Cloning of the pelA gene from Bacillus licheniformis 14A and biochemical characterization of recombinant, thermostable, high-alkaline pectate lyase

Berensmeier, S and Sridevi Annapurna, Singh and Meens, J and Buchholz, K (2004) Cloning of the pelA gene from Bacillus licheniformis 14A and biochemical characterization of recombinant, thermostable, high-alkaline pectate lyase. Applied Microbiology and Biotechnology, 64 (4). pp. 560-567.

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Abstract

The pectate lyase gene pelA from alkaliphilic Bacillus licheniformis strain 14A was cloned and sequenced. The nucleotide sequence corresponded to an open reading frame of 1,026 bp that codes for a 39 amino acid signal peptide and a mature protein with a molecular mass of 33,451 Da. The mature PelA showed significant homology to other pectate lyases belonging to polysaccharide lyase family 1, such as enzymes from different Bacillus spp. and Erwinia chrysanthemi. The pelA gene was expressed in Escherichia coli as a recombinant fusion protein containing a C-terminal His-tag, allowing purification to near homogeneity in a one-step procedure. The values for the kinetic parameters Km and Vmax of the fusion protein were 0.56 g/l and 51 μmol/min, respectively. The activity of purified PelAHis was inhibited in the presence of excess substrate. Characterization of product formation revealed unsaturated trigalacturonate as the main product. The yields of unsaturated trigalacturonic acids were further examined for the substrates polygalacturonic acid, citrus pectin and sugar-beet pectin.

Item Type:	Article
Uncontrolled Keywords:	Erwinia chrysanthemi; Escherichia coli; trigalacturonic acids
Subjects:	600 Technology > 08 Food technology > 09 Food Microbiology
Divisions:	Protein Chemistry and Technology
Depositing User:	Users 197 not found.
Date Deposited:	01 Jun 2011 05:56
Last Modified:	28 Dec 2011 10:23
URI:	http://ir.cftri.res.in/id/eprint/10049

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