Carica papaya latex is a rich source of a class II chitinase.
A class II chitinase is present in the latex of the tropical species Carica papaya. The enzyme may be readily purified by using a combination of hydrophobic interaction- and cation-exchange chromatography. This enzyme preparation is homogeneous with respect to the three physico-chemical criteria of charge, M(r) (28,000) and hydrophobicity. It is also completely free of any proteolytic and bacteriolytic activities. The enzyme was classified as a class II chitinase on the basis of its N-terminal amino acid sequence up to the 30th residue. In agreement with this classification, the enzyme preparation hydrolyses chitinase substrates only very slowly and several free thiol functions are present in the polypeptide chain. These free thiol functions are buried, and to be available for titration with 2,2'-dipyridyldisulphide, the enzyme must be denatured. Unfolding of papaya chitinase requires particularly drastic conditions, not less than 4 M guanidinium hydrochloride at 25 degrees and pH 6.8.
Azarkan
M
M
Protein Chemistry Department, Faculty of Medicine, Free University of Brussels, Belgium.
Amrani
A
A
Nijs
M
M
Vandermeers
A
A
Zerhouni
S
S
Smolders
N
N
Looze
Y
Y
D000595:Amino Acid Sequence; D002688:Chitinases / Q000737:chemistry / Q000302:isolation & purification*; D002852:Chromatography, Ion Exchange; D019791:Guanidine; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification; D007840:Latex / Q000737:chemistry*; D008969:Molecular Sequence Data; D010944:Plants / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D013050:Spectrometry, Fluorescence
D007527:Isoenzymes; D007840:Latex; D002688:Chitinases; D019791:Guanidine