title: A crosslinked inclusion body process for sialic acid synthesis.
abstract: The propensity of a recombinant protein produced in bacteria to aggregate has been assumed to be unpredictable, and inclusion bodies have been thought of as wasted cell material. However, a target protein can be purposely driven to inclusion bodies, which demonstrate full cell tolerable activity. Sialic acid aldolase, N-terminally fused with the cellulose-binding module of Clostridium cellulovorans, was almost quantitatively physiologically aggregated into active inclusion bodies. These inclusion bodies were entrapped in alginate beads and crosslinked by glutaraldehyde. The immobilized biocatalyst generated by this crosslinked inclusion bodies (CLIB) technology was used in a repetitive batch protocol for sialic acid production that was monitored on-line by flow calorimetry. The required substrate, N-acetyl-D-mannosamine, was obtained by partially improved alkaline epimerization.
authors.affiliation: Institute of Chemistry, Center for Glycomics, Slovak Academy of Sciences, Bratislava, Slovak Republic. nahalka@savba.sk
authors.forename: Jozef
authors.initials: J
authors.lastname: Nahálka
authors.forename: Alica
authors.initials: A
authors.lastname: Vikartovská
authors.forename: Eva
authors.initials: E
authors.lastname: Hrabárová
mesh_terms: D015374:Biosensing Techniques / Q000379:methods; D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents / Q000737:chemistry; D004800:Enzymes, Immobilized / Q000378:metabolism; D005434:Flow Cytometry; D005976:Glutaral / Q000737:chemistry; D006595:Hexosamines / Q000378:metabolism; D002479:Inclusion Bodies / Q000737:chemistry / Q000378:metabolism*; D019158:N-Acetylneuraminic Acid / Q000096:biosynthesis*; D007652:Oxo-Acid-Lyases / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D006595:Hexosamines; D007652:Oxo-Acid-Lyases; C018728:N-acetylneuraminate lyase; D019158:N-Acetylneuraminic Acid; D005976:Glutaral; C002022:N-acetylmannosamine