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Inclusion bodies of fuculose‐1‐phosphate aldolase as stable and reusable biocatalysts
Author(s) -
Sans Cristina,
GarcíaFruitós Elena,
Ferraz Rosa M.,
GonzálezMontalbán Núria,
Rinas Ursula,
LópezSantín Josep,
Villaverde Antonio,
Álvaro Gregorio
Publication year - 2012
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.1518
Subject(s) - aldolase a , solubility , chemistry , inclusion bodies , biocatalysis , phosphate , combinatorial chemistry , escherichia coli , enzyme , chemical engineering , biochemistry , catalysis , organic chemistry , reaction mechanism , engineering , gene
Fuculose‐1‐phosphate aldolase (FucA) has been produced in Escherichia coli as active inclusion bodies (IBs) in batch cultures. The activity of insoluble FucA has been modulated by a proper selection of producing strain, culture media, and process conditions. In some cases, when an optimized defined medium was used, FucA IBs were more active (in terms of specific activity) than the soluble protein version obtained in the same process with a conventional defined medium, supporting the concept that solubility and conformational quality are independent protein parameters. FucA IBs have been tested as biocatalysts, either directly or immobilized into Lentikat ® beads, in an aldolic reaction between DHAP and (S)‐Cbz‐alaninal, obtaining product yields ranging from 65 to 76%. The production of an active aldolase as IBs, the possibility of tailoring IBs properties by both genetic and process approaches, and the reusability of IBs by further entrapment in appropriate matrices fully support the principle of using self‐assembled enzymatic clusters as tunable mechanically stable and functional biocatalysts. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012