In recent years, diabetes patients have gained more therapeutic options thanks to the rise of next-generation GLP-1 analogs. Among them, Semaglutide—a modified glucagon-like peptide-1 (GLP-1) analog—has become a market leader for its powerful effects on glycemic control, weight reduction, and metabolic health improvement.
Clinically proven to work synergistically with other oral antidiabetic drugs, Semaglutide improves blood glucose control while promoting weight loss, reducing systolic blood pressure, and supporting pancreatic β-cell function. Its three formulations—Ozempic™ (injectable for diabetes), Rybelsus™ (oral tablet), and Wegovy™ (injectable for weight management)—achieved over $20 billion in global sales in 2023. By 2024, worldwide GLP-1 drug sales surpassed $50 billion, reinforcing the unprecedented momentum in obesity and diabetes therapeutics. With demand continuing to surge, more pharmaceutical companies are accelerating their investments in Semaglutide production technologies.
Semaglutide is a long-acting GLP-1 receptor agonist, structurally optimized from human GLP-1 to extend half-life and enhance stability. It is now a leading therapy for type 2 diabetes and obesity.
Peptide Backbone: Arg34-GLP-1(9–37), a 29–amino acid peptide: EGTFTSDVSSYLEGQAAKEFIAWLVRGRG
Scalable Production
Traditionally produced by chemical synthesis, semaglutide can also be manufactured through advanced fermentation. With synthetic biology driving efficiency and cost reduction, fermentation-based production is emerging as the future of large-scale GLP-1 drug manufacturing.
Table 1. Comparison of Semaglutide Production Methods
|
Method |
Advantages |
Limitations |
|
Chemical Synthesis |
High purity and consistent quality; Flexible; Suitable for small-scale production |
High cost; Lower production efficiency |
|
Synthetic Biology Fermentation |
Lower cost; Higher yield |
High technical requirements Production technology requires precision. |
Biological Production
Figure 1. Schematic Diagram of the Main Process Flow for the Biological Production Route of Semaglutide
Single-Enzyme Process: Recombinant Enterokinase (rEK)
Recombinant Enterokinase (rEK), a highly purified light-chain subunit of bovine enterokinase, exhibits the same substrate specificity as the native enzyme. It precisely recognizes the Asp-Asp-Asp-Asp-Lys cleavage site, removing N-terminal fusion tags or carrier sequences with minimal nonspecific activity.
This single-enzyme cleavage approach simplifies the manufacturing workflow:
- Streamlined process with fewer steps
- High catalytic efficiency and rapid reaction rates
- Ideal for large-scale industrial production due to robustness and ease of control
As a result, recombinant enterokinase has become the enzyme of choice for efficient and scalable Semaglutide intermediate peptide processing.
Features
High Specificity – Precisely cleaves at the Asp-Asp-Asp-Asp-Lys recognition site.
High Purity – Three-step purification((Ni column, DEAE anion exchange, and Phenyl hydrophobic chromatography) ensures no contaminating proteases or non-specific cleavage.
Animal-Free – Recombinant production, virus- and serum-free
Stable & Consistent: Reliable batch-to-batch quality.
Scalable Supply – 5 L–1500 L fermentation capacity to meet diverse needs.
Multiple Grades – Available in R&D (Cat#20395ES) and GMP (Cat#20396ES, ISO 13485 compliant with full documentation).
Product Properties
|
Product Number |
20395ES |
20396ES |
|
Product Name |
||
|
Product grade |
RUO grade |
GMP grade |
|
Source |
Recombinant Expression from Pichia pastoris |
Recombinant Expression from Pichia pastoris |
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Enzyme Concentration |
5 U/μL |
5 U/μL |
|
Purity |
≥95% |
≥95% |
|
Endotoxin |
<1 EU/μg |
<1 EU/μg |
|
Host protein |
N/A |
≤0.01% |
|
Host DNA |
N/A |
≤10 ng/mg |
|
Activity Definition |
One unit of enzyme activity is defined as the amount of enzyme required to cleave 95% of a 50 µg fusion protein containing an enterokinase recognition site (Yeasen enterokinase cleavage positive substrate, MW ~64.6 kDa, Cat#20391ES) after incubation at 25 °C for 12–16 hours in a reaction buffer (20 mM Tris-HCl, 50 mM NaCl, 2 mM CaCl₂, pH 8.0). |
|
|
Product Application |
Semaglutide: Supports large-scale peptide drug production with precise cleavage. IL-11: Enables efficient tag removal for active recombinant IL-11. hEGF: Facilitates purification of recombinant hEGF with high bioactivity. |
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Performance
A. High Purity
Figure 2. SDS-PAGE analysis showing >95% purity(Cat#20395ES)
B. Recombinant Enterokinase Activity Test
Figure 3. Enterokinase (Yeasen, Cat#20395ES) shows cleavage performance comparable to Supplier N*.
C: Positive substrate(Cat#20391).
C. High Specificity
Figure 4. Yeasen Enterokinase (Cat#20395ES) demonstrates superior cleavage performance compared to Supplier N*.
D. Strong Stability
Figure 5. Stability of Yeasen enterokinase (Cat#20396ES) under freeze–thaw and accelerated storage.
Enzyme activity remained stable after 10–20 freeze–thaw cycles and during storage at 25°C (7–32 days) and 37°C (7–14 days).
Dual-Enzyme (Tandem) Process: Recombinant Kex2 Enzyme + Carboxypeptidase B
In contrast, the dual-enzyme (tandem) system—combining recombinant Kex2 protease and carboxypeptidase B (CPB)—offers higher yields and precise cleavage control. This method enables fine-tuned processing of precursor peptides, achieving high product purity and enzymatic efficiency.
However, the tandem approach also introduces complexity:
- Process optimization challenges due to differing optimal pH (Kex2: ~9.0; CPB: 7.5–9.0) and metal ion sensitivities
- Higher production costs, as both Kex2 and CPB are recombinant proteins
- Scale-up limitations, especially when maintaining consistent enzyme activity in large fermentations
Therefore, while tandem cleavage remains valuable for research and high-precision applications, the single-enzyme enterokinase route offers a more practical, scalable, and cost-effective solution for industrial Semaglutide manufacturing.
Product Properties
|
Product Number |
20418ES |
20417ES |
|
Product Name |
||
|
Product grade |
GMP grade |
GMP grade |
|
Source |
Recombinant Expression from Pichia pastoris |
Recombinant Expression from E. coli |
|
Activity |
≥10.0 units/mg pro |
≥170 USP units/mg pro |
|
Cleavage Site |
Specifically recognizes and cleaves peptide bonds at the carboxylic acid terminal of dibasic amino acids such as Arg-Arg, Lys-Arg. |
Specifically hydrolyzes proteins at the amino-terminal side of basic amino acids (lysine, arginine, histidine) at the C-terminus. |
|
Product Application |
1. Process enzyme cleavage in peptide drug production. 2. Enzymatic digestion of proteins and peptide mapping, sequencing, etc. |
1. Production of recombinant insulin and its analogs. 2. Determination of amino acids at the C-terminus of proteins. 3. Removal of histidine tags at the C-terminus of proteins. 4. Production of other recombinant peptide substances. 5. Enzymatic synthesis of certain special compounds.
|
Yeasen recombinant Kex2 and Carboxypeptidase B exhibit high cleavage accuracy and enzyme activity. Through large-scale production, YeaSen has significantly reduced the cost of Kex2 enzyme, eliminating cost barriers in tandem enzymatic processes and overcoming production capacity bottlenecks.
Perfomance
HPLC Analysis of P29-Arg34 GLP-1(9–37) Cleavage by Kex2 and Carboxypeptidase B
Figure 6. Yeasen Kex2 protease and carboxypeptidase B efficiently cleave P29-Arg34 GLP-1 (9–37), supporting a tandem enzymatic process with high yield and high purity.
Semaglutide Intermediate: 29-mer Peptide P29-Arg34 GLP-1(9–37)
A recombinant E. coli-expressed backbone peptide for semaglutide, consisting of a 29-amino acid intermediate. This peptide serves as the starting raw material for semaglutide synthesis. YeaSen offers this intermediate in stock, eliminating the need for customers to establish in-house biological production, thereby shortening process development timelines and accelerating time-to-market.
Perfomance
P29-Arg34 GLP-1(9–37) Identification
Figure 7. The main peak retention time of Yeasen P29-Arg34 GLP-1 (9–37) matches that of the reference standard in liquid chromatography.
Purity Analysis of P29-Arg34 GLP-1(9–37)
Figure 8. Yeasen P29-Arg34 GLP-1 (9–37) exhibits a purity greater than 98%.
Related Products
|
Catgory |
Name |
Cat. No. |
Size |
|
Recombinant enterokinase |
20395ES60/76/90/92/94 |
100 U/500 U/5000 U/100 KU/1 MU（1000 KU） |
|
|
|
20396ES90/92/94 |
5000 U/100 KU/1 MU（1000 KU） |
|
|
|
20391ES03/11 |
1 mg/4 mg |
|
|
Recombinant Kex 2 & Carboxypeptidase |
20418ES80/90/91/92 |
1 mg/10 mg/100 mg/1 g |
|
|
|
UCF.ME™ Recombinant Kex2 Protease(1 mg/mL，Solution) |
20385ES60/80/90 |
100 μg/1 mg/10 mg |
|
|
20417ES03/10 |
1 mg/10 mg |
|
|
Intermediate peptide |
Semaglutide intermediate P29-Arg34GLP-1(9-37)(Powder) |
20381ES01/03/08/25/60 |
100 mg/1 g/5 g/25 g/ 100 g |
|
In vitro activity detection |
11411ES/11404ES/11412ES60/80 |
100 T/10× 100 T |
|
|
|
Firefly Glo Luciferase Reporter Gene Assay Kit |
11404ES60/80 |
100 T/1000 T |
|
|
11412ES60/80 |
100 T/10×100 T |