A biosensor based on co-immobilized L-glutamate oxidase and L-glutamate dehydrogenase for analysis of monosodium glutamate in food.
A monosodium glutamate (MSG) biosensor made by co-immobilized L-glutamate oxidase (L-GLOD) and L-glutamate dehydrogenase (L-GLDH) as the bio-component based on substrate recycling for highly sensitive MSG or L-glutamate determination, has been developed. Regeneration of MSG by substrate recycling provided an amplification of the sensor response. Higher signal amplification was found in the presence of ammonium ion. The sensor was standardized to determine MSG in the range of 0.02-3.0 mg/L. Linearity was obtained from 0.02 to 1.2 mg/L in presence of ammonium ion (10 mM) and NADPH (reduced nicotinamide adenine dinucleotide phosphate) (2 mM), but in absence of L-GLDH, the detection limit of MSG is confined to 0.1 mg/L. The apparent Km for MSG with L-GLOD-L-GLDH coupled reaction was 0.4451 mM but 1.9222 mM when only L-GLOD was immobilized. Cross linking with glutaraldehyde in the presence of bovine serum albumin (BSA) as a spacer molecule has been used for the method of immobilization. The response time of the sensor was 2 min. The optimum pH and temperature of the biosensor has been determined as 7+/-2 and 25+/-2 degrees C, respectively. The enzyme immobilized on the membrane was used for over 50 measurements. The standard error of the sample measurement was 4-5%. The activity of the enzyme-immobilized membrane was tested over a period of 60 days.
Basu
Anjan Kumar
AK
Department of Food Technology and Biochemical Engineering, Jadavpur University, Kolkata 700032, India.
Chattopadhyay
Parimal
P
Roychudhuri
Utpal
U
Chakraborty
Runu
R
D000594:Amino Acid Oxidoreductases; D015374:Biosensing Techniques; D005503:Food Additives / Q000032:analysis; D005504:Food Analysis; D005969:Glutamate Dehydrogenase; D010100:Oxygen; D011076:Polycarboxylate Cement; D012970:Sodium Glutamate / Q000032:analysis*
D005503:Food Additives; D011076:Polycarboxylate Cement; C005506:polycarbonate; D000594:Amino Acid Oxidoreductases; D005969:Glutamate Dehydrogenase; C023061:L-glutamate oxidase; D010100:Oxygen; D012970:Sodium Glutamate