Microbial detoxification of cyanide offered an inexpensive, safe, and viable alternative to physiochemical processes for the treatment of cyanide in industrial effluents or contaminated sites. This study involved isolation of novel strain with high resistance against cyanide toxicity and able to degrade the cyanide radical. The strain was isolated from rocky area and identified as Sphingobacterium multivorium using 16S ribosomal RNA. Resting pregrown cells were used in simple reaction mixture to avoid the complication associated with the media. One-gram fresh weight of this bacteria was able to remove 98.5% from 1.5 g/L cyanide which is a unique result. Factor affecting the biochemical process such as pH, temperature, agitation, glucose concentration was examined. The optimum conditions were, pH 6–7, 30–40°C, and 100–150 rpm shaking speed and 0.25% glucose. Furthermore, the cells were used after immobilization in polytetrafluoroethylene (PTFE) polymer. The PTFE is very safe carrier and the cells withstand the entrapment process and were able to remove 92% (1 g/L cyanide). The immobilized cells were used for six successive cycles with about 50% removal efficiency. The storage life extended to 14 days. No previous work studied the cyanide removal by Sphingobacterium spp. The strain showed good applicable characters.