Chymosin B from Aspergillus niger var. awamori

CAS 85713-24-2

Listed as a thickener, a stabiliser and an enzyme by the US.

Listed as thickeners (US) · stabilisers (US) · enzymes (US) · processing aids (US)

The B form of chymosin, the milk-clotting enzyme of calf rennet, made by fermentation rather than taken from calves. It is used in cheesemaking and is sold as a clear aqueous solution.

Where it is and isn’t permitted

Status of Chymosin B from Aspergillus niger var. awamori by jurisdiction. Every jurisdiction this site covers is listed, including those where the substance does not appear.
JurisdictionStatusScope and limitsCitation
Australia and New ZealandFood Standards Australia New ZealandNo listed food uses foundThis site has not matched this substance to any food use on this jurisdiction’s list of authorised food additives. A food additive may be used only in foods that list names. Flavourings, enzymes, processing aids and added nutrients are regulated separately and do not appear on it.
CanadaHealth CanadaNo listing matchedHealth Canada’s lists name additives without E or INS numbers, so this site matches them by exact name. A substance listed there under a name this site does not recognise has no row here.
European UnionEuropean Commission / EFSANo listed food uses foundThis site has not matched this substance to any food use on this jurisdiction’s list of authorised food additives. A food additive may be used only in foods that list names. Flavourings, enzymes, processing aids and added nutrients are regulated separately and do not appear on it.
Great BritainFood Standards Agency / Food Standards ScotlandNo listed food uses foundThis site has not matched this substance to any food use on this jurisdiction’s list of authorised food additives. A food additive may be used only in foods that list names. Flavourings, enzymes, processing aids and added nutrients are regulated separately and do not appear on it.
JapanConsumer Affairs AgencyNo listing matchedThis site imports only Japan’s list of designated additives (Appended Table 1 to the Regulations for Enforcement of the Food Sanitation Act), matched by English name. Existing additives, natural flavouring agents and ordinary foods used as additives are permitted outside that list and are not imported, so an empty row does not show that Japan does not permit the substance.
South KoreaMinistry of Food and Drug SafetyNo listing matchedThis site imports Part III of South Korea’s Standards and Specifications for Food Additives, matched by CAS number, then INS number, then exact name. Mixed preparations and sanitisers are not imported, and an entry whose identifiers disagreed with a page’s is held for review, so an empty row does not show that South Korea does not permit the substance.
United StatesFood and Drug AdministrationPermitted
Named in 21 CFR 184.1685, a direct food substance the regulation affirms as generally recognized as safe: chymosin preparation derived, via fermentation, from a nonpathogenic and n …

Named in 21 CFR 184.1685, a direct food substance the regulation affirms as generally recognized as safe: chymosin preparation derived, via fermentation, from a nonpathogenic and nontoxigenic strain of Aspergillus niger van Tieghem variety awamori containing the prochymosin gene (paragraph (a)(4)). The section is held on this site as the page rennet-animal-derived-and-chymosin-preparation-fermentation-derived; read it for the conditions.

21 CFR 184.1685retrieved 2026-10-18
Codex AlimentariusCodex Alimentarius Commission / JECFANot listedDoes not appear on this jurisdiction’s permitted list. That is not the same as being prohibited.

How much is permitted

A listing usually limits the quantity as well as the food. Only one of the forms that takes is a figure: quantum satis and good manufacturing practice permit the amount the technological effect requires and no more, which is a limit rather than the absence of one. Figures below are the range across every food category this page is listed in, and categories differ between regulators — see how much is permitted.

United States—1 no quantity stated

No regulator sets a figure for Chymosin B from Aspergillus niger var. awamori. That is not an absence of control: where the listing says quantum satis or good manufacturing practice, the quantity is limited to what the effect requires.

What it is made from

Made by fermenting Aspergillus niger var. awamori carrying the calf prochymosin gene, then converting the prochymosin to chymosin and purifying it.

CAS registry number
85713-24-2

How JECFA defines it

The international specification for identity and purity, prepared by the Joint FAO/WHO Expert Committee on Food Additives and published by FAO. It is a different document from any above, written by a different body and revised on its own cycle; it is advisory, and a country adopts it or does not.

Chymosin B from Aspergillus niger var. awamori containing the Prochymosin B Gene

Synonyms: Rennin, milk-clotting enzyme, chymosin, chymosin B, aspartyl protease Aspartyl protease; Chymosin; Chymosin B; Milk-clotting enzyme; Rennin

Assay, identification and purity criteria

Description and assay

Specification history
CHYMOSIN B from ASPERGILLUS NIGER var. AWAMORI containing the PROCHYMOSIN B GENE Prepared at the 53rd JECFA (1999) and published in FNP 52 Add 7 (1999), superseding tentative specifications prepared at the 37th JECFA (1990), published in FNP 52 (1992). ADI "Not specified" established at the 37th JECFA in 1990.
C.A.S. number
85713-24-2
SOURCES
Produced extracellularly by the controlled fermentation of Aspergillus niger var. awamori containing the bovine prochymosin B gene. The strain is non-pathogenic and non-toxicogenic (for example, NRRL 3112). After inactivation of the production organism, the cellular material is removed by centrifugation or filtration. The enzyme is purified by extraction with polyethylene glycol, followed by clarification with activated carbon and separation from the solvent by cation exchange chromatography.
Active principles
Chymosin
Systematic names and numbers
None (EC 3.4.23.4)
Reactions catalyzed
Cleaves a single bond in kappa-casein
DESCRIPTION
Clear, colourless or slightly coloured aqueous solutions containing the active enzyme
FUNCTIONAL USES
Enzyme preparation Used in clotting of milk for cheese production
GENERAL SPECIFICATIONS
Must conform to the General Specifications for Enzyme Preparations used in Food Processing (see Volume Introduction)

Identification

Milk clotting activity (Vol. 4)
The sample shows milk clotting activity

FAO JECFA Monograph 1 (2006)

Every name it appears under

7 recorded names. A label may use any of them.

Other names

  • Aspartyl protease
  • Chymosin
  • Chymosin B
  • Chymosin B from Aspergillus niger var. awamori
  • Chymosin B from Aspergillus niger var. awamori containing the Prochymosin B Gene
  • Milk-clotting enzyme
  • Rennin

Hazard reference values

No reference value for this substance in the sources we hold.

Evidence and how bodies read it

Studies, and the differing conclusions regulators drew from them, are curated rather than machine-read, and none has been written for this substance yet.

Common questions

Assembled from what is above, in the words the questions are usually asked in. Each answer is the same statement the relevant section makes, with the same citations behind it.

What is Chymosin B from Aspergillus niger var. awamori?
The B form of chymosin, the milk-clotting enzyme of calf rennet, made by fermentation rather than taken from calves. It is used in cheesemaking and is sold as a clear aqueous solution.
What is Chymosin B from Aspergillus niger var. awamori used for in food?
It is listed as a thickener, a stabiliser and an enzyme — the role the US assigns it. This page shows every function each regulator records, with the citation for each.
Is Chymosin B from Aspergillus niger var. awamori banned in Europe?
No. Chymosin B from Aspergillus niger var. awamori does not appear on the EU's list of authorised food additives, which is not the same as being banned: the EU permits an additive only where it has authorised it, and usually the reason for an absence is that no one applied. The EU has made no finding against this substance. It does mean it may not be used as a food additive in the EU today.
Where is Chymosin B from Aspergillus niger var. awamori permitted?
Of the seven regulators this site reads, the US permits it. Each permission is for named foods at named levels, not a blanket one; the table on this page gives them with citations.

Sources for this page

  • us-listings retrieved 2026-10-18
  • jecfaspecs retrieved 2026-09-23

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