Expression of a bacterial serine acetyltransferase in transgenic potato plants leads to increased levels of cysteine and glutathione.
The coding sequence of the wild-type, cys-sensitive, cysE gene from Escherichia coli, which encodes an enzyme of the cysteine biosynthetic pathway, namely serine acetyltransferase (SAT, EC 2.3.1. 30), was introduced into the genome of potato plants under the control of the cauliflower mosaic virus 35S promoter. In order to target the protein into the chloroplast, cysE was translationally fused to the 5'-signal sequence of rbcS from Arabidopsis thaliana. Transgenic plants showed a high accumulation of the cysE mRNA. The chloroplastic localisation of the E. coli SAT protein was demonstrated by determination of enzymatic activities in enriched organelle fractions. Crude leaf extracts of these plants exhibited up to 20-fold higher SAT activity than those prepared from wild-type plants. The transgenic potato plants expressing the E. coli gene showed not only increased levels of enzyme activity but also exhibited elevated levels of cysteine and glutathione in leaves. Both were up to twofold higher than in control plants. However, the thiol content in tubers of transgenic lines was unaffected. The alterations observed in leaf tissue had no effect on the expression of O-acetylserine(thiol)-lyase, the enzyme which converts O-acetylserine, the product of SAT, to cysteine. Only a minor effect on its enzymatic activity was observed. In conclusion, the results presented here demonstrate the importance of SAT in plant cysteine biosynthesis and show that production of cysteine and related sulfur-containing compounds can be enhanced by metabolic engineering.
Harms
K
K
Max-Planck-Institut für Molekulare Pflanzenphysiologie, Abt. Willmitzer, Am Mühlenberg 1, D-14476 Golm, Germany.
von Ballmoos
P
P
Brunold
C
C
Höfgen
R
R
Hesse
H
H
D000123:Acetyltransferases / Q000235:genetics* / Q000378:metabolism; D017360:Arabidopsis / Q000235:genetics; D001483:Base Sequence; D003545:Cysteine / Q000378:metabolism*; D017931:DNA Primers; D004926:Escherichia coli / Q000201:enzymology*; D005978:Glutathione / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000201:enzymology; D012333:RNA, Messenger / Q000235:genetics; D051050:Serine O-Acetyltransferase; D011198:Solanum tuberosum / Q000235:genetics*
D017931:DNA Primers; D012333:RNA, Messenger; D000123:Acetyltransferases; D051050:Serine O-Acetyltransferase; D005978:Glutathione; D003545:Cysteine