Efficient molecular evolution to generate enantioselective enzymes using a dual-channel microfluidic droplet screening platform.
Nat Commun
; 9(1): 1030, 2018 03 12.
Article
em En
| MEDLINE
| ID: mdl-29531246
ABSTRACT
Directed evolution has long been a key strategy to generate enzymes with desired properties like high selectivity, but experimental barriers and analytical costs of screening enormous mutant libraries have limited such efforts. Here, we describe an ultrahigh-throughput dual-channel microfluidic droplet screening system that can be used to screen up to ~107 enzyme variants per day. As an example case, we use the system to engineer the enantioselectivity of an esterase to preferentially produce desired enantiomers of profens, an important class of anti-inflammatory drugs. Using two types of screening working modes over the course of five rounds of directed evolution, we identify (from among 5 million mutants) a variant with 700-fold improved enantioselectivity for the desired (S)-profens. We thus demonstrate that this screening platform can be used to rapidly generate enzymes with desired enzymatic properties like enantiospecificity, chemospecificity, and regiospecificity.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Evolução Molecular Direcionada
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Archaeoglobus fulgidus
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Proteínas Arqueais
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Microfluídica
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Esterases
Tipo de estudo:
Diagnostic_studies
/
Evaluation_studies
/
Screening_studies
Idioma:
En
Ano de publicação:
2018
Tipo de documento:
Article