Part:BBa_K1658006

CAD1 Enzyme

Involved in lignin biosynthesis. Catalyzes the final step specific for the production of lignin monomers. Catalyzes the NADPH-dependent reduction of coniferaldehyde, 5-hydroxyconiferaldehyde, sinapaldehyde, 4-coumaraldehyde, caffeyl aldehyde, cinnamyl alcohol to their respective aldehyde and alcohols

The predicted three-dimensional structure of CAD1. Data were analysed by the SWISS-MODEL software, and the figure was prepared with the program MOLMOL. a-helices are indicated in red and yellow, b-sheets by arrows, and turns and coils by curled lines. The NADPH binding and substrate binding sites are also indicated.

Design Notes

This enzyme belongs to the family of oxidoreductases, specifically those acting on the CH-OH group of donor with NAD+ or NADP+ as acceptor. The systematic name of this enzyme class is cinnamyl-alcohol:NADP+ oxidoreductase. Other names in common use include cinnamyl alcohol dehydrogenase, and CAD. This enzyme participates in phenylpropanoid biosynthesis. Chemical Reaction of CAD: Cinnamyl alcohol + NADP+ = cinnamaldehyde + NADPH. Cofactor :Zn2+ Note: Binds 2 Zn2+ ions per subunit. Involved in lignin biosynthesis. Catalyzes the final step specific for the production of lignin monomers. Catalyzes the NADPH-dependent reduction of coniferaldehyde, 5-hydroxyconiferaldehyde, sinapaldehyde, 4-coumaraldehyde, caffeyl aldehyde and cinnaml alcohol to their respective alcohols Functional expression of the Ocimum CAD1in E.coli PCR was used to introduce EcoRI , Xbal, Spel, Notl and Pstl at side ends of CAD1 coding region. The amplified product was single and double digested with EcoRl and Notl and the resultant fragment was cloned into the same sides ofthe puC57 vector. The engineered plasmid was transformed into E.coli BL 2l (DE3) cells which has lack of nudeases and proteinoses and ideal for expression.

Source

Ocimum basilicum is sweet basil (Eukaryota, Viridiplantae, Streptophyta, Embryophyta, Tracheophyta, Spermatophyta, Magnoliophyta, eudicotyledons, Gunneridae, Pentapetalae, asterids, lamiids, Lamiales, Lamiaceae,Nepetoideae, Ocimeae, Ocimum) AY879285.1 Cinnamyl alcohol dehydrogenase (CAD, EC 1.1.1.195) is an NADP(H) specific oxidoreductase catalysing the reversible conversion of cinnamyl aldehydes to the corresponding alcohols or monolignols (Mansel et al., 1974; Wyrambik and Griesebach, 1975; Boudet et al., 1995). Polymerization of the three monolignols: /?-coumaryl, sinapyl and coniferyl alcohols lead to lignin. The enzyme is closely related to lignification, and chemical inhibition of its activity reduces the synthesis of lignins (Moesbacher et al., 1990; Mauch-Mani and Slusarenko, 1996).

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

References

Iijima, Yoko, and Guodong Wang. "Analysis of the Enzymatic Formation of Citral in the Glands of Sweet Basil." Archives of Biochemistry and Biophysics 21 July 2005: 145. Print Ma, Q.(2010). Functional analysis of a cinnamyl alcohol dehydrogenase involved in biosynthesis in wheat. Journal of Experiment Botany, 2735-2744.