PubMed · 3259408
The source did not provide an abstract. Follow the original record for more information.
Explore connections, maps & timelines
M R Fuh, L W Burgess, G D Christian. 1988-03-01. Single fiber-optic fluorescence enzyme-based sensor.. https://doi.org/10.1021/ac00156a012
Cite the original work for its findings. Save a collection to share your selection of sources.
KEEP EXPLORING
An aqueous two-phase system of polyethylene glycol (PEG) and potassium phosphate provided a favorable environment for bioconversion of penicillin G to 6-aminopenicillanic acid (6-APA). The recombinant E. coli cells containing penicillin acylase were partitioned in the phosphate-rich bottom phase, and the product 6-APA in the PEG-rich top phase, which protected the enzyme from deactivation for a longer period of time. The continuous production of 6-APA by recycling the bottom phase showed a slight decrease in enzyme activity from initial 80 mM of 6-APA production to 61 mM during the eight days of operation at a space velocity of 0.12 hours-1. This work showed a possibility that the aqueous two-phase whole-cell enzyme bioconversion could serve as an alternative to immobilized cells on solid matrices.
A flow injection system is described for the parallel determination of pullulan and glucose during a fermentation of the fungus Aureobasidium pullulans. The polysaccharide was hydrolyzed by pullulanase and amyloglucosidase, immobilized to controlled-pore glass (CPG). The glucose produced was oxidized by glucose dehydrogenase and the NADH formed determined photometrically. The pullulan concentration was calculated from the difference to the response obtained for free glucose. The calibration curves for monomer and polymer were both linear between 2 mg dm-3 and 20 mg dm-3. Analysis of one sample for the determination of glucose and pullulan took about 10 min.
Glucose oxidase (GOx) and glucoamylase (GA) were immobilized and coimmobilized through their carbohydrate moieties onto polyethyleneimine-coated magnetite crosslinked with glutaraldehyde and derivatized with adipic dihydrazide. The carbohydrates were oxidized with sodium periodate, and at optimal concentration, their Vm increased up to 18% for GOx and up to 16% for GA. After immobilization, a remaining activity as high as 88% and 70% for GA with maltose and maltodextrin respectively as substrates was obtained, independently of the particle loading. On the contrary, the remaining activity of GOx strongly decreased at high particle loading. Nevertheless, half of its initial activity was recovered at low loading and was not significantly affected when GA was coimmobilized by saturating the reactive groups left on the particle. The Vm of both immobilized enzymes was improved by crosslinking their carbohydrates with adipic dihydrazide, a treatment which allows further coimmobilization of the other enzyme on a second layer.