The substrate specificity of LipBL and its variants toward triglycerides with different fatty acid chain lengths was determined using a pH-stat device (Methrom). Hydrolysis reactions with each substrate were carried out for 30 min and released fatty acids were neutralized with 0.1 M NaOH at 30 uC to maintain constant pH. Tributyrin (tri-C4), tricaproin (tri-C6), tricaprin (triC10), trilaurin (tri-C12), tripalmitin (tri-C16) and triolein (triC18 : 1) were used as substrates (20 mM) in Tris/HCl, pH 7.0, containing 100 mM CaCl2 and gum arabic (0.5 %, w/v)