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Serine protease (EC 3.4.21.-) - 4a13

Serine protease (EC 3.4.21.-) produced by Bacillus licheniformis DSM 19670

Details

Publication date
4 November 2019 (Last updated on: 27 January 2020)

Description

RONOZYME® ProAct is the trade name of a feed additive containing as active substance serine protease (EC 3.4.21. - not yet fully classified) produced by Bacillus licheniformis (DSM 19670). This feed additive is currently authorized under the category/functional 4(a) "zootechnical additives"/"digestibility enhancers" according to the classification system of Annex I of Regulation (EC) No 1831/2003 (feed additive identification number 4a13). In the current application authorisation is sought under Article 4(1) for the use of the feed additive for chickens for fattening

The serine protease activity is expressed in PROT units, where "one PROT is the amount of serine protease that liberates one micromol per minute of para-nitroaniline (pNA) from 1 mM Suc-Ala-Ala-Pro-Phe-pNA substrate at pH 9.0 and 37 oC". The feed additive is intended to be marketed in a solid (RONOZYME® ProAct (CT)) and a liquid (RONOZYME® ProAct (L)) form with the same minimum serine protease activity (75 000 PROT/g). RONOZYME® ProAct is intended to be incorporated directly or through premixtures at a minimum activity of serine protease of 15 000 PROT/kg feedingstuffs. 

For the quantification of the activity of serine protease in the feed additive, premixtures and feedingstuffs, the Applicant submitted a single-laboratory validated and further verified colourimetric method, based on the enzymatic reaction of serine protease on the Suc-Ala-Ala-Pro-Phe-pNA substrate. 

Based on the performance characteristics available the EURL recommends for official control this method for the quantification of the serine protease activity in the three target matrices. 

Further testing or validation of the methods to be performed through the consortium of National Reference Laboratories as specified by Article 10 (Commission Regulation (EC) No 378/2005, as last amended by Regulation (EU) 2015/1761) is not considered necessary.

Recommended Method

as of:

Commission Implementing Regulation (EU) 2021/1426

 

For the quantification of the serine protease activity in the feed additive, premixtures, compound feed and feed materials:

colourimetric method based on the enzymatic reaction of serine protease on the Suc-Ala-Ala- Pro-Phe-pNA substrate

Files

  • 27 JANUARY 2020
finrep_fad-2019-0010_ronozyme_proact.pdf