English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Stereodivergent Protein Engineering of a Lipase To Access All Possible Stereoisomers of Chiral Esters with Two Stereocenters

Xu, J., Cen, Y., Singh, W., Fan, J., Wu, L., Lin, X., et al. (2019). Stereodivergent Protein Engineering of a Lipase To Access All Possible Stereoisomers of Chiral Esters with Two Stereocenters. Journal of the American Chemical Society, 141(19), 7934-7945. doi:10.1021/jacs.9b02709.

Item is

Basic

show hide
Genre: Journal Article

Files

show Files

Locators

show

Creators

show
hide
 Creators:
Xu, Jian1, Author
Cen, Yixin1, 2, Author
Singh, Warispreet3, Author
Fan, Jiajie1, Author
Wu, Lian2, Author
Lin, Xianfu1, Author
Zhou, Jiahai2, Author
Huang, Meilan3, Author
Reetz, Manfred T.4, 5, Author           
Wu, Qi1, Author
Affiliations:
1Department of Chemistry, Zhejiang University, Hangzhou 310027, PR China, ou_persistent22              
2State Key Laboratory of Bio-organic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, PR China, ou_persistent22              
3School of Chemistry and Chemical Engineering, Queen’s University, David Keir Building, Stranmillis Road, Belfast BT9 5AG, Northern Ireland, U.K., ou_persistent22              
4Research Department Reetz, Max-Planck-Institut für Kohlenforschung, Max Planck Society, ou_1445588              
5Chemistry Department, Philipps-University, Hans-Meerwein-Str. 4, 35032 Marburg, Germany, ou_persistent22              

Content

show
hide
Free keywords: -
 Abstract: Enzymatic stereodivergent synthesis to access all possible product stereoisomers bearing multiple stereocenters is relatively undeveloped, although enzymes are being increasingly used in both academic and industrial areas. When two stereocenters and thus four stereoisomeric products are involved, obtaining stereodivergent enzyme mutants for individually accessing all four stereoisomers would be ideal. Although significant success has been achieved in directed evolution of enzymes in general, stereodivergent engineering of one enzyme into four highly stereocomplementary variants for obtaining the full complement of stereoisomers bearing multiple stereocenters remains a challenge. Using Candida antarctica lipase B (CALB) as a model, we report the protein engineering of this enzyme into four highly stereocomplementary variants needed for obtaining all four stereoisomers in transesterification reactions between racemic acids and racemic alcohols in organic solvents. By generating and screening less than 25 variants of each isomer, we achieved >90% selectivity for all of the four possible stereoisomers in the model reaction. This difficult feat was accomplished by developing a strategy dubbed “focused rational iterative site-specific mutagenesis” (FRISM) at sites lining the enzyme’s binding pocket. The accumulation of single mutations by iterative site-specific mutagenesis using a restricted set of rationally chosen amino acids allows the formation of ultrasmall mutant libraries requiring minimal screening for stereoselectivity. The crystal structure of all stereodivergent CALB variants, flanked by MD simulations, uncovered the source of selectivity.

Details

show
hide
Language(s): eng - English
 Dates: 2019-03-112019-04-262019-05-15
 Publication Status: Published in print
 Pages: 12
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1021/jacs.9b02709
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Journal of the American Chemical Society
  Other : J. Am. Chem. Soc.
  Abbreviation : JACS
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: Washington, DC : American Chemical Society
Pages: - Volume / Issue: 141 (19) Sequence Number: - Start / End Page: 7934 - 7945 Identifier: ISSN: 0002-7863
CoNE: https://pure.mpg.de/cone/journals/resource/954925376870