6-phytase (EC 3.1.3.26)

In the current application, an authorisation of a preparation of 6-phytase (EC 3.1.3.26) is sought under Article 4(1) for all poultry, all pig and all fish species under the category/functional group 4 (a) "zootechnical additives"/"digestibility enhancers" according to Annex I of Regulation (EC) No 1831/2003.

According to the Applicant, the active agent of the product (HiPhoriusTM) is a 6-phytase, produced by Aspergillus oryzae DSM33737. Another preparation of 6-phytase from a different Aspergillus oryzae strain is currently authorised as feed additive.

The product is intended to be marketed in several solid and liquid formulations with a guaranteed minimum 6-phytase activities ranging from of 10000 to 50000 FYT / g depending on the formulation. According to the Applicant, the solid formulations are intended to be used for poultry and swine while the liquid formulations are intended to be used also for aquaculture. HiPhoriusTM is intended to be included through premixtures or directly in feedingstuffs to obtain a minimum activity of 200 FYT / kg feedingstuffs for pigs and poultry and a minimum phytase activity of 1000 FYT / kg feedingstuffs for fish.

The activity of 6-phytase is expressed in phytase units (FYT). One FYT unit is defined as "the amount of enzyme that releases 1 μmol of inorganic phosphate from phytate per minute under reaction conditions with a phytate concentration of 5.0 mM at pH 5.5 and 37 °C". This definition is in agreement with the phytase activity unit as stated in the EN ISO 30024.

For the quantification of the phytase activity in feedingstuffs the Applicant submitted the ring-trial validated colorimetric standard method EN ISO 30024 and also applied it, with minor experimental modifications, for the quantification of the phytase activity in the feed additive (HiPhoriusTM) and premixtures. The Applicant confirmed the equivalence of these modified methods to the ring-trial validated VDLUFA 27.1.3 and VDLUFA 27.1.4 methods.

Based on the performance characteristics available the EURL recommends for official control the ring-trial validated EN ISO and VDLUFA colorimetric methods mentioned above for the quantification of the phytase activity in the feed additive (HiPhoriusTM), 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) 2024/2177

For the quantification of phytase activity in the feed additive:

• Colorimetric method based on the enzymatic reaction of phytase on the phytate – VDLUFA 27.1.4

For the quantification of phytase activity in premixtures:

• Colorimetric method based on the enzymatic reaction of phytase on the phytate – VDLUFA 27.1.3

For the quantification of phytase activity in compound feed: 

• Colorimetric method based on the enzymatic reaction of phytase on the phytate – EN ISO 30024