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Strain Engineering for Itaconic Acid

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CD Biosynsis is committed to reducing the cost of itaconic acid production and increasing the yield of itaconic acid by regulating gene expression, eliminating by-products, and enhancing the strain's resistance to adversity.

Advantages of Synthetic Biology-Driven Itaconic Acid Production

Itaconic acid, also known as methylene dibutyric acid, is considered to be the cleanest alternative to petroleum-based acrylic acid. Itaconic acid can be synthesized chemically or biosynthetically. However, the chemical synthesis of itaconic acid requires multiple stages and low production efficiency, so chemical synthesis has never been used for commercial production of itaconic acid. Microbial fermentation is the most commonly used method for itaconic acid production, mainly obtained by fermentation using filamentous fungi. With the development of genetic engineering, metabolic engineering, and other technologies, synthetic biology-driven itaconic acid production is not only high yielding but also easy to control and improve.

Figure 1. Schematic representation of life cycle of rapidly renewable itaconic acid. (Bafana R, et al., 2018)Figure 1. Schematic representation of life cycle of rapidly renewable itaconic acid. (Bafana R, et al., 2018)

What We Provide

Our experienced scientists are dedicated to helping our customers solve problems in the production of itaconic acid using advanced technology. We want to provide you with high-quality and efficient one-stop service.

Metabolic Engineering

We expect to use metabolic engineering tools to help our customers improve the productivity of microorganisms for itaconic acid, reduce the productivity of by-products, and develop strains that can produce itaconic acid from inexpensive raw materials.

Genetic Engineering

We are able to use genetic engineering tools to increase the productivity of natural producers of itaconic acid and also to modify non-natural producing strains to have the ability to produce itaconic acid.

Phenotype Engineering

The cellular morphology of microbial chassis may affect the production of itaconic acid. We are able to use morphological engineering tools to improve the production of itaconic acid.

Transcriptome Analysis

We are able to find key enzymes affecting the microbial itaconic acid synthesis pathway through transcriptome analysis and establish optimal parameters for itaconic acid biosynthesis.

Deliverables

  • Itaconic acid production microorganism.
  • Itaconic acid.

How We Can Help

Development of Synthetic Biology Chassis for Itaconic Acid Production

Many microorganisms use cis-aconitate as a substrate to produce itaconic acid outside the mitochondria via cis-aconitate decarboxylase. The following microbial chassis have been successfully implemented for the production of itaconic acid. If you would like to learn about other microbial chassis development services, please contact us directly.

Aspergillus terreus Ustilago zeae Ustilago maydis
Candida sp. Pseudozyma antarctica Rhodotorula species
Ustilago maydis Yarrowia lipolytica

Itaconic Acid Producing Strain Improvement

Natural itaconic acid-producing strains have clusters of genes that specifically encode secretase and transporter proteins involved in itaconic acid production. We are able to use genetic engineering tools to regulate the expression of this gene cluster for efficient accumulation of itaconic acid production. We are able to provide the following strain improvement services, including but not limited to.

  • Random mutagenesis. We are able to induce the production of mutants with increased itaconic acid production by physically or chemically treating the microorganism.
  • Site-specific mutagenesis. Primary metabolite enhancement is an important approach to enhancing secondary metabolites. Therefore, enhanced glycolytic flux can increase the production of itaconic acid. We are able to disrupt the phosphorylation site of a key rate-limiting enzyme in the glycolytic pathway, remove the feedback inhibition of glycolysis by the enzyme, and introduce the mutant enzyme into itaconic acid producing strains to increase the yield of the glycolytic pathway, thereby increasing the yield of itaconic acid and shortening the fermentation cycle.
  • Genetic improvement of the adversity resistance. Enhancing the adversity resistance of the strain facilitates the fermentation intensity of the strain. We are able to overexpress resistance-related genes in itaconic acid production strains to ensure metabolic fluxes in the itaconic acid synthesis pathway. For example, the continuity of agitation and aeration during fermentation significantly affects the final yield of itaconic acid, and overexpression of the gene encoding vitreoscilla hemoglobin can avoid the altered metabolic flow caused by insufficient dissolved oxygen.
  • Systematic metabolic engineering. We are able to overexpress the gene encoding a key enzyme in the itaconic acid synthesis pathway using a strong promoter or achieve heterologous expression of the gene. The following genes are genes encoding key enzymes involved in the itaconic acid synthesis.
CadA Tad1 Adi1 MttA MfsA Itp1 Reg Cyp3 Rdo1 AcoA AcnB GltA CitA CitB Icd PfkA

Multi-Enzyme Self-Assembly for the Improvement of Itaconic Acid Production

Multi-enzyme cascade reactions can synthesize simple compounds with low cost into complex compounds with high added value through multi-step enzymatic reactions. We are able to use multi-enzyme self-assembly technology to achieve the self-assembly of multiple enzymes by intracellular co-expression of fusion proteins to improve the yield of itaconic acid.

Optobiochemical Control of Protein Activities

We are able to design and assemble light-sensing structural domains into the microbial photoregulatory system to control the function of proteins through light. We are able to help our customers establish optimal light control strategies to improve the production of itaconic acid.

Applications of Itaconic Acid

CD Biosynsis can develop tailored tools and customized approaches to harness the power of synthetic biology to drive itaconic acid production and meet the needs of customers in a variety of industries.

  • Used in the manufacture of paint emulsifiers.
  • Used in the production of synthetic resins.
  • Used in the production of synthetic fibers.
  • Used in the production of surface binders.
  • Used in the production of herbicides.
  • Used in the production of artificial glass.
  • Used in deodorant production.
  • In clinical applications, itaconic acid activates macrophages, regulates macrophage polarization, regulates tumor growth, and inhibits osteoclastogenesis.

Want to Learn More?

CD Biosynsis provides the most comprehensive and efficient solutions for synthetic biology workflows. We are committed to helping our customers solve all problems encountered in itaconic acid production to advance their applications in a wide range of fields. Each of our deliverables will undergo a rigorous quality inspection test to ensure the reliability and accuracy of the results. If you are interested in our services or have any further questions, please do not hesitate to contact us.

Reference

  1. Bafana R, Pandey RA. New approaches for itaconic acid production: bottlenecks and possible remedies. Crit Rev Biotechnol. 2018 Feb; 38(1): 68-82.

Please note that all services are for research use only. Not intended for any clinical use.

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