Safety evaluation of the food enzyme phospholipase A1 from the genetically modified Trichoderma reesei strain DP‐Nzk98

TL;DR

A genetically modified enzyme from Trichoderma reesei is evaluated for safety in food processing.

Contribution

The study confirms the safety of a genetically modified phospholipase A1 for use in food processing.

Findings

Genotoxicity tests showed no safety concerns.

The no observed adverse effect level was 1000 mg TOS/kg bw per day.

Allergen homology search found no matches to known allergens.

Abstract

The food enzyme phospholipase A1 (phosphatidylcholine 1‐acylhydrolase; EC 3.1.1.32) is produced with the genetically modified Trichoderma reesei strain DP‐Nzk98 by Genencor International B.V. The genetic modifications do not give rise to safety concerns. The food enzyme is free from viable cells of the production organism and its DNA. The food enzyme is intended to be used in the processing of cereals and other grains for the production of baked products. Dietary exposure was estimated to be up to 0.972 mg TOS/kg body weight (bw) per day in European populations. Genotoxicity tests did not indicate a safety concern. The systemic toxicity was assessed by means of a repeated dose 90‐day oral toxicity study in rats. The Panel identified a no observed adverse effect level of 1000 mg TOS/kg bw per day, the highest dose tested, which when compared with the estimated dietary exposure, results in a margin of exposure of at least 1029. A search for the homology of the amino acid sequence of the phospholipase A1 to known allergens was made and no match was found. The Panel considered that a risk of allergic reactions upon dietary exposure to the food enzyme cannot be excluded, but that the likelihood is low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns, under the intended conditions of use.

Tables

TABLE 1. Composition of the food enzyme.

Parameters	Unit	Batches
		1	2	3	4 a
Phospholipase A1 activity	U/g b	62,558	85,696	94,622	93,299
Protein	%	14.0	16.7	20.5	23.8
Ash	%	0.1	0.3	0.3	0.1
Water	%	83.2	77.8	73.8	70.9
Total organic solids (TOS) c	%	16.7	21.9	25.9	29.0
Activity/TOS ratio	U/mg TOS	374.6	391.3	365.3	321.7

TABLE 2. Intended use and recommended use level of the food enzyme as provided by the applicant.

Food manufacturing process a	Raw material (RM)	Recommended use level (mg TOS/kg RM) a
Processing of cereals and other grains
Production of baked products	Flour	10–87.72

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

Population group	Estimated exposure (mg TOS/kg body weight per day)
	Infants	Toddlers	Children	Adolescents	Adults	The elderly
Age range	3–11 months	12–35 months	3–9 years	10–17 years	18–64 years	≥ 65 years
Min–max mean (number of surveys)	0–0.203 (12)	0.010–0.497 (15)	0.006–0.489 (19)	0.002–0.245 (21)	0.072–0.166 (22)	0.081–0.150 (23)
Min–max 95th percentile (number of surveys)	0–0.592 (11)	0.047–0.912 (14)	0.025–0.972 (19)	0.009–0.499 (20)	0.163–0.426 (22)	0.166–0.291 (22)

TABLE 4. Qualitative evaluation of the influence of uncertainties on the dietary exposure estimate.

Sources of uncertainties	Direction of impact
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)	+
Possible national differences in categorisation and classification of food	+/−
Model assumptions and factors
Selection of broad FoodEx categories for the exposure assessment	+
Exposure to food enzyme–TOS always calculated based on the recommended maximum use level	+
Use of recipe fractions to disaggregate FoodEx categories	+/−
Use of technical factors in the exposure model	+/−

Full text

INTRODUCTION

1

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

‘Food enzyme’ means a product obtained from plants, animals or micro‐organisms or products thereof including a product obtained by a fermentation process using micro‐organisms: (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 EU 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.

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 Union 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.

On 5 December 2023, a new application has been introduced by the applicant “Genencor International B.V.” for the authorization of the food enzyme Phospholipase A1 from a genetically modified strain of Trichoderma reesei (strain DP‐Nzk98).

Terms of Reference

1.1.2

The European Commission requests the European Food Safety Authority to carry out the safety assessment and the assessment of possible confidentiality requests of the following food enzyme: Phospholipase A1 from a genetically modified strain of Trichoderma reesei (strain DP‐Nzk98), in accordance with Regulation (EC) No 1332/2008 estabilishing a common authorization 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 phospholipase A1 from a genetically modified T. reesei strain DP‐Nzk98.

Additional information was requested from the applicant during the assessment process on 28 November 2024 and received on 28 January 2025 (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 ‘Scientific Guidance for the submission of dossiers on food enzymes’ (EFSA CEP Panel, 2021) and the ‘Food manufacturing processes and technical data used in the exposure assessment of food enzymes’ (EFSA CEP Panel, 2023) have been followed for the evaluation of the application.

Public consultation

2.3

According to Article 32c(2) of Regulation (EC) No 178/20023 and to the Decision of EFSA's Executive Director laying down the practical arrangements on pre‐submission phase and public consultations, EFSA carried out a public consultation on the non‐confidential version of the technical dossier from 19 March to 9 April 2025 for which no comments were received.

ASSESSMENT

3

IUBMB nomenclaturePhospholipase A1Systematic namePhosphatidylcholine 1‐acylhydrolaseSynonymsNAIUBMB NoEC 3.1.1.32CAS No9043‐29‐2EINECS No618‐552‐1

Phospholipases A1 catalyse the hydrolysis of the fatty acyl ester bond at the sn‐1 position of the glycerol moiety of phospholipids, resulting in the formation of 2‐acyl‐1‐lysophospholipids and free fatty acids. The food enzyme under assessment is intended to be used in the processing of cereals and other grains for the production of baked products.

Source of the food enzyme

3.1

The phospholipase A1 is produced with the genetically modified filamentous fungus T. reesei strain DP‐Nzk98 (■■■■■),4 which is deposited at the Westerdijk Fungal Biodiversity Institute (the Netherlands) culture collection with deposition number ■■■■■.5 The production strain was identified as T. reesei ■■■■■.6

The genome of the production strain was searched for gene clusters with known functions and no cluster was found involved in the synthesis of compounds with known toxicity.7

Characteristics of the parental and recipient microorganisms

3.1.1

The parental strain T. reesei RL‐P37 has been derived from QM6a (ATCC 13631) by classical mutagenesis and selection for high cellulase activity (Sheir‐Neiss & Montenecourt, 1984).

The recipient strain ■■■■■.

■■■■■.

■■■■■.8

Characteristics of introduced sequences

3.1.2

■■■■■

■■■■■.

■■■■■

■■■■■

■■■■■

■■■■■

■■■■■

■■■■■.9

Description of the genetic modifications

3.1.3

The purpose of the genetic modification was to enable the production strain to overproduce the phospholipase A1 ■■■■■.

■■■■■

■■■■■

■■■■■.10

■■■■■.11

Safety aspects of the genetic modification

3.1.4

The technical dossier contains all necessary information on the recipient microorganism, the donor organism and the genetic modification process.

The production strain T. reesei DP‐Nzk98 differs from the recipient strain in its capacity to overproduce the phospholipase A1 ■■■■■

■■■■■.12

No issues of concern arising from the genetic modifications were identified by the Panel.

Production of the food enzyme

3.2

The food enzyme is manufactured according to the Food Hygiene Regulation (EC) No 852/200413 with food safety procedures based on Hazard Analysis and Critical Control Points, and in accordance with current Good Manufacturing Practice.14

The production strain is grown as a pure culture using a typical industrial medium in a submerged, batch or fed‐batch fermentation system with conventional process controls in place. After completion of the fermentation, the solid biomass is removed from the fermentation broth by filtration. The filtrate containing the enzyme is then further purified and concentrated, including an ultrafiltration step in which enzyme protein is retained, while most of the low molecular mass material passes the filtration membrane and is discarded.15 The applicant provided information on the identity of the substances used to control the fermentation and in the subsequent downstream processing of the food enzyme.16

The Panel considered that sufficient information has been provided on the manufacturing process and the quality assurance system implemented by the applicant to exclude issues of concern.

Characteristics of the food enzyme

3.3

Properties of the food enzyme

3.3.1

The phospholipase A1 is a single polypeptide chain of ■■■■■ amino acids.17 The molecular mass of the mature protein, calculated from the amino acid sequence, is ■■■■■ kDa.18 The food enzyme was analysed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis.19 A consistent protein pattern was observed across all batches. The gel showed a major protein band with an apparent molecular mass of about ■■■■■ kDa, consistent with the expected mass of the enzyme.20

No other enzyme activities were reported.21

The applicant's in‐house determination of phospholipase A1 activity is based on hydrolysis of phosphatidyl choline (reaction conditions: pH 7.0, 30°C, 10 min) by measuring the released free fatty acids from l‐α‐phosphatidyl choline using a colorimetric method. The enzyme activity is quantified relative to an internal enzyme standard and expressed in U/g.22

The food enzyme has a temperature optimum around 47°C (pH 7.0, 13 min) and a pH optimum at 8.0 (37°C, 10 min). Thermostability was tested after pre‐incubation of the food enzyme for 15 min (pH 7.0) at different temperatures. The enzyme activity decreased above 40°C showing no residual activity at 60°C.23

Chemical parameters

3.3.2

Data on the chemical parameters of the food enzyme were provided for three batches used for commercialisation and one batch used for the toxicological tests (Table 1).24 The mean total organic solids (TOS) of the three batches for commercialisation was 21.5% and the mean enzyme activity/TOS ratio was 377.1 U/mg TOS.

Purity

3.3.3

The lead content in the three commercial batches and in the batch used for toxicological studies was below 0.04 mg/kg which complies with the specification for lead as laid down in the general specifications for enzymes used in food processing (FAO/WHO, 2006).25 ^,^ 26

The food enzyme complies with the microbiological criteria for total coliforms, Escherichia coli and Salmonella, as laid down in the general specifications for enzymes used in food processing (FAO/WHO, 2006).27 No antimicrobial activity was detected in any of the tested batches.28

The presence of the aflatoxins (B1, B2, G1 and G2), ochratoxin A, fumonisins (B1 and B2), zearalenone, T2‐toxin and sterigmatocystin was examined in the food enzyme batch used for the toxicological studies. All concentrations were below the limits of quantitation (LoQs).29 ^,^ 30 Adverse effects caused by the possible presence of other secondary metabolites are addressed by the toxicological examination of the food enzyme–TOS.

The Panel considered that the information provided on the purity of the food enzyme was sufficient.

Viable cells and DNA of the production strain

3.3.4

The absence of viable cells of the production strain in the food enzyme was demonstrated in three independent batches analysed in triplicate. ■■■■■ No colonies of the production strain were detected.31 A positive control was included.32

The absence of recombinant DNA in the food enzyme was demonstrated by PCR analysis of three batches in triplicate. No DNA was detected ■■■■■ with a limit of detection of 10 ng spiked DNA/mL food enzyme.33

Toxicological data

3.4

A battery of toxicological tests including a bacterial reverse mutation test (Ames test), an in vitro mammalian cell micronucleus test, and a repeated dose 90‐day oral toxicity study in rats has been provided.

The Batch 4 used in these studies has a similar composition as the batches used for commercialisation and a lower activity per mg TOS ratio than the commercial batches, and thus is considered suitable as the test item.

Genotoxicity

3.4.1

Bacterial reverse mutation test

3.4.1.1

A bacterial reverse mutation test (Ames test) was performed according to the Organisation for Economic Co‐operation and Development (OECD) Test Guideline 471 (OECD, 2020) and following Good Laboratory Practice (GLP).34

Four strains of Salmonella Typhimurium (TA98, TA100, TA1535 and TA1537) and Escherichia coli WP2uvrA(pKM101) were used with or without metabolic activation (S9‐mix), applying the ‘treat and wash method’. A preliminary toxicity study and two main studies were carried out in triplicate.

The preliminary toxicity study was carried out using eight concentrations of the food enzyme ranging from 50 to 5000 μg TOS/plate. No cytotoxicity was observed at any concentration of the test substance.

The main experiments were carried out using five concentrations of the food enzyme of 50, 158, 500, 1581 and 5000 μg TOS/plate. No cytotoxicity was observed at any concentration of the test substance. Upon treatment with the food enzyme, there was no biologically relevant increase in the number of revertant colonies above the control values, in any strain tested, with or without S9‐mix.

The study was considered reliable without restrictions and the results were of high relevance.

The Panel concluded that the food enzyme phospholipase A1 did not induce gene mutations under the test conditions applied in this study.

In vitro mammalian cell micronucleus test

3.4.1.2

The in vitro mammalian cell micronucleus test was carried out according to OECD Test Guideline 487 (OECD, 2016) and following GLP.35

Three separate experiments were performed with duplicate cultures of human peripheral whole blood lymphocytes. The cell cultures were treated with the food enzyme with or without metabolic activation (S9‐mix).

A range finding test was carried out at seven concentrations of food enzyme from 20 to 5000 μg TOS/mL in a short‐term treatment with and without S9‐mix and in a long‐term treatment without S9‐mix. No cytotoxicity above 50% (expressed as decrease in replicative index (RI)) was seen at any concentrations tested.

In the first and second experiment, cells were exposed to the food enzyme and scored for the frequency of binucleated cells with micronuclei (MNBN) at concentrations of 313, 1250 and 5000 μg TOS/mL in a short‐term treatment (3‐h exposure and 21‐h recovery period) either with or without S9‐mix. In the third experiment, cells were exposed to the food enzyme and scored for MNBN at concentrations of 313, 1250 and 5000 μg TOS/mL in a long‐term treatment (24‐h exposure without recovery period) without S9‐mix.

Cytotoxicity of 35%, 32% and 37% (decrease of RI) was reported at the highest concentration tested in the short‐term treatment with S9‐mix, without S9‐mix and in the long‐term treatment without S9‐mix, respectively.

The frequency of MNBN was not statistically significantly different from the negative controls at all concentrations tested and conditions of treatment.

The study was considered reliable without restrictions and the results were of high relevance.

The Panel concluded that the food enzyme phospholipase A1 did not induce an increase in the frequency of MNBNs under the test conditions applied in this study.

Repeated dose 90‐day oral toxicity study in rodents

3.4.2

The repeated dose 90‐day oral toxicity study was performed under GLP and according to OECD Test Guideline 408 (OECD, 2018).36

Groups of 10 male and 10 female Sprague–Dawley (Crl: CD(SD)) rats received the food enzyme by gavage in doses of 250, 500 and 1000 mg TOS/kg bw per day. Controls received the vehicle (sterile water).

No mortality was observed.

The feed consumption was statistically significantly decreased on Days 1–9 of administration in low‐ and mid‐dose males (−2%, −3%), on Days 78–84 in low‐ and high‐dose males (−6%, −6%), increased on Days 29–36 in high‐dose males (+3%), on Days 22–29 in mid‐dose females (+7%) and on Days 36–43 in low‐ and mid‐dose females (+6%, +9%). The Panel considered the changes as not toxicologically relevant, as they were only recorded at a single time interval, the changes were small (< 10%), there was no dose–response relationship (on Days 1–9, 78–84; 22–29, 36–43) and there were no statistically significant changes in the final feed consumption, body weight and/or body weight gain.

In the functional observations, a statistically significant decrease in hind limb grip strength in mid‐dose males (−4%), a decrease in ambulatory time (−58% and −51%) and distance travelled (−64% and −54%) (at Interval 3) in mid‐ and high‐dose males, an increase in resting time (at Interval 3) in mid‐dose males (+144%), an increase in distance travelled (at Interval 1) in low dose males (+38%), and an increase in ambulatory time (at Interval 1) (+9%) and distance travelled (at Interval 3) (+57%) in high‐dose females were observed. In addition, lower resting times at Interval 1 (−72%) and Interval 2 (−58%) in high‐dose females were noted. The Panel considered the changes as not toxicologically relevant, as they were only recorded at single time intervals (all parameters), they were only observed in one sex (hind limb grip strength), there was no consistency between the changes in males and females (distance travelled at Interval 3) and there was no dose–response relationship (all parameters in males).

Haematological investigations revealed a statistically significant decrease in red cell distribution width (RDW) in low‐, mid‐ and high‐dose males (−9%, −5%, −5%), a decrease in haemoglobin distribution width (HDW) in low‐ and high‐dose males (−12%, −8%), an increase in red blood cell (RBC) count in mid‐dose males (+4%), a decrease in reticulocytes in low‐dose males (−18%), an increase in platelets in low‐dose males (+16%), a decrease in RBC (−5%) and haematocrit (Hct) (−5%) in high‐dose females, a decrease in mean corpuscular haemoglobin concentration (MCHC) (−3%), RDW (−3%) and HDW (−5%) in low‐dose females, and a decrease in RBC Ghosts in mid‐ and high‐dose females (−9%, −10%). The Panel considered the changes as not toxicologically relevant, as they were only observed in one sex (reticulocytes, platelets, MCHC, Hct, RBC Ghosts), there was no consistency between the change in males and females (RBC), there was no dose–response relationship (RDW, HDW, RBC, reticulocytes, platelets in males and MCHC, RDW, HDW in females), there were no histopathological changes in haematopoietic organs and the changes were within the historical control values (all parameters).

Clinical chemistry investigations revealed a statistically significant decrease in plasma calcium (Ca) concentration (−6%) and an increase in sodium (Na) concentration (+1%) in high‐dose males and an increase in Ca‐concentration (+4%) and Na‐concentration (+1%) in mid‐dose females. The Panel considered the changes as not toxicologically relevant, as there was no consistency between the changes in males and females (Ca‐concentration), there was no dose–response relationship (Ca‐ and Na‐concentrations in females), there were no changes in other relevant parameters (other electrolytes), there were no histopathological changes in kidneys or parathyroid gland and the changes were within the historical control values (both parameters).

Statistically significant changes detected in organ weights were a decrease in absolute brain weight in low‐ and mid‐dose males (both −4%). The Panel considered the change as not toxicologically relevant as it was only observed in one sex, there was no dose–response relationship, the change was small (< 10%) and there were no histopathological changes in the brain.

No other statistically significant or biologically relevant differences from controls were reported.

The Panel identified a no observed adverse effect level (NOAEL) of 1000 mg TOS/kg bw per day, the highest dose tested.

Allergenicity

3.4.3

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

The potential allergenicity of the phospholipase A1 produced with the Trichoderma reesei strain DP‐Nzk98 was assessed by comparing its amino acid sequence with those of known allergens as described in the EFSA GMO Scientific Opinion (EFSA GMO Panel, 2010). Using higher than 35% identity in a sliding window of 80 amino acids as the criterion, no match was found in the AllergenOnline database.37

No reports on oral and respiratory sensitisation or elicitation reactions of the phospholipase A1 under assessment have been published.

Phospholipases are implicated in allergic reactions due to insect bites (Perez‐Riverol et al., 2019). However, a literature search provided by the applicant did not find reports on allergic reactions to phospholipases after oral exposure.38

The Panel considered that the results of the sequence homology search and the available literature do not indicate a risk of allergic reactions upon dietary exposure to the phospholipase A1 under assessment.

■■■■■, a product from ■■■■■ that may cause allergies or intolerances (listed in the Regulation (EU) No 1169/201139), is used as raw material. In addition, ■■■■■, a known source of allergens, is present in the culture medium.40 During the fermentation process, these products will mostly be degraded and utilised by the production strain.

The Panel considered that residual amounts of allergenic proteins could be present: in the food enzyme. Taking into account the level of dietary exposure (see Section 3.5.2), this would result in minute amounts in the final foods, from which allergic reactions are usually not expected.

In conclusion, the Panel considered that under the conditions of use, a risk of allergic reactions upon dietary exposure to this food enzyme cannot be excluded, but that the likelihood is low.

Dietary exposure

3.5

Intended use of the food enzyme

3.5.1

The food enzyme is intended to be used in one food manufacturing process at the recommended use level summarised in Table 2.

In this process, the food enzyme is added to flour during the preparation of dough or batter.41 The phospholipase A1 is used to hydrolyse phospholipids present in the flour to facilitate the handling of the dough.42 The food enzyme–TOS remain in baked foods.

Based on data provided on the temperature profile and thermostability (see Section 3.3.1), the Panel considered that the food enzyme is inactivated during baking.

Dietary exposure estimation

3.5.2

Chronic exposure to the food enzyme–TOS was calculated using the FEIM webtool43 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). Exposure from all FoodEx categories was subsequently summed up, averaged over the total survey period (days) and normalised for body weight. This was done for all individuals across all surveys, resulting in distributions of individual average exposure. Based on these distributions, the mean and 95th percentile exposures were calculated per survey for the total population and per age class. Surveys with only 1 day per subject were excluded and high‐level exposure/intake was calculated for only those population groups in which the sample size was sufficiently large to allow calculation of the 95th percentile (EFSA, 2011).

Table 3 provides an overview of the derived exposure estimates across all surveys. 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 48 dietary surveys (covering infants, toddlers, children, adolescents, adults and the elderly), carried out in 26 European countries (Appendix B). The highest dietary exposure was estimated to be 0.972 mg TOS/kg bw per day in children at the 95th percentile.

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 4.

The conservative approach applied to estimate the exposure to the food enzyme–TOS, in particular assumptions made on the occurrence and use levels of this specific food enzyme, is likely to have led to an overestimation of the exposure.

Margin of exposure

3.6

A comparison of the NOAEL (1000 mg TOS/kg bw per day) identified from the 90‐day rat study with the derived exposure estimates of 0–0.497 mg TOS/kg bw per day at the mean and from 0 to 0.972 mg TOS/kg bw per day at the 95th percentile resulted in a margin of exposure (MOE) of at least 1029.

CONCLUSIONS

4

Based on the data provided and the derived margin of exposure, the Panel concluded that the food enzyme phospholipase A1 produced with the genetically modified T. reesei strain DP‐Nzk98 does not give rise to safety concerns under the intended conditions of use.

The Panel considered the food enzyme free from viable cells of the production organism and recombinant DNA.

DOCUMENTATION AS PROVIDED TO EFSA

5

Application for the authorisation of phospholipase A1 from a genetically modified Trichoderma reesei strain DP‐Nzk98 as a new food enzyme. May 2024. Submitted by Genencor International B.V.

Additional information. January 2025. Submitted by Genencor International B.V.

ABBREVIATIONSbwbody weightCASChemical Abstracts ServiceCEPEFSA Panel on Food Contact Materials, Enzymes and Processing AidsEINECSEuropean Inventory of Existing Commercial Chemical SubstancesFAOFood and Agricultural Organization of the United NationsGLPGood Laboratory PracticeGMOgenetically modified organismHcthaematocritHDWhaemoglobin distribution widthIUBMBInternational Union of Biochemistry and Molecular BiologyJECFAJoint FAO/WHO Expert Committee on Food AdditiveskDakiloDaltonLODlimit of detectionLoQlimit of quantitationMCHCmean corpuscular haemoglobin concentrationMNBNbinucleated cells with micronucleiMOEmargin of exposureOECDOrganisation for Economic Cooperation and DevelopmentPCRpolymerase chain reactionQPSqualified presumption of safetyRBCred blood cellRDWred cell distribution widthTOStotal organic solidsWGSwhole genome sequencingWHOWorld Health Organization

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2024‐00065

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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, Monika Sramkova, Henk Van Loveren, Laurence Vernis and Holger Zorn.

LEGAL NOTICE

The scientific output published implements EFSA's decision on the confidentiality requests submitted on specific items. As certain items have been awarded confidential status by EFSA they are consequently withheld from public disclosure by redaction.

Supporting information

Dietary exposure estimates to the food enzyme–TOS in details