Abstract: Objective: The study aimed to characterize lactic acid bacteria (LAB) in Kenyan spontaneously fermented milk, Amabere amaruranu for identification of starter cultures. Materials and Methods: Twenty four samples were collected from Kisii County, Kenya and cultured on MRS agar to isolate LAB. Thirty seven isolates were selected for phenotypic (Gram staining and catalase test) and physiological [growth at 15 and 45°C, on 4 and 6.5% (w/v) NaCl] characterization and identification by 16S rRNA gene sequencing. The obtained sequences were compared to DNA sequences in the GenBank by the BLASTN program on NCBI website. The sequences were used to generate phylogenetic tree on MEGA 6.0 software. Lactic acid production and pH change capability in milk were determined as technological characteristics of the isolates. Results: All isolates were Gram positive and catalase negative with shapes of rods or cocci. The isolates clustered into three groups; (1) those didn’t grow at 45°C but grew at 15°C 24 were considered mesophilic Lactobacilli, (2) One cocci that grew at 15°C but not 45°C and 6.5% (w/v) NaCl was considered mesophilic Lactococci and (3) Those 7 which grew at 45°C, 15°C and 6.5% NaCl, were considered Enterococci. DNA sequencing identified 5 isolates; two Lactobacillus plantarum spp., two Epicoccum spp. and one (1) Staphylococcus warneri spp. L. plantarum spp. coagulated milk and attained 0.73% titratable acidity and pH of 4.48 in 24 h. Conclusion: Lactobacillus plantarum spp. was identified from Amabere amaruranu and its technological characteristics demonstrated, hence it could be used as a starter culture for product development.