Endo-1,4-β-mannanase (E.C. 3.2.1.78) (Hemicell®)

In the current application authorisation is sought under article 4 for endo-1,4-β-mannanase (EC 3.2.1.78), under the category / functional group 4(a) "zootechnical additives / digestibility enhancers", according to the classification system of Annex I of Regulation (EC) No 1831/2003. Specifically, the authorisation is sought for the use of the feed additive for chickens and turkeys for fattening, chickens reared for laying, turkeys reared for breeding, minor poultry species, piglets (weaned), pigs for fattening and minor porcine species.
According to the Applicant, the feed additive contains the active substance endo-1,4-β-mannanase (EC 3.2.1.78) produced by the genetically modified Paenibacillus lentus CMG3376 (DSM 33618) strain.
The feed additive is intended to be marketed as solid (Hemicell®HT) or liquid (Hemicell®HT-L) enzyme formulations with a minimum activity of endo-1,4-β-mannanase in the products of 1.6 x 108 U / kg and 5.9 x 108 U / L, respectively.
According to the Applicant, the activity of endo-1,4-β-mannanase is expressed in units (U) or in 106 units (MU), where 1 U is defined as the amount of endo-1,4--mannanase enzyme which generates 0.72 micrograms of reducing sugars per minute from a mannose containing substrate at pH 7.0 and 40 °C.
The feed additive is intended to be used in premixtures and feedingstuffs at a proposed minimum endo-1,4-β-mannanase activity of 32000 U / kg or 48000 U / kg complete feedingstuffs, depending on the animal species.
For the quantification of the endo-1,4-β-mannanase activity in the feed additive and feedingstuffs, the Applicant submitted several single-laboratory validated and further verified colorimetric methods based on the enzymatic hydrolysis of mannose containing substrate (locust bean gum, LBG) by the action of endo-1,4-β-mannanase followed by the reaction of the released reducing sugars (mannose equivalent) with 3,5-dinitrosalicylic acid (DNS).
For the quantification of endo-1,4-β-mannanase activity in premixtures the Applicant diluted premixture samples with blank feed and applied one of the above mentioned method for feedingstuffs.
Based on the performance characteristics obtained, the EURL recommends for official control the above mentioned single-laboratory validated and further verified colorimetric methods, based on the enzymatic hydrolysis and the reaction of the reducing sugars (mannose equivalent) with 3,5-dinitrosalicylic acid (DNS), for the quantification of endo-1,4-β-mannanase total activity in the feed additive, premixtures and feedingstuffs. 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.