Literature DB >> 19509064

Directed evolution of Candida antarctica lipase A using an episomaly replicating yeast plasmid.

Anders G Sandström1, Karin Engström, Jonas Nyhlén, Alex Kasrayan, J-E Bäckvall.   

Abstract

We herein report the first directed evolution of Candida antarctica lipase A (CalA), employing a combinatorial active-site saturation test (CAST). Wild-type CalA has a modest E-value of 5.1 in kinetic resolution of 4-nitrophenyl 2-methylheptanoate. Enzyme variants were expressed in Pichia pastoris by using the episomal vector pBGP1 which allowed efficient secretory expression of the lipase. Iterative rounds of CASTing yielded variants with good selectivity toward both the (S)- and the (R)-enantiomer. The best obtained enzyme variants had E-values of 52 (S) and 27 (R).

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Year:  2009        PMID： 19509064     DOI： 10.1093/protein/gzp019

Source DB:  PubMed          Journal:  Protein Eng Des Sel        ISSN： 1741-0126            Impact factor:   1.650


  9 in total

1.  Combinatorial reshaping of the Candida antarctica lipase A substrate pocket for enantioselectivity using an extremely condensed library.

Authors:  Anders G Sandström; Ylva Wikmark; Karin Engström; Jonas Nyhlén; Jan-E Bäckvall
Journal:  Proc Natl Acad Sci U S A       Date:  2011-12-16       Impact factor: 11.205

2.  Validating computer simulations of enantioselective catalysis; reproducing the large steric and entropic contributions in Candida Antarctica lipase B.

Authors:  Patrick Schopf; Arieh Warshel
Journal:  Proteins       Date:  2014-01-25

3.  Towards quantitative computer-aided studies of enzymatic enantioselectivity: the case of Candida antarctica lipase A.

Authors:  Maria P Frushicheva; Arieh Warshel
Journal:  Chembiochem       Date:  2011-12-21       Impact factor: 3.164

4.  Rhodococcus sp. strain CR-53 LipR, the first member of a new bacterial lipase family (family X) displaying an unusual Y-type oxyanion hole, similar to the Candida antarctica lipase clan.

Authors:  Arnau Bassegoda; F I Javier Pastor; Pilar Diaz
Journal:  Appl Environ Microbiol       Date:  2012-01-06       Impact factor: 4.792

5.  Strains and Molecular Tools for Recombinant Protein Production in Pichia pastoris.

Authors:  Claudia Rinnofner; Michael Felber; Harald Pichler
Journal:  Methods Mol Biol       Date:  2022

Review 6.  Thermostable lipases and their dynamics of improved enzymatic properties.

Authors:  Siti Hajar Hamdan; Jonathan Maiangwa; Mohd Shukuri Mohamad Ali; Yahaya M Normi; Suriana Sabri; Thean Chor Leow
Journal:  Appl Microbiol Biotechnol       Date:  2021-09-06       Impact factor: 5.560

7.  Combinatorial library based engineering of Candida antarctica lipase A for enantioselective transacylation of sec-alcohols in organic solvent.

Authors:  Ylva Wikmark; Maria Svedendahl Humble; Jan-E Bäckvall
Journal:  Angew Chem Int Ed Engl       Date:  2015-02-09       Impact factor: 15.336

8.  Comprehensive clone screening and evaluation of fed-batch strategies in a microbioreactor and lab scale stirred tank bioreactor system: application on Pichia pastoris producing Rhizopus oryzae lipase.

Authors:  Johannes Hemmerich; Núria Adelantado; José Manuel Barrigón; Xavier Ponte; Astrid Hörmann; Pau Ferrer; Frank Kensy; Francisco Valero
Journal:  Microb Cell Fact       Date:  2014-03-07       Impact factor: 5.328

9.  Development of simple random mutagenesis protocol for the protein expression system in Pichia pastoris.

Authors:  Mikako Tachioka; Naohisa Sugimoto; Akihiko Nakamura; Naoki Sunagawa; Takuya Ishida; Taku Uchiyama; Kiyohiko Igarashi; Masahiro Samejima
Journal:  Biotechnol Biofuels       Date:  2016-09-19       Impact factor: 6.040
  9 in total

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