This study explored biodiesel production from a low cost, abundant, non-edible lignocellulosic biomass from aqueous extract of Cassia fistula L. (CAE) fruit pulp. The CAE was utilized as substrate for cultivating novel oleaginous yeast Rhodosporidium kratochvilovae HIMPA1. This oleaginous yeast accumulates high amount of triacylglycerides as large intracellular lipid droplets (4.35 +/- 0.54 mu m) using CAE as sole nutritional source. Total lipids (4.86 +/- 0.54 g/l) with lipid content of 53.18% (w/w) were produced by R. kratochvilovae HIMPA1 on CAE. The FAME profile obtained revealed palmitic acid (C-16:0) 43.06%, stearic acid (C-18:0) 28.74%, and oleic acid (C-18:1) 17.34% as major fatty acids. High saturated fatty acids content (72.58%) can be blended with high PUFA feedstocks to make it an industrially viable renewable energy product. (C) 2015 Elsevier Ltd. All rights reserved.