Joint Dossiers on Enzymes from Microbial Sources

16 March 2020

Joint dossiers on enzymes from microbial sources

EFSA FIP Unit – food enzyme team Legal basis of joint dossiers

Regulation EU N. 562/2012 amending Commission Regulation (EU) N. 234/2011

Food enzymes may be grouped, provided that:

. Same catalytic activity . Substantially the same manufacturing process . Produced by the same source material . microorganism from the same species (excl. GM) . QPS status, or . Already evaluated by France or Denmark . Edible parts of plants or animal (excl. GM)

2 Joint dossiers overview

16 joint dossiers on enzymes obtained from microorganisms . 10 production microorganisms (species) . ? numbers of production strain . 27 producers/interested parties . could all of them potentially be an individual applicant? . Potentially up to 99 food enzymes (2-12 per dossier) . 16 different declared enzyme activities . 17 intended uses . Not every enzyme in the same dossier can be relevant for all intended uses 3 Challenges/data gaps found in submitted joint dossiers

. Production strain => starting point for the assessment . Data on identification, characterization and details on deposition number not provided.

. Specific chemical composition of each food enzyme => necessary for the calculation of TOS, comparability of the commercial and tox batches . Not provided for all enzymes under assessment in the same dossier . Enzyme activity measured with different methods and expressed in different units . Enzyme activity analysed in tox studies different from the activities claimed in the commercial batches

4 Challenges/data gaps found in submitted joint dossiers

. Assessment must be specific for declared activities and

intended uses •- . Some dossiers pool different declared activities and uses, •- data on only one activity provided •- •-

•- •- . Exact amino acid sequence is necessary for the allergenicity assessment . Specific amino acid sequence not reported

5 Example: 2015-00038-42 / Pectinase, poly-galacturonase, pectin-esterase, pectin lyase and arabanase from Aspergillus niger.

PECTINASE POLY-GALACTURONASE PECTIN-ESTERASE PECTIN-LYASE ARABANASE

Hydrolysis of pectin Random hydrolysis of Hydrolysis of methyl Cleavage of (1→4)- Endo-hydrolysis of Producer (1→4)-α-D- groups on methylated α-D-galacturonan (1→5)-α- galactosiduronic linkages galacturonic acid chain methyl ester arabinofuranosidic in pectate and other linkages in (1→5)- galacturonans arabinans

A x

B x

C x x x x

D x x x x x

E x x x

F x x x x x

G x

H x

I x x x

J x 6 Example of the variety of data provided

Enzyme A B C D E F G H I J activity

Pectinase 972 720,266 590 13 6,000,000 61,633 17,900 307,125

Unit PE/g AVJP/g ASV-U/ml GPS/g µ/ml µ/ml Not U/g provided

Poly- 2,986 3,316 153 98 8,373 galacturonase Unit µ/ml PGU/g endoPG/ml PGNU/mg TOS PG/g

Pectin- 5,545 4,167 0,09 2,670 2,690 esterase Unit PEU/g PE-U/ml PEU/mg TOS PE/g Not provided Pectin-lyase 400 77,333 224 950

Unit PEL/g PL-U/ml PECTU/mg TOS PL/g

Arabanase 10 131 232

Unit endo-Arab-U/ml ARA/g Not provided

7 Splitting dossiers

Enzymes with different activities obtained from the same microbe species A single yes A single yes blended no Keep the production manufacturing catalytic strain? process? activities? dossier as it is

Enzymes no no yes with the same catalytic activities

Split the Split the Split the dossier by dossier by dossier production manufacturing by food strain process enzyme

8 Proposed way forward

EFSA-Q-2015-xxx P

E To be closed? F M A

Data package 1 (from A) EFSA Opinion 1 (EFSA-Q-2020-xx1)

Joint Dossier EC mandate Data package 2 (from B) (from AMFEP) EFSA Opinion 2 Union list (EFSA-Q-2020-xx2)

remains remains Data package 3 (from C) EFSA Opinion 3 (EFSA-Q-2020-xx3)

Individual data package, individual timeline

Data package n (from Z) EFSA Opinion n (EFSA-Q-2020-xxn)

send to [[email protected]] Lack of submission of data package from D is acceptable 9 Proposal way forward

Advantages: . Simplicity: Each data package will have its own opinion, as usual for “regular” dossiers; no issues with confidentiality . Flexibility: Non-responsive manufacturers will not condition the opinions of responsive manufacturers

EFSA-Q-2015-xxx P

E To be closed F M A

Data package 1 (from ABC) EFSA Opinion 1 (EFSA-Q-2020-xx1)

Joint Dossier EC mandate Data package 2 (from BCD) (from AMFEP) EFSA Opinion 2 Union list (EFSA-Q-2020-xx2)

remains remains Data package 3 (from DEF) EFSA Opinion 3 (EFSA-Q-2020-xx3)

Data package n (from ZZZ) EFSA Opinion n (EFSA-Q-2020-xxn)

10 Could a dossier remain joint?

Cases in which the dossier could remain joint

Enzymes from the same production Only one of the interested parties strain obtained by the same is the applicant manufacturing process and with the same declared activities

- Clarity on the role of each - Unlikely? interested party - To be treated as a regular - To be treated as a regular dossier dossier

11 How it works

. Each enzyme applicant will send a full data package to EFSA . A QN will be allocated to each . Existing QN may be closed . Each applicant will indicate a contact point to EFSA

. For each dossier, EFSA will deal with the contact point individually . Additional information requests . Teleconferences, extension of deadlines . AMFEP will arrange with contact points to be notified as the official applicant

. EFSA will issue a separate opinion for each QN . Opinions will be published as soon as adopted, even separately in time . Opinions will include an explanation about which concrete substance is under assessment among the joint dossier

12 16 March 2020

Joint dossiers on food enzymes from animal sources

EFSA FIP Unit – food enzyme team Enzyme activities and source tissues

EFSA-Q-2015-00131 Triacylglycerol lipase

Source material Species Enzyme activities

Pre-gastric tissues: “gullet” • cattle • pregastric lipase • goat • the oro-pharyngeal tissues • sheep (De Caro et al., 1995) • pregastric esterase ?

• Esophagus (Ramsey, 1961, 1962)

EFSA-Q-2015-00237 Rennet

Source material Composition ranges of batches (from 9 producers) Abomasum of: • Enzyme activity: 79–2,543 IMCU/mL • cattle • Protein: 0.2–6.2% • buffalo • Ash: 7–19% • goat • TOS: 0.48–7.2% • sheep • Activity/mg TOS: 3.0–54.8 IMCU/mg TOS • different types of rennet and its physical state (liquid, powder or paste) (EFSA, 2014)

• the ratio between chymosin and pepsin activity2 Challenges

. Source tissue: some dossiers include food enzymes from different tissues/species of animals

. Have all food enzymes the same catalytic activity?

. Chemical composition, including TOS, significantly vary among food enzymes from different producers

. Methods to ensure the absence of infectivity vary among producers . Details are lacking on the concrete manufacturing process of each producer

3 Proposed way forward

Food enzyme obtained from animal tissue

Enough A single yes yes Keep the technical applicant? dossier as it is details?

no no

SOURCE MATERIAL

CHEMICAL COMPOSITION Split the Enrich the dossier by dossier with NEED OF TOXICOLOGICAL manufacturer data DATA? ALLERGENICITY

DIETARY EXPOSURE

4 Information to be provided per manufacturer/ food enzyme

. Are interested parties equal to applicants? . Specific tissues/organs used as a source of the food enzyme . Documented evidence of human consumption . Quantity of consumption in the EU or elsewhere . Documented compliance with meat inspection requirements and in accordance with good hygienic practice . Data on the absence of infectious gents in the source tissue and methods to ensure the absence of any risk of infectivity . Detailed description of the manufacturing process and list of specific raw materials . Characteristics of the enzyme(s) (e.g. identification, amino acid sequence, molecular weight, pH and temperature optimum – literature or experimental data) . Chemical composition of the food enzyme, impurities and by- products from the source tissue/manufacturing process 5 Information to be provided per manufacturer/ food enzyme

. Batch-to-batch variation (e.g. SDS-PAGE) . Intended/unintended reaction products of the food enzyme with food constituents or from the degradation of the food enzyme . Allergenicity assessment – a comprehensive literature search for possible adverse reactions, allergy after consumption of source material, published in the last 10 years . Intended uses of the food enzyme aligned with the ‘EC working document describing the food processes in which food enzymes are intended to be used’ . Specify the intended uses per applicant . Flowcharts for each intended food process and indication at which step(s) the food enzyme is added and yield factor . Use levels to be expressed as mg TOS/kg raw material

6 Waiving of toxicological studies

. will be discussed together with the joint dossiers from plants

7 16 March 2020

Joint dossiers on food enzymes from plant sources

EFSA FIP Unit – food enzyme team Interested parties and enzyme activities

2015-00559 Activity

Papain sensu stricto Chymopapain Glycyl endopeptidase Caricain Value Interested parties Preference for an amino acid Similar to that of Preferential cleavage: Gly, Similar to those of (applicant: AMFEP) bearing a hydrophobic side papain in proteins and small papain and chain at the P2 position. molecule substrates chymopapain A X X X X 859 TU/mg

B X X X X 833 TU/mg

C X X X X 1,059,633 U/g D (?) No data No data No data No data

2016-00867 Activity

Bromelain sensu stricto Ananain Value Interested parties Cleavage of proteins. Strong Broader specificity than preference for Z-Arg-ArgNHMec fruit bromelain amongst small molecule substrates E X X 2,381 GDU/g

F (?) No data No data

2 Challenges

. Source tissue: not always specified, edibility not obvious . How many different food enzymes? => Role of interested parties unclear . Claimed activities are those "sensu stricto" or a complex with different main activities? . Enzyme activities measured with different methods . Chemical composition/TOS vary among enzymes from different producers . Details are lacking on the concrete manufacturing process/raw materials of each producer

3 Proposed way forward

Enzymes obtained from plant tissue

Enough A single yes yes Keep the technical applicant? dossier as it is details?

no no

SOURCE MATERIAL

CHEMICAL COMPOSITION Split the Enrich the dossier by dossier with NEED OF TOXICOLOGICAL applicant data DATA? ALLERGENICITY

DIETARY EXPOSURE

4 Information to be provided per manufacturer/ food enzyme

. Are interested parties equal to applicants? . Specific tissues/organs used as a source of the food enzyme . Documented evidence of human consumption . Quantity of consumption in the EU or elsewhere . Identity of the food enzyme: “strico sensu” or as the enzymatic complex with different main activities? . Information on pesticide residues in the source tissue and analytical data on the food enzyme • Detailed description of the manufacturing process and list of specific raw materials . Characteristics of the enzyme(s) (e.g. identification, amino acid sequence, molecular weight, pH and temperature optimum – literature or experimental data) . Chemical composition of the food enzyme, impurities and by-products from the source tissue/manufacturing process

5 Information to be provided per manufacturer/ food enzyme

. Batch-to-batch variation (e.g. SDS-PAGE) . Conversion between different enzyme activity units . Intended/unintended reaction products of the food enzyme with food constituents or from the degradation of the food enzyme . Allergenicity assessment – a comprehensive literature search for possible adverse reactions, allergy after consumption of source material, published in the last 10 years . Intended uses of the food enzyme aligned with the ‘EC working document describing the food processes in which food enzymes are intended to be used’ . Specify the intended uses per applicant . Flowcharts for each intended food process and indication at which step(s) the food enzyme is added and yield factor. . Use levels to be expressed as mg TOS/kg raw material

6 Waiving of toxicological studies

Commission Implementing Regulation (EU) No 562/2012

Explanation to the Article

(4) With regard to the toxicological properties of enzyme preparations, the SCF guidelines indicated that food enzymes which are derived from edible parts of (non-genetically modified) plants and animals are generally considered as posing no health problems. According to the guidelines no special documentation for safety needs to be supplied provided that the potential consumption under normal use does not lead to an intake of any components which is larger than can be expected from normal consumption of the source as such, and provided that satisfactory chemical and microbiological specifications can be established.

(5) The European Food Safety Authority ("the Authority") has also indicated in its guidance on data requirements for the evaluation of food enzyme applications that the justification for not supplying toxicological data for food enzymes from edible parts of animals and non genetically modified plants may include a documented history on the safety of the source of the food enzymes, the composition and the properties of the food enzyme as well as its use in food which demonstrates no adverse effects on human health when consumed in a comparable way, supported by any existing toxicological studies. Therefore, the enzyme application for food enzymes from such edible sources should not be required to include toxicological data. 7 “consumed in a comparable way”

Two sets of intake estimate – in a nutshell

Intake of food enzyme - TOS from processed foods Intake of plant components from foods

Concentration data TOS in raw material (e.g. barley grain ) Comparable portion of e.g. barley grain Data source Dossier Edibility from literature, recipe, etc Food coverage Foods produced from intended processes Foods containing or produced from barley Assessment Method EFSA comprehensive Database (Individual food consumption data, 6 age groups) Food selection FoodEx nomenclature / original food descriptor / food labels / recipes/ expert knowledge Factors needed Conversion of food groups to raw material, or food ingredients to raw material Open call-for-data* Dossier specific . FAO technical conversion factor (e.g. from grain to malt) . Enzyme yield factor . Cooking recipes (e.g. blood sausage) Source of factors . Ingredient list (e.g. 9% wholegrain barley in muesli) . Instruction of beverage preparation (e.g. 25g of powder in 200 ml of milk)

* to ensure transparency, consistency and equal treatment 8 Stay connected

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