title: Preparation of a whole-cell biocatalyst of mutated Candida antarctica lipase B (mCALB) by a yeast molecular display system and its practical properties.
abstract: To prepare a whole-cell biocatalyst of a stable lipase at a low price, mutated Candida antarctica lipase B (mCALB) constructed on the basis of the primary sequences of CALBs from C. antarctica CBS 6678 strain and from C. antarctica LF 058 strain was displayed on a yeast cell surface by alpha-agglutinin as the anchor protein for easy handling and stability of the enzyme. When mCALB was displayed on the yeast cell surface, it showed a preference for short chain fatty acids, an advantage for producing flavors; although when Rhizopus oryzae lipase (ROL) was displayed, the substrate specificity was for middle chain lengths. When the thermal stability of mCALB on the cell surface was compared with that of ROL on a cell surface, T (1/2), the temperature required to give a residual activity of 50% for heat treatment of 30 min, was 60 degrees C for mCALB and 44 degrees C for ROL indicating that mCALB displayed on cell surface has a higher thermal stability. Furthermore, the activity of the displayed mCALB against p-nitrophenyl butyrate was 25-fold higher than that of soluble CALB, as reported previously. These findings suggest that mCALB-displaying yeast is more practical for industrial use as the whole-cell biocatalyst.
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan. mk@kais.kyoto-u.ac.jp
authors.forename: Michiko
authors.initials: M
authors.lastname: Kato
authors.forename: Jun
authors.initials: J
authors.lastname: Fuchimoto
authors.forename: Takanori
authors.initials: T
authors.lastname: Tanino
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kondo
authors.forename: Hideki
authors.initials: H
authors.lastname: Fukuda
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Ueda
mesh_terms: D002175:Candida / Q000201:enzymology* / Q000235:genetics; D002473:Cell Wall / Q000201:enzymology; D004795:Enzyme Stability; D005656:Fungal Proteins; D008049:Lipase / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D009154:Mutation; D012233:Rhizopus / Q000201:enzymology; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D005656:Fungal Proteins; D008049:Lipase; C431349:lipase B, Candida antarctica