At the forefront of biotechnology, our team has made a breakthrough in the construction of a highly efficient exogenous protein expression system based on Saccharomyces cerevisiae GS115 and realized the precise synthesis of the glucose-lowering peptide P37, a 37-amino-acid peptide from legume plants containing six semi-optical cystine residues at sites 3, 7, 15, 20, 22 and 32, and the formation of three pairs of disulfide bonds within the molecule, which is resistant to degradation by gastrointestinal proteases. The three pairs of disulfide bonds can resist the degradation of gastrointestinal proteases, and has the effect of oral hypoglycemia.

This innovation not only reflects our deep understanding and precise application of yeast genetic engineering technology, but also opens up new pathways for the biosynthesis of complex proteins and peptides through the optimization of gene sequences, the design of highly efficient expression vectors, and the mastery of advanced transformation techniques.

Our work has not only accelerated the development of P37 glucose-lowering peptide as a potential therapeutic drug for diabetes, but also demonstrated far-reaching impacts on cost-effectiveness, industry-university-research cooperation, health and well-being, and environmental protection, contributing to the green and sustainable development of the pharmaceutical industry.

1、Optimization of gene sequences

2、Design of efficient expression carriers

We designed four plasmids

① We firstly completed the plasmid construction by EcoR I and Not I enzymatic cleavage of the vector and ligation with the target fragment. The molecular weight of the target protein product is about 16.9 kd.(EK-enterokinase cleavage site, his protein tag)

② In order to increase the expression amount, we expanded the copy number of the gene and tandemly linked the 3 vg together, and at the same time, we replaced the enterokinase with an acid-sensitive site (Asp-Pro), which is more convenient for later purification. The molecular weight of the target protein product was about 25.4 kd.

③ We inserted the NK gene between the α-factor signal peptide and the vg gene, which is a nattokinase gene with an open reading frame of 1146 bp. It encodes 381 amino acids and has a molecular weight of 27.7 kDa. Nattokinase is a protein that regulates glycolipid metabolism. We incorporated it into our recombinant protein to increase the molecular weight of the recombinant protein. The molecular weight of the target protein product is about 55.5 kDa.

By finely regulating the metabolic pathway and expression conditions of Pichia yeast, the efficient and stable expression of P37 glucose-lowering peptide was realized, which set a new benchmark for the biosynthesis of other complex proteins or peptide drugs.

The development process of P37 glucagon peptide, a novel drug candidate, is usually long and complex. However, the development cycle from gene sequence to active drug molecule has been greatly shortened by microbial synthesis technology, especially by utilizing Picrosporum yeast, an efficient expression system. This not only accelerates the preclinical study of the drug, but also lays a solid foundation for subsequent clinical trials and commercial production, which is expected to meet the needs of patients more quickly.

Compared to traditional protein production methods, such as chemical synthesis or plant extraction, the production of P37 hypoglycemic peptides using Picrosporum fermentation offers significant cost advantages. Picot yeast grows rapidly, has simple culture conditions, is easy to realize high-density fermentation, and its expressed proteins usually have high purity and biological activity. These features enable the production cost to be significantly reduced, while guaranteeing the quality and safety of the drug, which is conducive to the large-scale production and market popularization of the drug.

We have also cooperated with enterprises, we provide the technology and they produce it. The cooperation between our team and enterprises not only promotes the translation and application of research results, but also strengthens the communication and interaction between academia and industry. This mode of cooperation helps both parties share resources, complement each other's strengths, and jointly promote the development process of P37 glucose-lowering peptide drugs. Through the cooperation, enterprises can obtain cutting-edge biotechnological support and accelerate the launch of new products, while academic institutions can obtain more R&D funds and market feedback to promote the in-depth development of scientific research.

The successful development and potential application of P37 glucose-lowering peptide brings new therapeutic hope and options for diabetic patients.It has the following two advantages:

This drug is expected to effectively control the occurrence and development of diabetes and its complications by regulating blood glucose levels and improving insulin sensitivity. With the wide application of P37 glucose-lowering peptide, the quality of life of patients will be significantly improved, and the medical burden on society and families will be reduced.

The application of biotechnological means in drug production reflects the concept of green chemistry and sustainable development. Compared with traditional chemical synthesis methods, the biosynthesis process produces less waste and is easy to handle. In addition, as a renewable resource, the cultivation process of Bicarbonate yeast does not depend on limited natural resources, which helps to reduce the damage to the environment and the waste of resources. Therefore, the production of P37 glucose-lowering peptide using Picrosylla fermentation not only meets the environmental requirements, but also provides a strong support for realizing the sustainable development of the pharmaceutical industry.