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

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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

All Science Journal Classification (ASJC) codes

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

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    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, 64(8), 1656-1663. https://doi.org/10.1271/bbb.64.1656