Safety Assessment of the Food Enzyme D-Allulose 3-Epimerase from Genetically Modified Corynebacterium

If you buy or formulate with D-allulose, the enzyme behind it matters as much as the sweetener itself. This EFSA opinion explains why regulators cleared a D-allulose 3-epimerase produced by a genetically modified strain of Corynebacterium. It also gives you a concrete toxicological benchmark to use when reviewing supplier specs.

Why this paper exists

D-allulose is a rare sugar. It tastes like sucrose but delivers fewer calories, which makes it attractive for low-sugar formulations. To make it commercially, manufacturers use an enzyme that converts fructose into D-allulose. That enzyme must pass a food safety review before it can be used in Europe.

The paper is an EFSA safety assessment. EFSA’s Panel on Food Enzymes evaluated the enzyme preparation from a genetically modified Corynebacterium production strain. The enzyme, D-allulose 3-epimerase, catalyzes the conversion of fructose to D-allulose. The Chinese title translates directly to this same safety evaluation, and the DOI is 10.2903/j.efsa.2021.6870.

What the evaluation covered

The panel looked at the enzyme preparation itself, not at D-allulose as a finished ingredient. That’s an important distinction. The enzyme is used in processing, and the final sweetener is what ends up in your product.

The core question was simple. Does the enzyme preparation cause toxicological harm when consumed? To answer that, EFSA reviewed data on the production strain, the enzyme’s identity, and the potential for residues to carry over into the final product.

Two facts stand out from the abstract. First, the panel determined a no-observed-adverse-effect level, or NOAEL, of 1796 mg TOS per kg body weight per day. That was the highest dose tested. Second, during D-allulose production, TOS is removed, and no recombinant DNA is present in the finished D-allulose.

Key findings

Assessment point	Finding
Enzyme source	Genetically modified Corynebacterium
Function	Converts fructose into D-allulose
Highest dose tested	1796 mg TOS/kg body weight/day
Adverse effects at that dose	None observed
NOAEL	1796 mg TOS/kg body weight/day
TOS in final D-allulose	Removed during processing
Recombinant DNA in D-allulose	Not detected

TOS stands for total organic solids. It is how EFSA expresses the amount of enzyme preparation in feeding studies. A NOAEL at the highest dose tested is a strong outcome. It gives regulators a wide safety margin when calculating acceptable exposures.

What this means for manufacturers

For ingredient buyers, this opinion offers a clean regulatory path. It tells you that a recognized food safety authority reviewed the production strain, the enzyme preparation, and the carryover risk. That makes it easier to justify the ingredient in your supply chain.

It also gives you a specific number to check. If a supplier provides toxicological data, look for a NOAEL of 1796 mg TOS per kg body weight per day. That number came from the top dose tested, and no adverse effects appeared at that level. That is a solid reference point.

The recombinant DNA finding matters just as much from a labeling and consumer trust perspective. The final D-allulose contains no detectable process DNA. Combine that with TOS removal, and the story is straightforward: the enzyme does its job, and the finished sweetener is clean.

FAQ

Is D-allulose made with this enzyme a GMO ingredient?
No. The production strain is genetically modified, but the recombinant DNA does not survive into the final D-allulose. EFSA’s evaluation confirms that no recombinant DNA was detected in D-allulose.

What does the NOAEL mean for my ingredient spec?
It is the no-observed-adverse-effect level from the feeding study. At 1796 mg TOS per kg body weight per day, the highest dose tested, no adverse effects were seen. That gives toxicologists a strong margin when estimating safe intake.

Does this opinion cover D-allulose itself?
It covers the food enzyme used to process fructose into D-allulose. The panel evaluated the enzyme preparation and confirmed that TOS is removed and recombinant DNA is not present in the finished D-allulose. The sweetener itself was not the test substance.

EFSA published the opinion in 2021, and it remains a useful reference for anyone sourcing D-allulose. You now have the enzyme source, the function, and the toxicological endpoint in one place. That is exactly the kind of information you need to build a supplier qualification file.