Screening Assay for Directed Evolution of Chitin Deacetylases: Application to Vibrio cholerae Deacetylase Mutant Libraries for Engineered Specificity

Sergi Pascual, Antoni Planas

Producció científica: Article en revista indexadaArticleAvaluat per experts

8 Cites (Scopus)

Resum

Not only the degree of acetylation but also the pattern of acetylation of chitosans and chitooligosaccharides (COS) appear to be critical for their biological activities. Protein engineering may expand the toolbox of chitin deacetylases (CDAs) with defined specificities for the enzymatic production of partially deacetylated COS for biotech and biomedical applications. A high-throughput screening (HTS) assay for screening directed evolution libraries is reported. It is based on a fluorescence monitoring assay of the deacetylase activity on COS substrates after capturing the expressed enzyme variants fused to a chitin binding module with chitin-coated magnetic beads. The assay is applied to the screening of random libraries of a Vibrio cholera CDA for increased activity on longer COS substrates.

Idioma originalAnglès
Pàgines (de-a)10654-10658
Nombre de pàgines5
RevistaAnalytical Chemistry
Volum90
Número18
DOIs
Estat de la publicacióPublicada - 18 de set. 2018

Fingerprint

Navegar pels temes de recerca de 'Screening Assay for Directed Evolution of Chitin Deacetylases: Application to Vibrio cholerae Deacetylase Mutant Libraries for Engineered Specificity'. Junts formen un fingerprint únic.

Com citar-ho