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Title

Molecular characterization of α-amylase from Staphylococcus aureus

 

Authors

Hanumanthu Prasanna Lakshmi1, Uppu Venkateswara Prasad1, Sthanikam Yeswanth1, Vimjam Swarupa1, Osuru Hari Prasad1, Mangamoori Lakshmi Narasu2, Potukuchi Venkata Gurunadha Krishna Sarma1*

 

Affiliation

1Department of Biotechnology, Sri Venkateswara Institute of Medical Sciences, Tirupati-517 507, AP, India; 2Centre for Biotechnology, Institute of science and technology, JNTU, Hyderabad-500085, India.

 

Email

sarmasvims@gmail.com; *Corresponding author

 

Article Type

Hypothesis

 

Date

Received February 02, 2013; Accepted February 04, 2013; Published March 19, 2013

 

Abstract

Staphylococcus aureus is one of the prominent Gram positive human pathogen secretes many surface and secretary proteins including various enzymes and pathogenic factors that favour the successful colonization and infection of host tissue. α-amylase is one of the enzymes secreted by S. aureus which catalyses the breakdown of complex sugars to monosaccharides, which are required for colonization and survival of this pathogen in any anatomical locales. In the present study we have cloned, sequenced, expressed and characterized α-amylase gene from S. aureus ATCC12600. The recombinant enzyme has a molecular weight of 58kDa and the kinetics showed Vmax 0.0208±0.033 (mg/ml)/mg/min and Km 10.633±0.737mg/ml. The multiple sequence analysis showed α-amylase of S. aureus exhibited large differences with Bacillus subtilis and Streptococcus bovis. As the crystal structure of S. aureus α-amylase was unavailable, we used homology modelling method to build the structure. The built structure was validated by Ramachandran plot which showed 90% of the residues in the allowed region while no residue was found in the disallowed region and the built structure was close to the crystal structure with Z-Score: -6.85. The structural superimposition studies with α-amylases of Bacillus subtilis and Streptococcus bovis showed distinct differences with RMSD values of 18.158Åand 7.091Å respectively which correlated with enzyme kinetics, indicating α-amylase is different among these bacteria. 

 

Keywords

α-amylase, Km, Vmax, Z-score, RMSD.

 

Citation

Lakshmi et al.   Bioinformation 9(6): 281-285 (2013)

 

Edited by

P Kangueane

 

ISSN

0973-2063

 

Publisher

Biomedical Informatics

 

License

This is an Open Access article which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. This is distributed under the terms of the Creative Commons Attribution License.