FAQ - Enzymes

Enzymes are naturally occurring biological macromolecules made up of proteins. They accelerate, or catalyze, chemical reactions in biological systems.

Enzymes convert substrates — molecules that the enzyme reacts with — into different molecules known as products. They do this through a highly specific "binding site" where the substrate fits like a lock and key. Once the substrate is released, it can react with other chemicals to allow quantification of the amount of substrate in a sample.

Enzymes are present in every species of animal, plant, and fungi. NECi began by extracting Nitrate Reductase from corn seedlings and purifying it for commercial sale — this is known as a native enzyme.

Since the DNA sequence of nitrate reductase is known, a recombinant form was later designed 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 and speed up chemical reactions under gentle biological conditions. They are present in our bodies and in the food we eat, and are completely non-toxic to handle, ship, and dispose of.

Because enzymes only react with their specific substrate, they are excellent candidates for quantifying specific molecules in a sample — even in complex mixtures. This selectivity, combined with their sensitivity and non-toxic nature, makes enzymes ideal reagents for accurate analytical chemistry without hazardous materials or methods.

Enzyme reactions occur under gentle biological conditions — no organic solvents, heavy metals, high heat, or pressure involved. Unlike traditional nitrate analysis methods that rely on toxic metals like cadmium or zinc, NECi's enzyme-based systems are just as accurate (if not more so), while being safe for both users and the environment. Cadmium disposal, in particular, is difficult and hazardous — our enzyme-based approach eliminates that concern entirely.

Nitrate is a negatively charged ion composed of one nitrogen atom and three oxygen atoms (formula weight 62 g/mol). It is very soluble in water.

Nitrite is a negatively charged ion composed of one nitrogen atom and two oxygen atoms (formula weight 46 g/mol). It is also very soluble in water.

Nitrate is naturally occurring and essential for plant growth and amino acid production. However, human activity has significantly altered nitrate concentrations in the environment. Sources include agricultural fertilizer inputs, industrial processing waste, and municipal waste.

Concentrations in undisturbed ambient water sources are generally at or below 2 ppm nitrate-N.

Nitrate may be reported in different units depending on region. Most of the U.S. reports in nitrate-N (measuring only the nitrogen in the molecule), while California and Europe report in units of nitrate (the entire molecule), resulting in higher numerical values.

The scientific community typically uses molarity (mM) or moles per liter. The conversion is: 1 ppm nitrate-N = 4.4 ppm nitrate = 71 mM nitrate.

Testing for nitrate allows you to maximize crop yields, protect water quality, protect livestock from nitrate toxicity, and ensure regulatory compliance with mandated safe nitrate levels.

The US EPA sets the maximum contaminant level at 10 ppm nitrate-N for safe drinking water. In the ocean, levels above 2 ppm nitrate-N can begin killing coral species and contribute to harmful algal blooms. Private well owners should note that the EPA does not regulate private wells — testing is the homeowner's responsibility.

There are three main areas of concern: drinking water, food, and aquatic ecosystems.
The US EPA sets the maximum contaminant level at 10 ppm nitrate-N for safe drinking water. Livestock and other ruminants may tolerate slightly higher levels, though recommendations vary by species.
Nitrate occurs naturally in many vegetables — especially leafy greens and root crops like spinach, celery, beets, and carrots — and is used to prevent bacterial growth in cured meats. The acceptable level of nitrate in food remains a subject of ongoing scientific discussion.
In the ocean, levels above 2 ppm nitrate-N can begin killing coral species and contribute to algal growth, which may cause toxic blooms and dead zones. Once nitrate reaches excess levels in water, it is very difficult to remove due to its high solubility.

Nitrate affects different individuals depending on health, age, and exposure level. Most nitrate consumed through food or water is quickly eliminated through urine — it is water soluble and does not accumulate in body fat. However, chronic exposure (such as from a contaminated well) can lead to health issues.
When nitrate is consumed, bacteria in the body convert it to nitrite, which can bind to hemoglobin to form methaemoglobin, reducing the blood's ability to carry oxygen. This is especially a concern for infants and can cause a condition known as methaemoglobinaemia, or "blue-baby syndrome." The elderly and individuals with gastrointestinal infections are also more susceptible.
Research published in Environmental Health Perspectives (April 2016) also suggests nitrate may affect thyroid function by blocking iodide uptake, which may adversely affect certain stages of child development.
Note: Nitrate must be ingested to have a negative health impact — it is not absorbed through the skin.

Phosphate is a molecule in which one phosphorus atom is bound to four oxygen atoms. It is the naturally occurring form of the element phosphorus, which is mined for agriculture and other industrial processes.

Phosphorus is a finite resource used in agriculture to stimulate plant growth. World phosphate rock reserves are believed to be in steep decline, driving fertilizer prices up. Monitoring phosphate levels helps prevent crop deficiency, reduces excess runoff into aquatic systems, and conserves resources.

Excess phosphate in waterways contributes to toxic algal blooms and dead zones in surface waters. NECi's kits can detect concerning levels of phosphate in environmental and industrial effluent.

The levels of phosphate measured in environmental systems depend on a number of factors, including proximity to human influence.

Phosphate is a macro-nutrient vital to plant growth. It is a component of nucleic acids and plays an important role in plant reproduction. For certain crops such as grains, it is essential that the plant has the ability to reproduce effectively, making adequate phosphate levels critical to a successful harvest.

There are two main types to be aware of: organic and inorganic phosphate. Inorganic phosphate (also called orthophosphate) is the only form available to plants. Organic phosphate exists in plant or animal tissue and must first be decomposed by microorganisms before plants can use it.

Both forms can also shift between soluble (dissolved in water, available for uptake) and insoluble (attached to soil or sand particles) states. When measuring phosphate, it is important to know whether it is organic or inorganic, soluble or insoluble.

Traditional methods include ion chromatography, the ascorbic acid method, the stannous chloride method, and digestion methods. These typically involve expensive equipment, unstable or dangerous reagents, and high temperatures, and can be inaccurate.

NECi's enzymatic phosphate method is accurate, fast, and involves no harmful reagents. It is formatted for both on-site and laboratory use.