Strategic proteome analysis of Candida magnoliae with an unsequenced genome.
Erythritol is a noncariogenic, low calorie sweetener. It is safe for people with diabetes and obese people. Candida magnoliae is an industrially important organism because of its ability to produce erythritol as a major product. The genome of C. magnoliae has not been sequenced yet, limiting the available proteome database. Therefore, systematic approaches were employed to construct the proteome map of C. magnoliae. Proteomic analysis with systematic approaches is based on two-dimensional electrophoresis, matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS), tandem mass spectrometry (MS/MS) and database interrogation. First, 24 spots were analyzed using peptide mass fingerprinting along with MALDI-TOF MS with high mass accuracy. Only four spots were reliably identified as carbonyl reductase and its isoforms. The reason for low sequence coverage seemed to be that these identification strategies were based on the presence of the protein database obtained from the publicly accessible genome database and the availability of cross-species protein identification. MS/MS (MS/MS ion search and de novo sequencing) in combination with similarity searches allowed successful identification of 39 spots. Several proteins including transaldolase identified by MS/MS ion searches were further confirmed by partial sequences from the expressed sequence tag database. In this study, 51 protein spots were analyzed and then potentially identified. The identified proteins were involved in glycolysis, stress response, other essential metabolisms and cell structures.
Kim
Hyo-Jin
HJ
School of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul, Korea.
Lee
Do-Yup
DY
Lee
Dae-Hee
DH
Park
Yong-Cheol
YC
Kweon
Dae-Hyuk
DH
Ryu
Yeon-Woo
YW
Seo
Jin-Ho
JH
D000465:Algorithms; D000595:Amino Acid Sequence; D002175:Candida / Q000235:genetics / Q000378:metabolism*; D019295:Computational Biology; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D004896:Erythritol / Q000378:metabolism; D020224:Expressed Sequence Tags; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism*; D016678:Genome; D008969:Molecular Sequence Data; D020543:Proteome / Q000737:chemistry / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D005656:Fungal Proteins; D020543:Proteome; D004896:Erythritol