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OwnerPadma Commuri, Peter L. Keeling +4inventors
InventorsPadma Commuri, Peter L. Keeling, Nona Ramirez, Angela McKean, Zhong Gao, Hanping Guan
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The invention relates to a method for changing the glucan chain lengths using fusion protein domains of various starch synthase enzymes in any starch or starch granule producing organism. The invention relates to identification of a GLucan ASSociation domain (herein after referred to as “GLASS” domain) of granule bound starch synthase (GBSS) used in combination with any other GLYcosyl TRransferase domain otherwise referred to as pfam00534-catalytic domain (herein after referred to as “GLYTR” domain) of one or more of any of the other starch synthase enzymes. The invention relates to identifying and using the new and surprising discovery that starch synthases are composed of at least two distinct functional domains herein after labeled as “GLASS” and “GLYTR”. More specifically, this invention relates to the genetic constructs that encode the fusions of the above domains and to the plants transformed with said constructs. The method of invention can thus be used in particular to provide a modified profile of starch granule associated starch synthase (SS) enzymes and by which modified glucan chain lengths of amylopectin and hence, modified starches and or complexes will be generated. This can be done in any organism and more particularly any plant that stores or synthesizes starch in any of its parts, such as potato, sweet potato, cassaya, pea, taro, banana, yam and cereal crops such as rice, maize, wheat, barley, oats, and sorghum.
An isolated DNA molecule encoding a fusion protein consisting of four different functional domains selected from the group consisting of GLASS, LINKR, GLYTR, and CTEND which are operably linked to one another.
An isolated DNA molecule of any of the claims in 2, 4, 6 or 8, in which the LINKR domain is a GBSS-LINKR, SSI-LINKR, SSII-LINKR or SSIII-LINKR.
An isolated DNA molecule encoding a fusion peptide comprising a GBSS GLASS domain operably linked to a LINKR and a catalytic domain from a functional protein that synthesizes an α-1,4 glucan or an α-1,3 glucan, or an α-1,6 glucan, said fusion peptide being capable of modifying the glucan structure of a starch producing organism when starch is produced by said organism or part thereof in the presence of said fusion peptide.
An isolated DNA molecule encoding a polypeptide with glucan association properties of a maize GBSS enzyme capable of modification of starch metabolism in a plant or plant cell, said DNA comprising a molecule selected from the group consisting of:
An isolated DNA molecule encoding a polypeptide with a glycosyl transferase function of a soluble or granule bound maize SS enzymes capable of modifying starch metabolism in a plant or plant cell, said DNA molecule being selected from the group consisting of:
A recombinant DNA molecule comprising a DNA molecule of any of the above claims comprising a maize GBSS nucleotide coding region encoding for an amino acid sequence of SEQ. ID NOs. 101-146 fused with a corresponding coding region of a maize SS enzyme that encode for an amino acid sequence of SEQ. ID NOs:
A recombinant DNA molecule comprising a DNA molecule of any of the above claims comprising a GLYTR, LINKR or CTEND domain DNA sequence selected from any one of SEQ ID NOs:
A recombinant DNA molecule comprising a DNA molecule of any of the above claims comprising a DNA sequence differing from the sequence of any of the DNA molecules of SEQ ID NOs:
A recombinant DNA molecule comprising a DNA molecule of any of the above claims comprising a wherein the DNA sequence is selected from the group consisting of a coding region of a glucan association domain of SEQ ID NOs:75-120;
Method of expressing a starch synthase fusion proteins or polypeptides in a plant, in which the starch synthase protein or polypeptide domains are expressed as a fusion with a glucan association domain of granule bound starch synthase.
39 A method according to any one of the preceding claims, in which the protein or polypeptide is heterologous with respect to the plant in which the fusion is expressed.
Method according to any one of the preceding claims, comprising the steps of:
Method according to any one of the preceding claims in which the protein or polypeptide or recombinant protein or recombinant polypeptide is an enzyme.
A vector comprising a DNA molecule according to any of the preceding claims.
A plant cell comprising a DNA molecule according to any one of the preceding claims linked to a heterologous promoter.
A modified starch derived from cells of a plant of any of the preceding claims.
Dependencies are inferred from the claim text.