Premium
Purification of chondroitin precursor from Escherichia coli K4 fermentation broth using membrane processing
Author(s) -
Schiraldi Chiara,
Carcarino Immacolata Loredana,
Alfano Alberto,
Restaino Odile Francesca,
Panariello Andrea,
De Rosa Mario
Publication year - 2011
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201000266
Subject(s) - escherichia coli , fermentation , chemistry , microbiology and biotechnology , membrane , food science , biochemistry , chromatography , biology , gene
Recently the possibility of producing the capsular polysaccharide K4, a fructosylated chondroitin, in fed‐batch experiments was assessed. In the present study, a novel downstream process to obtain chondroitin from Escherichia coli K4 fermentation broth was developed. The process is simple, scalable and economical. In particular, downstream procedures were optimized with a particular aim of purifying a product suitable for further chemical modifications, in an attempt to develop a biotechnological platform for chondroitin sulfate production. During process development, membrane devices (ultrafiltration/diafiltration) were exploited, selecting the right cassette cut‐offs for different phases of purification. The operational conditions (cross‐flow rate and transmembrane pressure) used for the process were determined on an ÄKTA cross‐flow instrument (GE Healthcare, USA), a lab‐scale automatic tangential flow filtration system. In addition, parameters such as selectivity and throughput were calculated based on the analytical quantification of K4 and defructosylated K4, as well as the major contaminants. The complete downstream procedure yielded about 75% chondroitin with a purity higher than 90%.