RECOMBINANT ALGAE HAVING HIGH BIOMASS AND LIPID PRODUCTIVITY
The invention provides a recombinant algal organism that has been genetically modified in a gene encoding a protein kinase-like protein. The recombinant organism exhibits higher biomass productivity and higher lipid productivity versus a corresponding control algal organism not having the genetic modification. The recombinant organism is therefore useful in applications requiring biomass and/or lipid productivity, e.g. in the production of biofuels or other lipidic matter. Methods of using the organism and biomass containing or produced by the organism are also provided.
1 . A recombinant algal organism comprising a deletion, disruption, or inactivation of:
a gene encoding a protein kinase-like domain that has a polypeptide sequence having at least 80% sequence identity to SEQ ID NO: 1; or
a gene encoding a protein kinase-like domain having a nucleic acid sequence having at least 80% sequence identity to SEQ ID NO: 2 or 3;
wherein the recombinant algal organism exhibits higher biomass productivity and higher lipid productivity versus a corresponding control algal organism not having the deletion, disruption, or inactivation.
2 . The recombinant algal organism of claim 1 , having a gene encoding a protein kinase-like domain that has a polypeptide sequence having at least 85% sequence identity to SEQ ID NO: 1.
3 . The recombinant algal organism of claim 1 , having a gene encoding a protein kinase-like domain having a nucleic acid sequence having at least 85% sequence identity to SEQ ID NO: 3.
4 . The recombinant algal organism of claim 1 , wherein the organism is a Chlorophyte alga.
5 . The recombinant algal organism of claim 4 , wherein the organism is of the Class Trebouxiophyceae.
6 . The recombinant algal organism of claim 1 , wherein the deletion, disruption, or inactivation is to a regulatory sequence of the gene encoding the protein kinase-like domain.
7 . The recombinant algal organism of claim 6 , wherein the regulatory sequence is a promoter.
8 . The recombinant algal organism of claim 1 , wherein the deletion, disruption, or inactivation comprises a deletion of one or more amino acids of the encoded protein kinase-like domain.
9 . The recombinant algal organism of claim 1 , wherein the deletion, disruption, or inactivation comprises an insertion in the gene encoding the kinase-like protein.
10 . The recombinant algal organism of claim 9 , wherein the insertion comprises insertion of a stop codon in a sequence encoding the kinase-like domain.
11 . The recombinant algal organism of claim 1 , wherein the recombinant alga has at least 20% higher lipid productivity versus a control algae.
12 . The recombinant algal organism of claim 1 , wherein the recombinant alga has at least 25% higher lipid productivity versus a control algae.
13 . The recombinant algal organism of claim 1 , wherein the recombinant alga has at least 35% higher biomass productivity per unit time versus the corresponding control algal cell or organism.
14 . The recombinant algal organism of claim 1 , wherein the recombinant alga has a FAME/TOC ratio of at least 0.4 after two days of cultivation.
15 . The recombinant algal organism of claim 1 , wherein the recombinant alga has higher biomass productivity under nitrogen deficient conditions.
16 . The recombinant algal organism of claim 1 , wherein the recombinant alga has higher total organic carbon production under nitrogen deficient conditions.
17 . The recombinant algal organism of claim 1 , of a family selected from the group consisting of: Oocystaceae, Chlorellaceae, and Eustigmatophyceae.
18 . The recombinant algal organism of claim 1 , wherein the recombinant alga is of a genus selected from the group consisting of: Chlorella, Parachlorella, Picochlorum, Tetraselmis , and Oocystis.
19 . The recombinant algal organism of claim 18 , wherein the recombinant alga is from the genus Oocystis.
20 . The recombinant algal organism of claim 1 , wherein the protein kinase-like domain comprises a polypeptide sequence having at least 90% sequence identity to SEQ ID NOs: 1 or 9.
21 . The recombinant algal organism of claim 1 , wherein the gene encoding the protein kinase-like domain has a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 2 or SEQ ID NO: 3.
22 . A biomass product comprising the recombinant alga of claim 1 .
23 . A recombinant algal organism comprising a genetic modification to:
a gene encoding a protein kinase-like domain that has a polypeptide sequence having at least 90% sequence identity to SEQ ID NO: 9; or
a gene encoding a protein kinase-like domain having a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 8; and
wherein the recombinant algal organism exhibits higher biomass productivity and higher lipid productivity versus a corresponding control algal organism not having the genetic modification.
24 . The recombinant algal organism of claim 23 , wherein the genetic modification is a deletion, disruption, or inactivation.
25 . The recombinant algal organism of claim 23 , wherein the recombinant alga has at least 25% higher lipid productivity versus a control algae.
26 . The recombinant algal organism of claim 23 , wherein the recombinant alga has at least 35% higher biomass productivity per unit time versus the corresponding control algal cell or organism.
27 . A method of producing a composition containing lipids comprising performing a genetic modification in an algal organism to:
a gene encoding a protein kinase-like domain having a polypeptide sequence having at least 80% sequence identity to SEQ ID NO: 1 or 9; or
a gene encoding a protein kinase-like domain having a nucleic acid sequence having at least 80% sequence identity to SEQ ID NO: 2-3 or 8;
wherein the recombinant algal organism exhibits higher biomass productivity and higher lipid productivity versus a corresponding control algal organism not having the genetic modification; and
cultivating the organism, and thereby producing a composition containing lipids.
28 . The method of claim 27 , further comprising harvesting a lipidic composition from the algal organism.
29 . The method of claim 27 , wherein the genetic modifications to the sequence encoding the protein-kinase like domain is a deletion, disruption, or inactivation.
30 . The method of claim 29 , wherein the genetic modification is a disruption.
31 . The method of claim 27 , wherein the recombinant alga has at least 50% greater lipid productivity versus a control alga.