Mannan-penicillin G acylase neoglycoproteins and their potential applications in biotechnology.
Mannan-penicillin G acylase neoglycoproteins were prepared by the conjugation of Saccharomyces cerevisiae mannan with enzyme penicillin G acylase using the reductive amination method. Eight neoglycoproteins preparations were obtained after gel chromatography. The preparations contained from 42 to 67% (w/w) saccharides and their molar masses varied from 283 to over 1000 kDa. Significant biospecific interaction of separated fractions with the lectin concanavalin A was evaluated by the precipitation and sorption method (equilibrium constants) and further characterized using surface plasmon resonance to determine kinetic association and dissociation constants. K (D) was determined over the range 10(-7) M. High-molar-mass preparations appeared to be more suitable for preparation of stable and active complexes with concanavalin A for prospective use as a penicillin G acylase biocatalyst in enzyme reactors. The enzyme stability of such complexes was significantly increased compared with the original neoglycoprotein. Lower-molar-mass preparations were more suitable for applications such as biocatalysts in bioanalytical devices.
Masárová
Jana
J
Institute of Chemistry, Slovak Academy of Sciences, SK-84238 Bratislava, Slovakia. jana.masarova@savba.sk
Mislovicová
Danica
D
Mendichi
Raniero
R
Svitel
Juraj
J
Gemeiner
Peter
P
Danielsson
Bengt
B
D000327:Adsorption; D000586:Amination; D001426:Bacterial Proteins / Q000378:metabolism*; D001709:Biotechnology; D002384:Catalysis; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D003208:Concanavalin A / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000737:chemistry; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology; D006023:Glycoproteins / Q000378:metabolism*; D007700:Kinetics; D037102:Lectins / Q000378:metabolism; D008351:Mannans / Q000737:chemistry / Q000378:metabolism*; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010399:Penicillin Amidase / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000737:chemistry; D020349:Surface Plasmon Resonance
D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; D006023:Glycoproteins; D037102:Lectins; D008351:Mannans; D003208:Concanavalin A; D010399:Penicillin Amidase