scholarly journals Preparation of some immobilized linked enzyme systems and their use in the automated determination of disaccharides

1974 ◽  
Vol 137 (1) ◽  
pp. 25-32 ◽  
Author(s):  
D. J. Inman ◽  
W. E. Hornby
Keyword(s):  
Glucose Oxidase ◽  
Enzyme System ◽  
Inside Surface ◽  
Enzyme Systems ◽  
Nylon Tube ◽  
In Series ◽  
Automated Determination

1. Glucose oxidase (EC 1.1.3.4), amyloglucosidase (EC 3.2.1.3), invertase (EC 3.2.1.26) and β-galactosidase (EC 3.2.1.23) were covalently attached via glutaraldehyde to the inside surface of nylon tube. 2. The linked enzyme system, comprising invertase immobilized within a nylon tube acting in series with glucose oxidase immobilized in a similar way, was used for the automated determination of sucrose. 3. The linked enzyme system, comprising β-galactosidase immobilized within a nylon tube acting in series with glucose oxidase immobilized in a similar way, was used for the automated determination of lactose. 4. The linked enzyme system, comprising amyloglucosidase immobilized within a nylon tube acting in series with glucose oxidase immobilized in a similar way, was used for the automated determination of maltose. 5. Mixtures of glucose oxidase and amyloglucosidase were immobilized within the same piece of nylon tube and used for the automated determination of maltose. 6. Mixtures of glucose oxidase and invertase were immobilized within the same piece of nylon tube and used for the automated determination of sucrose.

scholarly journals The immobilization of enzymes on nylon structures and their use in automated analysis

1972 ◽  
Vol 129 (2) ◽  
pp. 255-262 ◽  
Author(s):  
D. J. Inman ◽  
W. E. Hornby
Keyword(s):  
Glucose Oxidase ◽  
Automated Analysis ◽  
Nylon Membrane ◽  
Nylon Tube ◽  
Immobilization Of Enzymes ◽  
The Matrix ◽  
Immobilized Glucose Oxidase ◽  
Derivatives Of ◽  
Automated Determination

1. Glucose oxidase (EC 1.1.3.4) and urease (EC 3.5.1.5) were covalently attached through glutaraldehyde to low-molecular-weight nylon powder. 2. Immobilized derivatives of glucose oxidase and urease were prepared by cross-linking the respective enzymes within the matrix of a nylon membrane. 3. An improved process is described for the immobilization of glucose oxidase and urease on the inside surface of partially hydrolysed nylon tube. 4. Automated analytical procedures are described for the determination of glucose with each of the three immobilized glucose oxidase derivatives and for the determination of urea with each of the three immobilized urease derivatives. 5. The efficiencies of the three immobilized enzyme structures as reagents for the automated determination of their substrates were compared.

Single- and coupled-enzyme nylon-tube reactors for plasma glucose determination with glucose oxidase and aldehyde dehydrogenase.

1980 ◽  
Vol 26 (12) ◽  
pp. 1652-1655 ◽  
Author(s):  
W Hinsch ◽  
A Antonijewić ◽  
P V Sundaram
Keyword(s):  
Glucose Oxidase ◽  
Aldehyde Dehydrogenase ◽  
Plasma Glucose ◽  
Continuous Flow ◽  
Flow System ◽  
Low Cost ◽  
Immobilized Enzymes ◽  
Glucose Determination ◽  
Nylon Tube

Abstract We describe routine methods for determining glucose in plasma with use of aldehyde dehydrogenase or glucose oxidase-aldehyde dehydrogenase immobilized in a nylon tube that is integrated into a continuous-flow system. Although the coupled-enzyme nylon-tube reactors require the presence of a third enzyme, catalase, in solution, the kinetics are not so complicated as to preclude reliable routine determination of glucose at very low cost. Precision is good, and results correlate well with those by the method involving glucose oxidase in solution. More than 3000 tests may be carried out with one reactor. The immobilized enzymes are stable for several months at 4 degrees C when not in use.

scholarly journals The preparation of nylon-tube-supported hexokinase and glucose 6-phosphate dehydrogenase and the use of the co-immobilized enzymes in the automated determination of glucose.

1975 ◽  
Vol 147 (3) ◽  
pp. 593-603 ◽  
Author(s):  
D L Morris ◽  
J Campbell ◽  
W E Hornby
Keyword(s):  
Thermal Stability ◽  
Automated Analysis ◽  
Immobilized Enzymes ◽  
Nylon Tube ◽  
The Individual ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Thermal Stability Of ◽  
Automated Determination

Triethyloxonium tetrafluoroborate was used to O-alkylate nylon-tube thus producing the imidate salt of the nylon which was further made to react with 1,6-diaminohexane. 2. Hexokinase (EC 2.7.1.1) and glucose 6-phosphate dehydrogenase (EC 1.1.1.49) were immobilized on the amino-substituted nylon tube through glutaraldeyde and bisimidates. 3. The effect of varying the conditions of O-alkylation and the amount of enzyme immobilized on the activity of nylon tube-hexokinase derivatives was determined. 4. The effect of varying the amount of enzyme immobilized on the activity of nylon-tube-glucose 6-phosphate dehydrogenase derivatives was determined. 5. The thermal stability of nylon-tube-hexokinase and nylon-tube-glucose 6-phosphate dehydrogenase derivatives was studied. 6. Different ratios of hexokinase and glucose 6-phosphate dehydrogenase were co-immobilized on nylon tube, and the rate of conversion of glucose into 6-phosphogluconolactone was compared with the individual activities of the immobilized enzymes. 7. Hexokinase and glucose 6-phosphate dehydrogenase co-immobilized on nylon tube were used in the automated analysis of glucose.

An Improved Automated Determination of Serum Total Cholesterol with a Single Color Reagent

1966 ◽  
Vol 12 (10) ◽  
pp. 681-689 ◽  
Author(s):  
Walter D Block ◽  
K John Jarrett ◽  
Jacob B Levine
Keyword(s):  
Total Cholesterol ◽  
Ferric Chloride ◽  
Flow Cell ◽  
Serum Total Cholesterol ◽  
Color Reagent ◽  
In Series ◽  
Tubular Flow ◽  
Automated Determination ◽  
Heating Bath

Abstract An anhydrous color reagent containing ferric chloride is pumped through Tygon tubing to a glass coil in a 95° heating bath. Manually prepared serum extracts (1:20 isopropanol dilution) are presented to the stream of preheated color reagent, and the two are passed through three mixing coils, connected in series. The absorbance of the resultant color is determined at 550 mµ in a 15 mm. tubular flow-cell. The improved N-automated procedure gave values of greater precision (5.6%) for total cholesterol determined in replicate samples from sera pools than the N-automated procedure which lacked precision (27.7%). The values found in 60 individual serums are within 6.0% of total cholesterol values determined by the Abell et al. method.

Enzymatic Determination of Starch in Feeds, Feed Ingredients, and Feces

1969 ◽  
Vol 52 (5) ◽  
pp. 956-958
Author(s):  
W W Turner
Keyword(s):  
Glucose Oxidase ◽  
Constant Temperature ◽  
Enzyme System ◽  
Aqueous Alcohol ◽  
Feed Ingredients ◽  
Enzymatic Determination ◽  
Oxidase Enzyme

Abstract Starch is determined in various materials such as feeds, grains, cereals, grasses, and feces by the combination of some existing techniques plus new methodology. Samples are extracted with aqueous alcohol and hydrolyzed with HCl; the resulting glucose is measured with a glucose oxidase enzyme system. The actual handling operations, including troublesome nitrations and constant temperature baths, are minimized or eliminated.

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