,The EFSA FAF Panel has issued a positive scientific opinion on the safety of a modified manufacturing process for the food additive enzymatically produced steviol glycosides (E 960c). The revised process involves enzymatic bioconversion of purified steviol glycosides extracted from Stevia rebaudiana leaves, resulting in two new preparations predominantly composed of specific steviol glycosides.
Key Changes in the Manufacturing Process
Under the proposed modification, purified steviol glycosides are converted through enzymatic bioconversion catalysed by glucosyltransferase and sucrose synthase enzymes. These enzymes are produced using three newly developed genetically modified strains of Escherichia coli:
CDX-044 W3110-TKO
CDX-045 W3110-TKO
CDX-047 W3110-TKO
The modified process yields two distinct preparations:
SBP1, composed predominantly of rebaudioside M
SBP2, composed predominantly of rebaudioside D
As a result of the new process, changes are introduced to the definition of the food additive, as well as to specifications related to residual protein levels, residual solvents, microbiological criteria, and particle size.
Regulatory Implications
The FAF Panel agreed with the applicants proposal to introduce two new entries in Commission Regulation (EU) No. 231/2012, corresponding to:
SBP1 (predominantly rebaudioside M), and
SBP2 (predominantly rebaudioside D).
These entries would sit alongside existing specifications for enzymatically produced steviol glycosides under E 960c.
Safety Assessment
EFSA concluded that the modified manufacturing process does not raise safety concerns, based on the following considerations:
No viable cells or DNA from the genetically modified E. coli production strains remain in the final products.
Food enzymetotal organic solids (TOS) used during bioconversion are removed by at least 99%, resulting in negligible dietary exposure.
Rebaudioside M and rebaudioside D produced via this new process have the same physicochemical characteristics as those already authorised under E 960c(i), (ii) and (iii).
Consequently, existing biological and toxicological data from previous EFSA evaluations are considered applicable to SBP1 and SBP2.
Conclusion
The EFSA FAF Panel concluded that there is no safety concern associated with the proposed modification of the manufacturing process for enzymatically produced steviol glycosides (E 960c), including the use of new genetically modified E. coli strains for the production of SBP1 (rebaudioside M) and SBP2 (rebaudioside D)