Production of lipids and terpenoids in Auxenochlorella protothecoides
Inventors
Moseley, Jeffrey • Lee, Byung-hee • IM, Chung Soon • Kim, Jane • Kim, Dayoung • Bhat, Riyaz
Assignees
Phycoilbiotech Korea Inc • Phycoilbiotech Korea Inc • Phycoil Biotechnology International Inc
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-12037630-B2
Publication Date
2024-07-16
Expiration Date
Abstract
Methods to produce oils with modified profiles of fatty acid, carotenoids and/or terpenoids in microalgal mutants are provided. Microalgal mutants produce the oil containing fatty acids, carotenoids and/or terpenoids of a modified profile with a disruption or ablation of one or more alleles of an endogenous polynucleotide or comprising an exogeneous gene are also provided.
Core Innovation
The invention provides a method for producing oils with modified profiles of omega-3 fatty acids and omega-6 fatty acids, and optionally lutein, zeaxanthin, astaxanthin, 4-keto-lutein, or squalene, by growing a microalgal mutant and isolating the oil from the microalgal mutant. The microalgal mutant is a mutant of Auxenochlorella protothecoides and is made by knock-out, knock-in, or homologous recombination.
The engineered microalgal mutant includes genetic changes related to lycopene cyclase epsilon and squalene epoxidase, including a knock-out of at least one allele of lycopene cyclase epsilon LCYE-1 and/or lycopene cyclase epsilon LYCE-2, or a knock-out of at least one allele of squalene epoxidase SQE-1 and/or squalene epoxidase SQE-2. These genetic modifications are associated with altered carotenoid profiles including lutein and zeaxanthin, with zeaxanthin enrichment shown by HPLC in the described supporting content.
The invention also describes promoter replacement of a native FAD3 promoter by a FATA gene promoter from Auxenochorella protothecoides or by a stearoyl-ACP desaturase (SAD2) promoter from Auxenochorella protothecoides to alter omega-3 and omega-6 fatty acid profiles. The supporting content reports increased omega-3 including α-linolenic acid and a reduced omega-6:omega-3 ratio, and disruption of squalene epoxidase alleles results in squalene accumulation.
Claims Coverage
The independent claim covers a method of producing oils with modified omega-3/omega-6 profiles and optionally modified lutein, zeaxanthin, astaxanthin, 4-keto-lutein, or squalene, using an Auxenochlorella protothecoides microalgal mutant made by knock-out, knock-in, or homologous recombination, followed by oil isolation. Main inventive features are driven by specific genetic modifications: lycopene cyclase epsilon allele knock-outs, squalene epoxidase allele knock-outs, and optional FAD3 promoter replacement using promoters associated with FATA or SAD2.
Auxenochlorella mutant producing oils with modified omega profiles
Growing a microalgal mutant of Auxenochlorella protothecoides made by knock-out, knock-in or homologous recombination, wherein the microalgal mutant is used to produce oils with modified profiles of omega-3 fatty acids, omega-6 fatty acids, and/or lutein, zeaxanthin, astaxanthin, 4-keto-lutein, or squalene, followed by isolating the oil from the microalgal mutant.
Lycopene cyclase epsilon allele knock-out for carotenoid profile modification
A microalgal mutant comprising a knock-out of at least one allele of lycopene cyclase epsilon LCYE-1 and/or lycopene cyclase epsilon LYCE-2.
Squalene epoxidase allele knock-out for squalene accumulation
A microalgal mutant comprising a knock-out of at least one allele of squalene epoxidase SQE-1 and/or squalene epoxidase SQE-2.
FAD3 promoter replacement using FATA or SAD2 promoter
A microalgal mutant wherein a replacement of a native FAD3 promoter is performed by a FATA gene promoter from Auxenochlorella protothecoides or by a stearoyl-ACP desaturase (SAD2) promoter from Auxenochlorella protothecoides.
Overall, the claim coverage focuses on producing oils by growing an Auxenochlorella protothecoides microalgal mutant engineered by knock-out, knock-in, or homologous recombination, with inventive differentiation based on lycopene cyclase epsilon allele knock-outs, squalene epoxidase allele knock-outs, and optional FAD3 promoter replacement using FATA or SAD2 promoters.
Stated Advantages
Produces oils with modified profiles of omega-3 fatty acids and omega-6 fatty acids.
Produces oils with optionally modified lutein, zeaxanthin, astaxanthin, 4-keto-lutein, or squalene profiles.
Documented Applications
Functional oils/compositions are described in the document context, including uses aligned with the modified oil compositions.
Interested in licensing this patent?