Kinetin-induced caffeic acid O -methyltransferases in cell suspension cultures of Vanilla planifolia Andr. and isolation of caffeic acid O -methyltransferase cDNAs
Identifieur interne : 003B52 ( Main/Exploration ); précédent : 003B51; suivant : 003B53Kinetin-induced caffeic acid O -methyltransferases in cell suspension cultures of Vanilla planifolia Andr. and isolation of caffeic acid O -methyltransferase cDNAs
Auteurs : Zhong-Tian Xue [Suède] ; Peter E. Brodelius [Suède]Source :
- Plant Physiology and Biochemistry [ 0981-9428 ] ; 1998.
English descriptors
- KwdEn :
- Teeft :
- Acid, Alfalfa, Amino, Amino acid residues, Amino acid sequences, Bacterial extracts, Biochem, Biosynthesis, Blue cells, Brodelius, Brodelius phenylpropanoid metabolism, Caffeic, Caffeic acid, Caffeic acid activity, Caomt, Caomt edna clones, Caomt genes, Caomts, Ccoaomt, Cdna, Cdna clone, Cell cultures, Cell suspension cultures, Clone, Coding region, Coli, Edna, Encoding, Enzyme, Ferulic, Ferulic acid, General phenylpropanoid pathway, High homology, Homology, Iptg, Isoferulic, Isoferulic acid, Kinetin, Kinetin treatment, Lignin, Lignin biosynthesis, Lignin precursors, Methylation, Nucleotide, Omts, Pathway, Pcomtl, Phenylpropanoid, Phenylpropanoid pathway, Physiol, Planifolia, Plant cell, Plant omts, Plant physiol, Populus, Primary structure, Second type, Sequencing, Sinapic acid, Suspension cultures, Vanilla, Vanilla cells, Vanilla planifolia, Vanilla planifolia andr, Vanillic, Vanillic acid.
Abstract
Abstract: Two different S-adenosyl-L-methionine:caffeic acid methyltransferases are induced in kinetin-treated cell cultures of Vanilla planifolia. Upon addition of kinetin to the culture, maximum caffeic acid 4-O-methyltransferase and caffeic acid 3-O-methyltransferase activity is obtained after around 2 and 40 h, respectively. The former enzyme is involved in the biosynthesis of vanillic acid and the latter enzyme is co-induced with other enzymes of the general phenylpropanoid pathway and is involved in the biosynthesis of lignin precursors. A number of cDNA clones encoding S-adenosyl-L-methionine:caffeic acid-methyltransferases were isolated by heterologous probe screening of a λZapII cDNA library constructed from mRNA of a kinetin-treated cell suspension culture of V. planifolia. A full-length cDNA clone, Vpomt35, contains a 1 089-bp open reading frame coding for 363 amino acid residues, a 25-bp 5′-end sequence and a 214-bp 3′-end non-coding sequence. The deduced amino acid sequence of Vpomt35 revealed 56 to 80 % sequence identity when compared to those of other plant caffeic acid O-methyltransferases. Detection of methyltransferase activity in extracts of Escherichia coli XL-1 Blue cells transformed with a pBluescript phagemid construct containing Vpomt35 confirmed that this cDNA clone encodes a caffeic acid 3-O-methyltransferase. The enzyme also exhibits a low caffeic acid 4-O-methyltransferase activity (2 % of the caffeic acid 3-O-methyltransferase activity). A number of other cDNA clones encoding caffeic acid O-methyltransferases was also sequenced. Comparison of nucleotide and deduced amino acid sequences of these cDNAs indicated two types of caffeic acid O-methyltransferase cDNAs which showed 94 % sequence identity in the coding region, but only 55 % homology with several gaps in the 3′ untranslated end. Results from Southern blot analysis suggest that the caffeic acid O-methyltransferase gene is organized as a small family with at least two genes, which are expressed in Vanilla cell suspension cultures.
Url:
DOI: 10.1016/S0981-9428(99)80014-X
Affiliations:
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Le document en format XML
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<term>CCoAOMT</term>
<term>IPTG</term>
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<term>S-Adenosyl-L-methionine:caffeic acid methyltransferase</term>
<term>Vanilla planifolia</term>
<term>cDNA</term>
<term>in vitro expression</term>
<term>induction</term>
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<term>suspension cultures</term>
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<term>Second type</term>
<term>Sequencing</term>
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<term>Suspension cultures</term>
<term>Vanilla</term>
<term>Vanilla cells</term>
<term>Vanilla planifolia</term>
<term>Vanilla planifolia andr</term>
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<front><div type="abstract" xml:lang="en">Abstract: Two different S-adenosyl-L-methionine:caffeic acid methyltransferases are induced in kinetin-treated cell cultures of Vanilla planifolia. Upon addition of kinetin to the culture, maximum caffeic acid 4-O-methyltransferase and caffeic acid 3-O-methyltransferase activity is obtained after around 2 and 40 h, respectively. The former enzyme is involved in the biosynthesis of vanillic acid and the latter enzyme is co-induced with other enzymes of the general phenylpropanoid pathway and is involved in the biosynthesis of lignin precursors. A number of cDNA clones encoding S-adenosyl-L-methionine:caffeic acid-methyltransferases were isolated by heterologous probe screening of a λZapII cDNA library constructed from mRNA of a kinetin-treated cell suspension culture of V. planifolia. A full-length cDNA clone, Vpomt35, contains a 1 089-bp open reading frame coding for 363 amino acid residues, a 25-bp 5′-end sequence and a 214-bp 3′-end non-coding sequence. The deduced amino acid sequence of Vpomt35 revealed 56 to 80 % sequence identity when compared to those of other plant caffeic acid O-methyltransferases. Detection of methyltransferase activity in extracts of Escherichia coli XL-1 Blue cells transformed with a pBluescript phagemid construct containing Vpomt35 confirmed that this cDNA clone encodes a caffeic acid 3-O-methyltransferase. The enzyme also exhibits a low caffeic acid 4-O-methyltransferase activity (2 % of the caffeic acid 3-O-methyltransferase activity). A number of other cDNA clones encoding caffeic acid O-methyltransferases was also sequenced. Comparison of nucleotide and deduced amino acid sequences of these cDNAs indicated two types of caffeic acid O-methyltransferase cDNAs which showed 94 % sequence identity in the coding region, but only 55 % homology with several gaps in the 3′ untranslated end. Results from Southern blot analysis suggest that the caffeic acid O-methyltransferase gene is organized as a small family with at least two genes, which are expressed in Vanilla cell suspension cultures.</div>
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