Aims: Short-chain fructo-oligosaccharides (scFOSs) are good prebiotics that enhance the growth of probiotic bacteria. The aim of this study is to determine the effect of scFOSs produced by levan hydrolysis using recombinant endo-levanase from B. lehensis G1 on the growth of probiotics isolated from commercially cultured milk drinks. Methodology and results: Two probiotic bacteria, Lactobacillus casei and L. rhamnosus, were isolated from commercially cultured milk drinks. ScFOSs were produced by levan hydrolysis using recombinant endo-levanase from B. lehensis G1. The scFOS and levan (control) were added independently to the growth medium, and the growth rates of the probiotic bacteria were determined. Results showed that the growth rate of L. casei decreased in the presence of levan compared with the control medium but increased by approximately 20% when supplemented with scFOS produced by Levblg1-N28S. Similarly, the growth rate of L. rhamnosus increased by approximately 20% when supplemented with scFOS produced by Levblg1 and Levblg1-N28S. Conclusion, significance and impact of study The scFOSs produced by the enzymatic hydrolysis of levan using a recombinant endo-levanase from B. lehensis G1 have significant potential as prebiotics because they were able to promote the growth of the probiotic bacteria.