Co-expression of the recombined alcohol dehydrogenase and glucose dehydrogenase and cross-linked enzyme aggregates stabilization. (January 2017)

• Record Type: Journal Article
• Authors: Hu, Xiaozhi; Liu, Liqin; Chen, Daijie; Wang, Yongzhong; Zhang, Junliang; Shao, Lei
• Is Part Of: Bioresource technology. Volume 224(2017)
• Page Start: 531
• Page End: 535
• Publication Date: 2017-01
• Subjects: Recombinant alcohol dehydrogenase -- Recombinant glucose dehydrogenase -- co-CLEAs -- Crizotinib -- (S)-1-(2, 6-dichloro-3-fluorophenyl) phenethyl alcohol; Biomass -- Periodicals; Biomass energy -- Periodicals; Bioremediation -- Periodicals; Agricultural wastes -- Periodicals; Factory and trade waste -- Periodicals; Organic wastes -- Periodicals; Bioénergie -- Périodiques; Déchets agricoles -- Périodiques; Déchets industriels -- Périodiques; Déchets organiques -- Périodiques; Déchets (Combustible) -- Périodiques; 662.88
• DOI: 10.1016/j.biortech.2016.10.076
• Languages: English
• ISSNs: 0960-8524

Abstract

Highlights: The chiral precursor of Crizotinib can be prepared by the coupling enzyme reaction. Alcohol dehydrogenase (ADH) and glucose dehydrogenase (GDH) were co-expressed. The ADH/GDH co-crosslinked enzyme aggregates were prepared. The ADH/GDH co-CLEAs shows increased stability and reusability. Abstract: As the key chiral precursor of Crizotinib (S)-1-(2, 6-dichloro-3-fluorophenyl) phenethyl alcohol can be prepared from 1-(2, 6-dichloro-3-fluorophenyl) acetophenone by the reductive coupling reactions of alcohol dehydrogenase (ADH) and glucose dehydrogenases (GDH). In this work the heterologous expression plasmids harbouring the encoding genes of ADH and GDH were constructed respectively and co-expressed in the same E. coli strain. After optimization, a co-cross-linked enzyme aggregates (co-CLEAs) of both ADH and GDH were prepared from crude enzyme extracts by cross-linking with the mass ratio of Tween 80, glutaraldehyde and total protein (0.6:1:2) which rendered immobilized biocatalysts that retained 81.90% (ADH) and 40.29% (GDH) activity retention. The ADH/GDH co-CLEAs show increased thermal stability and pH stability compared to both enzymes. The ADH/GDH co-CLEAs also show 80% (ADH) and 87% (GDH) residual activity after seven cycles of repeated use. These results make the ADH/GDH co-CLEAs a potential biocatalyst for the industrial preparation of (S)-1-(2, 6-dichloro-3-fluorophenyl) phenethyl alcohol.