Production of cytidine 5'-monophospho-N-acetyl-beta-D-neuraminic acid (CMP-sialic acid) using enzymes or whole cells entrapped in calcium pectate-silica-gel beads.
The present study focuses on the application of immobilization technology to enzymic sugar syntheses. The paper describes an improved silica-alginate matrix established for entrapment and encapsulation. The replacement of alginate with pectate provided enhanced chemical resistance of the matrix, which allows the use of 1% (w/v) polyphosphate in reaction mixtures. Polylysine, a reagent for silica condensation, was replaced by a much cheaper alternative, namely polyethyleneimine. The proposed design was applied in the production of cytidine 5'-monophospho-N-acetyl-beta-D-neuraminic acid (CMP-sialic acid) by immobilized recombinant enzymes or Escherichia coli cells containing overexpressed enzymes. A comparison between these two strategies was made. On the basis of the results we conceptualized a system to synthesize sialyloligosaccharides by using a biocatalyst entrapped in calcium pectate-silica gel beads.
Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, SK-84538 Bratislava, Slovak Republic. nahalka@savba.sk
Jozef
J
Nahálka
Bingyuan
B
Wu
Jun
J
Shao
Peter
P
Gemeiner
Peng George
PG
Wang
D018929:Cell Culture Techniques / Q000379:methods; D018914:Cells, Immobilized / Q000201:enzymology*; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid / Q000138:chemical synthesis / Q000378:metabolism*; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D007652:Oxo-Acid-Lyases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010368:Pectins / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D058428:Silica Gel; D012822:Silicon Dioxide / Q000737:chemistry*
D020099:Coated Materials, Biocompatible; D004800:Enzymes, Immobilized; D011994:Recombinant Proteins; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid; D058428:Silica Gel; D012822:Silicon Dioxide; D010368:Pectins; C018730:N-acetylneuraminate synthase; D007652:Oxo-Acid-Lyases; C018728:N-acetylneuraminate lyase; C003181:polygalacturonic acid