BioChem - DOS

Technical Considerations for Scale-Up of Imine-Reductase-Catalyzed Reductive Amination: A Case Study

Organic process research & development

Bornadel, Amin; Bisagni, Serena; Pushpanath, Ahir; Montgomery, Sarah L.; Turner, Nicholas J.; Dominguez, Beatriz

<P>Imine reductases (IREDs) have attracted increasing attention as novel biocatalysts for the synthesis of various cyclic and acyclic amines. Herein a number of guidelines and considerations toward the development and scale-up of IRED catalyzed reactions have been determined based on the reductive amination of cyclohexanone (<B>1</B>) with cyclopropylamine (<B>2</B>). A Design of Experiments (DoE) strategy has been followed to study the different reaction parameters, facilitating resource-efficient and informative screening. Enzyme stability was identified to be the limiting factor. By moving from batch to fed-batch, it was possible to double the concentration of the substrate and turnover number (TON). Kinetic studies revealed that IRED-33 was the best enzyme for the reaction with respect to both activity and stability. Under the optimal reaction conditions, it was possible to react <B>1</B> and <B>2</B> at 750 mM concentration and reach 100% conversion to the desired amine (>90% isolated yield) in the space of 8 h. Hence, excellent volumetric productivity of 12.9 g L<SUP>-1</SUP> h<SUP>-1</SUP> and TON above 48 000 were achieved.</P><BR>[FIG OMISSION]</BR>

2019

American Chemical Society

1083-6160

1520-586x

23

6

pp.1262-1268

10.1021/acs.oprd.9b00123

http://click.ndsl.kr/servlet/OpenAPIDetailView?keyValue=05787966&target=NART&cn=NART97805995

IRED-catalyzed reaction; reductive amination; enzyme kinetics; fed-batch reaction; biocatalytic process development