9001-75-6

Basic information

• Product Name: Pepsin
• Synonyms: IMMOBILIZED PEPSIN;e.c.3.4.1;elixerlactateofpepsin;lactatedpepsin;lactatedpepsinelixir;pepsinfortior;PEPSIN 1:10000 NF POWDER;PEPSIN FROM HOG STOMACH 2XCRYST.LYOPH. S T-FR.POWD. ~150U/G
• CAS NO: 9001-75-6
• Molecular Formula: n.a.
• Molecular Weight: 0
• EINECS: 232-629-3
• Product Categories: Enzymes;API;#N/A
• Mol File: 9001-75-6.mol
• Article Data: 0

Chemical Properties

• Appearance: slightly beige/powder
• Density: 1.287 g/cm3
• Storage Temp.: 2-8°C
• Solubility: 10 mM HCl: soluble1.0mg/mL, clear to faintly turbid, colorless
• Water Solubility: Soluble in water and dilute aqueous buffers
• Sensitive: Hygroscopic
• Stability: Stable. Incompatible with strong bases, alkalies, metallic salts, alcohols.
• Merck: 13,7225
• CAS DataBase Reference: Pepsin(CAS DataBase Reference)
• NIST Chemistry Reference: Pepsin(9001-75-6)
• EPA Substance Registry System: Pepsin(9001-75-6)

Safety Data

• Hazard Codes: Xn
• Statements: 36/37/38-42-42/43
• Safety Statements: 22-24-26-36/37-36-45
• WGK Germany: 1
• RTECS: SC6132000
• F: 3-10
• TSCA: Yes
• Hazardous Substances Data: 9001-75-6(Hazardous Substances Data)

Usage

Description

Pepsin is an enzyme that is secreted in the digestive tract of mammals. It plays a crucial role in the breakdown of proteins into smaller peptides, which can then be easily absorbed by the small intestine.

Uses

Used in Digestive Health Applications:
Pepsin is used as a digestive aid for enhancing the process of protein digestion. It helps break down proteins into smaller peptides, making them more accessible for absorption by the small intestine. This is particularly beneficial for individuals with digestive issues or those who need additional support in breaking down proteins for better nutrient absorption.
Used in Pharmaceutical Industry:
Pepsin is utilized as an active ingredient in various digestive enzymes supplements and formulations. It is incorporated into these products to support the body's natural protein digestion process and to provide relief from indigestion, bloating, and other gastrointestinal discomforts associated with protein malabsorption.
Used in Food and Beverage Industry:
Pepsin is employed as a processing aid in the food and beverage industry, particularly in the production of fermented foods and beverages. It helps in breaking down proteins, improving the texture, and enhancing the flavor profile of these products.
Used in Research and Development:
Pepsin is used as a research tool in various scientific studies and experiments. It is valuable for understanding the mechanisms of protein digestion and for developing new methods and products related to protein breakdown and absorption.

Biological location and functions

Pepsin is synthesized and secreted in the gastric membrane in an inactive state called pepsinogen (PG) (molecular weight of 40kDa). Compared with pepsin, pepsinogen contains an additional 44 amino acids and is stable in neutral and weak alkaline environments, but when exposed to the hydrochloric acid (HCl) present in gastric juice (pH of 1.5 – 2.0), the 44 amino acids are proteolytically removed in an autocatalytic way to activate it to pepsin[11]. Its main role in protein proteolysis is to cleave aromatic amino acids (such as phenylalanine and tyrosine) from the N–terminus of proteins[11]. Pepsin is one of three principal proteases in the human digestive system with the other two being chymotrypsin and trypsin. During the process of digestion in the digestive system, these enzymes, each of which is specialized in severing links between particular types of amino acids, collaborate to break down dietary proteins into their components, i.e., peptides and amino acids, which can be readily absorbed by the small intestine. Pepsin is most efficient in cleaving peptide bonds between hydrophobic and preferably aromatic amino acids such as phenylalanine, tryptophan, and tyrosine12. Pepsin's proenzyme, pepsinogen, is released by the chief cells in the stomach wall, and upon mixing with the hydrochloric acid of the gastric juice, pepsinogen activates to become pepsin13.

Factor affecting the activity of pepsin

The activity of pepsin can be strongly affected by pH, temperature and inhibitors. pH Both the optimum pH (the pH value giving the highest enzymatic activity) and pH stability (the pH range giving good enzyme stability) have significant effects on the activity of pepsin. When the pH deviates from the optimum value, the activity of pepsin drops. Generally, pepsin stability is mentioned at lower pH values. Pepsin is a type of acidic protease and depression of its stability is attributed to the denaturation of proteins above the pH of 6.0. Study has found that the stability and activity of pepsin showed that 70% of maximal peptic activity was present at pH4.5 and that pepsin was irreversibly inactivated at pH 8[14]. Pepsin exhibits maximal activity at pH 2.0 and is inactive at pH 6.5 and above[15, 16]. Temperature Temperature has a great influence on the activity of pepsin. Like pH, the optimum temperature and thermal stability range are very important. Pepsin is most active in acidic environments between 37 °C and 42 °C[17]. Inhibitors Pepstatin A (a typical aspartic proteinase inhibitor) can combine with pepsin and prevents the binding of the enzyme to substrate, thereby resulting in a complete inhibition of its activity. Pepstatin A is a peptide isolated from several species of actinomyces, such as Streptomyces spp. It has one of the lowest known inhibitions constant (Ki= 45 pM) for pepsin[18, 19].

References

Jongjareonrak A, Benjakul S, Visessanguan W, Nagai T, Tanaka M (2005) Isolation and characterisation of acid and pepsin–solubilised collagens from the skin of brownstripe red snapper (Lutjanus vitta). Food Chem 93: 475-484. Zhang Y, Liu W, Li G, Shi B, Miao Y, et al. (2007) Isolation and partial characterization of pepsin-soluble collagen from the skin of grass carp (Ctenopharyngodon idella). Food Chem 103: 906-912. Nalinanon S, Benjakul S, Visessanguan W, Kishimura H (2007) Use of pepsin for collagen extraction from the skin of bigeye snapper (Priacanthustayenus). Food Chem104: 593-601. Nalinanon S, Benjakul S, Visessanguan W, Kishimura H (2008) Improvement of gelatin extraction from bigeye snapper skin using pepsin-aided process in combination with protease inhibitor. Food Hydrocolloids 22: 615-622. Aehle W (2007) Enzymes in industry: production and applications. (3rdedn), Wiley–VCH, Weinheim, Germany, 136-137. Thorne Research (2010) Betaine HCl and Pepsin; Dover, USA. Sogawa K, Fujii–Kuriyama Y, Mizukami Y, Ichihara Y, Takahashi T (1983) Primary structure of human pepsinogen gene. J Biol Chem 258: 5306-5311. Fruton JS (1971) Hydrolysis: peptide bonds, (3rd edn), Academic Press Inc: New York 3: 120-152. Effront J, Prescott SC, Venable CS (2007) Biochemical catalysts in life and industry: proteolytic enzymes. Kessinger Publishing: 151-289. Shahidi F, Janak Kamil YVA (2001) Enzymes from fish and aquatic invertebrates and their application in the food industry. Trends Food Sci Technol 12: 435-464. https://enzyme.expasy.org/EC/3.4.23.1 Raufman J (2004) Pepsin. In Encyclopedia of Gastroenterology, Johnson L R. Ed., Academic Press, Amsterdam 3: 147-148. https://www.britannica.com/science/pepsin Kageyama, T, and K. Takahashi. "Occurrence of two different pathways in the activation of porcine pepsinogen to pepsin." Journal of Biochemistry 93.3(1983):743-754. Piper, D. W., and B. H. Fenton. "pH stability and activity curves of pepsin with special reference to their clinical importance." Gut 6.5(1965): 506-508. Squires EJ, Haard NF, Feltham LA (1986) Gastric proteases of the Greenland cod Gadus ogac. I. Isolation and kinetic properties. Biochem Cell Biol 64: 205-214. Johnston N, Dettmar PW, Bishwokarma B, Lively MO, Koufman JA (Jun 2007). "Activity/stability of human pepsin: implications for reflux attributed laryngeal disease". The Laryngoscope. 117 (6): 1036–9. http://www.worthington-biochem.com/introbiochem/effectspH.html Umezawa H, Aoyagi T, Morishima H, Matsuzaki M, Hamada M (1970) Pepstatin, a new pepsin inhibitor produced by agtinomygetes. J Antibiot (Tokyo) 23: 259-262. Zollner H (1999) Handbook of enzyme inhibitors. (3thedn), Wiley–VCH, Chichester, UK, 440-445. Gorgas FJS (2009) Dental medicine. A manual of dental material medica and therapeutics, Nabu Press, Washington, 48-50. Murado MA, González MDP, Vázquez JA (2009) Recovery of proteolytic and collagenolytic activities from viscera by-products of rayfish (Raja clavata). Mar Drugs 7: 803-815. Alm?s KA (1990) Utilization of marine biomass for production of microbial growth media and biochemicals. In Advances in Fisheries Technology and Biotechnology for increased profitability, Voigt MN, Botta JR, Eds., Technomic Publishing Company, Lamcaster, 361-372. Falkenburg WJ, van Schaardenburg D, Ooijevaar-de Heer P, Tsang-A-Sjoe MW, Bultink IE, Voskuyl AE, Bentlage AE, Vidarsson G, Wolbink G, Rispens T (2016). "Anti-Hinge Antibodies Recognize IgG Subclass- and Protease-Restricted Neoepitopes". Journal of Immunology. Kun LY (2006). Microbial Biotechnology: Principles And Applications. World Scientific Publishing Company.? Jongjareonrak A, Benjakul S, Visessanguan W, Nagai T, Tanaka M (2005) Isolation and characterisation of acid and pepsin–solubilised collagens from the skin of brownstripe red snapper (Lutjanus vitta). Food Chem 93: 475-484. Roe SD (2001) Purification strategy. In Protein purification techniques (2ndedn), Roe S, Eds, Oxford University Press Inc, New York, 1-10.

Preparation

Pepsin is a stomach enzyme that serves to digest proteins found in ingested food. Production method of pepsin: using porcine gastric mucosa as raw material, after extraction with phosphate buffer solution, use DEAE-cellulose column chromatography, and after acidification, use Sephadex C-25 and Sephadex G-25 column chromatography again to refine the product.

Biochem/physiol Actions

Unlike many other peptidases, pepsin hydrolyzes only peptide bonds, not amide or ester linkages. The cleavage specificity includes peptides with an aromatic acid on either side of the peptide bond, especially if the other residue is also an aromatic or a dicarboxylic amino acid. Increased susceptibility to hydrolysis occurs if there is a sulfur-containing amino acid close to the peptide bond, which has an aromatic amino acid. Pepsin will also preferentially cleave at the carboxyl side of phenylalanine and leucine, and to a lesser extent at the carboxyl side of glutamic acid residues. It does not cleave at valine, alanine, or glycine linkages. Z-L-tyrosyl-L-phenylalanine, Z-L-glutamyl-L-tyrosine, or Z-L-methionyl-L-tyrosine may be used as substrates for pepsin digestion. Pepsin is inhibited by several phenylalanine-containing peptides.

Purification Methods

Pepsin is re-chromatographed on a column of Amberlite CG-50 using a pH gradient prior to use. Crystallise it from EtOH. [Richmond et al. Biochim Biophys Acta 29 453 1958, Huang & Tang, J Biol Chem 244 1085 1969, 245 2189 1970.]