Our company exhibited at "ifia JAPAN 2024 - The 29th International Food Ingredients/Additives Exhibition & Conference" (organized by Food Chemical News Co., Ltd.), held at Tokyo Big Sight from May 22nd (Wednesday) to 24th (Friday), 2024.

We would like to express our sincere gratitude to everyone who visited our exhibition booth.

In this article, we will introduce and answer the questions we received from all of you during the exhibition period, focusing on the ones that were particularly frequent. Please stay tuned until the end.

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Q: What does your company do?

A: In response to our clients' needs, we conduct new enzyme exploration and enzyme modification. By employing our unique bioinformatics technology, which differs from conventional methods, we facilitate rapid enzyme development. We believe that this innovation accelerator can benefit both enzyme manufacturers and food manufacturers.

Q: Do you have any specific examples?

A: In chemical applications, we have successfully explored new enzymes needed by users and achieved significant improvements in enzyme activity. In food applications, we are currently addressing specific themes requested by multiple clients and actively working on them.

Q: What properties of enzymes can be modified in the digzyme Spotlight (enzyme modification program)?

A: Potential modifications include enhancing activity, improving heat resistance, and altering optimal pH. Modifications to substrate specificity are addressed using the digzyme Moonlight (enzyme exploration program) as needed.

Q: What is the development process like?

A: Depending on the client's situation, we set the start and end goals, but the main process typically follows these steps:

1. Development Consultation: We listen to the client's challenges and select the target enzymes.
2. Enzyme Design: Using supercomputers, we design the target enzymes.
3. Enzyme Library Provision: Enzymes designed on the computer are produced at the lab scale using microorganisms, and the suitability of the enzymes for the intended purpose is verified and confirmed.
4. Enzyme Production Provision: We scale up production from the lab to the plant, ensuring stable enzyme supply as a product.

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That concludes the Q&A for this article. Thank you very much for reading until the end. If you have any further questions or inquiries, please contact us using the following contact form.

[Contact Form] https://www.digzyme.com/contact/

Introduction

I am Isozaki from the Business Development Department. Our company conducts explorations of artificial synthetic routes from "raw materials" to "target products" using enzymatic reactions. By simply inputting the compound structure data of the "target products" and "raw materials", we can output potential synthetic route candidates for producing the target product from the starting compound. In this blog, I will introduce a specific example where we predict a route to synthesize 4-amino-cinnamic acid, a which is used in the production of high-strength polymers  for high-strength polymers, from glucose and the enzymes involved in the reactions.

Materials Used for Synthetic Pathway Exploration

In Tateyama et al. (2016), 4-amino-cinnamic acid is used as a which is used in the production of high-strength polymers for producing high-strength polymers. The pathway used to synthesize this 4-amino-cinnamic acid is shown in Figure 1. Glucose serves as the raw material, and 4-amino-phenylalanine is produced using Escherichia coli engineered with Aminodeoxychorismate synthase (PapA) derived from Streptomyces venezuelae and Aminodeoxychorismate synthase (PapBC) derived from S. pristinaespiralis. Furthermore, this 4-amino-phenylalanine is used as a raw material, along with E. coli engineered with Phenylalanine ammonia-lyase (RgPAL) derived from Rhodotorula glutinis, to produce 4-amino-cinnamic acid.

Results

1. Biosynthetic Pathway Exploration

By inputting glucose as the Starting compound and 4-amino cinnamic acid as the product, an artificial synthesis pathway, as shown in Figure 1, was output. The output pathway was identical to the known synthesis pathway of chorismate from glucose, leading to the synthesis of 4-amino cinnamic acid via 4-amino phenyl alanine.

2. Similar Reaction Exploration

Among the artificial synthesis pathways identified in Result 1, the similar reaction from 4-amino phenyl alanine to 4-amino cinnamic acid was explored.

Through the exploration of similar reactions, a reaction that removes an amino group and generates a double bond was identified. Some of the similar reactions with a high degree of similarity to the target reaction and their rankings are shown in Figure 2. Similar reactions were extracted, including those that match the target reaction exactly.

3. Exploration of Corresponding Enzymes for Similar Reactions

In Result 2, similar reactions for the target reaction were extracted. The enzyme sequences responsible for these similar reactions were extracted by taxon. The filtered sequences were then compared with the enzymes used in the paper. Sequences were extracted at three levels: Rhodotorula genus, Eukaryota domain, and all taxa (Table 1). The extracted sequences included those that exhibited over 90% sequence homology with the sequences used in the paper.

Conclusion

In this blog, we demonstrated the exploration of artificial synthetic pathways. We explored an artificial route to synthesize the compound 4-amino cinnamic acid, which serves as a raw material for high-strength polymers, from glucose. We aimed to determine whether we could find enzymes that synthesize 4-amino cinnamic acid from 4-amino phenyl alanine using similar reaction enzyme exploration techniques. For the above reactions, we extracted sequences by taxon and presented the number of sequences for each. We successfully extracted multiple sequences that included several with high similarity to the enzymes used in the paper.

Acknowledgments

We utilized data from the following paper for this synthetic pathway exploration:

Tateyama et al. (2016). Ultrastrong, Transparent Polytruxillamides Derived from Microbial Photodimers. Macromolecules.