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Publication Number
US-12692473-B2
Publication Date
2026-07-28
Expiration Date
Abstract
Provided is an alga belonging to Cyanidiophyceae having a diploid cell and a haploid cell form. Also provided is a nutrient composition containing an alga belonging to Cyanidiophyceae or an extract thereof.
Core Innovation
The invention relates to novel Cyanidiophyceae microalgae having both diploid and haploid cell forms, including Cyanidium sp. YFU3 (FERM BP-22334) and HKN1 (FERM BP-22333). The haploid cell form lacks a rigid cell wall, and this property enables cell rupture under a pH 7 condition for nutrient extraction or nutrient absorption from the haploid cell form.
The invention also addresses production of haploid algae by culturing diploid cells until the alga enters the stationary phase, isolating haploid cells that have no rigid cell wall from the obtained culture, and culturing the isolated haploid cells. It further includes production of diploids from mixed haploids, and the disclosure is framed around cell form transitions between diploid and haploid states within algae belonging to Galdieria or Cyanidium.
The invention defines a drug-generating gene concept for Cyanidiophyceae that are used to produce a therapeutic and/or preventive drug. In particular, the drug-generating gene is introduced into Cyanidiophyceae so that the cells produce drug substances, including antigenic proteins for vaccines against infectious diseases or enzymes that catalyze drug synthesis.
The disclosure further presents Cyanidiophyceae producing nutrient-related compounds, including γ-aminobutyric acid (GABA), and describes transformation and confirmation of introduced genes to produce increased intracellular amounts proportional to introduced gene copies.
Claims Coverage
The partial content provides one independent claim (clm-00001) with dependent refinements. The inventive features in the independent claim and its refinements focus on producing haploid algae from diploid cells by stationary-phase culturing and isolating a haploid cell lacking a rigid cell wall, with further limitations on rupturing, gene definition, strain selection, and transformant selection.
Producing a haploid alga from diploids via stationary-phase culturing and wall-free haploid isolation
A method for producing a haploid alga comprising culturing a diploid cell of an alga belonging to Galdieria or Cyanidium until the alga enters the stationary phase, isolating a haploid cell which has no rigid cell wall from the culture obtained, and culturing the haploid cell isolated.
Rupturing wall-free haploid cells under a pH 7 condition
The method includes rupturing a haploid cell under a pH condition of 7.
Using a ribulose 1,5-bisphosphate carboxylase/oxygenase large subunit gene defined by SEQ ID NO: 1 or 2
The method is performed using an alga ribulose 1,5-bisphosphate carboxylase/oxygenase large subunit gene having a base sequence shown in SEQ ID NO: 1 or 2.
Using specified Cyanidium strains as the alga source
The method for producing a haploid alga is performed using an alga selected from Cyanidium sp. strains FERM BP-22334 or FERM BP-22333.
Selecting a transformant using an auxotrophy-related selection marker
The method further specifies that the transformant is obtained using an auxotrophy-related gene as a selection marker.
Overall claim coverage in the provided material centers on producing haploid algae from diploid algae in Galdieria or Cyanidium by stationary-phase culturing and isolating haploid cells that have no rigid cell wall, with dependent limitations that further specify a pH 7 condition for rupturing, define a ribulose 1,5-bisphosphate carboxylase/oxygenase large subunit gene by SEQ ID NO: 1 or 2, narrow the alga to stated Cyanidium strains (FERM BP-22334 or FERM BP-22333), and define selection of a transformant using an auxotrophy-related gene as a selection marker.
Stated Advantages
Enables nutrient extraction or nutrient absorption from the haploid cell form because haploid cells lack a rigid cell wall and can undergo rupture under a pH 7 condition.
Enables Cyanidiophyceae to produce a therapeutic and/or preventive drug by introduction of a drug-generating gene.
Enables production of vaccine-related antigenic proteins against infectious diseases using Cyanidiophyceae.
Enables production of drug-enzyme products by introducing an enzyme that catalyzes drug synthesis.
Provides intracellular increases in target compounds corresponding to introduced gene copies, including γ-aminobutyric acid (GABA).
Documented Applications
Nutrient composition and nutrient-related products, including functional foods.
Nutritional supplement.
Feed.
Pet food.
Cosmetic.
Production of therapeutic and/or preventive drugs using engineered Cyanidiophyceae.
Vaccine use cases involving antigenic proteins for infectious diseases.
Production of nutrient-related products such as γ-aminobutyric acid (GABA) from Cyanidiophyceae.
Industrial applicability describing nutrient compositions and products such as supplements and feeds, and use in cosmetics.
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