Biotechnologically Relevant Enzymes and Proteins

Applied Microbiology and Biotechnology

, Volume 75, Issue 6, pp 1319-1325

First online:

Development of yeast cells displaying Candida antarctica lipase B and their application to ester synthesis reaction

  • Takanori TaninoAffiliated withDivision of Molecular Science and Material Engineering, Graduate School of Science and Technology, Kobe University
  • , Takumi OhnoAffiliated withDepartment of Chemical Science and Engineering, Faculty of Engineering, Kobe University
  • , Tohru AokiAffiliated withCentral Research Laboratories, Dainippon Ink and Chemicals
  • , Hideki FukudaAffiliated withDivision of Molecular Science and Material Engineering, Graduate School of Science and Technology, Kobe University
  • , Akihiko KondoAffiliated withDepartment of Chemical Science and Engineering, Faculty of Engineering, Kobe University Email author 

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Abstract

We isolated the lipase B from Candida antarctica CBS 6678 (CALB CBS6678) and successfully constructed CALB-displaying yeast whole-cell biocatalysts using the Flo1p short (FS) anchor system. For the display of CALB on a yeast cell surface, the newly isolated CALB CBS6678 exhibited higher hydrolytic and ester synthesis activities than the well-known CALB, which is registered in GenBank (Z30645). A protease accessibility assay using papain as a protease showed that a large part of CALB, approximately 75%, was localized on an easily accessible part of the yeast cell surface. A comparison of the lipase hydrolytic activities of yeast whole cells displaying only mature CALB (CALB) and those displaying mature CALB with a Pro region (ProCALB) revealed that mature CALB is preferable for yeast cell surface display using the Flo1p anchor system. Lyophilized yeast whole cells displaying CALB were applied to an ester synthesis reaction at 60°C using adipic acid and n-butanol as substrates. The amount of dibutyl adipate (DBA) produced increased with the reaction time until 144 h. This indicated that CALB displayed on the yeast cell surface retained activity under the reaction conditions.

Keywords

Cell surface display Candida antarctica lipase B Ester synthesis