Abstract

The ability of Commercial Native Linamarase (CNLIN) and Engineered Linamarase (GELIN) extracts from Saccharomyces cerevisiae to hydrolyse cassava linamarin was challenged. CNLIN acting as control was used together with GELIN extracts from Saccharomyces cerevisiae to evaluate the kinetic data for test enzymes at pH 3.5, 6.8 and 10.5, respectively and ambient temperature (35°C). Data obtained from the varying activity versus substrate concentrations were fitted with the Michaelis-Menten plots and Lineweaver-Burk model to obtain maximum velocity (V_max), affinity coefficient (K_m), physiological efficiency (K_m/V_max) and r² (linear regressing coefficient). The results indicated that the engineered linamarase conferred different enzyme kinetic data showing degradation of cassava linamarin by CNLIN and GELIN from Saccharomyces cerevisiae at the optimum pH and temperature. The relation was best described by the characteristic sigmoid Michaelis-Menten plots and Lineweaver- Burk model evidence from the high coefficient of linear regression (r²>0.976). V_max and K_m derived from the Lineweaver-Burk model varied from 10.0 to 13.0 μmol/min and 0.5 to 0.9 μM respectively for engineered enzymes and 0.0-10.0 mol/min and 0.0 to 0.9 μM respectively for CNLIN. The kinetics profiles of the studied enzymes showed their actions on cassava linamarin were influenced by degree of genetic manipulation, purification and pH at ambient temperature. The wide pH tolerance in the degradation of linamarin suggests a possible use of the engineered linamarase from Saccharomyces cerevisiae in detoxifying linamarin in cassava for the production of cyanide-free cassava-based food products.

Keywords:

Enzyme kinetics, linamarin, lineweaver-burk, michaelis-menten, pH, Saccharomyces cerevisiae,

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