Designation of L -methionine as a feed additive  
 
Ministry of Agriculture, Forestry and Fisheries (MAFF) will designate Muramidase as a  
feed additive and establish its standards and specifications in the ministerial ordinance.  
Outline of standards and specifications is as follows.  
 
Muramidase  
 
Specifications for method  of manufacture  of feed in general  
Muramidase  is allowed to be used as feed additives for broilers only.  
 
Specifications for feed additives  
Active Substance  
Compositional specifications  
Unit  ≥ 30,000 peptidoglycan  degradation /g 
Physical and chemical properties  
(1) It comes in  light-dark brown liquid . 
(2) pH of aqueous solution  or aqueous  suspension  (1 part solute/100 parts solution ) is 3.0 to 5.0. 
(3) It has the best enzyme  activity when pH is between 4.0 and 7.5. 
Purity test:  
(1) Lead: 0. 5 g (0.45~0.54 g) of this product is weighed. When lead is tested using this sample by 
the lead test method (Method No. 1 of atomic absorption spectrophotometry test method), 
concentration  of lead in the solution shall not exceed  20 µg/g.  
(2) Arsenic: 1.0 g (0.95~1.04 g) of this product is weighed. When arsenic is tested using this 
sample by Method No. 3 of arsenic test method (using the apparatus A), the concentration of 
arsenic in the solution shall not exceed 2 µg/g.  
(3) Antibacterial activity:  1 g (0. 5~1.4 g) of this product is weighed.  When antibacterial activity is 
tested using this sample by the antibacterial activity test method,  no antibacterial activity sh all 
be exhibited for Micrococcus luteus ATCC 9341 and Escherichia coli ATCC 27166.   
Ignition residue : ≤ 5.0% ( 1 g) 
Strength test of enzyme:  Method of Strength test of decomposing peptidoglycan . 
  
Standard  for method  of manufacture  
Genetically modified Trichoderma reesei  should be cultured. T he cultured solution  should be 
filtered  or extracted with water  and then filtered to remove bacteria . the filtrate  must be  
concentrated to produce the product . 
Standard  for method  of storage  
The products  must be stored in light-shielding  and airtight  containers.   Standard for method of labelling  
  Containers or packaging of t he products must be labeled that pH  value  (up to the first decimal 
place)  showing the maximum  enzyme  activity.  
 
Product  No. 1 
Compositional specifications  
This product comes in the form of pieces, particles or powder s and is produced by mixing with 
excipient substances  to the Active Substance of Muramidase . If necessary, it is mixed with sodium 
sulfate . 
Unit  This product contains the amount of Muramidase  corresponding to 85 to 17 0% of the 
peptidoglycan  degradation  unit of activity shown  on the label.  
Strength test of enzyme: Method of Strength test of decomposing peptidoglycan . 
Standard for method of storage   
Same as the standard for method of storage  by Active Substance  of Muramidase . 
Standard for method of labelling  
Same as the standard for method  of labelling by Active Substance of Muramidase . 
 
Product  No. 2 
Compositional specifications  
This product comes in the form of liquid and is produced by adding sorbitol to the Active 
Substance of Muramidase.  
Unit This product contains the amount of Muramidase  corresponding to 85 to 17 0% of the 
peptidoglycan degradation  unit of activity shown on the label.  
Strength test of enzyme: Method of Strength test of decomposing peptidoglycan . 
Preparation of Sample solution:  Strength test of enzyme  for Active Substance of Muramidase . 
Standard for method of storage  
Same as the standard for method  of storage  by Active Substance  of Muramidase . 
Standard for method of labelling  
Same as the standard for method of labelling by Active Substance of Muramidase.  
 
Analytical method  
 Strength test of enzyme  
  Method of Strength test of decomposing peptidoglycan  
   (See Japanese attachment. )