Processes for preparing and purifying protein from plant material, and compositions and uses comprising the same, are provided.

The present disclosure provides recombinant microorganisms and methods for the anaerobic production of 2,4-furandicarboxylic acid from one or more carbon sources. The microorganisms and methods provide redox-balanced and ATP positive pathways for co-producing 2,4-furandicarboxylic acid with ethanol and for co-producing 2,4-furandicarboxylic acid with ethanol and 1-propanol. The method provides recombinant microorganisms that express endogenous and/or exogenous nucleic acid molecules encoding polypeptides that catalyze the conversion of a carbon source into 2,4-furandicarboxylic acid and that coupled the 2,4-furandicarboxylic acid pathway with an additional metabolic pathway.

The disclosure describes methods for the purification of protein-enriched extracts to provide concentrates and isolates and methods for incorporation of such materials into products. The purification methods are adapted for removal of, e.g., chlorophyll and may thus provide lightening the color of the protein-enriched extracts. The methods generally include treatment with peracetic acid or hydrogen peroxide and filtrations. A protein composition in the form of a concentrate or isolate is provided, the protein composition including RuBisCO, F2 fraction proteins, or combination thereof extracted from a plant material.

A method of improving a ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCo) to have a higher protein score is disclosed. The method includes the steps of: making a modified RbcL of the RuBisCo, by, on an RbcL unit of the RuBisCo, either substituting Met for Leu, Phe, Val, or Ile or combinations thereof; substituting Lys for Arg, Thr, or His or combinations thereof; or both of these substitutions. The modified RbcL consequently modifies the RuBisCo and is added to a biomass host where it is stable for homologous recombination. Plastid and nucleus integration was observed. Example RbcL sequences are disclosed with the desirable substitutions. The improved RuBisCo can be used as an improved proteinaceous food source for humans and animals.

The disclosure describes methods for the purification of protein-enriched extracts to provide concentrates and isolates and methods for incorporation of such materials into products. The purification methods are adapted for removal of, e.g., chlorophyll and may thus provide lightening the color of the protein-enriched extracts. The methods generally include treatment with peracetic acid or hydrogen peroxide and filtrations. A protein composition in the form of a concentrate or isolate is provided, the protein composition including RuBisCO, F2 fraction proteins, or combination thereof extracted from a plant material.

The invention concerns a genetically modified microorganism expressing a functional type I or II RuBisCO enzyme and a functional phosphoribulokinase (PRK), and in which the glycolysis pathway is at least partially inhibited, said microorganism being genetically modified so as to produce an exogenous molecule and/or to overproduce an endogenous molecule. According to the invention, the oxidative branch of the pentose phosphate pathway may also be at least partially inhibited. The invention also concerns the use of such a genetically modified microorganism for the production or overproduction of a molecule of interest and processes for the synthesis or bioconversion of molecules of interest.

Provided are methods for plastid genome editing and development of plants, plant cells, plant parts, and seeds comprising edited plastid genomes. Compositions for transformation of plastid genomes are further provided. Specifically, the plastid genome is modified to comprise the replacement of a targeted endogenous plastid genome sequence with a modified version of the target plastid sequence, where the modified plastid genome sequence has been designed to deliberately reduce its homology to the targeted endogenous plastid genome sequence and, in some cases, encode a protein containing one or more mutations.

The invention relates to a recombinant cell, preferably a yeast cell comprising: a) one or more heterologous genes encoding a glycerol dehydrogenase activity; b) one or more genes encoding a dihydroxyacetone kinase (E.C. 2.7.1.28 and/or E.C. 2.7.1.29); c) one or more heterologous genes encoding a ribulose-1,5-biphosphate carboxylase oxygenase (EC 4.1.1.39, RuBisCO); and d) one or more heterologous genes encoding a phosphoribulokinase (EC 2.7.1.19, PRK); and optionally e) one or more heterologous genes encoding for a glycerol transporter. This cell can be used for the production of ethanol and advantageously produces little or no glycerol.

The present disclosure relates to processes for recovering valuable products from Fabaceae family plant fractions, in particular from Medicago sativa ssp. The processes disclosed herein include processes for obtaining macrofibers, microfibers, a saponin precursor, chloroplast liquid and dry compositions and a Rubisco precursor. There is also disclosed herein processes for extracting from Fabaceae family plants valuable compounds such as proteins, enzymes, peptides, amino acids, fatty acids, fatty alcohols, terpenes, phenols and pigments. The processes may comprise at least one of separating plant fibers while attenuating shear forces, maintaining the temperature at or below 45° C., maintaining the pH above 4 and adding antioxidant and/or antimicrobial agents. Compositions comprising these recovered Fabaceae family plant products and uses thereof are also disclosed.

The present invention provides a plant expressing a target protein, a method of preparing the same and a method of preparing a composition for oral administration of a biopharmaceutical using the same. A target protein expression system using plant cells according to the present invention solves conventional problems in plant cell culture, provides a method of producing a large quantity of target proteins including a biopharmaceutical protein and allowing a target protein to have a resistance to a protease present in a digestive organ, and therefore is very effective to enable commercialization of plant-derived biopharmaceuticals.