Screening for new hydroxynitrilases from plants.
We established a simple HPLC method to determine the activity and stereochemistry of the chiral mandelonitrile synthesized from benzaldehyde and cyanide, and applied it to screen for hydroxynitrile lyase (HNL) activity of plant origin. A total of 163 species of plants among 74 families were examined for (R)- and (S)-HNL activities using the method. We discovered that homogenate of leaves of Baliospermum montanum shows (S)-HNL activity, while leaves and seeds from Passiflora edulis, and seeds from Eriobotrya japonica, Chaenomles sinensis, Sorbus aucuparia, Prunus mume, and Prunus persica show (R)-HNL activity. Partially purified (R)-HNLs from Passiflora edulis and Eriobotrya japonica acted not only on benzaldehyde but also on aliphatic ketone. The enantiomeric excess of (R)-methylpropylketone cyanohydrin synthesized from 2-pentanone using homogenate from leaves of Passiflora edulis was 87.0%, and that of (R)-mandelonitrile synthesized by homogenate from seeds of Eriobotrya japonica was 85.0%.
Asano
Yasuhisa
Y
Biotechnology Research Center, Toyama Prefectural University, Japan. asano@pu-toyama.ac.jp
Tamura
Ken'ichirou
K
Doi
Nobutaka
N
Ueatrongchit
Techawaree
T
H-Kittikun
Aran
A
Ohmiya
Tohru
T
D000097:Acetonitriles / Q000378:metabolism; D000619:Aminohydrolases / Q000737:chemistry* / Q000302:isolation & purification; D001547:Benzaldehydes / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification; D009682:Magnetic Resonance Spectroscopy; D009570:Nitriles / Q000378:metabolism; D010422:Pentanones / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry; D010944:Plants / Q000201:enzymology*; D011190:Potassium Cyanide / Q000378:metabolism; D011487:Protein Conformation; D027861:Prunus / Q000737:chemistry; D012639:Seeds / Q000737:chemistry; D013237:Stereoisomerism
D000097:Acetonitriles; D001547:Benzaldehydes; D007527:Isoenzymes; D009570:Nitriles; D010422:Pentanones; D010936:Plant Extracts; C430593:cyanohydrin; C045516:mandelonitrile; D000619:Aminohydrolases; C021867:nitrilase; C076402:2-pentanone; D011190:Potassium Cyanide; C032175:benzaldehyde