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The slope of the response was measured and used as an indication of the amount of serine protease, activity. Suitable polymeric dye transfer inhibiting agents include, but are not limited to, polyvinylpyrrolidone polymers, polyamine N-oxide polymers, copolymers of N-vinylpyrrolidone and N-vinylimidazole, polyvinyloxazolidones and polyvinylimidazoles or mixtures thereof. In some embodiments, the required level of enzyme is achieved by the addition of one or more species of neutral metalloprotease provided by the present invention. Indeed, it is contemplated that any suitable builder will find use in various embodiments of the present invention. Most effort has been focused on increasing the stability of the protease by altering structural determinants identified through the use of molecular modeling suggested to prevent local unfolding processes that would result in autolysis of the protein and cause the neutral protease to denature at high temperatures See e.

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Blank controls, as well as a control containing two microswatches and detergent but no enzyme were also included. Considering the bleeding risk, physician may not prescribe aspirin unless for other needed. The present invention also provides methods of cleaning, the comprising the steps of:.

Note that if a convenient flanking restriction site is available, the above method need be used only in connection with the flanking region which does not contain a site. In this Example, methods used to assess the citrate-induced autolysis of wild-type and recombinant variant nprE e.

After identification of plasmids containing the correct DNA construct sequence for plasmid pEL, transformed into competent B. In some preferred embodiments, the composition comprises a sufficient amount of a pH modifier to provide the composition with a neat pH of from about 3 to about 5, the composition being essentially free of materials that hydrolyze at a pH of from about 3 to about 5.

In some embodiments, these necessary elements are supplied as the gene's own homologous promoter if it is recognized, i.

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As used herein, the term "polymerase chain reaction" "PCR" refers to the methods of U. Indeed, much research has been focused on the engineering Bacillus subtilis neutral proteases to increase the thermal stability of the enzyme See e. The term "introduced" in the context of inserting a nucleic acid sequence into a cell, means transformation, transduction or transfection.

This property is defined herein as the "primary property. Also, as used herein, the singular terms "a," "an," and "the" include the plural reference unless the context clearly indicates otherwise.

Third, since the duration of database, we may lose a number of patients who had AA diagnosis before and still alive, but lost to follow-up during to Fist all buffers, stock, and working solutions were prepared. Patients who were diagnosed with AA and also received computed tomography CT were included in this study.

Citrate is a detergent component that rapidly causes the autolysis of NprE, in the absence of calcium. Since all samples were desalted to remove any exogenous metals, these weakly bounding calcium sites are expected to be unoccupied.

It is not intended that the present invention be limited to any particular detergent formulation or composition. The pUBnprE expression vector, primers, synthesized at Invitrogen desalted, 50 nmol scale were used to generate the libraries.

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Thus, the addition of zinc may increase the storage life of neutral metalloprotease in the presence of detergent components. Please enter a valid email address. Enzyme substrate specificities vary, depending upon the substrate tested. Moreover, many stains are composed of complex mixtures of fibrous material, mainly incorporating carbohydrates and carbohydrate derivatives, fiber, and cell wall components e. Similarly, the binding of FluoZin-3 was monitored by excitation at nm and the emission spectra were collected from to nm.

Posted on Jul 30, The filtered culture broth was diluted with dilution buffer 1. In particular, the better wash performance of PMN or neutral metalloprotease from B. Polyethylene glycol, of a molecular weight of x. The intact nprE is 32 kDa. The possible reason of the phenomena might be that there is no routine CT screening program in Taiwan, and asymptomatic AA is an imperceptible disease for general population.

EP1934340B1 - Use and production of storage-stable neutral metalloprotease - Google Patents

In some embodiments, the fabric comprises any fabric capable of being laundered in normal consumer use conditions. Following this incubation, the cell pellet was harvested by sufficient centrifugation to provide a cell pellet.

As used herein, the term "target," when used in reference to amplification methods e. In this Example, experiments conducted to assess the performance of various nprE variants in the BMI assay outlined above are described.

Before cutting of the swatches, the fabric EMPA was washed with water. The assay was mixed for 5 minutes and the kinetic change at nm was read over 2 to 5 minutes.