Rennet () is a complex set of enzymes produced in the stomachs of ruminant mammals. Its key component is chymosin, a protease enzyme that curdles the casein proteins in milk. Rennet also contains other enzymes such as pepsin and a lipase. Rennet has traditionally been used to separate milk into solid curds and liquid whey, used in the production of cheeses. Rennet from calves has become less common for this use, to the point that less than 5% of cheese in the United States is made using animal rennet today. Most cheese is now made using chymosin derived from bacterial sources.
Molecular action of rennet enzymes One of the main actions of rennet is its protease chymosin cleaving the kappa casein chain. Casein is the main protein of milk. Cleavage removes the slightly negatively charged glycomacropeptide (GMP) from the surface of the casein micelle. Because negative charges repel other negative charges, the GMP prevents casein micelles from adhering to each other. With the GMP removed, the casein micelles can begin to cluster and lose their polar charge, causing them to rise out of the polar water molecules and join non-polar milk fat as a portion of the cheese curd. This action is enhanced in the presence of strong ions like those formed from calcium and phosphate. As such, those chemicals are occasionally added to supplement pre-existing quantities in the cheese making process, especially in calcium phosphate-poor goat milk. The solid truncated casein protein network traps other components of milk, such as fats and minerals, to create cheese.
Extraction of calf rennet Calf rennet is extracted from the inner mucosa of the fourth stomach chamber (the abomasum) of young, nursing calves as part of livestock butchering. These stomachs are a byproduct of veal production. Rennet extracted from older calves (grass-fed or grain-fed) contains less or no chymosin, but a high level of pepsin and can only be used for special types of milk and cheeses. As each ruminant produces a special kind of rennet to digest the milk of its own species, milk-specific rennets are available, such as kid goat rennet for goat's milk and lamb rennet for sheep's milk.
Traditional method Dried and cleaned stomachs of young calves are sliced into small pieces and then put into salt water or whey, together with some vinegar or wine to lower the pH of the solution. After some time (overnight or several days), the solution is filtered. The crude rennet that remains in the filtered solution can then be used to coagulate milk. About 1 gram of this solution can normally coagulate 2 to 4 litres of milk.
Modern method Deep-frozen stomachs are milled and put into an enzyme-extracting solution. The crude rennet extract is then activated by adding acid; the enzymes in the stomach are produced in an inactive form and are activated by the stomach acid. The acid is then neutralized and the rennet extract is filtered in several stages and concentrated until reaching a typical potency of about 1:15,000; meaning 1 g of extract can coagulate 15 kg of milk. One kilogram of rennet extract has about 0.7 g of active enzymes – the rest is water and salt and sometimes sodium benzoate (E211), 0.5%–1.0% for preservation. Typically, 1 kg of cheese contains about 0.3 mg of rennet enzymes.
Alternative sources Because of the limited availability of mammalian stomachs for rennet production, cheese makers have sought other ways to coagulate milk since at least Roman times. The many sources of enzymes that can be a substitute for animal rennet range from plants and fungi to microbial sources. Cheeses produced from any of these varieties of rennet are suitable for lactovegetarians, as well as those keeping Kosher. Fermentation-produced chymosin is used more often in industrial cheesemaking in North America and Europe today because it is less expensive than animal rennet.
Vegetable Many plants have coagulating properties. Homer suggests in the Iliad that the Greeks used an extract of fig juice to coagulate milk. Other examples include several species of Galium, dried caper leaves, nettles, thistles, mallow, Withania coagulans (also known as Paneer Booti, Ashwagandh and the Indian Cheesemaker), and ground ivy. Some traditional cheese production in the Mediterranean uses enzymes from thistle or Cynara (artichokes and cardoons). Phytic acid, derived from unfermented soybeans, or fermentation-produced chymosin (FPC) may also be used. Vegetable rennet might be used in the production of kosher and halal cheeses, but nearly all kosher cheeses are produced with either microbial rennet or FPC. Commercial so-called vegetable rennets usually contain an extract from the mold Rhizomucor miehei described below.
Microbial Some molds such as Rhizomucor miehei are able to produce proteolytic enzymes. These molds are produced in a fermenter and then specially concentrated and purified to avoid contamination with unpleasant byproducts of the mold growth. The traditional view is that these coagulants result in bitterness and low yield in cheese, especially when aged for a long time. Over the years, microbial coagulants have improved greatly, largely due to the characterization and purification of secondary enzymes responsible for bitter peptide formation/non-specific proteolytic breakdown in cheese aged for long periods. Consequently, it has become possible to produce several high-quality cheeses with microbial rennet. It is also suitable for the elaboration of vegetarian cheese, provided no animal-based ingredients are used in its production.
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