Title : Purification, characterization, and cloning of alpha-hydroxynitrile lyase from cassava (Manihot esculenta Crantz) - Hughes_1994_Arch.Biochem.Biophys_311_496 |
Author(s) : Hughes J , Carvalho FJ , Hughes MA |
Ref : Archives of Biochemistry & Biophysics , 311 :496 , 1994 |
Abstract :
alpha-Hydroxynitrile lyase (HNL, acetone cyanohydrin lyase, EC 4.1.2.37) was purified to homogeneity from young leaves of the cyanogenic tropical crop plant cassava (Manihot esculenta Crantz). The purified protein is a homo-trimer with a subunit relative molecular mass of 28,500 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The active protein is not glycosylated and does not contain a flavin group. HNL exhibits complex kinetics which vary according to substrate concentration and may be related to aggregation of the enzyme. HNL activity against two natural substrates, acetone cyanohydrin and 2-butanone cyanohydrin, and one nonphysiological substrate, 2-pentanone cyanohydrin, was demonstrated. N-terminal and internal peptide sequences, obtained from HNL digested with the endoproteinase Glu-C, were used to design degenerate oligonucleotide primers for polymerase chain reaction with single-strand cDNA, using purified mRNA from cotyledons as template. The resulting DNA fragment was used to probe a cassava cotyledon cDNA library. Four cDNA clones were isolated, sequenced, and shown to contain derived amino acid sequences identical to those obtained from the purified protein. |
PubMedSearch : Hughes_1994_Arch.Biochem.Biophys_311_496 |
PubMedID: 8203915 |
Gene_locus related to this paper: manes-hnl |
Gene_locus | manes-hnl |
Hughes J, Carvalho FJ, Hughes MA (1994)
Purification, characterization, and cloning of alpha-hydroxynitrile lyase from cassava (Manihot esculenta Crantz)
Archives of Biochemistry & Biophysics
311 :496
Hughes J, Carvalho FJ, Hughes MA (1994)
Archives of Biochemistry & Biophysics
311 :496