A soluble beta-cyanoalanine synthase from the gut of the variegated grasshopper Zonocerus variegatus (L.).
Beta-cyanoalanine synthase (beta-cyano-l-alanine synthase; l-cysteine: hydrogen sulphide lyase (adding hydrogen cyanide (HCN)); EC 4. 4.1.9) was purified from the cytosolic fraction of the gut of grasshopper Zonocerus variegatus (L.) by ion-exchange chromatography on DEAE-Cellulose and gel filtration on Sephadex G-100 columns. The crude enzyme had a specific activity of 2.16nmol H2S/min/mg. A purified enzyme with a specific activity, which was seventeen times higher than that of the crude extract, was obtained. A molecular weight of about 55.23+/-1.00Kd was estimated from its elution volume on Sephadex G-100. The fraction when subjected to sodium dodecyl sulphate-polyacrylamide elel electrophoresis revealed the presence of a protein band with Mr of 23.25+/-0.25Kd. The enzyme exhibited Michaelis-Menten kinetics having Km of 0.38mM for l-cysteine and Km of 6.25mM for cyanide. The optimum temperature and pH for activity were determined to be at 30 degrees C and pH 9.0, respectively. This enzyme might be responsible for the ability to detoxify cyanide in this insect pest and hence its tolerance of the cyanogenic cassava plant. Biophysical, biochemical and kinetic properties of this enzyme, which will reveal how this ability can possibly be compromised by enzyme inhibition, may lead, in the long term, to the potential use of this enzyme as drug target for pest control.
Ogunlabi
Olugbenga Owolabi
OO
Department of Biochemistry, Olabisi Onabanjo University Remo Campus, Ikenne, Ogun State, Nigeria.
Agboola
Femi Kayode
FK
D000818:Animals; D041981:Gastrointestinal Tract / Q000737:chemistry; D006110:Grasshoppers / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008190:Lyases / Q000737:chemistry / Q000302:isolation & purification*; D002365:Manihot / Q000469:parasitology; D008970:Molecular Weight; D013696:Temperature
D008190:Lyases; C019955:beta-cyanoalanine synthase