Linamarase production by some microbial isolates and a comparison of the rate of degradation of cassava cyanide by microbial and cassava linamarases

Abstract

Production of linamarase and the effects of media composition on enzyme production were studied. A total of eight linamarase-producing bacteria were isolated from fermenting cassava tubers and soil samples. Selection of the isolates was based on their fast growth in media containing 800 mg/L potassium cyanide solution. Eight of the isolates which showed very fast growth in the growth medium as demonstrated by increase in their optical density readings to at least 0.6 in the cyanide containing media were selected for further studies. The isolates were identified as Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus amylovorus, Lactobacillus cellobiosus, Leuconostoc mesenteroides, Pseudomonas stutzeri, Bacillus pumilus and Bacillus subtilis. All the isolates were grown in media containing Tween 80 solution and in control media without the surfactant. Best enzyme activity of 6.82 U/mL was obtained in the medium containing Tween 80 solution and Lactobacillus fermentum as the test bacterium. Comparatively, linamarase production by the isolates in media without Tween 80 showed lower enzyme productivity. Cassava endogenous and microbial enzymes were tested for their abilities to hydrolyze cyanide in cassava flour samples pretreated to either remove the endogenous or microbial enzyme. Residual cyanide in cassava flour samples treated with linamarase of Lactobacillus plantarum was undetected in 30 h, while in contrast, the residual cyanide in cassava flour samples treated with endogenous linamarase was 0.39 mg/10 g cassava flour after 80 h. Residual cyanide in the untreated control sample was 1.98 mg HCN /10 g cassava flour after 80 h. The results from this finding demonstrated improved cassava cyanide degradation with microbial linamarase as compared to endogenous cassava linamarase. Massive inoculation of fermenting cassava tubers with the isolates reported in this study would enable better control of the cassava fermentation process and may lead to the production of standardized and non-toxic cassava food products.