Production of Candida antarctica Lipase B Gene Open Reading Frame Using Automated PCR Gene Assembly Protocol on Robotic Workcell and Expression in an Ethanologenic Yeast for Use as Resin-Bound Biocatalyst in Biodiesel Production

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Kenneth M. Bischoff ◽  
Marjorie A. Jones ◽  
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John M. Woodley ◽  
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Csaba Paizs ◽  
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Maisa Pessoa Pinheiro ◽  
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Denise Maria Guimarães Freire ◽  
Ivanildo José da Silva ◽  
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Ferenc Fülöp ◽  
Liisa T Kanerva

The Candida antarctica lipase B-catalyzed reactions of five β-amino esters with neat butyl butanoate and with 2,2,2-trifluoroethyl butanoate in diisopropyl ether were studied, as were the reactions of the same β-amino esters and their N-butanamides with neat butanol. The possibility for sequential resolution, where the amino and ester functions of the substrate both react with an achiral butanoate, became less likely with increasing size of the substrate from ethyl 3-aminobutanoate (1a) to pentanoate (1b) or larger. On the other hand, the alcoholyses of N-acylated β-amino esters successfully proceeded in butanol with E > 100. Gram-scale resolution of the N-butanoylated 1a was performed to demonstrate the usefulness of the method. Key words: lipase, interesterification, acylation, alcoholysis, resolution, β-amino esters.


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