Metabolic engineering and synthetic biology have developed rapidly in recent years. One of the most important applications is metabolic pathway engineering. By building cell factories, products with high added value can be produced. Metabolic channel engineering is to connect metabolic networks to developed strains; Metabolic pathway engineered studies often require the introduction of multiple genes from one or more different organisms, either by enhancing various endogenous and exogenous metabolic pathways or by directly introducing genes borrowed from other host organisms to achieve a high yield production of products. Metabolic pathway engineering research has essential application value in many fields, such as chemical synthesis, medicine, agriculture, and the environment. Several teams have reconstituted and optimized the biosynthetic pathways of compounds such as paclitaxel and artemisinin in E. coli, and Large-scale industrialization has been achieved through functional verification of metabolic pathways and optimization of host selection.

Creative Enzyme has rich experience in metabolic pathway research, biosynthesis, and strain modification. Based on our professional scientist teams and advanced technical equipment, we can provide integrated services of metabolic pathway engineering to fully meet the needs of customers.

Fig.1 Metabolic engineering strategies at different levels^[1]

Highlighted Features:

We provide several methods of Metabolic pathway engineering:

• Multi-modular optimization

The Multi-modular optimization divides the enzymes in the metabolic pathway into several modules according to the metabolic pathway nodes and the enzymes' catalytic efficiency. Metabolic pathways can be optimized by adjusting these modules at the level of transcription (e.g., promoter, gene copy number), translation (e.g., ribosome binding site), or catalytic properties of enzymes. This method is a simple, efficient, general optimization tool for metabolic pathway engineering.

• Optimization of metabolic pathway scaffolds

Metabolic pathway intermediates may be toxic to the host, consumed by competing pathways, or lost through secretion. The best way to solve this problem is to organize the enzymes in the metabolic pathway into multienzyme complexes. Multi-enzyme complexes need to be realized by synthetic scaffolding technology, which provides a new method for the co-localization of two or more enzymes in the pathway. Proteins, DNA, and RNA, can all be used as scaffolds for forming enzyme complexes, and optimizing the scaffolds for forming enzyme complexes plays an important role in studying metabolic pathway engineering.

• Large-scale gene editing technology

The CRISPR-CAS9 system is also an essential tool in the research of metabolic pathway engineering. By using the modified technology, critical enzymes in the metabolic pathway can be directionally modified without affecting other enzymes in the pathway. High-yielding metabolic strains can be obtained while avoiding the risk of recombination events that alter host behavior and phenotype.

• Metabolic flux analysis

Conventional metabolomics detects static metabolite concentrations, but the static content sometimes does not fully explain the problem. Metabolic flux analysis can calculate the flow direction and distribution of the compound in the metabolic pathway. By analyzing the metabolic flux of the organism, the activity of the specific metabolic pathway of the organism can be obtained. Metabolic pathways and fermentation processes can be optimized through metabolic flux analysis.

• Metabolic pathway library construction

Creative Enzyme has rich experience in library construction. We provide metabolic pathway synthesis services and customized metabolic pathway library assembly services. The library capacity can be customized through various library assembly technologies. High-yielding metabolic pathway strains were obtained by high-throughput screening.

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