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Indian Journal of Biochemistry and Biophysics (IJBB) >
IJBB Vol.48 [2011] >
IJBB Vol.48(2) [April 2011] >
| Title: | Isolation and characterization of a metal ion-dependent alkaline protease from a halotolerant Bacillus aquimaris VITP4 |
| Authors: | Shivanand, Pooja
Jayaraman, Gurunathan |
| Keywords: | Alkaline protease
Antimicrobial activity
Bacillus aquimaris VITP4
Gelatin zymography
Halostability
Metal ion binding |
| Issue Date: | Apr-2011 |
| Publisher: | NISCAIR-CSIR, India |
| Abstract: | A halotolerant bacterium Bacillus acquimaris VITP4 was used for
the production of extracellular protease. Fractional precipitation using
ammonium chloride was used to obtain the enzyme. The protease exhibited optimum
activity at pH 8.0 and 40°C
and retained 50% of its optimal proteolytic activity even in the presence of 4 M NaCl, suggesting that it is
halotolerant. The molecular mass of protease, as revealed by SDS-PAGE was found
to be 34 kDa and the homogeneity of the enzyme was confirmed by gelatin
zymography and reverse-phase HPLC. Upon purification, the specific activity of
th enzyme increased from 533 U/mg to 1719 U/mg. Protease inhibitors like phenyl
methane sulphonyl fluoride and
2-mercaptoethanol did not affect the activity of the enzyme, but EDTA inhibited
the activity, indicating the requirement of metal ions for activity. Cu2+,
Ni2+ and Mn2+ enhanced the enzyme activity, but Zn2+,
Hg2+ and Fe2+ decreased the activity, while Mg2+,
Ca2+ and K+ had no effect on the enzyme activity. The
protease was quite stable in the presence of cationic (CTAB), anionic (SDS) and
neutral detergents (Triton X-100 and Tween-20) and exhibited antimicrobial
activity against selected bacterial and fungal strains. The stability characteristics
and broad spectrum antimicrobial activity indicated the potential use of this
protease in industrial applications. |
| Page(s): | 95-100 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-0959 (Online); 0301-1208 (Print) |
| Source: | IJBB Vol.48(2) [April 2011]
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