A potential non-Saccharomyces yeast species, identified as Wickerhamomyces anomalus SZ1 strain ( Gupta, et al., 2018), which gave even higher (33%) mannan oligosaccharides (MOS) than that obtained from the traditionally used Saccharomyces cerevisiae strain were selected for optimization of suitable media study for maximum yield of MOS by the one factor at a time (OFAT) method. Mannose was found to the best carbon source for optimum production of MOS, which significantly enhanced the yield by folds of MOS at 2% mannose concentration as in place of dextrose in YEPD media. Higher concentration of Mannose cannot significantly (p˂) enhance the MOS production further. 2% peptone and 1% yeast extract in combination were found to be the best nitrogen source. | Optimization of culture media and conditions enhances mannan oligosaccharides production of Wickerhamomyces anomalus SZ1 strain