title: Synthesis of functional dipeptide carnosine from nonprotected amino acids using carnosinase-displaying yeast cells.
abstract: Carnosine (beta-alanyl-L-histidine) is one of the bioactive dipeptides and has antioxidant, antiglycation, and cytoplasmic buffering properties. In this study, to synthesize carnosine from nonprotected amino acids as substrates, we cloned the carnosinase (CN1) gene and constructed a whole-cell biocatalyst displaying CN1 on the yeast cell surface with alpha-agglutinin as the anchor protein. The display of CN1 was confirmed by immunofluorescent labeling, and CN1-displaying yeast cells showed hydrolytic activity for carnosine. When carnosine was synthesized by the reverse reaction of CN1, organic solvents were added to the reaction mixture to reduce the water content. The CN1-displaying yeast cells were lyophilized and examined for organic solvent tolerance. Results showed that the CN1-displaying yeast cells retained their original hydrolytic activity in hydrophobic organic solvents. In the hydrophobic organic solvents and hydrophobic ionic liquids, the CN1-displaying yeast cells catalyzed carnosine synthesis, and carnosine was synthesized from nonprotected amino acids in only one step. The results of this research suggest that the whole-cell biocatalyst displaying CN1 on the yeast cell surface can be used to synthesize carnosine with ease and convenience.
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo, Kyoto, 606-8502, Japan.
authors.forename: Chiaki
authors.initials: C
authors.lastname: Inaba
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Higuchi
authors.forename: Hironobu
authors.initials: H
authors.lastname: Morisaka
authors.forename: Kouichi
authors.initials: K
authors.lastname: Kuroda
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Ueda
mesh_terms: D055162:Biocatalysis; D002336:Carnosine / Q000096:biosynthesis*; D003001:Cloning, Molecular; D004150:Dipeptidases / Q000235:genetics / Q000378:metabolism*; D005612:Freeze Drying; D006639:Histidine / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D057927:Hydrophobic and Hydrophilic Interactions; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D012997:Solvents; D013379:Substrate Specificity; D014170:Transformation, Genetic; D015091:beta-Alanine / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D012997:Solvents; D015091:beta-Alanine; D006639:Histidine; D002336:Carnosine; D004150:Dipeptidases; C019673:aminoacyl-histidine dipeptidase