Direct production of a genetically-encoded immobilized biodiesel catalyst

Scientific Reports
Bradley S HeaterMichael K Chan

Abstract

The use of immobilized enzymes as biocatalysts has great potential to improve the efficiency and environmental sustainability of many industrial processes. Here, we report a novel approach that allows for the direct production of a highly active immobilized lipase within the bacterium Bacillus thuringiensis. Cry3Aa-lipA crystals were generated by genetically fusing Bacillus subtilis lipase A to Cry3Aa, a protein that naturally forms crystals in the bacteria. The crystal framework significantly stabilized the lipase against denaturation in organic solvents and high temperatures, resulting in a highly efficient fusion crystal that could catalyze the conversion of triacylglycerols to fatty acid methyl ester biodiesel to near-completion over 10 cycles. The simplicity and robustness of the Cry-fusion crystal (CFC) immobilization system could make it an appealing platform for generating industrial biocatalysts for multiple bioprocesses.

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Citations

Nov 28, 2019·World Journal of Microbiology & Biotechnology·Takuya MatsumotoHiroyasu Ogino
Jul 12, 2020·Applied Microbiology and Biotechnology·Vera D JägerUlrich Krauss
May 15, 2020·Journal of the American Chemical Society·Bradley S HeaterMichael K Chan
May 25, 2021·Chemical Society Reviews·Roger A SheldonDean Brady
Jul 3, 2021·Toxins·Guillaume TetreauJacques-Philippe Colletier
Dec 21, 2021·Biomaterials Science·Mariko KojimaTakafumi Ueno

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Methods Mentioned

BETA
electron microscopy
electrophoresis
lipA
PCR
gel filtration

Software Mentioned

KaleidaGraph
FlexControl
Data Analysis
Synergy
GraphPad Prism
ProteinScape

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