Synthesis of plant triosephosphate isomerase in Escherichia coli.

Sato, Fumihiko; Fitchen, John H.; Takeshita, Norimatsu; Hashimoto, Takashi; OKADA, Naosuke; Yamada, Yasuyuki

doi:10.1271/bbb1961.54.2189

Cited by 4 publications

(5 citation statements)

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“…4 The extremely high turnover rate of TPI has led to its description as a nearly perfect catalyst. 5 Genes encoding TPI have been cloned and sequenced from 11 species, including chicken, 6 human 7,8 maize, 9 mouse, 10 Aspergillus nidulans, 11 Coptis japonica, 12 Escherichia coli, 13 Macaca mulatta, 14 T. brucei, 15 Saccharomyces cerevisiae, 16 and Schizosaccharomyces pombe. 17 The high degree of sequence conservation of TPI suggests a low rate of evolutionary divergence, a common feature of all the glycolytic enzymes.…”

Section: Introductionmentioning

confidence: 99%

Characterization of stress and methylglyoxal inducible triose phosphate isomerase (OscTPI) from rice

Sharma¹,

Mustafiz²,

Singla‐Pareek³

et al. 2012

Plant Signaling & Behavior

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As compared with plant system, triose phosphate isomerase (TPI), a crucial enzyme of glycolysis, has been well studied in animals. In order to characterize TPI in plants, a full-length cDNA encoding OscTPI was cloned from rice and expressed in E. coli. The recombinant OscTPI was purified to homogeneity and it showed Km value of 0.1281 ± 0.025 µM, and the Vmax value of 138.7 ± 16 µmol min (-1) mg (-1) which is comparable to the kinetic values studied in other plants. The OscTPI was found to be exclusively present in the cytoplasm when checked with the various methods. Functional assay showed that OscTPI could complement a TPI mutation in yeast. Real time PCR analysis revealed that OscTPI transcript level was regulated in response to various abiotic stresses. Interestingly, it was highly induced under different concentration of methylglyoxal (MG) stress in a concentration dependent manner. There was also a corresponding increase in the protein and the enzyme activity of OscTPI both in shoot and root tissues under MG stress. Our result shows that increases in MG leads to the increase in TPI which results in decrease of DHAP and consequently decrease in the level of toxic MG.

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Section: Introductionmentioning

confidence: 99%

Characterization of stress and methylglyoxal inducible triose phosphate isomerase (OscTPI) from rice

Sharma¹,

Mustafiz²,

Singla‐Pareek³

et al. 2012

Plant Signaling & Behavior

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“…The genes for cytosolic TPI (cTPI) have been isolated and sequenced from several sources, including maize (5), Coptis japonica (6), rice (4), Stellaria longipes (7), lettuce (8), rye (9), Arabidopsis (10), and petunia corollas (11), but expression is limited (12,13). Contrary, chloroplast TPIs (cpTPIs) have been purified from rye, spinach, and lettuce (14,15), but studied mainly at the biochemical level.…”

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confidence: 99%

Overexpression in Escherichia coli and Characterization of the Chloroplast Triosephosphate Isomerase from Spinach

Tang

Wang

Bao

et al. 1999

Protein Expression and Purification

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“…Recently, Chen et al (2012) reported the functional expression of THBO isolated from Corydalis saxicola using a Coptis triosephosphate isomerase cDNA (CjTPI) sequence. However, we did not detect THBO activity using recombinant CjTPI expressed in E. coli (Sato et al 1990). In fact, it is very difficult to measure THBO activity since the substrate THB is easily oxidized and THBO is very unstable.…”

Section: Discussionmentioning

confidence: 72%

“…In our previous report (Okada et al 1989), we purified THBO 10.8-fold from cultured C. japonica cells, but this purified fraction was contaminated with triosephosphate isomerase (TPI; Sato et al 1990). Thus, we re-purified THBO from cultured C. japonica cells as described in the Materials and methods.…”

Section: Isolation Of Cdna For Thbomentioning

confidence: 99%

“…All of the cDNAs for these reactions have been isolated and functionally expressed in microbial cells to characterize their enzymological properties, except for the enzyme in the last step THBO. While a previous report described the isolation of cDNA of THBO from cultured Coptis japonica cells (Okada et al 1989), we later found that the purified THBO was contaminated by triosephosphate isomerase (TPI; Sato et al 1990). Here, we report the re-isolation of THBO cDNA from cultured Coptis japonica cells and its functional characterization in transgenic California poppy (Eschscholzia californica) cells (Figure 1).…”

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confidence: 92%

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Pathway engineering of benzylisoquinoline alkaloid biosynthesis in transgenic California poppy cells with ectopic expression of tetrahydroberberine oxidase from <i>Coptis japonica</i>

Matsushima

Minami

Hori

et al. 2012

Plant Biotechnology

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(S)-Tetrahydroberberine oxidase is the enzyme in the last step of berberine biosynthesis. While a previous report described the isolation of cDNA of tetrahydroberberine oxidase (THBO) from cultured Coptis japonica cells, we later found that purified THBO was heavily contaminated by triosephosphate isomerase. Here, we report the re-isolation of THBO cDNA from cultured C. japonica cells and its functional characterization in transgenic California poppy cells.A cDNA clone for (S)-tetrahydroberberine oxidase was isolated from an EST library prepared from high berberineproducing cultured C. japonica cells based on the partial amino acid sequence of the purified enzyme. Analyses of the nucleotide sequences of the cloned cDNA inserts of 1728 base pairs revealed an open reading frame that encoded a 540-amino acid polypeptide with putative 28-amino acid signal peptides and a mature polypeptide with a molecular mass of 57,748. A protein blast search also shows that CjTHBO belongs to the FAD-containing berberine bridge enzyme oxidoreductase family. Since all attempts to produce active recombinant CjTHBO in Escherichia coli and Saccharomyces cerevisiae cells failed, we tried to express CjTHBO in California poppy (Eschscholzia californica) cells. When transgenic California poppy cells that ectopically expressed CjTHBO under the control of Cauliflower mosaic virus 35S promoter were established, the transgenic cells showed small but evident new alkaloid peaks, which were scarcely detected in control cells that did not express CjTHBO. LC-MS analyses showed that these peaks were coptisine and dehydrocheilanthifoline, which were expected to be generated by the reaction of CjTHBO from pathway intermediates, i.e., cheilanthifoline and stylopine. The usefulness of CjTHBO for metabolic engineering in transgenic California poppy cells is discussed.

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Synthesis of plant triosephosphate isomerase in Escherichia coli.

Cited by 4 publications

References 0 publications

Characterization of stress and methylglyoxal inducible triose phosphate isomerase (OscTPI) from rice

Characterization of stress and methylglyoxal inducible triose phosphate isomerase (OscTPI) from rice

Overexpression in Escherichia coli and Characterization of the Chloroplast Triosephosphate Isomerase from Spinach

Pathway engineering of benzylisoquinoline alkaloid biosynthesis in transgenic California poppy cells with ectopic expression of tetrahydroberberine oxidase from <i>Coptis japonica</i>

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