Dual bioimprinting of Thermomyces lanuginosus lipase for synthesis of biodiesel

•High activity TLL is made by dually bioimprinting with substrate alcohol and a surfactant.•TLL (28 U/g of oil) bioimprinted with only the surfactants could yield 99% biodiesel from soybean oil in about 4 h.•Dually bioimprinted TLL (only 1.4 U/g of oil) was able to yield 99% biodiesel within 48 h

Use of biodiesel as an alternative to non-renewable sources of energy has become an attractive option in recent years. The enzymatic synthesis of biodiesel by transesterification of fats/oils with an alcohol is a much more sustainable route than the chemical method. However, cost effectiveness of the enzymatic route is a major barrier in its commercialization. In this work, a high activity biocatalyst design of Thermomyces lanuginosus lipase is made by dually bioimprinting it with substrate and a surfactant (which is believed to open up the lid covering the active site of the lipase) during precipitation of the lipase in organic solvent. When the lipase was bioimprinted with only the surfactants, 28 U of the enzyme/g of oil could yield 99% biodiesel from soybean oil in about 4 h. However, when dually bioimprinted even very low enzyme load 1.4 U/g of oil, yielded 99% biodiesel within 48 h.

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