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@ARTICLE{Bollinger:888809,
author = {Bollinger, Alexander and Molitor, Rebecka and Thies,
Stephan and Koch, Rainhard and Coscolín, Cristina and
Ferrer, Manuel and Jaeger, Karl-Erich},
title = {{O}rganic-{S}olvent-{T}olerant {C}arboxylic {E}ster
{H}ydrolases for {O}rganic {S}ynthesis},
journal = {Applied and environmental microbiology},
volume = {86},
number = {9},
issn = {1098-5336},
address = {Washington, DC [u.a.]},
publisher = {Soc.},
reportid = {FZJ-2020-05223},
pages = {e00106-20},
year = {2020},
abstract = {Biocatalysis has emerged as an important tool in synthetic
organic chemistry enabling the chemical industry to execute
reactions with high regio- or enantioselectivity and under
usually mild reaction conditions while avoiding toxic waste.
Target substrates and products of reactions catalyzed by
carboxylic ester hydrolases are often poorly water soluble
and require organic solvents, whereas enzymes are evolved by
nature to be active in cells, i.e., in aqueous rather than
organic solvents. Therefore, biocatalysts that withstand
organic solvents are urgently needed. Current strategies to
identify such enzymes rely on laborious tests carried out by
incubation in different organic solvents and determination
of residual activity. Here, we describe a simple assay
useful for screening large libraries of carboxylic ester
hydrolases for resistance and activity in water-miscible
organic solvents. We have screened a set of 26 enzymes, most
of them identified in this study, with four different
water-miscible organic solvents. The triglyceride tributyrin
was used as a substrate, and fatty acids released by
enzymatic hydrolysis were detected by a pH shift indicated
by the indicator dye nitrazine yellow. With this strategy,
we succeeded in identifying a novel highly
organic-solvent-tolerant esterase from Pseudomonas
aestusnigri. In addition, the newly identified enzymes were
tested with sterically demanding substrates, which are
common in pharmaceutical intermediates, and two enzymes from
Alcanivorax borkumensis were identified which outcompeted
the gold standard ester hydrolase CalB from Candida
antarctica},
cin = {IMET},
ddc = {570},
cid = {I:(DE-Juel1)IMET-20090612},
pnm = {581 - Biotechnology (POF3-581)},
pid = {G:(DE-HGF)POF3-581},
typ = {PUB:(DE-HGF)16},
pubmed = {32111588},
UT = {WOS:000529303800005},
doi = {10.1128/AEM.00106-20},
url = {https://juser.fz-juelich.de/record/888809},
}