Endo-1,4-beta-mannanase (EC 3.2.1.78)

In the current application an authorisation is sought under Article 4(1) for endo-1,4-beta-D-mannanase 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 chickens for fattening, turkeys for fattening, other poultry for fattening, minor growing poultry species and ornamental birds. 

According to the Applicant, the feed additive contains as an active substance endo-1,4-beta-D-mannanase produced by the genetically modified Thermothelomyces thermophilus strain LU19287 (DSM 33149). 

The feed additive is intended to be marketed under the trade name Natupulse® as solid (Natupulse®TS) or liquid (Natupulse®TS L) formulations with a minimum activity of endo-1,4-beta-D-mannanase of 8000 thermostable mannanase units (TMU) / g feed additive. 

According to the Applicant, one TMU is defined as the amount of enzyme that produces reducing carbohydrates, having a reducing power corresponding to one μmol of mannose, from locust bean gum (0.3 g/100 ml buffer solution) in one minute at 50 °C and pH 3.5. 

The feed additive is intended to be used directly into feedingstuffs or through premixtures at a proposed minimum endo-1,4-beta-D-mannanase activity of 800 TMU/kg complete feedingstuffs. 

For the quantification of the endo-1,4-beta-D-mannanase activity in the feed additive, premixtures and feedingstuffs, the Applicant submitted two single-laboratory validated and further verified methods based on the enzymatic hydrolysis of dyed carob galactomannan with the endo-1,4-beta-D-mannanase, where the amount of released low-molecular weight dyed fragments are measured by photometry. Based on this measurement, the enzyme activity of the sample is then quantified against a standard enzyme with known activity. 

Based on the overall available performance data, the EURL recommends for official control the two above mentioned single-laboratory validated and further verified methods based on the enzymatic hydrolysis of dyed carob galactomannan with the endo-1,4-beta-D-mannanase, followed by photometric determination of dyed fragments, for official control for the determination of endo-1,4-beta-D-mannanase 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.

Recommended Method

as of:

- Commission Implementing Regulation (EU) 2024/1743

For the determination of the endo-1,4-beta-mannanase activity in the feed additive, premixtures and compound feed:

• Enzymatic hydrolysis of dyed carob galactomannan with endo-1,4-beta-mannanase followed by photometry