Analysis of the chalcone synthase from Humulus lupulus L. and biotechnology aspects of medicinal hops

Matoušek, J.; Novák, Petr; Patzak, Josef; Břı́za, J.; Krofta, Karel

doi:10.17221/4200-pse

Cited by 4 publications

(1 citation statement)

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“…Partial sequences of VPS and CHS2 from the hop cone's secretory glands (also called lupulin glands) were obtained. It is known that VPS and CHS1 are expressed in lupulin glands ( Okada and Ito, 2001 ; Matousek et al , 2002a , b ) and the presence of a gene family of VPS, as well as one of CHS, has been suggested. Supplementary Figure S2 shows the strategy to obtain the full-length cDNAs of the likely PKS gene(s) differing from the earlier CHS-type PKS gene.…”

Section: Resultsmentioning

confidence: 99%

In silicio expression analysis of PKS genes isolated from Cannabis sativa L.

2010

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Cannabinoids, flavonoids, and stilbenoids have been identified in the annual dioecious plant Cannabis sativa L. Of these, the cannabinoids are the best known group of this plant's natural products. Polyketide synthases (PKSs) are responsible for the biosynthesis of diverse secondary metabolites, including flavonoids and stilbenoids. Biosynthetically, the cannabinoids are polyketide substituted with terpenoid moiety. Using an RT-PCR homology search, PKS cDNAs were isolated from cannabis plants. The deduced amino acid sequences showed 51%-73% identity to other CHS/STS type sequences of the PKS family. Further, phylogenetic analysis revealed that these PKS cDNAs grouped with other non-chalcone-producing PKSs. Homology modeling analysis of these cannabis PKSs predicts a 3D overall fold, similar to alfalfa CHS2, with small steric differences on the residues that shape the active site of the cannabis PKSs.

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Section: Resultsmentioning

confidence: 99%

In silicio expression analysis of PKS genes isolated from Cannabis sativa L.

2010

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

Novák¹,

Krofta²,

Matoušek³

2006

Biol. plant.

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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 timelimited expression of CHS homologues in cultivars with low bitter acids content.

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