1.Introduction

It and other proteolytic enzymes decompose proteins through hydrolysis, which involves breaking peptide bonds between specific amino acids (building blocks of proteins) by adding water, thereby decomposing proteins. There is ample literature showing that oral proteolytic enzymes (including sherardinase) are fully absorbed into the bloodstream and exert systemic effects. Studies have shown that serpentinase is first delivered to tissues that are damaged or under pressure. It has been used for over 50 years in Europe and Japan, and according to literature, it is beneficial for the human body's response to injury, as well as for the health of the immune system, blood vessels, and respiratory tract.

Like many other enzymes, its dosage is not based on weight (milligrams), but on the enzymatic activity of the enzyme, expressed in units of the enzyme or SPU. In most applications, the dosage ranges from 40000 to 100000 units, with an additional three times a day. The dosage range depends on the size of the person and how much help is needed. To achieve optimal results, serpentinase should be taken on an empty stomach. To speed up the repair after surgery, the general plan is to use serapeptidase one day before surgery, one night after surgery (once), and five days after surgery (three times a day at most).

2.Flow Chart

Serratia marcescens E15---->Fermentation---->Extraction---->Purification--->

3.Quality Standard



	NMT 5% of that of the solution A.
	Greater than or equal to 2000SPU units/mg	2010SPU units/mg
	＜5 ppm	＜5 ppm
	＜3 ppm	＜3 ppm
	Less than or equal to 8.0%	3.0%
	Less than or equal to 5.0%	2.30%
	100% pass 80mesh
	Less than or equal to 10000CFU/g Negative Absence/25g ＜100cfu/g	1000CFU/g Negative Absent

4.Testing Methods

Assay (i) Sample solution: Dissolve exactly {{0}}.1{{10}}0g of Serrapeptase in a solution of ammonium sulfate(1 in 20) to make exactly 100 mL. Pipet 1 mL of this solution, add borate-hydrochloric acid buffer solution (pH 9.0) to make exactly 200 mL, and use this solution as the sample solution.(ii) Tyrosine standard solution: Dissolve exactly 0.160g of Tyrosine RS, previously dried at 105C degree for 3 hours, in 0.2mol/L hydrochloric acid TS to make exactly 1000 mL. Pipet 10mL of this solution, and add 0.2 mol/L hydrochloric acid TS to make exactly 100mL. Prepare before use.

<2.41> (60 degree , reduced pressure not exceeding 0.67kPa, 3 hours) of milk casein, previously dried. To exactly 1.20g of the milk casein, calculated based on the loss on drying, add 160mL of a solution of sodium borate (19 in 1000),

and heat in a water bath to dissolve. After cooling, adjust the pH to exactly 9.0 with 1mol/L hydrochloric acid TS,and add borate-hydrochloric acid buffer solution (pH 9.0) to make exactly 20 mL. Use after warming to 37 t 0.5C.Prepare before use.

(iv) Precipitation reagent: Trichloroacetic acid TS for serrapeptase. Use after warming to 37± 0.5 degree .

(v) Procedure: Pipet 1 mL of the sample solution, put in a glass-stoppered tube (15 x 130 mm), allow to stand at 37±{{10}}.5% degree for 5 minutes, add exactly 5mL of the substrate solution, and mix well immediately. Allow to stand at 37±0.5 degree for exactly 20 minutes, add exactly 5mL otrichloroacetic acid TS for serrapeptase, mix, allow to standat 37±0.5 degree for 30 minutes, and filter through a dried filter paper. Pipet 2mL of the filtrate, add exactly 5mL of solution of anhydrous sodium carbonate (3 in 50), mix, add exactly 1mL of diluted Folin's TS (1 in 3), mix well, and allow to stand at 37±0.5% degree for 30 minutes. Determine the absorbance of this solution at 660nm, A, as directed under Ultraviolet-visible Spectrophotometry <2.24> using water as the blank. Separately, pipet 1mL of the sample solution,add exactly 5mL of trichloroacetic acid TS for serrapeptase,mix, add exactly 5mL of the substrate solution, allow tostand at 37±0.5% degree for 30 minutes, and proceed in the same manner as directed above to determine the absorbance A₂.Separately, pipet 2 mL of the tyrosin standard solution, addexactly 5mL of a solution of anhydrous sodium carbonate (3in 50), mix, add exactly 1mL of diluted Folin's TS(1 in 3),mix well, and proceed in the same manner as directed above to determine the absorbance A₃. Separately, pipet 2mL of 0.2mol/L hydrochloric acid TS, and proceed in the same manner as directed above to determine the absorbance A₄.

20: Reaction time (minute)

200: Dilution rate

176: Conversion factor (Total volume of enzyme reaction solution/volume of filtrate taken x amount of tyro-sine in 2mL of tyrosine standard solution)

One srrapeptase Unit corresponds to the amount of serrapeptase which produces 1ug of tyrosine per minute from 5mL of the substrate solution under the above conditions.

Sererepeptidase exerts its wide-ranging therapeutic effects through multiple pathways:

Anti inflammatory and anti-inflammatory: inhibits protein breakdown. It can inhibit protein degrading enzyme activity, reduce tissue damage and inflammatory response.

Relieve pain: reduce nerve sensitivity: alleviate pain by reducing nerve sensitivity through anti-inflammatory effects.

In summary, it exerts a wide range of therapeutic effects by inhibiting the release of inflammatory mediators, promoting wound healing, reducing viscosity, and facilitating antibiotic transfer. It is suitable for various inflammation and swelling related diseases, such as postoperative, post-traumatic, chronic sinusitis, bronchitis, etc.

6.Application

7.Stability and Safety Study

We did a lot of stability and safety study about this product, according to accelerated and long-term stability data, it shows stable characteristics; according to animal clinical study, it shows safe results.

8.Third Party Test Report

We did various 3rd test reports for this product in past years from organizations like SGS, Eurofins, Pony ect. Any of your test requirements can be realized here.Pls email me if you need details.

9.Certifications

Kono Chem Co.Ltd has been certified by ISO22000 by an authorized certification body

KONO'ISO certificate