Selective Biosensing of l-Lysine by a Low-Temperature Flow-Injection Technique Using an Immobilized Lysine Oxidase Reactor

Abstract

An automated flow-injection analytical system, equipped with an immobilized L-lysine-a-oxidase reactor (stable for over 6 months) and having a hydrogen peroxide electrode in the manifold, was developed for biosensing L-lysine. A substantial improvement in the selectivity for lysine was achieved by lowering the system temperature to 10°C; the sensor response was linearly dependent on the L-lysine concentration (from tens of µM to mM order). The analytical results were verified by HPLC using protein hydrolyzates as test solutions comprising various interfering amino acids. By the aid of an in-line filter, the leaching of lysine from L-lysine-enriched fish feed into an aqueous buffer was automatically monitored.

Application of Amperometric Biosensors to the Determination of Vitamins and α-Amino Acids

“…Glutaraldehyde was also used with collagen, , gelatin, , avidin, or egg albumin . In other cases, some supports were activated with glutaraldehyde: controlled-pore glass beads, ,,− polyion complex single or double membrane constituted by poly( l -lysine) and poly(4-styrene sulfonate), , cellulose membranes, ,, polycarbonate membrane, silica gel, , or nylon membrane. ,, A packed enzyme bead electrode is inherently more sensitive than an electrode with enzyme localized by a membrane of functionalization of the nylon by immersing it in boiling dimethyl sulfoxide; after washing, a lysine arm is linked to the membrane, which is again reacted with glutaraldehyde.…”

“…Three different enzymes are known to be applicable to analytical systems for l -lysine quantitation: l -lysine-2-monooxygenase (LMO), l -lysine-α-oxidase (LO), and l -lysine decarboxylase (LD) catalyze the reactions A flow-injection system based on LMO showed high specificity, assay rate and precision and good reproducibility, but the kinetic behavior of LMO did not allow the determination of l -lysine at concentrations lower than 5.5 mM . Selective biosensing by a low-temperature flow-injection technique using an immobilized LO reactor has been developed . When the system temperature is 10 °C, a substantial improvement in the selectivity for l -lysine was achieved …”

“…Aging consisted of exposure of the stock glutaraldehyde solution to air at room temperature for at least 24 h. 85 Glutaraldehyde was also used with collagen, 28,86 gelatin, 87,88 avidin, 89 or egg albumin. 29 In other cases, some supports were activated with glutaraldehyde: controlled-pore glass beads, 33,74,[90][91][92][93][94][95][96] polyion complex single or double membrane constituted by poly(Llysine) and poly(4-styrene sulfonate), 59,60 cellulose membranes, 62,97,98 polycarbonate membrane, 99 silica gel, 100,101 or nylon membrane. 86,95,102 A packed enzyme bead electrode is inherently more sensitive than an electrode with enzyme localized by a membrane.…”

“…182 Selective biosensing by a lowtemperature flow-injection technique using an immobilized LO reactor has been developed. 92 When the system temperature is °C, a substantial improvement in the selectivity for L-lysine was achieved. 92 In general, arginine, 35,86,100,105 phenylalanine, 35,100 ornithine, 66,86,100 and cysteine 83,86,114,130 were the main interferents.…”

“…92 When the system temperature is °C, a substantial improvement in the selectivity for L-lysine was achieved. 92 In general, arginine, 35,86,100,105 phenylalanine, 35,100 ornithine, 66,86,100 and cysteine 83,86,114,130 were the main interferents.…”