biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 50:48-54, 2006 | DOI: 10.1007/s10535-005-0073-y

Chalcone synthase homologues from Humulus lupulus: some enzymatic properties and expression

P. Novak1,2, K. Krofta3, J. Matousek1,*
1 Institute of Plant Molecular Biology AS CR, Ceske Budejovice, Czech Republic
2 Biological Faculty, South Bohemian University, Ceske Budejovice, Czech Republic
3 Hop Research Institute, Zatec, Czech Republic

The enzymatic properties of four chalcone synthase homologues CHS_H1, VPS, CHS 2 and CHS 4 from Humulus lupulus L. were investigated after heterologous expression in Escherichia coli. It was found that both VPS and CHS_H1 can utilize isovaleryl-CoA and isobutyryl-CoA as substrates producing compounds with positions in thin layer chromatography characteristic for phloroisovalerophenone and phloroisobutyrophenone. These reactions are accompanied by the formation of associated byproducts. The formation of naringenin chalcone can be catalyzed primarily by CHS_H1. Comparatively the ability of VPS to perform chalcone synthase reaction is very limited. Since only CHS_H1 has true chalcone synthase activity, this enzyme can be considered a key enzyme in prenylflavonoid biosynthesis. Both CHS 2 and CHS 4 utilize isovaleryl-CoA and isobutyryl-CoA as substrates, but the reactions were prematurely terminated. In comparison with VPS and CHS_H1, the optimum pH of CHS 2 was shifted to lower value. High expression of chalcone synthase-like genes were found in maturating hop cones of cultivars with high bitter acid content (Agnus, Magnum, Target) by Northern and Western blotting using probes specific for vps, chs_H1, chs 4 and polyspecific serum risen against recombinant protein CHS4, respectively. It was also found that these cultivars maintained expression of CHS homologues for a longer period of time during cone development in contrast to time-limited expression of CHS homologues in cultivars with low bitter acids content.

Keywords: bitter acids; hop; prenylflavonoids; valerophenone synthase
Subjects: bitter acids; chalcone synthase; hop; Humulus lupulus; lupulin; Northern blot analysis; prenylflavonoids; valerophenone synthase

Received: June 2, 2004; Accepted: October 27, 2004; Published: March 1, 2006  Show citation

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Novak, P., Krofta, K., & Matousek, J. (2006). Chalcone synthase homologues from Humulus lupulus: some enzymatic properties and expression. Biologia plantarum, 50(1), 48-54. doi: 10.1007/s10535-005-0073-y
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