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Purification and Characterization of a Trehalase–Invertase Enzyme with Dual Activity from Candida Utilis

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Title Purification and Characterization of a Trehalase–Invertase Enzyme with Dual Activity from Candida Utilis
 
Creator Lahiri, Sagar
Basu, Arghya
Sengupta, Shinjinee
Banerjee, Shakri
Dutta, Trina
Soren, Dhananjay
Chattopadhyay, Krishnananda
Ghosh, Anil K
 
Subject Drug Development/Diagnostics & Biotechnology
Structural Biology & Bioinformatics
 
Description Trehalose and sucrose, two important anti-stress non-reducing natural disaccharides, are catabolized by
two enzymes, namely trehalase and invertase respectively. In this study, a 175 kDa enzyme protein active
against both substrates was purified from wild type Candida utilis and characterized in detail. Substrate
specificity assay and activity staining revealed the enzyme to be specific for both sucrose and trehalose.
The ratio between trehalase and invertase activity was found to be constant at 1:3.5 throughout the
entire study. Almost 40-fold purification and 30% yield for both activities were achieved at the final step
of purification. The presence of common enzyme inhibitors, thermal and pH stress had analogous effects
on its trehalase and invertase activity. Km values for two activities were similar while Vmax and Kcat also
differed by a factor of 3.5. Competition plot for both substrates revealed the two activities to be occurring
at the single active site. N-terminal sequencing and MALDI-TOF data analysis revealed higher similarity of
the purified protein to previously known neutral trehalases. While earlier workers mentioned independent
purification of neutral trehalase or invertase from different sources, the present study reports the
purification of a single protein showing dual activity.
 
Publisher Elsevier
 
Date 2012
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/1491/1/ARCHIVES_OF_BIOCHEMISTRY_AND_BIOPHYSICS__522__(2)90%2D994;2012[3].pdf
Lahiri, Sagar and Basu, Arghya and Sengupta, Shinjinee and Banerjee, Shakri and Dutta, Trina and Soren, Dhananjay and Chattopadhyay, Krishnananda and Ghosh, Anil K (2012) Purification and Characterization of a Trehalase–Invertase Enzyme with Dual Activity from Candida Utilis. Archives of Biochemistry and Biophysics, 522 (2). pp. 90-99.
 
Relation http://dx.doi.org//10.1016/j.abb.2012.03.026
http://www.eprints.iicb.res.in/1491/