Part:BBa_K4395013
NADPH-dependent aldehyde reductase YahK catalyzes the reduction of a wide range of aldehydes into their corresponding alcohols. Has a strong preference for NADPH over NADH as the electron donor. Cannot use a ketone as substrate. Is a major source of NADPH-dependent aldehyde reductase activity in E.coli[1]. In addition, this enzyme has been actively expressed in E.coli[2].These characteristics of RsAs attract us and thus we choose it as aldehyde reductase of our experiment.
Reference: 1.Pick A, Rühmann B, Schmid J, Sieber V. Novel CAD-like enzymes from Escherichia coli K-12 as additional tools in chemical production. Appl Microbiol Biotechnol. 2013;97(13):5815-5824. doi:10.1007/s00253-012-4474-5 2.Kramer L, Le X, Rodriguez M, Wilson MA, Guo J, Niu W. Engineering Carboxylic Acid Reductase (CAR) through a Whole-Cell Growth-Coupled NADPH Recycling Strategy. ACS Synth Biol. 2020;9(7):1632-1637. doi:10.1021/acssynbio.0c00290
Contribution From NJTech-China-B 2023
Group:iGEM NJTech-China-B
Author: Yao Yao
Characterization from iGEM23-NJTech-China-B
1.1 The transformation of palsmid pRSFDuet-YahK into E. coli BL21(DE3)
The gene of YahK was amplified from the genome of E. coli MG1655, and integrated into pRSFDuet-1 vector to obtain the plasmid pRSFDuet-YahK. By sequencing, the correct plasmid pRSFDuet-YahK was then transformed to E. coli BL21(DE3) (Figure 1).
YahK
1.1 The transformation of palsmid pRSFDuet-YahK into E. coli BL21(DE3)
The gene of YahK was amplified from the genome of E. coli MG1655, and integrated into pRSFDuet-1 vector to obtain the plasmid pRSFDuet-YahK. By sequencing, the correct plasmid pRSFDuet-YahK was then transformed to E. coli BL21(DE3) (Figure 1).
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