Fumonisin esterase produced by Komagataella phaffii DSM 33835

In the current application an authorisation is sought under article 4(1) for Fumonisin esterase produced by Komagataella phaffii DSM 33835 under the category/functional group 1(m) "technological additives" / "substances for reduction of the contamination of feed by mycotoxins: substances that can suppress or reduce the absorption, promote the excretion of mycotoxins or modify their mode of action", according to the classification system of Annex I of Regulation (EC) No 1831/2003. The authorisation is sought for the use of the feed additive for all animal species. According to the Applicant, the product contains fumonisin esterase (EC 3.1.1.87) as active substance which is added to a maltodextrin carrier to result in a minimum guaranteed enzyme activity of 3000 U / g. Fumonisin esterase is used to degrade fumonisin B₁; it detoxifies this mycotoxin by cleavage of the toxin's diester bonds and removal of the tricarballylic acid side chains. The Applicant defined the enzyme activity unit (U) as the threefold enzymatic activity that releases 1 μmol tricarballylic acid (TCA) per minute from 100 μM fumonisin B₁ (FB1) in 20 mM Tris-Cl buffer pH 8.0 with 0.1 mg / ml bovine serum albumin at 30 °C. Fumonisin esterase produced by Komagataella phaffii DSM 33835 is intended to be used in premixtures and compound feed, at a minimum recommended enzyme activity of 15 U / kg compound feed. For the determination of the fumonisin esterase activity in the feed additive and compound feed the Applicant submitted a single-laboratory validated and further verified method based on High Performance Liquid Chromatography coupled with tandem mass spectrometry (LC-MS/MS). The method is based on the quantification of the tricarballylic acid released from the action of the enzyme on fumonisin B₁. For the determination of the enzyme activities in premixtures, the Applicant suggested diluting them with corn meal or with compound feed (according to the recommended inclusion rate) and applying the above-mentioned method for compound feed.
Based on the experimental evidences available the EURL recommends for official control the LC-MS/MS method submitted by the Applicant for the determination of the fumonisin esterase activity in the feed additive, premixtures and compound feed. 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.