Protease from B. subtilis CBS 148232 with spores of three Bacillus strains for pigs for fattening: colorimetric activity on dyed casein (pH 10, 50 °C), strains by PFGE to CEN/TS 17697, counts to EN 15784 — minimum 5000 U and 1.5 × 10⁸ CFU/kg
Commission Implementing Regulation (EU) 2025/2502, published on 12 December 2025, authorises until 1 January 2036 a Genencor International B.V. preparation (4a63, in the EFSA opinion: Syncra® SWI 201 TPT) — protease (EC 3.4.21.62) from the genetically modified strain B. subtilis CBS 148232 and viable spores of B. velezensis NRRL B-50508, B. velezensis NRRL B-50509 and B. subtilis NRRL B-50510 — as a digestibility enhancer for pigs for fattening of all porcine species and weaned piglets of minor porcine species. The applicant withdrew the application for suckling piglets on 27 June 2025.
- Methods in the annex: protease in the additive, premixtures and feed — a colorimetric method based on hydrolysis of a dyed, cross-linked casein substrate at pH 10.0 and 50 °C; strain identification — pulsed-field gel electrophoresis (PFGE) to CEN/TS 17697 or DNA sequencing; total Bacillus spp. — spread plate on tryptone soya agar to EN 15784. One activity unit is the amount of enzyme releasing 2.3 µg of phenolic compounds (as tyrosine) from casein per minute under these conditions.
- Composition: at least 50 000 U/g protease and 1.5 × 10⁹ CFU/g Bacillus spp. (strains at 1:1:1), mineral oil ≤ 0.4 %, polyvinyl alcohol ≤ 3.5 %. Five batches according to EFSA: 67 120–73 560 U/g (mean 70 340) and 4.6–8.0 × 10⁹ CFU/g; in three batches arsenic 0.06–0.10, cadmium 0.03–0.04, lead 0.16–0.26 mg/kg, mercury < 0.005 mg/kg, B. cereus < 10 CFU/g; dusting potential 106–133 mg/m³.
- Stability: after 15 months at 25 °C a loss of 0.6 log bacteria and 18 % activity, after 12 months at 40 °C 0.6 log and 38 %; in a premixture −9 % activity after 6 months; pelleting at 95 °C −7.6 % activity with no bacterial loss. Homogeneity in feed: CV 1.7 % for bacteria and 8 % for protease activity.
- The applicant proposed a minimum of 1250 U and 3.75 × 10⁷ CFU/kg feed, while EFSA (opinion of 29 January 2025) indicated possible efficacy at 5000 U and 1.5 × 10⁸ CFU/kg — these values became the minimum content in the act. Asked by the Commission, EFSA confirmed on 5 May 2025 that “other minor porcine species at the same physiological stage” means weaned piglets or pigs for fattening of minor species. The preparation is not an eye or skin irritant but is a skin and respiratory sensitiser.