Enzymes are naturally occurring biological macromolecules that are made up of proteins. They are responsible for accelerating, or catalyzing, chemical reactions in many biological systems.

Enzymes convert substrates, which are molecules that the enzyme reacts with into different molecules, known as products.

Using enzymes to test for nitrate content can allow people to: maximize crop yields, protect water quality, protect livestock from nitrate toxicity, and also make sure that organizations stay in regulatory compliance with the mandated safe nitrate levels.

Enzymes only react with their specific substrate, which makes them excellent candidates for quantifying specific molecules in a sample, even in complex mixtures.

They do this by having a very specific "binding-site", where the substrate will fit, like a lock-and-key or two puzzle pieces would. When the substrate is released, it can react with other chemicals to quantify the amount of substrate in the sample.

Enzymes are present in nearly every species of animal, plant, and fungi. NECi started by extracting Nitrate Reductase from corn seedlings, then purifying it for commercial sale. This is known as a native enzyme, since it was extracted from its natural source.

Since the DNA sequence of nitrate reductase is known, a recombinant form was designed in order to harness desired properties and allow for more efficient production using a vector yeast, Pichia pastoris. Recombinant protein technology allows for modification of native DNA for optimized properties and the ability to produce the enzyme outside of its native source.

No! Enzymes are essential for almost every organism to perform routine functions. They speed up chemical reactions in all sorts of biological systems, and react under those gentle conditions. They're in our bodies and in the food that we eat, and are completely non-toxic to handle, ship, and dispose of, but please don't eat them!

Enzymes are catalysts that drive complex biological reactions. They happen to be excellent reagents for analytical chemistry because they are reliable, accurate, sensitive, selective, and non-toxic.

Enzyme reactions occur in gentle, biological conditions with no organic solvents, heavy metals, high heat or pressure involved.

They are sensitive, which means they have low detection limits in complex mixtures.

They will ONLY react with a specific substrate, using their highly specialized binding site that only fits THEIR substrate . Once their substrate is bound, catalysis changes the substrate into a new product. After this is complete, the enzyme releases the newly formed product and is ready to begin the process again!

Analyzing samples with enzymes will result in accurate and consistent data without the need for any hazardous materials or methods, unlike methods employing cadmium or zinc for nitrate analysis.

NECi Superior Enzymes has developed a variety of products to fit the needs of users with various backgrounds and experience. Each product has been formatted for its specific application so that anyone, from elementary students to laboratory scientists, can find a product that fits their application.

On-Site test kits are designed for taking quick measurements of nitrate or phosphate in soil, water, or plant samples where they are collected. They are simplified versions of the lab kits, providing accurate and reliable results without the need for laboratory equipment. However, we've designed an affordable portable, smartphone linked photometer for these kits to provide digital data in the field, increasing accuracy of the results.

Laboratory test kits are for users with knowledge of basic laboratory procedures and access to laboratory equipment (test tubes, spectrophotometer, pipettes, etc.)

Enzyme reagent packs are designed for experienced users that prefer to supply their own buffers and color reagents (if necessary), and are available with an optimized amount of co-factor included or as stand-alone enzyme.

All reagents have been designed and developed to be used with spectrophotometers, automated liquid handling equipment, discrete analyzers, etc.

Most importantly, all test kits were designed for ease of use and reliability, to get the best results with the safest and most environmentally friendly reagents in any commercial nitrate test kit available today!

NECi's nitrate test kits use an enzyme called nitrate reductase to convert the nitrate in a sample to nitrite. Once the Griess color reagents are added, nitrite becomes visible as a pink solution. This visible pink color is analyzed by comparison to a color chart, or by measuring absorbance at 540 nm in a spectrophotometer, or NECi's handheld dual wavelength photometer. The darker the shade of pink is, the more nitrate is in the sample.

NECi's enzyme based phosphate test kits work in a similar manner: the enzyme reacts with all of the phosphate in a sample to convert it to a new molecule, however there are no color reagents added, and therefore no color change occurs. Instead, absorbance is measured in the UV range at 340 nm using a spectrophotometer or NECi's handheld dual wavelength photometer, detecting the amount of the newly formed product, which correlates to the amount of phosphate in a sample.

We recommend that laboratory users begin by purchasing a complete test kit for convenience of pre-made buffers and detailed instructions. This limits error and helps with troubleshooting. Once users are familiar with the assay, economical reagent packs are available which includes only the enzyme, co-factor, and enzyme reconstitution buffer.

All on-site kits are simplified, pre-calculated versions of the laboratory methods and are the best choice for fast and accurate analysis anywhere on-site. These kits require no laboratory equipment or chemistry experience and can be used to analyze water, plant, soil, forage, and aqueous process effluent samples.

For high throughput automated laboratory analysis, we have developed reagent packs complete with instructions customized to the users equipment. These reagent packs include enzyme, co-factor, and enzyme reconstitution buffer, complete with instructions for instruments such as discrete analyzers, flow through analyzers, and more. Contact NECi today to develop a custom kit for your automated laboratory instrument.

NECi offers on-site phosphate kits formatted for Water and Soil Samples, with the testing range of between 0 and 20 ppm phosphate for standard-range kits. There is also a low-range water option good between 0.5 and 5 ppm.

For the test tube formatted phosphate lab kits customers can choose from a low-range option measuring from 0.5 to 5 ppm phosphate and a standard-range formatted kit good from 0-20 ppm phosphate.

If you need a different testing range, you can contact NECi and, if possible, a kit can be formatted to meet that need.

Most components should be stored cold and dark: see below for outlines for each of the components’ cold-storage requirements.

With regards to storage equipment, it is important to note that NADH and Enzyme (AtNaR, YNaR) are sensitive to oxidizing agents and thus clear glass, PP (polypropylene), or PE (polyethylene) tubes and vials should be used.

The enzyme packs contain the enzyme and NADH, freeze-dried in cuvettes and containers, which are then vacuum-sealed in foil pouches.

There will be a 6-month expiration date on all kits, but packs consistently are stable for more than 6 months stored at any temperature below 22C; stability study samples have been effective even after multiple years. The enzyme can remain stable for years if stored in these packets at -20C.

Because NADH oxidizes to NAD+ over time it will eventually be unable to supply the electrons needed for the nitrate reduction step.

Use these precautions for NADH: store portioned into small aliquots - the volume you would use in a day's work. Store the aliquots in a fridge (at least one week) or frozen (one month). You may see longer activity retention - a month or more - but one or two weeks is safe.

The assay buffer, when stored cold, is stable for one month.

Both Color Reagent 1 and Color Reagent 2 are stable enough for use after being stored for up to one month cold, at or below 4C, and dark.

Color reagent 2 (NED reagent) will start to turn brownish after some weeks after the addition of water. This will cause a loss in sensitivity as the background in blanks becomes too high.

There will be a 6-month expiration date on all kits. Packs consistently are stable for more than 6 months stored at any temperature below 22C; stability study samples have been effective even after multiple years.

The enzyme can be stable for years if stored in these packets at -20C.

Once the kit process has been started you should aim to complete it in a timely fashion, being sure to FOLLOW STATED TIMES carefully. The most common errors are either stopping the reduction reaction too soon, or not proceeding to the next step in time.

DO NOT let the solutions you prepare sit for extended periods of time. Once started, the kit process should be completed that day.

It is best to keep components cold and sealed until use.

DO NOT open or reconstitute ANY component until ready to use your kit.

As supplied in freeze-dried form, PNP is stable for months when stored at -20 C or colder.

Once reconstituted, PNP should be used that day.

As supplied in freeze-dried form, MESG is stable for months when stored at -20 C or colder.

Once reconstituted, MESG should be used that day.

The buffer can be stable for months when stored at 4 C.

Do not open until ready to use.