Purification, characterization, and molecular cloning of a chitinase from the seeds of Benincasa hispida.
A chitinase was purified from the seeds of Benincasa hispida, a medicinal plant also called white gourd, and a member of the Cucurbitaceae family. Purification was done by using a procedure consisting of only two fractionation steps: an acid denaturation step followed by ion-exchange chromatography. The sequence of the N-terminal forty amino acid residues was analyzed and the sequence indicated that the enzyme is a class III chitinase. The enzyme, which is a basic chitinase, is one of at least five chitinases detected in the seed extract of B. hispida. Like other class III chitinases, this enzyme also has lysozyme activity. A genomic clone of the gene encoding the enzyme was isolated and sequenced. The gene has the potential to encode a protein of 301 amino acid residues. The deduced amino acid sequence of the protein, as expected from the N-terminal amino acid sequence, shares high degrees of similarity with other class III chitinases.
Shih
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Health Research Center, Department of Biology, Southern University, Baton Rouge, LA 70813, USA.
Khan
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Jia
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Wu
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Shih
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D000595:Amino Acid Sequence; D001483:Base Sequence; D002688:Chitinases / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D018744:DNA, Plant; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D009113:Muramidase / Q000378:metabolism; D010940:Plant Proteins; D010946:Plants, Medicinal / Q000201:enzymology*; D019685:Rosales / Q000201:enzymology*; D012639:Seeds
D018744:DNA, Plant; D010940:Plant Proteins; D002688:Chitinases; C110892:chitinase, class III; D009113:Muramidase