Hyaluronidase from sheep testes, CAS No.37326-33-3

CAS: 37326-33-3 Cat. No.: H754922 Número EC: 253-464-3

GRADE & PURITY Bioactive, ActiBioPure™, Native, Endotoxin tested, High Performance, EnzymoPure™, ≥1000 IU/mg dry weight

Accession #

Q8SQG7

Endotoxin Concentration

<0.2 EU/IU

Bioactivity

≥1000 IU/mg dry weight

Descripción general

Enzymes extracted from mammalian testes (e.g., sheep testes) can hydrolyze mucopolysaccharides of the hyaluronic acid type. They may contain a suitable stabilizer. Potency: Minimum 1000 IU of hyaluronidase activity per milligram (of dry substance).

Production

Animals used for producing hyaluronidase must meet the health requirements for animals intended for human consumption.

Characteristics

Appearance: White or yellowish-white, amorphous powder.

Solubility: Soluble in water, almost insoluble in acetone and absolute ethanol.

Identification

A solution containing 100 IU of hyaluronidase in 1 mL of 9 g/L sodium chloride solution depolymerizes a 10 g/L sodium hyaluronate BRP solution at 20°C, resulting in a significant decrease in viscosity. Heating the hyaluronidase at 100°C for 30 minutes destroys this effect.

Tests

1.Appearance of Solution: The solution should be clear. Dissolve 0.10 g in water and dilute to 10 mL with the same solvent.

2.pH: 4.5 to 7.5. Dissolve 30 mg in carbon dioxide-free water and dilute to 10 mL with the same solvent.

3.Loss on Drying: Maximum 5.0%. Determine by drying 0.500 g at 60°C under a pressure not exceeding 670 Pa for 2 hours.

4.Bacterial Endotoxins: ≤ 0.2 EU/IU.

Assay

The activity of hyaluronidase is determined using a slope-ratio assay, by comparing the rate at which it hydrolyzes sodium hyaluronate BRP with the rate obtained using the International Standard or a reference preparation calibrated in International Units.

Substrate Solution

In a 25 mL conical flask, add 0.10 g of sodium hyaluronate BRP, then slowly add 20.0 mL of water at 4°C. The addition rate must be slow enough to allow the substrate particles to swell (approximately 5 minutes). Maintain at 4°C and stir for at least 12 hours. Store at 4°C and use within 4 days.

For both the test solution and the reference solution, prepare the solutions and perform dilutions at 0°C to 4°C.

Test Solution: Dissolve an appropriate amount of the substance in hyaluronidase diluent to obtain a solution containing 0.6 ± 0.3 IU of hyaluronidase per mL.

Reference Solution: Dissolve an appropriate amount of hyaluronidase BRP in hyaluronidase diluent to obtain a solution containing 0.6 IU of hyaluronidase per mL.

In a reaction vessel, mix 1.50 mL of phosphate buffer solution (pH 6.4) and 1.0 mL of the substrate solution, and equilibrate at 37 ± 0.1°C. At time t₀ = 0 (using the first timer), add 0.50 mL of the test solution containing E milligrams of the enzyme to be tested, mix well. Maintain the mixture at 37 ± 0.1°C using a suitable viscometer, record the flow time t using a second timer (with 0.1-second intervals), and perform multiple measurements over approximately 20 minutes (monitoring with the first timer). Use the following viscometer: microviscometer (DIN 51 562, Part 2), capillary type MII, with a viscometer constant of approximately 0.1 mm²/s².

Repeat the above procedure using 0.50 mL of the reference solution containing hyaluronidase BRP. Calculate the viscosity ratio using the following expression:

K = Viscometer constant (in mm²/s², indicated on the viscometer);

t₂ = Flow time of the solution (in seconds);

0.6915 = Kinematic viscosity of the buffer solution at 37°C (in mm²/s).

Since the enzymatic reaction continues during the flow time measurement, the actual reaction time is equal to t₀ + t/2 (i.e., half of the flow time (t/2) is added to the initial measurement time t₀). Plot (ln η)⁻¹ as a function of the reaction time (t₀ + t/2) (in seconds); a linear relationship should be obtained. Calculate the slope (b) of the substance to be tested and the slope (bᵣ) of the reference preparation. Determine the specific activity in International Units per milligram using the following expression:

A = Specific activity of hyaluronidase BRP (in International Units per milligram).

Perform at least three complete sets of the procedure and calculate the average activity of the substance to be tested.

Storage

Store in a tightly closed container at a temperature of 2°C to 8°C. If the substance is sterile, the container should also be sterile and tamper-proof. 

Specifications

Product Name

Hyaluronidase from sheep testes, CAS No.37326-33-3

Sinónimos

Hyaluronate 4-glycanohydrolase | Hyaluronoglucosaminidase | HYAL2 | oHyal2

Grado

ActiBioPure™, Bioactive, Endotoxin tested, High Performance, Native, EnzymoPure™

Especificaciones y pureza

Bioactive, ActiBioPure™, EnzymoPure™, Native, High Performance, endotoxin tested, ≥1000 IU/mg dry weight

Mecanismos bioquímicos y fisiológicos

Catalyzes hyaluronan degradation into small fragments that are endocytosed and degraded in lysosomes by HYAL1 and exoglycosidases. Essential for the breakdown of extracellular matrix hyaluronan.

Bioactividad

≥1000 IU/mg dry weight

Concentración de endotoxinas

<0.2 EU/IU

Número de adhesión

Q8SQG7

CAS

37326-33-3

Número de la Comisión de Enzimas

3.2.1.35

Tipo de molécula

Enzyme

Almacenamiento y envío

Concentración

≥1000 IU/mg dry weight

Reconstitución

Reconstitute in water.

Condiciones de almacenamiento de almacenamiento

Store at 2-8°C

Enviado en

Wet ice

Estabilidad y almacenamiento

Store at 2-8℃ long term (2 years). Upon receipt, it is recommended to aliquot.

Definición de unidad

The activity of hyaluronidase is determined by comparing the rate at which it hydrolyzes sodium hyaluronate BRP with the rate obtained using an international standard, or by using a reference preparation calibrated in international units, using a slope-ra

Taxonomic Classification

Taxonomy Tree

• Kingdom Organic compounds
  • Superclass Organoheterocyclic compounds
    • Clase Indoles and derivatives
      • Subclass Indolyl carboxylic acids and derivatives

• Kingdom: Organic compounds
• Superclass: Organoheterocyclic compounds
• Clase: Indoles and derivatives
• Subclass: Indolyl carboxylic acids and derivatives
• Intermediate Tree Nodes: Not available
• Direct Parent: Indolyl carboxylic acids and derivatives
• Alternative Parents: Indoles Tertiary alkylarylamines Medium-chain fatty acids Heterocyclic fatty acids Amino fatty acids Benzenoids Amino acids Monocarboxylic acids and derivatives Enamines Carboxylic acids Azacyclic compounds Organic oxides Hydrocarbon derivatives Hydrobromides Carbonyl compounds
• Molecular Framework: Aromatic heteropolycyclic compounds
• Substituents: Indolyl carboxylic acid derivative - Indole - Medium-chain fatty acid - Tertiary aliphatic/aromatic amine - Amino fatty acid - Heterocyclic fatty acid - Fatty acyl - Benzenoid - Amino acid or derivatives - Amino acid - Tertiary amine - Carboxylic acid derivative - Carboxylic acid - Azacycle - Enamine - Monocarboxylic acid or derivatives - Amine - Organonitrogen compound - Organooxygen compound - Hydrobromide - Hydrocarbon derivative - Organic oxide - Carbonyl group - Organic oxygen compound - Organic nitrogen compound - Aromatic heteropolycyclic compound
• Descripción: This compound belongs to the class of organic compounds known as indolyl carboxylic acids and derivatives. These are compounds containing a carboxylic acid chain (of at least 2 carbon atoms) linked to an indole ring.
• External Descriptors: Not available

Data sources

1. Djoumbou Feunang Y, Eisner R, Knox C, Chepelev L, Hastings J, Owen G, Fahy E, Steinbeck C, Subramanian S, Bolton E, Greiner R, and Wishart DS. ClassyFire: Automated Chemical Classification With A Comprehensive, Computable Taxonomy. Journal of Cheminformatics, 2016, 8:61.

DOI: 10.1186/s13321-016-0174-y

Referencias

1. Yidan Chen, Tiantian Feng, Xiaohong Zhu, Yuting Tang, Yao Xiao, Xiuhua Zhang, Sheng-Fu Wang, Dong Wang, Wei Wen, Jichao Liang, Huayu Xiong.  (2024)  Ambient Synthesis of Porphyrin-Based Fe-Covalent Organic Frameworks for Efficient Infected Skin Wound Healing.  BIOMACROMOLECULES,      [PMID:38720431] [10.1021/acs.biomac.4c00261]

2. Mengyao Qi, Yi Ke, Yifei Li, Peng Li, Huakuo Zhou, Xia Zhang, Jieshi Chen, Jun Meng.  (2024)  Au@Pt nanozyme-based smart hydrogel for visual detection of hyaluronic acid.  SENSORS AND ACTUATORS B-CHEMICAL,      [PMID:] [10.1016/j.snb.2024.137144]

3. Tao Meng, Zhengyu Zhao, Mengqiang Song, Rui Wang, Xu Xu, Min Yang, Weihua Li, Shiyong Song.  (2025)  Carboxybetaine modified chitosan as viscosupplementation for osteoarthritis therapy.  INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES,      [PMID:40090277] [10.1016/j.ijbiomac.2025.142155]

4. Yu Pang, Jie Lv, Chengcai He, Chengda Ju, Yulong Lin, Cong Zhang, Meng Li.  (2024)  Covalent organic frameworks-derived carbon nanospheres based nanoplatform for tumor specific synergistic therapy via oxidative stress amplification and calcium overload.  JOURNAL OF COLLOID AND INTERFACE SCIENCE,      [PMID:38330663] [10.1016/j.jcis.2024.01.217]

5. Siying Huang, Simeng Xiao, Xuehao Li, Ranran Tao, Zhangwei Yang, Ziwei Gao, Junjie Hu, Yan Meng, Guohua Zheng, Xinyan Chen.  (2024)  Development of Dual-Targeted Mixed Micelles Loaded with Celastrol and Evaluation on Triple-Negative Breast Cancer Therapy.  Pharmaceutics,  16  (9): (1174).  [PMID:39339211] [10.3390/pharmaceutics16091174]

6. Shiyi Yao, Hui Yuan, Luqi Yang, Yin Zhang, Hanyu Wang, Renxuan Li, Tingjun Ye, Wenguo Cui, Lei Wang.  (2025)  Downregulation of the PI3K/AKT/mTOR/MMP-13 pathway for promoting interface healing via lubricating microspheres.  Acta Biomaterialia,      [PMID:39761787] [10.1016/j.actbio.2025.01.001]

7. Jing Zhang, Yunfeng Zeng, Yongyuan Heng, Yu Shen, Ximeng Sun, Yijia Wang, Haiping Zheng, Ming Zeng, Ziyi Yu.  (2024)  Enzyme-Assisted Activation Technique for Producing Versatile Hydrogel Microparticle Scaffolds with High Surface Chemical Reactivity.  ADVANCED FUNCTIONAL MATERIALS,      [PMID:] [10.1002/adfm.202400858]

8. Jingnan Li, Chengyan Luo, Tingyu Sun, Yinglin Zhou, Xinchang Huang, Dezhou Wu, Xirui Luo, Chao Zeng, Huanan Li.  (2024)  Hypoxia-Specific Metal–Organic Frameworks Augment Cancer Immunotherapy of High-Intensity Focused Ultrasound.  ACS Nano,      [PMID:38949962] [10.1021/acsnano.4c02921]

9. Huiping Yang, Yinning Lin, Qian Mo, Zhaoquan Li, Fan Yang, Xinchun Li.  (2024)  Monitoring Enzymatic Reaction Kinetics and Activity Assays in Confined Nanospace.  ANALYTICAL CHEMISTRY,      [PMID:39024010] [10.1021/acs.analchem.4c01901]

10. Shuo Yu, Lu Zhang, Yanshen Yang, Meijuan Wang, Tingting Liu, Wenwen Ji, Yang Liu, Hao Lv, Yang Zhao, Xi Chen, Tinghua Hu.  (2024)  Polydopamine-Based Resveratrol-Hyaluronidase Nanomedicine Inhibited Pancreatic Cancer Cell Invasive Phenotype in Hyaluronic Acid Enrichment Tumor Sphere Model.  ACS Pharmacology & Translational Science,      [PMID:38633596] [10.1021/acsptsci.3c00304]

11. Xin Jin, Chengxiong Wei, Kai Li, Peinan Yin, Chengwei Wu, Wei Zhang.  (2024)  Polyphenol-mediated hyaluronic acid/tannic acid hydrogel with short gelation time and high adhesion strength for accelerating wound healing.  CARBOHYDRATE POLYMERS,      [PMID:39048222] [10.1016/j.carbpol.2024.122372]

12. Dandan Wei, Guanhua Jiao, Yinghua Tao, Liuxin Yang, Tao Liu, Fengya Jing, Tianzhu Zhang.  (2025)  Polypropylene Mesh Coated with Dual Cross-Linked Hyaluronic Acid/Polyvinyl Alcohol Composite Hydrogel with Antiadhesion and Angiogenesis Properties for Abdominal Wall Repair.  Advanced Materials Technologies,      [PMID:] [10.1002/admt.202401786]

13. Dandan Wei, Yulin Huang, Min Liang, Liuxin Yang, Guanhua Jiao, Yinghua Tao, Li Xu, Tianzhu Zhang, Zhenling Ji.  (2024)  Polypropylene mesh coated with hyaluronic acid/polyvinyl alcohol composite hydrogel for preventing bowel adhesion.  INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES,      [PMID:38705326] [10.1016/j.ijbiomac.2024.132061]

14. Chao-fan He, Tian-hong Qiao, Xu-chao Ren, Mingjun Xie, Qing Gao, Chao-qi Xie, Peng Wang, Yuan Sun, Huayong Yang, Yong He.  (2025)  Printability in Multi-material Projection-Based 3-Dimensional Bioprinting.  Research,      [PMID:40041038] [10.34133/research.0613]

15. Zhu Dan, Zhong Hanyan, Shi Jingzhan, Liu Qiang, Wang Yiping.  (2025)  Ratiometric surface-enhanced Raman scattering quantification of extracellular matrix metalloproteinase-2 activity for tumor diagnosis.  ANALYTICAL AND BIOANALYTICAL CHEMISTRY,      [PMID:39966175] [10.1007/s00216-025-05792-5]

16. Yingying Chen, Dan Zhu, Hanyan Zhong, Zhixing Gan, Shenfei Zong, Zhuyuan Wang, Yiping Cui, Yiping Wang.  (2024)  Ultrasensitive Detection of Matrix Metalloproteinase 2 Activity Using a Ratiometric Surface-Enhanced Raman Scattering Nanosensor with a Core–Satellite Structure.  ACS Applied Materials & Interfaces,      [PMID:38204415] [10.1021/acsami.3c15344]

17. Shangting Du, Lingyu Li, Junhao Kou, Tianyi Zhu, Zhenyi Song, Yonghua Zhan, Daocheng Wu, Wenhua Zhan.  (2025)  Blackberry-Like Doxorubicin Loaded Hyaluronic Acid/Zinc Phthalocyanine Loaded Mesoporous Silica Nanocomposites for Long-Term Tumor Photodynamic and Chemotherapy Synergistic Therapy.  International Journal of Nanomedicine,      [PMID:40692537] [10.2147/IJN.S530041]

18. Jiancong Ni, Xiaochun Xu, Bifang Yang, Liyang Liu, Weiqiang Yang, Xiaoping Chen, Guiping Chen, Qingxiang Wang, Zhenyu Lin.  (2025)  Indicator-Free Detection of Hyaluronidase Based on a Conductivity-Regulated Bipolar Electrochemiluminescence Platform.  ANALYTICAL CHEMISTRY,      [PMID:40827572] [10.1021/acs.analchem.5c03409]

19. Haojie Qiu, Jingyan Zhu, Kaikai Zhang, Xinnan Wang, Yaoting Chu, Feng Zhang, Xuefeng Yang.  (2025)  Dual Drug-Loaded Hyaluronate-Based Self-Adaptable Hydrogel for Diabetic Infected Wound Healing.  BIOMACROMOLECULES,      [PMID:40845349] [10.1021/acs.biomac.5c00712]

20. Wenjun Xue, Xiaoyu Kuang, Xuanzhen Meng, Boyu Sun, Zijun Zhao, Guizhao Liang.  (2025)  Identification and inhibition mechanism of novel collagen-derived hyaluronidase inhibitory peptides.  Food Bioscience,      [PMID:] [10.1016/j.fbio.2025.106263]