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<span style="font-size:11.0pt;mso-bidi-font-size: 10.0pt;font-family:"Times New Roman";mso-fareast-font-family:"Times New Roman"; mso-ansi-language:EN-US;mso-fareast-language:EN-US;mso-bidi-language:AR-SA" lang="EN-US">Molecular cloning, characterization and expression of lipoxygenase 2 (<i>lox-2</i>) isozyme from Indian soybean [<i>Glycine max</i> (L.) Merrill] <i>cv.</i> Pusa 16</span>

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Title Molecular cloning, characterization and expression of lipoxygenase 2 (lox-2) isozyme from Indian soybean [Glycine max (L.) Merrill] cv. Pusa 16
 
Creator Mandal, Somnath
Sahana, Nandita
Rajarani, A P
Santha, I M
 
Subject Glycine max (L.) Merril
Soybean
Lipoxygenase
cDNA Cloning
Hydropathy profile
Prokaryotic expression
Recombinant Enzyme kinetics
Inhibitors
 
Description 54-63
The consumption of soybean is limited
worldwide, despite being highly nutritious and having versatile uses, due to the presence of grassy, beany and rancid
off-flavour. The lipoxygenase-2 (LOX-2) is the
key enzyme responsible for the production of volatiles released from the beans,
which cause off-flavour in soy products. In
this study, a 2.6-kb full-length lox2 gene (NCBI accession No.
JQ929619.1) was isolated and cloned
from soybean (Glycine max L. Merril) cv. Pusa 16. The cloned cDNA
sequence of lox2 gene showed the complete open reading frame (ORF) of a
putative protein, having 866 amino acids with start codon present at the
foremost position and stop codon at the end. The theoretical pI of predicted
protein was 6.22. A hydropathy profile calculated from the amino acid sequence
resembled those of dicot LOXs, suggesting conservation of the secondary
structure of these enzymes. The LOX-2 showed conserved six Histidine residues within
a span of 520 to 590 amino acid position, a signature element for the enzyme
activity. The lox2 gene was expressed using pET vector in prokaryotic
expression system. The recombinant LOX-2 protein was purified after induction
with IPTG (isopentyl thiogalactoside). A prominent band of 97 kDa was observed,
when affinity purified fractions were analyzed by SDS-PAGE. The purified
protein was characterized for the enzyme activity, substrate preference and Km.
Inhibitor studies with natural antioxidant molecules present in soybean revealed
α-tocopherol to be the most effective inhibitor of LOX-2.
 
Date 2013-02-23T11:27:28Z
2013-02-23T11:27:28Z
2013-02
 
Type Article
 
Identifier 0975-0959 (Online); 0301-1208 (Print)
http://hdl.handle.net/123456789/16066
 
Language en_US
 
Rights CC Attribution-Noncommercial-No Derivative Works 2.5 India
 
Publisher NISCAIR-CSIR, India
 
Source IJBB Vol.50(1) [February 2013]