Fermentation-Based Production for Enzyme and API
With the rapid development of the pharmaceutical industry, green pharmaceuticals represented by fermentation production, asymmetric catalysis, and continuous flow are increasingly sought after. Fermentation of active pharmaceutical ingredients (APIs) and enzymes is one of the important ways of the API production process, which mainly covers amino acids and antibiotics, and some biological products. These compounds are usually obtained by a semisynthetic method combining fermentation and chemical synthesis.
Due to the technical specificity of enzymes produced by fermentation and the fermentation method itself, the cost of fermented APIs is much lower than that of the synthetic method. Despite the cost advantages of fermentation-based production of enzymes and APIs, there may be some undesired by-products produced in the fermentation process, and enzyme or tissue residues may also remain in API products. Hereby, the control of impurities and microorganisms, as well as the treatment of enzyme and cell tissue residues, are discussed to help researchers bring more enzymes and APIs to the market.
- Control of impurities and microorganisms
The API and enzyme produced by the fermentation method have uniqueness, which is reflected in the congeners and side reaction impurities generated by fermentation with difficulty to separate and purify. For this type of API manufacturing, the development process may begin with a theoretical analysis of impurity and principal component homolog, and peaks greater than 0.1% in multiple batches should also be analyzed and identified to determine the substance component. At the same time, pharmacological and toxicological tests are performed to determine whether the element is an effective principal component homolog or an impurity.
Additionally, enzyme manufacturing and API production involved with enzymes should be formulated as measures to prevent microbial contamination in microbiology. For example, the manufacturing environment and production equipment need to be clean and disinfected, and the enzyme inactivation process should be carefully tackled. Especially in the appropriate production stage such as material exposure process should also be carried out to monitor the microbial load and exogenous microorganisms, and consider the product for bacterial endotoxin monitoring.
- Enzyme and tissue residue treatment
Even if the R&D department and the production department can theoretically explain that the enzyme and cell tissue can be completely removed in the preparation, separation, and purification stages of the fermentation-based production of API, there are still risks of residual enzyme and cell tissue the APIs. Therefore, to reduce enzyme and tissue residues, the following treatments are usually performed:
- In the production process stage, R&D personnel need to remove as many enzymes and cell tissues as possible through separation, ultrafiltration, nanofiltration, and other steps;
- Developing special test reagents for enzymes and cell tissues used in the production of the API, and conducting methodological verification on the specificity, precision, linearity, accuracy, the limit of detection and quantification, durability, and other items of the developed reagents;
- Conducting quantitative enzyme and cell tissue tests on intermediates, filtrates, and finished products of the above steps using specific methods after verification, and testing the transfer process of enzyme and cell tissue in production and the residual amount in finished products
In a word, the fermentation of enzyme and API can take the enzyme as the special starting material for API production to carry out comprehensive quality control, while considering the impurities generated in the process with enzyme involvement and the transfer process; On the other hand, researchers should scientifically control the activities of enzymes in the fermentation process and API production, such as the special environment for enzymes and fermentation strategies, to obtain high-quality products.
