Safety evaluation of the food enzyme papain, a cysteine endopeptidase complex from the latex of Carica papaya L

TL;DR

This study evaluates the safety of papain, an enzyme from papaya latex, used in food processing and finds it generally safe but notes potential allergen risks.

Contribution

The study provides a safety evaluation of a cysteine endopeptidase complex from Carica papaya L. and identifies potential allergenic and quality assurance concerns.

Findings

Estimated dietary exposure to the enzyme is up to 6.104 mg TOS/kg body weight per day.

Papain and chymopapain in the enzyme complex are known allergens with sequence homology to other allergens.

Presence of mycotoxins in all enzyme batches indicates quality assurance issues.

Abstract

The food enzyme is a cysteine endopeptidase complex, containing papain (EC 3.4.22.2), chymopapain (EC 3.4.22.6), caricain (EC 3.4.22.30) and glycyl endopeptidase (EC 3.4.22.25), obtained from the latex of unripe Carica papaya L. by Troplandis BVBA. Dietary exposure was evaluated for seven food manufacturing processes and was estimated to be up to 6.104 mg TOS/kg body weight per day. This exposure is in the same order of magnitude as the intake of the corresponding fraction from unripe C. papaya L. latex. Toxicological studies were not required according to the current guidance. Among the four proteins in the cysteine endopeptidase complex, papain and chymopapain are known food allergens. Homology searches of the amino acid sequences of the four proteins in the complex to known allergens identified matches with six food and eight respiratory allergens. The Panel considered that a risk of allergic reactions upon dietary exposure to the food enzyme cannot be excluded. Based on the data provided, the origin of the food enzyme being an edible plant source and the estimated dietary exposure, the Panel concluded that the food enzyme does not give rise to safety concerns under the intended conditions of use. However, the Panel notes the presence of multiple mycotoxins in all food enzyme batches indicating deficiencies in the quality assurance system.

Figures

Tables

TABLE 1. Composition of the food enzyme.

Parameters	Unit	Batches
		1	2	3
Proteolytic activity	TU/mg a	987	988	975
Protein	%	92.0	92.0	92.0
Ash	%	5.5	5.9	5.8
Water	%	2.6	2.8	3.0
Total organic solids (TOS) b	%	91.9	91.3	91.2
Proteolytic activity/TOS ratio	TU/mg TOS	1074	1082	1069

TABLE 2. Intended uses and recommended use levels of the food enzyme as provided by the applicant. 27

Food manufacturing process a	Raw material (RM)	Recommended use level (mg TOS/kg RM) b
Processing of dairy products
Production of cheese	Milk	6.8–68
Production of modified milk proteins	Milk protein	6.8–68 28
Processing of eggs and egg products	Egg white	65–260
Processing of meat and fish products
Production of modified meat and fish products	Meat	130–520
Production of protein hydrolysates from meat and fish proteins	Meat and fish proteins	Not provided 29 , 30
Processing of cereals and other grains
Production of baked products	Flour	12.5–50
Production of brewed products	Beer	60–240
Processing of plant‐ and fungal‐derived products
Production of protein hydrolysates from plants and fungi	Vegetable protein	Not provided 31 , 32
Processing of yeast and yeast products	Yeast biomass	73.5–294

TABLE 3. Summary of the estimated dietary exposure to the food enzyme–TOS in six consumer groups.

Consumer group	Estimated exposure (mg TOS/kg body weight per day)
	Infants	Toddlers	Children	Adolescents	Adults	The elderly
Age range	4–11 months	12–35 months	3–9 years	10–17 years	18–64 years	≥ 65 years
Actual consumers of the relevant food categories
Min–max mean (number of surveys)	0.424–1.800 (14)	1.307–2.825 (17)	1.280–2.534 (21)	0.702–1.411 (23)	0.526–1.099 (23)	0.378–0.826 (25)
Min–max 95th percentile (number of surveys)	1.724–5.439 (13)	2.963–6.104 (16)	2.310–5.530 (21)	1.476–3.224 (22)	1.209–2.635 (23)	1.025–1.867 (24)

TABLE 4. Summary of estimated dietary exposure to the SMT‐Equivalent resulting from eating raw and cooked unripe papaya.

TABLE 5. Qualitative evaluation of the influence of uncertainties on the dietary exposure estimate.

Sources of uncertainties	Direction of impact
	Exposure to FE–TOS	Exposure to SMT‐equivalent
Model input data
Consumption data: different methodologies/representativeness/underreporting/misreporting/no portion size standard	+/−	+/−
Use of data from food consumption surveys of a few days to estimate long‐term (chronic) exposure for high percentiles (95th percentile)	+	NA
Possible national differences in categorisation and classification of food	+/−	+/−
Use of the consumption data of unripe C. papaya L. from Thailand and the Philippines to calculate the exposure to the SMT‐Equivalent	NA	+/−
Model assumptions and factors
Selection of broad FoodEx categories to calculate the exposure to FE‐TOS	+	NA
Only green papaya salad and soup are included to calculate the intake of SMT‐Equivalent	NA	−
For yeast processing, although only yeast extract is produced, the food categories chosen for calculation cover also those containing mannoproteins resulting from the treatment of yeast cell walls.	+	NA
Egg white is the only raw material indicated by the applicant. However, for egg processing, the food categories chosen for calculation are egg yolk, egg white and raw eggs (without shells)	+	NA
Use of recipe fractions to disaggregate FoodEx categories	+/−	NA
Use of technical factors in the exposure model	+/−	NA
The applied food enzyme yield factor was the mean value	NA	+/−

Full text

INTRODUCTION

1

Article 3 of the Regulation (EC) No 1332/20081 provides definitions for ‘food enzyme’ and ‘food enzyme preparation’.

‘Food enzyme’ means a product obtained from plants, animals or microorganisms or products thereof including a product obtained by a fermentation process using microorganisms: (i) containing one or more enzymes capable of catalysing a specific biochemical reaction; and (ii) added to food for a technological purpose at any stage of the manufacturing, processing, preparation, treatment, packaging, transport or storage of foods.

‘Food enzyme preparation’ means a formulation consisting of one or more food enzymes in which substances such as food additives and/or other food ingredients are incorporated to facilitate their storage, sale, standardisation, dilution or dissolution.

Before January 2009, food enzymes other than those used as food additives were not regulated or were regulated as processing aids under the legislation of the Member States. On 20 January 2009, Regulation (EC) No 1332/2008 on food enzymes came into force. This Regulation applies to enzymes that are added to food to perform a technological function in the manufacture, processing, preparation, treatment, packaging, transport or storage of such food, including enzymes used as processing aids. Regulation (EC) No 1331/20082 established the European Union (EU) procedures for the safety assessment and the authorisation procedure of food additives, food enzymes and food flavourings. The use of a food enzyme shall be authorised only if it is demonstrated that:

• it does not pose a safety concern to the health of the consumer at the level of use proposed;
• there is a reasonable technological need;
• its use does not mislead the consumer.

All food enzymes currently on the European Union market and intended to remain on that market, as well as all new food enzymes, shall be subjected to a safety evaluation by the European Food Safety Authority (EFSA) and approval via an EU Community list.

The ‘Guidance on submission of a dossier on food enzymes for safety evaluation’ (EFSA, 2009a) lays down the administrative, technical and toxicological data required.

Background and terms of reference as provided by the requestor

1.1

Background as provided by the European Commission

1.1.1

Only food enzymes included in the European Union (EU) Community list may be placed on the market as such and used in foods, in accordance with the specifications and conditions of use provided for in Article 7 (2) of Regulation (EC) No 1332/2008 on food enzymes.

An application has been introduced by the applicant ‘Troplandis BVBA’ for the authorization of food enzyme papain from Carica papaya.

Following the requirements of Article 12.1 of Regulation (EC) No 234/2011 implementing Regulation (EC) No 1331/2008, the Commission has verified that the three applications fall within the scope of the food enzyme Regulation and contain all the elements required under Chapter II of that Regulation.

Terms of reference

1.1.2

In accordance with Article 29(1)(a) of Regulation (EC) No 178/2002, the European Commission requests the European Food Safety Authority to carry out the safety assessment on the following food enzyme: papain from Carica papaya, in accordance with the Regulation (EC) No 1331/2008 establishing a common authorisation procedure for food additives, food enzymes and food flavourings.

DATA AND METHODOLOGIES

2

Data

2.1

The applicant has submitted a dossier in support of the application for authorisation of the food enzyme papain from Carica papaya. The dossier was updated on 10 June 2021.

Additional information was requested from the applicant during the assessment phase on 16 November 2021 and on 1 March 2024 and received on 17 December 2021 and on 11 March 2024 (see ‘Documentation provided to EFSA’).

Methodologies

2.2

The assessment was conducted in line with the principles described in the EFSA ‘Guidance on transparency in the scientific aspects of risk assessment’ (EFSA, 2009a) and following the relevant guidance documents of the EFSA Scientific Committee.

The ‘Guidance on the submission of a dossier on food enzymes for safety evaluation’ (EFSA, 2009b) has been followed for the evaluation of the application. Additional information was requested in accordance with the updated ‘Scientific Guidance for the submission of dossiers on food enzymes’ (EFSA CEP Panel, 2021) and the guidance on the ‘Food manufacturing processes and technical data used in the exposure assessment of food enzymes’ (EFSA CEP Panel, 2023).

ASSESSMENT

3

The food enzyme under application is a cysteine endopeptidase complex, containing four proteolytic activities: papain, chymopapain, caricain and glycyl endopeptidase. Papain is the generally accepted name of this complex, and therefore, the term ‘papain’ is used throughout the text of the scientific opinion.IUBMB nomenclaturePapainSystematic name–SynonymsPapayotin; papaya peptidase IIUBMB NoEC 3.4.22.2CAS No9001‐73‐4EINECS No232‐627‐2IUBMB nomenclatureChymopapainSystematic name–SynonymsChymopapain A; chymopapain B; chymopapain SIUBMB NoEC 3.4.22.6CAS No9001‐09‐6EINECS No232‐580‐8IUBMB nomenclatureCaricainSystematic name–SynonymsPapaya peptidase A; papaya peptidase II; chymopapain SIUBMB NoEC 3.4.22.30CAS No39307‐22‐7EINECS No–IUBMB nomenclatureGlycyl endopeptidaseSystematic name–SynonymsPapaya peptidase B; papaya proteinase IV; chymopapain MIUBMB NoEC 3.4.22.25CAS No149719‐24‐4EINECS No–

Papains catalyse the hydrolysis of proteins with broad specificity for peptide bonds, with preference for amino acids with large hydrophobic side chains at the P2 position, resulting in the generation of peptides and amino acids.

The following seven food manufacturing processes were assessed in the current evaluation: processing of dairy products for the production of (1) cheese and (2) modified milk proteins; (3) processing of eggs and egg products; (4) processing of meat and fish products for the production of modified meat and fish; processing of cereals and other grains for the production of (5) baked and (6) brewed products; and (7) processing of yeast and yeast products.

Source of the food enzyme

3

3.1

The food enzyme papain is obtained from the latex harvested from the unripe fruit of non‐genetically modified Carica papaya L,4 a species belonging to the Caricaceae family (Badillo, 1971; OECD, 2010) and a perennial tree native to Mexico and South America (Garret, 1995; Siar et al., 1998).

The milky latex is stored in secretory structures known as laticifers, which are distributed throughout various parts of the papaya plant. The highest concentration of latex is present in the peels of unripe papaya fruit. During the ripening of the papaya fruit, the production of the latex by the laticifers gradually declines. Mechanical injury, such as longitudinal incisions on the surface of the unripe fruit, induces latex exudation (El Moussaoui et al., 2001).

The ripe papaya fruit is commonly consumed worldwide. Unripe papaya is also used as vegetable in south Asian countries such as Thailand, where it is cooked in soup or consumed raw as a salad ingredient (Sone et al., 1998), or canned in sugar syrup in Puerto Rico (Morton, 1987).

A literature search was conducted by EFSA to identify compounds in papaya peels or latex or extracts thereof that could be hazardous to human health upon oral exposure. The Panel did not identify any study both reliable and relevant to the source of the food enzyme, except for allergenicity.

No issues of concern were identified by the Panel from the source material.

Production of the food enzyme

3.2

The food enzyme is manufactured according to the Food Hygiene Regulation (EC) No 852/2004,5 with food safety procedures based on Hazard Analysis and Critical Control Points and in accordance with good manufacturing practice.6

The food enzyme is obtained from the papaya latex. The latex batches are routinely tested for the presence of pesticides, heavy metals, mycotoxins and enterotoxins. The latex is homogenised, and the insoluble material is separated by centrifugation and filtration. After separation, the liquid containing the enzyme is concentrated by an ultrafiltration step, in which enzyme protein is retained, while most of the low molecular mass material passes the filtration membrane and is discarded.7 Around 10 kg of fresh latex is needed to produce 1 kg of papain, corresponding to a yield factor of 0.1.8 The applicant provided information on the identity of the substances used in the extraction and in the subsequent downstream processing of the food enzyme.9

The Panel considered that sufficient information has been provided on the manufacturing process. However, the Panel noted that the quality assurance system implemented by the applicant does not guarantee to exclude issues of concern (see Section 3.3).

Characteristics of the food enzyme

3.3

Properties of the food enzyme

3.3.1

The papain activity of the food enzyme is attributed to four proteolytic enzymes of 345, 352, 348 and 348 amino acids,10 which include the signal peptide, propeptide and the mature protein. The molecular masses of the mature proteins, calculated from the amino acid sequences, are 23.3, 23.3, 23.4 and 23.7 kDa.11 The food enzyme was analysed by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis.12 A consistent protein pattern was observed across all batches. The gel showed a major protein band corresponding to an apparent molecular band of about ■■■■■ kDa, consistent with the expected mass of the enzyme. No other enzyme activities were reported.

The applicant's in‐house determination of papain activity is based on the hydrolysis of casein (reaction conditions: pH 7.2, 40°C, 30 min) and is determined by measuring the release of L‐tyrosine equivalents spectrophotometrically at 280 nm. The papain activity is expressed in tyrosine units (TU)/g. One TU is defined as the amount of enzyme that produces 1 μg of tyrosine equivalents per minute under the conditions of the assay.13

Temperature and pH profiles of the food enzyme under assessment have not been provided. According to the literature, the temperature optimum is between 50 and 65°C and the pH optimum between 5.0 and 8.0 (Fernández‐Lucas et al., 2017).14 Minor residual activity remains after 10 min of incubation at 80°C (pH 8.6) (Choudhury et al., 2010).

Chemical parameters

3.3.2

Data on the chemical parameters of the food enzyme were provided for three batches (Table 1).15 The mean total organic solids (TOS) is 91.5% and the mean enzyme activity/TOS ratio is 1075 TU/mg TOS.

Purity

3.3.3

The lead content in all batches was below 0.5 mg/kg16 which complies with the specification for lead as laid down in the general specifications for enzymes used in food processing (FAO/WHO, 2006). For arsenic, cadmium and mercury, the average concentrations determined in the commercial batches were 0.06, 0.02 and 0.35 mg/kg, respectively.17 ^,^ 18 These concentrations are not considered to raise concerns.

The food enzyme complies with the microbiological criteria for total coliforms, Escherichia coli, total Enterobacteriaceae and Salmonella, as laid down in the general specifications for enzymes used in food processing (FAO/WHO, 2006). Additionally, the presence of sulfite‐reducing anaerobes, Staphylococcus aureus and Pseudomonas aeruginosa, was examined and not detected in any of the batches. The presence of yeasts and filamentous fungi was < 10 CFU/g in 25 g.19 The presence of Bacillus cereus was examined in three food enzyme batches and it was found to be absent in 25 g.20

Data on pesticide residues were provided for an extensive list of pesticides analysed in three food enzyme batches, and no pesticide residues were detected, except for folpet (0.18, 0.22 and 0.21 mg/kg) in all batches.21

Using the highest estimated dietary exposure of 6.104 mg TOS/kg body weight (bw) per day (see Section 3.5.2), European consumers could be exposed to up to 1.33 ng/kg bw per day of folpet. This estimate is below the acceptable daily intake (ADI) of 0.1 mg/kg per day set by EFSA for folpet (EFSA, 2023). Therefore, the Panel considers that the amount detected in the food enzyme batch is of no safety concern.

The chloramphenicol content was determined in three food enzyme batches and was below the respective limit of detection (LoD).22

The presence of aflatoxins was examined in the three food enzyme batches and was below the LoD of the applied analytical method.23 Ochratoxin A, zearalenone, fumonisins (B1, B2 and B3), deoxynivalenol and citrinin were quantified in all batches with mean concentrations of 24.3, 89.5, 119.7, 741 and 481.2 μg/kg, respectively.24 Using the highest estimated dietary exposure of 6.104 mg TOS/kg bw per day as the reference (see Section 3.5.2), European consumers could be exposed up to 0.16, 0.6, 0.80, 4.94 and 3.21 ng/kg bw per day to ochratoxin A, zearalenone, fumonisins (B1, B2 and B3), deoxynivalenol and citrinin, respectively. These estimates are below the benchmark dose lower confidence limit (BMDL) for non‐neoplastic effects for ochratoxin A (4.73 μg/kg bw per day, EFSA Panel on Contaminants in the Food Chain (CONTAM), 2020), below the tolerable daily intake (TDI) for zearalenone (0.25 μg/kg bw per day, EFSA CONTAM Panel, 2011), below the TDI for fumonisins (1 μg/kg bw per day, EFSA CONTAM Panel, 2018), below the TDI for deoxynivalenol (1 μg/kg bw per day, EFSA CONTAM Panel, 2017) and below the level of no concern for nephrotoxicity for citrinin (0.2 μg/kg bw per day, EFSA CONTAM Panel, 2012).

These mycotoxins are produced by fungi and found in contaminated food products. Therefore, they are likely originating from contaminated raw materials used during the food enzyme manufacturing process. The presence of multiple mycotoxins in all food enzyme batches indicates deficiencies in the quality assurance system.25

Toxicological data

3.4

According to the EFSA ‘Scientific Guidance for the submission of dossiers on Food Enzymes’, for food enzymes derived from plants and animals that are consumed by the European population, three criteria must be met to waive toxicological studies: (i) the food enzyme is obtained from an edible plant source, (ii) no hazard is introduced through the manufacturing process and (iii) when it can be demonstrated that the dietary exposure to the food enzyme–TOS is within the same order of magnitude as the dietary intake of the fraction of the plant or animal material comparable to the food enzyme–TOS (EFSA CEP Panel, 2021).

The Panel considered that sufficient information has been provided on the plant source, its history of consumption as well as the food enzyme manufacturing process. Therefore, the need for toxicological data is waived.

Allergenicity

3.4.1

The allergenicity assessment considers only the food enzyme and not any additives, carriers or other excipients, which may be used in the final formulation.

The potential allergenicity of papain, chymopapain, caricain and glycyl endopeptidase was assessed by the Panel by separately comparing each amino acid sequence with those of known allergens as described in the EFSA GMO Scientific opinion (EFSA GMO Panel, 2010). The amino acid sequences of these four proteins present a high degree of homology (62%–81% sequence identity). Using higher than 35% identity in a sliding window of 80 amino acids as the criterion, matches with six food and eight respiratory allergens were found using the AllergenOnline and COMPARE databases.26

Papain and chymopapain (Cari p 2) are food allergens from papaya (Carica papaya L.). Cari p 2 was detected in pollen and pulp of raw and ripe papaya fruits (Bhowmik et al., 2021). Several studies reported occupational rhinitis and asthma in workers of industries where papain is handled (Baur et al., 1982; Baur & Fruhmann, 1979; Niinimaki et al., 1993; Soto‐Mera et al., 2000; Van Kampen et al., 2005). Administration of chymopapain for chemonucleolysis resulted in sensitisation in some patients (Garcia‐Ortega et al., 1991). Severe systemic reactions mediated by papain‐specific IgE were observed in some individuals that ingested papain‐containing meat tenderiser (Mansfield & Bowers, 1983). Caricain and glycyl endopeptidase showed 82.5%–87.5% sequence identity to papain and chymopapain, and IgE binding to caricain and glycyl endopeptidase was reported (Dando et al., 1995).

Matching food allergens were Ana c 2 (52.5%–62.5% sequence identity), a bromelain from pineapple (Ananas comosus); actinidins (56.3%–61.3% sequence identity) from kiwi fruits (Actinidia deliciosa, A. chinensis) and a cysteine protease (47.5%–50% sequence identity) from soybean (Glycine max).

Bromelain from pineapple was described as an occupational allergen eliciting allergic reactions after inhalation or dietary exposure (Gailhofer et al., 1988; Nettis et al., 2001).

Actinidins are major kiwi allergens (Grozdanovic et al., 2014). Kiwi allergic individuals also demonstrated IgE reactivity to bromelain and papain. In addition, the presence of cross‐reactive allergens in papaya and fig was shown by cross‐inhibition experiments (Hemmer et al., 2004).

IgE binding to the soybean cysteine protease was described in soybean‐allergic individuals with skin‐related symptoms (Ogawa et al., 1993; Morita et al., 2012).

The matching pollen allergen was Amb a 11 (53.8%–58.8% sequence identity), a cysteine protease from ragweed (Ambrosia artemisiifolia). Ragweed is associated with the pollen‐food allergy syndrome. Reactions within this syndrome are usually restricted to the buccal cavity and seldom lead to anaphylaxis (Sarkar et al., 2018).

The matching respiratory allergens were group 1 mite allergens (36.3%–48.8% sequence identity), cysteine proteases from Dermatophagoides pteronyssinus, Dermatophagoides farinae, Blomia tropicalis, Euroglyphus maynei, Tyrophagus putrescentiae and Sarcoptes scabiei. Der p 1 and Der f 1 are major mite allergens associated with rhinitis and asthma (Xu et al., 2025). No evidence of papain‐related allergic reactions upon dietary exposure in individuals sensitised to mites is available (Giangrieco et al., 2023).

The Panel considered that the results of the sequence homology search and the available literature indicate a risk of allergic reactions for papaya, ananas, kiwi, soy, fig and pollen‐allergic individuals upon dietary exposure to the papain under assessment.

The food enzyme is obtained from the latex of unripe papaya. Papaya also contains other food allergens, e.g. endo‐polygalacturonase (Cari p 1) and endochitinase, involved in the latex‐fruit syndrome (Rojas‐Mandujano et al., 2018). Cari p 1 was detected in papaya peel, pulp and pollen (Sarkar et al., 2018; Bhowmik et al., 2021). The Panel considered that these allergenic proteins could also be present in the food enzyme.

In conclusion, the Panel considered that, under the intended conditions of use, a risk of allergic reactions upon dietary exposure to this food enzyme, particularly for papaya, ananas, kiwi, soy, fig and pollen allergic individuals, cannot be excluded. However, the likelihood of such reactions will not exceed the risk of reactions after consumption of papaya, ananas, kiwi, soy and fig.

Dietary exposure

3.5

Intended use of the food enzyme

3.5.1

The food enzyme is intended to be used in seven food manufacturing processes at the recommended use levels summarised in Table 2. The intended uses evaluated by the Panel excluded two food manufacturing processes because the applicant did not provide their use levels, despite being requested by EFSA.

TABLE 2: Intended uses and recommended use levels of the food enzyme as provided by the applicant. 27

In the production of cheese, the food enzyme is added to milk during the coagulation step33 to hydrolyse κ‐casein. After coagulation, the food enzyme–TOS partition into the whey and the curd.

In the production of modified milk proteins, the food enzyme is added to milk proteins (e.g. whey and casein)34 to improve the solubility and taste. The food enzyme–TOS remains in the final products.

In the processing of eggs and egg products, the food enzyme is added to treat the egg white35 to improve the sensory and technological properties of these products. The food enzyme–TOS remains in these enzyme‐modified egg products.

In the production of modified meat and fish products, the food enzyme is added to meat to hydrolyse the fibrous proteins for tendering purposes.36 The food enzyme–TOS remains in the final processed foods.

In the production of protein hydrolysates from meat and fish proteins, despite being requested, the applicant did not provide the specific use level.37 ^,^ 38 Therefore, this food manufacturing process is excluded from the exposure evaluation.

In the production of baked products, the food enzyme is added to flour during the preparation of the dough39 to hydrolyse gluten proteins, facilitating the handling of the dough.40 The food enzyme–TOS remains in the baked products.

In the production of brewed products, the food enzyme is added to beer before the pasteurisation step41 to prevent haze formation.42 The food enzyme–TOS remains in the brewed products.

In the production of protein hydrolysates from plants and fungi, despite being requested, the applicant did not provide the specific use level.43 Therefore, this food manufacturing process is excluded from the exposure evaluation.

In the processing of yeast and yeast products, the food enzyme is added to the yeast culture during the lysis step44 to increase the yield and to enrich the savoury taste of the yeast extracts,45 in which the food enzyme–TOS remains.

Based on data provided on thermostability (see Section 3.3.1) and the downstream processing steps applied in the respective food manufacturing processes, the Panel considered that the food enzyme may remain in its active form in all food manufacturing processes, depending on the processing conditions.

Dietary exposure estimation

3.5.2

Following the EFSA Guidance Document on food enzymes (EFSA CEP Panel, 2021), a comparison was made between the chronic exposures:

• dietary exposure to the food enzyme–TOS, resulting from the intended uses as proposed by the applicant (herein referred as ‘FE‐TOS’) and
• dietary exposure to a fraction of C. papaya L. unripe fruit comparable to the food enzyme–TOS, resulting from the consumption of raw or cooked unripe C. papaya L. (herein referred to as source material TOS equivalent, ‘SMT‐Equivalent’).

Estimated dietary exposure to the food enzyme–TOS

3.5.2.1

Chronic exposure to the food enzyme–TOS was calculated using the FEIM webtool46 by combining the maximum recommended use level with individual consumption data (EFSA CEP Panel, 2021). The estimation involved selection of relevant food categories and application of technical conversion factors (EFSA CEP Panel, 2023).

Table 3 provides an overview of the exposure‐derived estimates across all surveys from actual consumers of the relevant food categories. Detailed mean and 95th percentile exposure to the food enzyme–TOS per age class, country and survey, as well as contribution from each FoodEx category to the total dietary exposure are reported in Appendix A – Tables 1 and 2. For the present assessment, food consumption data were available from 51 dietary surveys (covering infants, toddlers, children, adolescents, adults and the elderly), carried out in 27 European countries (Appendix B). The highest dietary exposure was estimated to be 6.104 mg TOS/kg bw per day in toddlers at the 95th percentile. As the food categories relevant to the intended uses of this food enzyme are commonly consumed by Europeans, the estimates calculated for survey respondents are the same as those calculated for actual consumers.

Estimated dietary exposure to the SMT‐equivalent

3.5.2.2

Unripe papaya (also known as green papaya) is rich in papain, especially in the latex. The food enzyme is obtained from the papaya latex. In Europe, papaya is consumed mainly as ripe fruit, while the consumption of unripe papaya is limited and consumption data are not available. In Southeast Asia, unripe papaya is commonly eaten raw as salad or cooked in soup. For the estimation of the SMT‐Equivalent, consumption data from actual consumers of papaya salad or soup made of unripe papaya in the Philippines47 and Thailand48 were used.

The chronic dietary exposure to the SMT‐Equivalent was calculated in two steps: firstly, the intake of unripe papaya was converted to the intake of latex, using a factor of 0.006,49 as provided by the applicant, to account for the amount of latex in unripe C. papaya L. Secondly, the intake of latex was converted into a fraction comparable to the food enzyme–TOS, by applying a yield factor (0.1)50 to take into account the yield of the food enzyme–TOS from fresh latex (Section 3.2).

Table 4 provides an overview of the estimated exposure to the SMT‐Equivalent for the actual consumers of raw and cooked unripe green papaya. The age ranges are presented as provided in the surveys from the Philippines and Thailand. The highest dietary exposure was estimated to be 3.159 mg/kg bw per day in children at the 95th percentile.

Comparison of the two exposure estimates

3.5.2.3

The intakes of the SMT‐Equivalent by the consumers from the Philippines and Thailand (Table 4) are in the same order of magnitude as the dietary exposure to the food enzyme–TOS for European consumers (Table 3).

Uncertainty analysis

3.5.3

In accordance with the guidance provided in the EFSA opinion related to uncertainties in dietary exposure assessment (EFSA, 2006), the following sources of uncertainties have been considered and are summarised in Table 5.

The estimation of the SMT‐Equivalent is based on a realistic scenario, while the estimation of the dietary exposure to the food enzyme–TOS is based on a conservative approach. In particular, assumptions made on the occurrence and use levels of this specific food enzyme have likely led to an overestimation of the dietary exposure to the food enzyme–TOS.

Margin of exposure

3.6

Since toxicological tests were not required for this food enzyme (see Section 3.4), the margin of exposure was not calculated.

CONCLUSION

4

Based on the data provided, the origin of the food enzyme being an edible plant source and the estimated dietary exposure, the Panel concluded that the food enzyme endopeptidase complex (papain) extracted from the latex of the unripe Carica papaya L. does not give rise to safety concerns under the intended conditions of use. However, the presence of multiple mycotoxins in all food enzyme batches indicated deficiencies in the quality assurance system.

DOCUMENTATION AS PROVIDED TO EFSA

5

Request for the authorisation of a papain from Carica papaya for use as a food processing aid. November 2020. Submitted by Troplandis BVBA.

Additional information. December 2021. Submitted by Troplandis BVBA

Additional information March 2024. Submitted by Troplandis BVBA.

ABBREVIATIONSCASChemical Abstracts ServiceCEFEFSA Panel on Food Contact Materials, Enzymes, Flavourings and Processing AidsCEPEFSA Panel on Food Contact Materials, Enzymes and Processing AidsEINECSEuropean Inventory of Existing Commercial Chemical SubstancesFAOFood and Agricultural Organization of the United NationsGMOgenetically modified organismIUBMBInternational Union of Biochemistry and Molecular BiologyJECFAJoint FAO/WHO Expert Committee on Food AdditiveskDakiloDaltonLoDlimit of detectionOECDOrganisation for Economic Cooperation and DevelopmentTOStotal organic solidsWHOWorld Health Organization

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2021‐00173

COPYRIGHT FOR NON‐EFSA CONTENT

EFSA may include images or other content for which it does not hold copyright. In such cases, EFSA indicates the copyright holder and users should seek permission to reproduce the content from the original source.

PANEL MEMBERS

José Manuel Barat Baviera, Claudia Bolognesi, Francesco Catania, Gabriele Gadermaier, Ralf Greiner, Baltasar Mayo, Alicja Mortensen, Yrjö Henrik Roos, Marize de Lourdes Marzo Solano, Henk Van Loveren, Laurence Vernis and Holger Zorn.

LEGAL NOTICE

The full opinion will be published in accordance with Article 12 of Regulation (EC) No 1331/2008 once the decision on confidentiality is received from the European Commission.

Supporting information

APPENDIX A: Dietary exposure estimates to the food enzyme–TOS and the SMT‐equivalent in details