Production, Purification, and Characterization of Extracellular Lipases from <i>Hyphopichia wangnamkhiaoensis</i> and <i>Yarrowia deformans</i>

The efficient production of microbial lipases from organic wastes has garnered great interest because of the diverse and potential biotechnological applications of these enzymes. However, the extracellular lipases from the novel yeast strains <i>Hyphopichia wangnamkhiaoensis</i> and <i>Yarrowia deformans</i> remain uncharacterized. Thus, this study aimed to investigate the characteristics and production of lipases from both yeasts. Lipases from <i>H. wangnamkhiaoensis</i> and <i>Y. deformans</i> were purified and biochemically characterized, and their production was measured in batch cultures with olive oil (reference), waste cooking oil, and glycerol as substrates. The purified lipases from <i>H. wangnamkhiaoensis</i> and <i>Y. deformans</i> had molecular weights of approximately 33 and 45 kDa, respectively. Their activities on <i>p</i>-nitrophenyl palmitate were optimal at pH 8.0 and 40 °C. Moreover, the activities of the lipases were inhibited by ethylenediaminetetraacetic acid, phenylmethylsulfonyl fluoride, and 4-(2-aminoethyl)benzenesulfonyl fluoride, and were reactivated by Ca²⁺ and Mg²⁺, indicating that both lipases are metalloenzymes and serine-type enzymes. The lipases were more tolerant to hydrophilic solvents than to hydrophobic solvents, and they followed Michaelis–Menten kinetics. Among the various substrates used, waste cooking oil yielded the highest lipase production and productivity. These results indicate that <i>H. wangnamkhiaoensis</i> and <i>Y. deformans</i> are suitable and potential candidates for lipase production.