Cancer is a growing reason for death and morbidity worldwide. L-methioninase (EC 4.4.1.11) has a possible application against many types of cancers like glioblastoma, kidney, breast, lung, and colon cancer. The aim of the present study screening of L-methioninase production by bacteria isolated from sewage samples obtained from Makkah city in Saudi Arabia. Twelve bacterial were isolate from sewage samples. The bacterial isolate's ability for L-methioninase production was tested on a modified mineral salt M9 L-methionine agar medium using phenol red as the pH indicator. Selected Colonies with pink-red and yellow zone around as L-methionine degrading bacteria. The highest L-methioninase produced isolate identified based on16S rDNA sequencing, along with the biochemical characterization, and lastly completed by BLAST analysis by the structure of a phylogenetic tree. It was Escherichia coli WSM2 strain. The rRNA sequence of Escherichia coli WW11 was deposited to the gene bank (NCBI accession number MK072729). Results revealed using of organic byproduct residues, the high amount of the enzyme produced 0.128U/min/ml in cultures supplemented with pomegranate juice wastes with L-methionine and yeast extract after 48h of incubation, while the maximum amount of L-methioninase 0.237U/min/ml was produced by E. coli when 75% Whey and 25% and was used as fermentation media, incubated in a shaking incubator (150rpm)