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. 1992 Jan;98(1):12–16. doi: 10.1104/pp.98.1.12

Purification and Characterization of Cinnamyl Alcohol Dehydrogenase from Tobacco Stems 1

Claire Halpin 1,2, Mary E Knight 1,2, Jacqueline Grima-Pettenati 1,2, Deborah Goffner 1,2, Alain Boudet 1,2, Wolfgang Schuch 1,2
PMCID: PMC1080143  PMID: 16668601

Abstract

Cinnamyl alcohol dehydrogenase (CAD) is an enzyme involved in lignin biosynthesis. In this paper, we report the purification of CAD to homogeneity from tobacco (Nicotiana tabacum) stems. The enzyme is low in abundance, comprising approximately 0.05% of total soluble cell protein. A simple and efficient purification procedure for CAD was developed. It employs three chromatography steps, including two affinity matrices, Blue Sepharose and 2′5′ ADP-Sepharose. The purified enzyme has a specific cofactor requirement for NADP and has high affinity for coniferyl alcohol (Km = 12 micromolar) and coniferaldehyde (Km = 0.3 micromolar). Two different sized polypeptide subunits of 42.5 and 44 kilodaltons were identified and separated by reverse-phase HPLC. Peptide mapping and amino acid composition analysis of the polypeptides showed that they are closely related, although not identical.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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