6 results about "Enzyme free" patented technology

The present application relates to the field of bioengineering and microbial technology, and relates to a yeast surface display construction method of a double-enzyme system. AGA1 The gene promoter is replaced by a strong constitutive promoter TEF1 and a glucose-responsive promoter HXT7 ; a double-enzyme free expression vector containing a galactose-inducible promoter GAL1 driven detoxification enzyme gene is constructed, and is transformed into a modified chassis cell to obtain a double-enzyme co-display engineering bacterium; the chassis protein AGA1 is expressed in a culture medium, and YtdepA and YtdepB are induced in a galactose-induced culture medium, so that the YtdepA and YtdepB are co-displayed on the yeast surface through the coupling mechanism of AGA1-AGA2. The strong constitutive promoter TEF1 and the glucose-repressed promoter HXT7 are coupled with the galactose-inducible promoter GAL1 respectively, so that the chassis protein and the detoxification enzyme are matched in the expression timing and intensity, and the detoxification enzyme is displayed on the yeast surface.

The present application relates to aldose reductase activity analysis technical field, specifically relates to a kind of aldose reductase activity and its inhibitory effect quantitative analysis method.Local substrate concentration regulation is constructed by dividing multiple detection segments, combining bypass sampling channel, electrochemical detection cavity and compensation injection point, and the substrate environment of each detection segment is maintained consistent;Enzyme-free reference channel parallel to microfluidic reaction channel is set, and the total absorbance change amount background is separated, to obtain the real enzymatic absorbance change amount;Based on sliding time window, extract local dynamic enzyme reaction rate, combined with wall adsorption correction to eliminate the influence of enzyme activity attenuation along the way, identify stable reaction interval through local fluctuation coefficient of variation, determine local half response time and local onset time;Local onset time is used as the integration starting point, to calculate comprehensive inhibition intensity, realize inhibitor onset order and inhibition strength spatial quantitative analysis, improve the accuracy, stability and spatial comparability of detection result.

Described herein is a one-pot, four-enzyme cascade of enzymes, three bound to quantum dots with one enzyme free in solution, for the conversion in vitro of fumarate to 1,3-diaminopropane. The cascade operates via two distinctly different enzymatic channeling mechanisms which simultaneously function to increase the overall rate. The first three enzymes of the pathway (AspB->LysC->Asd) were able to engage in channeling in a nanoparticle displayed format, but addition of the last two enzymes to this pathway in this format (AspB->LysC->Asd->Dat->Ddc) did not result in complete channeling through the entire pathway to the final diaminopropane product. Surprisingly, replacement of the last two enzymes (Dat->Ddc) with a naturally occurring fused Dat-Ddc hybrid (Daba) provided for full channeling in this system (AspB->LysC->Asd->Daba).

The application belongs to the field of glucose detection, and particularly relates to an enzyme-free glucose sensor based on a hydrophobic substrate and a preparation method and application thereof. The application can realize glucose detection in various pH solution environments, reduces the dependence on high OH ‑ concentration of a solution; and the oxygen reduction reaction can be carried out at a lower reduction potential, which provides convenience for low-energy consumption use of the sensor.

The utility model discloses an incubation module and full -automatic enzyme free instrument, wherein the incubation module includes the incubator cover, the incubation module is equipped with the placement area for placing the incubator cover, the incubator cover includes first cover, second cover, counterweight spare and heating assembly, second cover with first cover is connected to and is enclosed and forms the accommodation cavity, the counterweight spare is located in the accommodation cavity, the heating assembly includes heating spare and conducting structure, heating spare is located in the accommodation cavity, conducting structure with heating spare is connected, and at least part of conducting structure stretches out the accommodation cavity outside, when the incubator cover is located in the placement area, conducting structure can with external circuit guide to with heating spare heating. It is designed to improve the incubation efficiency while reducing the production of condensate during the incubation process, also avoid the risk of incubator cover being corroded.

This invention relates to the field of aldose reductase activity analysis technology, specifically to a quantitative analysis method for aldose reductase activity and its inhibitory effect. By dividing the detection into multiple segments and combining a bypass sampling channel, an electrochemical detection chamber, and a compensation injection point to construct a local substrate concentration control system, the substrate environment of each detection segment is maintained consistently. An enzyme-free reference channel parallel to the microfluidic reaction channel is set up to separate the background of the total absorbance change, obtaining the true enzymatic absorbance change. Based on a sliding time window, the local dynamic enzyme reaction rate is extracted, and the influence of enzyme activity decay along the process is eliminated by wall adsorption correction. The stable reaction interval is identified by the local fluctuation coefficient of variation, determining the local half-response time and local onset time. Using the local onset time as the integration starting point, the comprehensive inhibition intensity is calculated, achieving spatial quantitative analysis of the inhibitor's onset order and inhibitory strength, improving the accuracy, stability, and spatial comparability of the detection results.