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Methods for rapid screening and isolation of bacteria producing acidic lipase: feasibility studies and novel activity assay protocols
Authors:Chien-Hung Liu  Wen-Ming Chen  Jo-Shu Chang
Affiliation:(1) Department of Chemical Engineering , National Cheng Kung University , Tainan , 701 , Taiwan;(2) Department of Seafood Science, National Kaohsiung Marine University, Kaohsiung, Taiwan
Abstract:Acidic lipase finds its commercial values in medical applications and bioremediation of food wastes. In this work, approaches for rapid screening of lipase-producing bacteria were developed and the feasibility assessment of the screening methods was performed. From food waste samples, the proposed screening procedures allowed isolation of sixteen pure bacterial strains expressing higher lipase activity at acidic pH (pH 6.0) than at alkaline pH (pH 9.0). To enhance the accuracy of lipase activity determination under acidic conditions, a novel assay procedure was also developed by deactivating lipase activity by microwave treatment prior to back titration. This additional step could minimize interferences arising from residual lipase activity during conventional direct back-titration methods in measuring lipase activity at acidic pH. Using the four strategies proposed in this work, the best acidic-lipase-producing isolate was obtained by strategy C (SSC) and was identified as Aeromonas sp. C14, displaying an optimal lipase activity of 0.7 U/ml at an acidic pH of 6.0.
Keywords:Back-titration  Enzyme assay  Food wastes  Lipase  Microwave treatment  pH-stat titrimetry  Strain isolation
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