#Japan #Tokyo #Regenesome #Koji Mold #Fermented Sweeteners

Introduction to the Innovation in Koji Mold Selection

Regenosome Corporation, part of Space Seed Holdings and based in Minato, Tokyo, has recently collaborated with Kanazawa Institute of Technology in Ishikawa and Orize Inc. in Shibuya, Tokyo, to file a patent for a groundbreaking method of selecting koji mold suitable for producing fermented sweeteners such as amazake and koji. The patent was submitted to the Japan Patent Office on August 31, 2026.

Background on Koji Mold

Koji mold, specifically Aspergillus oryzae, is an essential microorganism in the production of many traditional Japanese fermented foods, including sake, miso, and soy sauce. In recent years, fermented sweeteners derived from rice starch converted into sugars by koji enzymes—such as amazake and rice malt sweeteners—have garnered significant attention both domestically and internationally as natural sweeteners.

The sweetness and quality of these fermented sweeteners are heavily influenced by enzymes secreted by the koji mold, particularly alpha-amylase. However, due to the presence of multiple alpha-amylase genes with over 99% homology in the koji mold genome, distinguishing between them using conventional PCR techniques has been challenging. Moreover, it is known that the number of gene copies does not directly determine enzyme production levels, making it difficult to select the most suitable strains without relying heavily on manufacturing experience or extensive whole-genome sequencing.

Overview of the Invention

The innovative method utilizes PCR amplification patterns of regions surrounding three alpha-amylase gene loci in the koji mold combined with enzyme activity related to saccharification after koji-making. This approach aims to simplify and objectify the selection process for strains suitable for fermented sweetener production compared to sequencing methods. The specific procedure involves:

1. Obtaining PCR amplification patterns for target strains' genomic DNA at the three alpha-amylase gene loci.
2. Narrowing down candidate strains that show different amplification patterns from the standard strain (RIB40).
3. Measuring phenotypic values related to saccharification, including alpha-amylase activity, in the koji made from the candidate strains.
4. Selecting strains that show higher phenotypic values than the reference strain for fermented sweetener production.

In experimental applications, this method successfully identified practical strains exhibiting approximately 2.2 times (218% increase) the alpha-amylase activity compared to control strains, leading to successful amazake production with the koji made from these strains.

Applications and Future Development

Additionally, the PCR amplification patterns can also be utilized for strain identification, ensuring batch consistency and quality control during production. The patent also encompasses a method for strain selection, a strain identification method, reagent sets, identification kits, and manufacturing methods for both rice koji and fermented sweeteners.

Features of the Invention

• - Simplicity: Selecting candidate strains through PCR and enzyme activity measurement avoids the need for whole-genome sequencing.
• - Objectivity: By combining molecular markers and phenotypic values, this method facilitates high reproducibility and less reliance on subjective experience.
• - Scalability: The approach can extend beyond alpha-amylase to encompass proteases and lipases, allowing for targeted selection of koji molds based on desired flavors and aromas.
• - Quality Control: Strain identification through amplification patterns can aid in confirming the consistency of koji batches and managing inventory.

The Patent Details

• - Title of Invention: Method for Selecting Koji Mold for the Production of Fermented Sweeteners, Koji Strain Identification Method, Reagent Sets, Identification Kits, Rice Koji Production Method, and Fermented Sweetener Manufacturing Method.
• - Filing Date: August 31, 2026.
• - Applicants: Kanazawa Institute of Technology, Orize Inc., Regenosome Corporation (Joint Application).
• - Inventors: Masayuki Machida (Kanazawa Institute of Technology), Motoaki Sano (Kanazawa Institute of Technology), Ai Kawahara (Orize), Kengo Suzuki (Regenosome).
• - International Patent Classification: C12Q 1/6895, C12Q 1/686, C12N 1/14.

Future Prospects

Regenosome aims to advance technology related to fermentation using this invention to realize longevity-enhancing technologies. Collaborating with Kanazawa Institute of Technology and Orize, they plan to leverage genomic information of koji molds to create higher-value fermented foods.

About Regenosome Corporation

Regenosome is committed to offering innovative solutions in anti-aging and regenerative medicine by utilizing nanoparticles. With their base combining research and development at the Takanawa Gateway City’s ‘LiSH (Link Scholars' Hub)’ and an active healthcare business under the 'Saisei Lab' brand, they are accelerating research for future advancements in the medical field. Their long-term goal is to contribute to human longevity and facilitate humanity’s exploration of space.
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About Space Seed Holdings

Space Seed Holdings operates as a deep-tech venture builder driven by the mission to transform science fiction into reality, focusing on investment, research, and business development. They strive to address societal challenges through programs like the ‘Fermentation and Longevity Fund,’ aimed at providing necessary technologies for human habitation in outer space by 2040.
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