE
  • 2010-06. Cel8H, a novel endoglucanase from the halophilic bacterium Halomonas sp. S66-4: Molecular cloning, heterogonous expression, and biochemical characterization in JOURNAL OF MICROBIOLOGY
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/s12010-015-1903-9

    DOI

    http://dx.doi.org/10.1007/s12010-015-1903-9

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1024330645

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/26494136


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