Method for increasing the yield of microalgae and products produced thereby, an altered cam1 gene and polypeptide, and a novel chlamydomonas sp.

Patent metadata

Field	Value
Publication number	US-2021198684-A1
Application number	US-202017135402-A
Country	US
Kind code	A1
Filing date	Dec 28, 2020
Priority date	Dec 31, 2019
Publication date	Jul 1, 2021
Grant date	—

Abstract

A method for increasing the yield of microalgae and the yield of a product produced by the microalgae is provided. The method includes performing a change procedure on CAM1 gene and/or calmodulin 1 encoded by the CAM1 gene in a microalga, such that a change occurs in a nucleotide and/or the nucleotide sequence of the CAM1 gene and/or an amino acid and/or the amino acid sequence of the calmodulin 1 encoded by the CAM1 gene in the microalga to obtain an altered microalga. The altered microalga has an altered CAM1 gene and/or an altered calmodulin 1. The altered microalga has a higher growth rate and a higher product production rate and/or yield than an unaltered microalga.

First claim

What is claimed is:

1 . A method for increasing the yield of microalgae and the yield of a product produced by the microalgae, comprising:
performing a change procedure on CAM1 gene and/or calmodulin 1 encoded by the CAM1 gene in a microalga, such that a change occurs in a nucleotide and/or the nucleotide sequence of the CAM1 gene and/or an amino acid and/or the amino acid sequence of the calmodulin 1 encoded by the CAM1 gene in the microalga to obtain an altered microalga which has an altered CAM1 gene and/or an altered calmodulin 1, wherein the altered microalga has a higher growth rate and a higher product production rate and/or yield than an unaltered microalga.

2 . The method for increasing the yield of microalgae and the yield of a product produced by the microalgae as claimed in claim 1 , wherein the change comprises substitution, insertion, frameshift, deletion and/or addition in a sequence, and/or comprises modification to a nucleotide and/or an amino acid.

3 . The method for increasing the yield of microalgae and the yield of a product produced by the microalgae as claimed in claim 1 , wherein the nucleotide sequence of the CAM1 gene comprises the sequence of SEQ ID NO. 1.

4 . The method for increasing the yield of microalgae and the yield of a product produced by the microalgae as claimed in claim 1 , wherein the amino acid sequence of the protein encoded by the CAM1 gene comprises the sequence of SEQ ID NO. 2.

5 . The method for increasing the yield of microalgae and the yield of a product produced by the microalgae as claimed in claim 1 , wherein the nucleotide sequence of the CAM1 gene comprises the sequence of SEQ ID NO. 1, and a region where the change occurs in the nucleotide sequence of the CAM1 gene comprises:
at least one of the regions of the sequence of SEQ ID NO. 1 in the following:
(i) a region between position 80 and position 90;
(ii) a region between position 160 and position 170;
(iii) a region between position 230 and position 240;
(iv) a region between position 310 and position 320;
(v) a region between position 330 and position 340; and
(vi) a region between position 335 and position 345.

6 . The method for increasing the yield of microalgae and the yield of a product produced by the microalgae as claimed in claim 1 , wherein the nucleotide sequence of the CAM1 gene comprises the sequence of SEQ ID NO. 1, and the change occurring in the nucleotide sequence of the CAM1 gene comprises:
at least one change occurring in the sequence of SEQ ID NO. 1 from among the following:
(i) a substitution of a nucleotide at position 82;
(ii) an insertion of at least one nucleotide between position 167 and position 168;
(iii) a substitution of a nucleotide at position 233;
(iv) a substitution of a nucleotide at position 313;
(v) a substitution of a nucleotide at position 335; and
(vi) a substitution of a nucleotide at position 340.

7 . The method for increasing the yield of microalgae and the yield of a product produced by the microalgae as claimed in claim 6 , wherein the substitution of a nucleotide at position 82 is a substitution from C to A, the insertion of at least one nucleotide between position 167 and position 168 is an insertion of a polynucleotide between position 167 and position 168, and the sequence of the polynucleotide is the sequence of SEQ ID NO. 3, the substitution of a nucleotide at position 233 is a substitution from G to C, the substitution of a nucleotide at position 313 is a substitution from T to G, the substitution of a nucleotide at position 335 is a substitution from C to G or the substitution of a nucleotide at position 340 is a substitution from A to G.

8 . The method for increasing the yield of microalgae and the yield of a product produced by the microalgae as claimed in claim 1 , wherein the nucleotide sequence of the CAM1 gene comprises the sequence of SEQ ID NO. 1, and the change occurring in the nucleotide sequence of the CAM1 gene comprises:
changes occurring in the sequence of SEQ ID NO. 1 in the following:
(i) an insertion of a polynucleotide between position 167 and position 168, wherein the sequence of the polynucleotide is the sequence of SEQ ID NO. 3; and
(ii) a substitution from A to G at position 340.

9 . The method for increasing the yield of microalgae and the yield of a product produced by the microalgae as claimed in claim 1 , wherein the nucleotide sequence of the CAM1 gene comprises the sequence of SEQ ID NO. 1, and the change occurring in the nucleotide sequence of the CAM1 gene comprises:
changes occurring in the sequence of SEQ ID NO. 1 among the following:
(i) a substitution from C to A at position 82;
(ii) a substitution from G to C at position 233;
(iii) a substitution from T to G at position 313;
(iv) a substitution from C to G at position 335; and
(v) a substitution from A to G at position 340.

10 . The method for increasing the yield of microalgae and the yield of a product produced by the microalgae as claimed in claim 1 , wherein the nucleotide sequence of the altered CAM1 gene comprises the sequence of SEQ ID NO. 4 or the sequence of SEQ ID NO. 5.

11 . The method for increasing the yield of microalgae and the yield of a product produced by the microalgae as claimed in claim 1 , wherein the amino acid sequence of the altered calmodulin 1 comprises the sequence of SEQ ID NO. 6 or the sequence of SEQ ID NO. 7.

12 . The method for increasing the yield of microalgae and the yield of a product produced by the microalgae as claimed in claim 1 , wherein the change procedure comprises clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 gene editing, zinc-finger nucleases (ZFN) gene editing, or transcription activator-like effector nucleases (TALENs) gene editing.

13 . The method for increasing the yield of microalgae and the yield of a product produced by the microalgae as claimed in claim 1 , wherein a product produced by the microalgae comprises chlorophyll, a lipid, astaxanthin, a protein, a fatty acid, an amino acid, a carbohydrate, a vitamin, a compound, a cellulose, an enzyme, a colloid, diatomaceous earth, or a pigment.

14 . The method for increasing the yield of microalgae and the yield of the microalgae as claimed in claim 1 , wherein the microalgae comprises Chlamydomonas sp., Haematococcus sp., Synechococcus sp., Volvox sp., Emiliania huxleyi or Heterocapsa triquetra.

15 . The method for increasing the yield of microalgae and the yield of the microalgae as claimed in claim 1 , wherein the altered microalga is Chlamydomonas reinhardii ITRI-ALG-3 which is deposited at Bioresource Collection and Research Centre (BCRC) of Food Industry Research and Development Institute (FIRDI) and whose deposit number is BCRC 980055 or Chlamydomonas reinhardtii ITRI-ALG-8 which is deposited at Bioresource Collection and Research Centre of Food Industry Research and Development Institute and whose deposit number is BCRC 980056.

16 . An altered CAM1 gene, of which the nucleotide sequence comprises the sequence of SEQ ID NO. 1, in which the nucleotide sequence of the CAM1 gene has a change, and a region having the change in the nucleotide sequence of the CAM1 gene comprises:
at least one of the regions of the sequence of SEQ ID NO. 1 in the following:
(i) a region between position 80 and position 90;
(ii) a region between position 160 and position 170;
(iii) a region between position 230 and position 240;
(iv) a region between position 310 and position 320;
(v) a region between position 330 and position 340; and
(vi) a r

Assignees

• Ind Tech Res Inst

Inventors

• Chen Chang-Chieh
• Wu Shao-I
• Chiang Jo-Di
• Lin yu-pei
• Lu Wen-Chang

Classifications

• Y02A40/146

  Genetically Modified [GMO] plants, e.g. transgenic plants · CPC title

• C12R2001/89

  Algae {; Processes using algae} · CPC title

• C12N1/125

  Unicellular algae isolates · CPC title

• C07K14/405 Primary

  from algae · CPC title

• C12N15/8261 Primary

  with agronomic (input) traits, e.g. crop yield · CPC title