Safety of the proposed amendment of the specifications for steviol glycosides (E 960) as a food additive: Rebaudioside M produced via enzyme‐catalysed bioconversion of purified stevia leaf extract

Abstract

The EFSA Panel on Food Additives and Flavourings (FAF) provides a scientific opinion on the safety of the proposed amendment of the specifications for steviol glycosides (E 960) as a food additive, in particular related to rebaudioside M produced via enzyme-catalysed bioconversion of purified stevia leaf extract. Rebaudioside M (95% on dry basis) is produced via enzymatic bioconversion of purified stevia leaf extract using uridine diphosphate (UDP)-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified yeasts K. phaffii UGT-a and K. phaffii UGT-b, that facilitates the transfer of glucose to purified stevia leaf extract via glycosidic bonds. The Panel considered that the parental strain K. phaffii ATCC 20864 qualifies for the qualified presumption of safety (QPS) approach for safety assessment and, therefore, is considered to be safe for production purposes. The Panel concluded that there is no safety concern for Rebaudioside M produced via enzymatic bioconversion of purified stevia leaf extract using UDP-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified yeasts K. phaffii UGT-a and K. phaffii UGT-b, to be used as a food additive. However, the Panel recommended that the European Commission considers establishing separate specifications for Rebaudioside M produced via enzymatic bioconversion of purified stevia leaf extract in Commission Regulation (EU) No 231/2012.

Safety of the proposed amendment of the specifications for enzymatically produced steviol glycosides (E 960c): Rebaudioside D produced via enzymatic bioconversion of purified stevia leaf extract

“…The Panel noted these results are consistent with those previously considered in other scientific opinions (EFSA FAF Panel, 2019, 2020, 2021 and obtained under the same experimental conditions with rebaudioside D and related steviol glycosides from stevia leaf extract.…”

Environmental life cycle assessment of production of the high intensity sweetener steviol glycosides from Stevia rebaudiana leaf grown in Europe: The SWEET project

“…Therefore, the results are also presented in terms of 1 kg SE RA60 (0.004 kg RA60). However, it should be noted that these results are used as a guide, because RA60 sweetness equivalence varies depending on use and concentration, and is typically cited as being between 200 and 300 (Cardello et al 1999;Wallin 2004). The effect of a varying sweetness equivalence is also presented.…”

“…In the EU, SGs are classified as a food additive E960 and allowed as an ingredient if certain criteria are met (European Union 2012 ; Younes et al 2019 ). One such criteria is that the maximum acceptable daily intake (ADI) is 4 mg/kg body weight.…”

“…The resulting molecules each have a much greater sweetness per unit mass than sugar. The sweetness of each Reb variant is different but tend to be in the region of 200-300 times that of sugar, depending on application in food or beverage (Cardello et al 1999;Wallin 2004;Prakash et al 2014). This means that 1 kg of sugar can be replaced by a much smaller mass of SGs.…”

Co-immobilized recombinant glycosyltransferases efficiently convert rebaudioside A to M in cascade

“…Steviol glycosides (SGs) in leaves of Stevia rebaudiana such as 1,2-stevioside, Rebaudioside A (Reb A), Rebaudioside D (Reb D) and Rebaudioside M (Reb M) (structures shown in Scheme 1 ) have attracted widespread attention for their zero-calorie energy content 1 and received the Generally Recognized as Safe (GRAS) status from the United States Food and Drug Administration (USFDA). 2 In S. rebaudiana , the 1,2-stevioside and Reb A are the most abundant steviol glucosides that constitute 5–10 and 2–4% w/w of the leaf dry weight, respectively. 3 Reb D and Reb M have attracted extensive interest due to their high sweetness, which is 350-fold sweeter than sucrose and a much less prolonged bitter taste than most other SGs and they thus have higher prices than Reb A.…”