Chloroplastic glucose 6-phosphate dehydrogenase from Chlorella vulgaris alleviates freezing and menadione-induced oxidative stresses in Saccharomyces cerevisiae

Takeshi Machida, Naoto Ohashi, Ayano Mimura, Ken-ichi Honjoh, Masayoshi Iio, Takahisa Miyamoto

Research output: Contribution to journalArticle

Abstract

Enhanced glucose 6-phosphate dehydrogenase (E.C.1.1.1.49, G6PDH) activity has been identified as a hardening-induced intracellular change of Chlorella vulgaris, which acquires freezing tolerance during hardening. In the present study, a full-length cDNA clone corresponding to a gene encoding a chloroplastic isoform of G6PDH, designated CvchG6PDH, was isolated from C. vulgaris C-27. By comparing the deduced amino acid sequence of CvchG6PDH with the N-terminal amino acid sequence of mature G6PDH 2 protein isolated previously, a DNA region encoding mature CvchG6PDH was determined and designated mCvchG6PDH. The deduced amino acid sequence of CvchG6PDH showed higher homology to those of plant plastidic G6PDH genes than those of cytosolic ones. A recombinant mCvchG6PDH protein expressed in Escherichia coli showed similar enzymatic properties to previously-isolated Chlorella G6PDH 2, suggesting that the gene encoded plastidic G6PDH 2 protein. Expression of CvchG6PDH was induced transcriptionally throughout 24-h hardening, while the translation was induced up to 9-h hardening and then decreased, and the change did not reflect the enhanced G6PDH activity during hardening. Furthermore, the mCvch-G6PDH alleviated both freezing and menadione-induced oxidative stresses in yeast. We showed the contribution of CvchG6PDH in menadione stress tolerance as one of its functions in the acquisition of freezing tolerance of Chlorella.

Original languageEnglish
Pages (from-to)29-38
Number of pages10
JournalJournal of the Faculty of Agriculture, Kyushu University
Volume55
Issue number1
Publication statusPublished - Feb 2010
Externally publishedYes

Fingerprint

Chlorella vulgaris
menadione
Vitamin K 3
Glucosephosphate Dehydrogenase
glucose-6-phosphate 1-dehydrogenase
Freezing
Saccharomyces cerevisiae
Chlorella
Amino Acid Sequence
freezing
Oxidative Stress
oxidative stress
Genes
Recombinant Proteins
Protein Isoforms
Proteins
amino acid sequences
Complementary DNA
Clone Cells
Yeasts

All Science Journal Classification (ASJC) codes

  • Agronomy and Crop Science
  • Biotechnology

Cite this

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title = "Chloroplastic glucose 6-phosphate dehydrogenase from Chlorella vulgaris alleviates freezing and menadione-induced oxidative stresses in Saccharomyces cerevisiae",
abstract = "Enhanced glucose 6-phosphate dehydrogenase (E.C.1.1.1.49, G6PDH) activity has been identified as a hardening-induced intracellular change of Chlorella vulgaris, which acquires freezing tolerance during hardening. In the present study, a full-length cDNA clone corresponding to a gene encoding a chloroplastic isoform of G6PDH, designated CvchG6PDH, was isolated from C. vulgaris C-27. By comparing the deduced amino acid sequence of CvchG6PDH with the N-terminal amino acid sequence of mature G6PDH 2 protein isolated previously, a DNA region encoding mature CvchG6PDH was determined and designated mCvchG6PDH. The deduced amino acid sequence of CvchG6PDH showed higher homology to those of plant plastidic G6PDH genes than those of cytosolic ones. A recombinant mCvchG6PDH protein expressed in Escherichia coli showed similar enzymatic properties to previously-isolated Chlorella G6PDH 2, suggesting that the gene encoded plastidic G6PDH 2 protein. Expression of CvchG6PDH was induced transcriptionally throughout 24-h hardening, while the translation was induced up to 9-h hardening and then decreased, and the change did not reflect the enhanced G6PDH activity during hardening. Furthermore, the mCvch-G6PDH alleviated both freezing and menadione-induced oxidative stresses in yeast. We showed the contribution of CvchG6PDH in menadione stress tolerance as one of its functions in the acquisition of freezing tolerance of Chlorella.",
author = "Takeshi Machida and Naoto Ohashi and Ayano Mimura and Ken-ichi Honjoh and Masayoshi Iio and Takahisa Miyamoto",
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T1 - Chloroplastic glucose 6-phosphate dehydrogenase from Chlorella vulgaris alleviates freezing and menadione-induced oxidative stresses in Saccharomyces cerevisiae

AU - Machida, Takeshi

AU - Ohashi, Naoto

AU - Mimura, Ayano

AU - Honjoh, Ken-ichi

AU - Iio, Masayoshi

AU - Miyamoto, Takahisa

PY - 2010/2

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N2 - Enhanced glucose 6-phosphate dehydrogenase (E.C.1.1.1.49, G6PDH) activity has been identified as a hardening-induced intracellular change of Chlorella vulgaris, which acquires freezing tolerance during hardening. In the present study, a full-length cDNA clone corresponding to a gene encoding a chloroplastic isoform of G6PDH, designated CvchG6PDH, was isolated from C. vulgaris C-27. By comparing the deduced amino acid sequence of CvchG6PDH with the N-terminal amino acid sequence of mature G6PDH 2 protein isolated previously, a DNA region encoding mature CvchG6PDH was determined and designated mCvchG6PDH. The deduced amino acid sequence of CvchG6PDH showed higher homology to those of plant plastidic G6PDH genes than those of cytosolic ones. A recombinant mCvchG6PDH protein expressed in Escherichia coli showed similar enzymatic properties to previously-isolated Chlorella G6PDH 2, suggesting that the gene encoded plastidic G6PDH 2 protein. Expression of CvchG6PDH was induced transcriptionally throughout 24-h hardening, while the translation was induced up to 9-h hardening and then decreased, and the change did not reflect the enhanced G6PDH activity during hardening. Furthermore, the mCvch-G6PDH alleviated both freezing and menadione-induced oxidative stresses in yeast. We showed the contribution of CvchG6PDH in menadione stress tolerance as one of its functions in the acquisition of freezing tolerance of Chlorella.

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