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Purification and Characterization of a Low Molecular Weight Endo-xylanase from Mushroom Termitomyces clypeatus

IR@IICB: CSIR-Indian Institute of Chemical Biology, Kolkata

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Title Purification and Characterization of a Low Molecular Weight Endo-xylanase from Mushroom Termitomyces clypeatus
 
Creator Soren, Dhananjay
Jana, Mohanlal
Sengupta, Subhabrata
Ghosh, Anil K
 
Subject Drug Development/Diagnostics & Biotechnology
 
Description A low molecular weight endo-xylanase (EC 3.2.1.8) was purified from an edible mushroom Termitomyces clypeatus grown in submerged medium with oat spelt xylan. Xylanase was purified to apparent homogeneity by ammonium sulfate fractionation and gel filtration chromatography. Its molecular weight was determined by gel filtration chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 12 kDa. The enzyme was found to be most active at 50°C and pH 5.0, being most stable at pH 6.5. The Km for oat spelt xylan was determined to be 10.4 mg/ml. The specificities of the enzyme was observed to be highly specific towards oat spelt xylan and was inhibited by mercuric chloride (HgCl2), N-bromosuccinimide, and trans-1,2-diaminocyclohexane-N′,N′,N′,N′-tetraacetic acid strongly. The inhibitory action of N-bromosuccinimide on enzyme confirmed the presence of one tryptophan residue in its substrate-binding site. Amino acid analysis for xylanase showed the presence of high amount of hydrophobic serine, glycine, threonine, and alanine residues. The N-terminal sequencing study for the previously purified and characterized 56 kDa xylanolytic amyloglucosidase reveal the presence of 33.30% identity with glucoamylase chain A from Aspergillus awamori. The N-terminal sequence analysis of the present 12 kDa enzyme showed highest similarity (72.22% identity) towards xylanase from Neurospora crassa.
 
Date 2010
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/22/1/APPLIED_BIOCHEMISTRY_AND_BIOTECHNOLOGY_162(2)_373%2D389;2010[44].pdf
Soren, Dhananjay and Jana, Mohanlal and Sengupta, Subhabrata and Ghosh, Anil K (2010) Purification and Characterization of a Low Molecular Weight Endo-xylanase from Mushroom Termitomyces clypeatus. Appl Biochem Biotechnol, 162. pp. 373-389.
 
Relation http://doi.org/10.1007/s12010-009-8763-0
http://www.eprints.iicb.res.in/22/