On-column enzyme-catalyzed microreactions using capillary electrophoresis: quantitative studies.
Capillary electrophoresis and on-column enzyme-catalyzed microreactor techniques were used to quantitate the reaction projects resulting from three model systems: i) the conversion of nicotinamide adenine dinucleotide (NAD) to nicotinamide adenine dinucleotide, reduced form (NADH) in the oxidation of glucose-6-phosphate (glc-6-p) to 6-phosphogluconate by glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49); ii) the conversion of adenosine triphosphate (ATP) to adenosine diphosphate (ADP) and adenosine monophosphate (AMP) by hexokinase (HK, EC 2.7.1.1) and apyrase (APY, EC 3.6.1.5), respectively, in the conversion of glucose to glucose-6-phosphate and inorganic phosphate, respectively, and; iii) the conversion of fructose-1,6-bisphosphate to dihydroxyacetone phosphate and glyceraldehyde-3-phosphate by fructose-biphosphate aldolase (ALD, EC 4.1.2.13). Single and double microreactor techniques employing direct or indirect detection were used to follow the conversion of substrate to product(s). In addition, electrophoresis conditions including voltage, enzyme concentration, and mixing time of the reaction, were correlated to product distribution profiles.
Zhang
Ying
Y
Department of Chemistry and Biochemistry, California State University, Los Angeles 90032-8202, USA. fgomez2@calstatela.edu
Kaddis
John
J
Silverio
Catherine
C
Zurita
Cecilia
C
Gomez
Frank A
FA
D000244:Adenosine Diphosphate / Q000032:analysis*; D000255:Adenosine Triphosphate / Q000032:analysis*; D001081:Apyrase / Q000378:metabolism; D019149:Bioreactors; D019075:Electrophoresis, Capillary / Q000379:methods*; D004798:Enzymes / Q000032:analysis*; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism; D006593:Hexokinase / Q000378:metabolism; D008832:Microchemistry / Q000379:methods; D009243:NAD / Q000032:analysis*; D010084:Oxidation-Reduction; D015203:Reproducibility of Results
D004798:Enzymes; D009243:NAD; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D005954:Glucosephosphate Dehydrogenase; D006593:Hexokinase; D001081:Apyrase; D005634:Fructose-Bisphosphate Aldolase