Analysis of the Biological Fermentation Process for the Synthesis of "Weight Loss Drug" semaglutide

How to synthesize semaglutide？

Semaglutide is a long-acting glucagon like peptide-1 receptor (GLP-1R) agonist, which is mainly used to treat type 2 diabetes and obesity. Semalutide physiologically simulates the function of the naturally existing GLP-1 hormone in the human body. This hormone is secreted by intestinal L cells after meals, which can promote insulin secretion, inhibit glucagon secretion, and also slow gastric emptying speed, thus regulating blood sugar levels and reducing appetite, helping patients control diet and weight. Therefore, Smegglutide has transformed into a "slash" and become a new favorite of "weight loss drugs" on the internet.

How is Semaglutide produced?

Semaglutide is composed of 31 amino acids, and its production mainly includes two methods: pure chemical synthesis method and synthetic biological fermentation method (also known as semi recombinant method). The synthetic biology fermentation method mainly uses yeast or Escherichia coli to ferment and synthesize the main peptide chain (also known as GLP-1 analog precursor peptide) of drugs. The precursor peptide in the fermentation broth is extracted and purified, and the 26th lysine position of the GLP-1 skeleton is modified by enzymatic digestion or other methods. Then, it is coupled with the N-terminal amino acid protrusion containing non protein amino acids to obtain active smeglutide molecules.

So, using synthetic biological fermentation to produce the main peptide chain of Smegglutide is the first and extremely crucial step. Its advantages are mainly reflected in two aspects: (1) Firstly, compared to traditional chemical synthesis methods, synthetic biology fermentation technology breaks through production capacity bottlenecks and is more suitable for large-scale continuous production. It can effectively overcome the yield limitations caused by complexity and scale expansion in chemical synthesis. (2) Secondly, this method demonstrates significant advantages in cost control. By optimizing the biological fermentation process and utilizing the efficient biosynthesis ability of microorganisms, production costs can be further reduced, resulting in more significant economic benefits. 

Specific production process of synthetic biology fermentation method

What is it like?

The synthetic biological fermentation process of Semaglutide combines genetic engineering technology, microbial fermentation, and chemical synthesis steps, mainly divided into the following steps:

Genetic design:

Firstly, a recombinant plasmid containing the coding sequence of the precursor peptide of Semaglutide, usually a human GLP-1 analogue, was designed and constructed through genetic engineering techniques. Import this recombinant plasmid into suitable host cells, such as Saccharomyces cerevisiae or Escherichia coli, to express the target peptide.

Fermentation process:

Inoculate engineering strains carrying recombinant genes into fermentation tanks and conduct large-scale cultivation under specific conditions, including temperature, pH value, dissolved oxygen concentration, nutritional composition, etc., to promote efficient expression and accumulation of target peptides. It is worth noting that during the fermentation process, as microorganisms consume nutrients and produce metabolic waste, the cultivation environment will become increasingly harsh, such as carbon dioxide accumulation, pH changes, temperature rise, and oxygen depletion. These conditions that are not conducive to microbial synthesis of target products can sometimes be sudden and cause irreversible damage to microorganisms, affecting the quality and yield of target products. It can be seen that effective biological process control is very necessary. Efficient biological process control cannot be achieved without direct and real-time monitoring of biological process parameters. Currently, most fermentation equipment can be equipped with sensors to monitor various parameters of the cultivation system in real time. The combination of sensors and process control software can achieve efficient control of the process. For example, the data acquisition and monitoring system software DASare developed by Eppendorf specifically for advanced biological process control can achieve efficient management of next-generation biological process information. It has an intuitive user interface, advanced programming and experimental design functions, and can efficiently complete process monitoring, process control, and data recording. And it can support seamless integration of Eppendorf bioprocess control system with laboratory sampling and analysis equipment, easily integrating nutrition analyzers, cell counters, biomass monitors, mass spectrometers, automated platforms and automatic sampling instruments, HPLC, Raman spectrometers and other equipment, achieving better process understanding and control. In addition, considering that microorganisms need to consume a large amount of oxygen during the peptide fermentation synthesis process, sufficient oxygen supply helps to correctly express and assemble the target peptide, reducing the probability of misfolding or incomplete maturation. In addition, sufficient oxygen can inhibit certain anaerobic metabolic pathways, making the metabolic flow more inclined towards the formation of the target product, thereby improving the synthesis efficiency of the target peptide. Therefore, the oxygen supply efficiency of the system is particularly important. In large-scale fermentation systems, the improvement or optimization of oxygen supply efficiency is of great significance for the increase of peptide product yield. Therefore, whether there is an efficient and flexible oxygen supply system is one of the important factors to consider when choosing a suitable bioreactor. Eppendorf's BioFlo 320 bioprocess reactor, equipped with intelligent sensors and advanced gas control systems, can quickly and intelligently respond to dissolved oxygen concentration, carbon dioxide concentration, pH, etc. It can help customers automatically and easily control the gas during the fermentation process through gas mixing algorithms, significantly improving flexibility.

Extraction and purification of precursor peptides:

In the stage of peptide extraction and purification, it is first necessary to collect bacterial cells from the fermentation broth. Eppendorf's CR30NX high-speed ground centrifuge can be equipped with a large capacity rotor for bacterial collection: the uniquely designed fixed angle rotation R9A2, equipped with four 1.5L triangular centrifuge bottles, can accelerate the sample processing process; In addition, the R10C2 continuous flow rotor can process tens of liters of samples through continuous injection and extraction, reducing inter batch differences and laying a solid foundation for large-scale continuous production.

Then, it is necessary to release the expressed precursor peptides through bacterial lysis and other methods, and then use various protein purification techniques such as centrifugation, affinity chromatography, ion exchange chromatography, size exclusion chromatography, etc. to separate and preliminarily purify the precursor peptides. The key point to note in this step is to choose the appropriate cell lysis and fragmentation method, which not only ensures the effective rupture of the cell wall and membrane structure, but also minimizes the damage and denaturation of the target protein as much as possible. Control the conditions for bacterial lysis, fragmentation, and purification, such as temperature, pH value, time, and pressure, to avoid excessive conditions that may cause protein inactivation or degradation.

Chemical modification and final product generation:

The obtained precursor peptides also need to be modified through chemical reactions (such as amino acid chemical modification, coupling reactions, etc.) to form the final structure of Smegglutide, ensuring the biological activity and stability of the product.

Structural confirmation and quality control:

After chemical modification, the molecular weight, purity, and structure of Smegglutide need to be confirmed through analytical techniques such as high-performance liquid chromatography (HPLC), mass spectrometry (MS), or nuclear magnetic resonance (NMR), and meet the quality requirements of pharmacopoeia and regulatory agencies.

In summary, the production process of Semaglutide is a precise and technically demanding process that involves multiple technical links. The meticulous control of each link is the key to obtaining high-quality target products. Learn from others' successful methods, enhance your platform advantages, and Eppendorf supports all your process development needs.