Cryoprotective activities of group 3 late embryogenesis abundant proteins from Chlorella vulgaris C-27

Ken-ichi Honjoh, Hiroko Matsumoto, Hideyuki Shimizu, Kanae Ooyama, Kageyuki Tanaka, Yuichi Oda, Ryoji Takata, Toshio Joh, Koushirou Suga, Takahisa Miyamoto, Masayoshi Iio, Shoji Hatano

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

The nucleotide sequence of hiC12, isolated as a cDNA clone of hardening-induced Chlorella (hiC) genes, was identified. The clone encodes a late embryogenesis abundant (LEA) protein having six repeats of a 11-mer amino acid motif, although in a slightly imperfect form. To overexpress the hiC61) and hiC12 genes, their coding regions were PCR amplified and subcloned into a pGEX-1λT vector. The HIC6 and HIC12 proteins were expressed as GST fusion proteins inE. coli, then purified. The two HIC proteinswere found to be effective in protecting a freeze-labile enzyme, LDH, against freeze-inactivation. On a molar concentration basis, they were about 3.1×106 times more effective in protecting LDH than sucrose and as effective as BSA. Cryoprotection tests with five kinds of chain-shortened polypeptides, synthesized based on the 11-mer amino acid motif of the HIC6 protein showed that the cryoprotective activity decreased with a decrease in the repeating units of the 11-mer motif. In fact, cryoprotective activities of three kinds of single 11-mer amino acids were very low even at high concentrations. All the results suggested that the sufficiently repeated 11-mer motif is required for the cryoprotective activities of Chlorella LEA proteins.

Original languageEnglish
Pages (from-to)1656-1663
Number of pages8
JournalBioscience, Biotechnology and Biochemistry
Volume64
Issue number8
DOIs
Publication statusPublished - Jan 1 2000

Fingerprint

Chlorella vulgaris
Embryonic Development
Chlorella
Amino Acid Motifs
Proteins
Amino Acids
Clone Cells
Genes
Sucrose
Hardening
Fusion reactions
Nucleotides
Complementary DNA
C 27
Polymerase Chain Reaction
Peptides
Enzymes

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

Cite this

Cryoprotective activities of group 3 late embryogenesis abundant proteins from Chlorella vulgaris C-27. / Honjoh, Ken-ichi; Matsumoto, Hiroko; Shimizu, Hideyuki; Ooyama, Kanae; Tanaka, Kageyuki; Oda, Yuichi; Takata, Ryoji; Joh, Toshio; Suga, Koushirou; Miyamoto, Takahisa; Iio, Masayoshi; Hatano, Shoji.

In: Bioscience, Biotechnology and Biochemistry, Vol. 64, No. 8, 01.01.2000, p. 1656-1663.

Research output: Contribution to journalArticle

Honjoh, K, Matsumoto, H, Shimizu, H, Ooyama, K, Tanaka, K, Oda, Y, Takata, R, Joh, T, Suga, K, Miyamoto, T, Iio, M & Hatano, S 2000, 'Cryoprotective activities of group 3 late embryogenesis abundant proteins from Chlorella vulgaris C-27', Bioscience, Biotechnology and Biochemistry, vol. 64, no. 8, pp. 1656-1663. https://doi.org/10.1271/bbb.64.1656
Honjoh, Ken-ichi ; Matsumoto, Hiroko ; Shimizu, Hideyuki ; Ooyama, Kanae ; Tanaka, Kageyuki ; Oda, Yuichi ; Takata, Ryoji ; Joh, Toshio ; Suga, Koushirou ; Miyamoto, Takahisa ; Iio, Masayoshi ; Hatano, Shoji. / Cryoprotective activities of group 3 late embryogenesis abundant proteins from Chlorella vulgaris C-27. In: Bioscience, Biotechnology and Biochemistry. 2000 ; Vol. 64, No. 8. pp. 1656-1663.
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