title: Isolation and characterization of hardening-induced proteins in Chlorella vulgaris C-27: identification of late embryogenesis abundant proteins.
abstract: Hardening-induced soluble proteins of Chlorella vulgaris Beijerink IAM C-27 (formerly Chlorella ellipsoidea Gerneck IAM C-27) were isolated and purified by two-dimensional high-performance liquid chromatography (2D-HPLC) on an anion-exchange column, with subsequent reversed-phase chromatography. Some of the proteins were resolved by SDS-PAGE, characterized by amino-terminal sequencing and identified by searching for homologies in databases. Separation of the soluble proteins during the hardening of Chlorella by a combination of 2D-HPLC and SDS-PAGE revealed that at least 31 proteins were induced or increased in abundance. Of particular interest was the induction after 12 h of a 10-kDa protein with the amino-terminal amino acid sequence AGNKPITEQISDAVGAAGQKVG and the induction after 6 h of a 14-kDa protein with the amino-terminal sequence ALGEESLGDKAKNAFEDAKDAVKDAAGNVKEAV. The amino-terminal sequences of these proteins indicated that they were homologous to late embryogenesis abundant (LEA) proteins. Furthermore, the level of a 22-kDa protein also increased after 12 h. The amino-terminal sequence of this protein, AAPLVGGPAPDFTAAAVFD, indicated that it was homologous to thioredoxin peroxidase.
authors.affiliation: Department of Food Science and Technology, Faculty of Agriculture, Kyushu University, Fukuoka, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Honjoh
authors.forename: M
authors.initials: M
authors.lastname: Yoshimoto
authors.forename: T
authors.initials: T
authors.lastname: Joh
authors.forename: T
authors.initials: T
authors.lastname: Kajiwara
authors.forename: T
authors.initials: T
authors.lastname: Miyamoto
authors.forename: S
authors.initials: S
authors.lastname: Hatano
mesh_terms: D000595:Amino Acid Sequence; D002478:Cells, Cultured; D002708:Chlorella / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012995:Solubility
chemical_list: D010940:Plant Proteins; C066246:late embryogenesis abundant protein, plant