KR20230151310A - Microorganism with improved lipid productivity into which glutathione peroxidase presumed gene is introduced, and method for manufacturing lipids using the same - Google Patents

Abstract

The present invention relates to a glutathione peroxidase-like gene, a recombinant vector containing the same, a microorganism with improved lipid productivity into which the gene has been introduced, a method for producing the microorganism, and a method for producing lipids using the same.

Description

Microorganism with improved lipid productivity into which glutathione peroxidase-like gene is introduced and lipid production method using the same {Microorganism with improved lipid productivity into which glutathione peroxidase presumed gene is introduced, and method for manufacturing lipids using the same}

The present invention relates to a glutathione peroxidase-like gene, a recombinant vector containing the same, a microorganism with improved lipid productivity into which the gene has been introduced, a method for producing the microorganism, and a method for producing lipids using the same.

Recently, in the face of depletion of fossil energy and global warming around the world, developed countries, including the United States, have been enacting policies to utilize and expand environmentally friendly renewable energy such as bio, hydrogen, and solar energy, as well as developing new renewable energy. Promoting supply and use. Although much effort is being made in Korea to develop new renewable energy, it is currently in the research stage. Bioenergy is renewable and has the advantage of reducing the acceleration of global warming by fixing carbon dioxide, so research on bioethanol, biobutanol, biodiesel, etc. is currently being actively conducted.

Among bioenergy, the one on which research is most actively underway is biodiesel. Biodiesel is produced by converting fatty acids contained in biomass into methyl ester or ethyl ester form through a transesterification process. It has a flash point of 150 ℃, making it cheaper than diesel, which has a flash point of 64 ℃. It is classified as a non-flammable liquid because it does not catch fire easily and is more stable at low temperatures than highly volatile gasoline (45°C), and because it catches fire at high temperatures, it can be said to have higher stability. In addition, unlike diesel, biodiesel is known to be a non-toxic energy that emits less harmful gases that cause carcinogens and mutations when burned and has significantly lower other emissions.

It is known that such biodiesel can be produced biologically using microalgae. Microalgae are used as feed or fertilizer, and in particular, green algae such as Spirulina sp., Chlorella sp., Dunaliella sp., and Nostoc sp. are health foods. It is also used. Since the solar energy utilization efficiency of these microalgae is about 25 times higher than that of plants, microalgae with excellent lipid content can be used as biomass for the production of biodiesel.

Botryococcus sp. and Schizochytrium sp. are known as microalgae with excellent lipid content. Microalgae do not have high lipid content in normal living environments, but when nutritional supply is interrupted, microalgae do not have high lipid content. After a certain period of time passes in this state, it exhibits the characteristic of accumulating lipids within cells.

Using these characteristics, microalgae can be used as biomass for the production of biodiesel. However, to increase the lipid content in microalgae, the first process of cultivating and proliferating microalgae and supplying nutrients to the proliferated microalgae are required. A second process must be performed to increase the lipid content within the cells by stopping the cultivation for a certain period of time. However, because the second-stage process requires an excessive amount of time, it has not yet been used in the industrial production of biodiesel. There is an urgent need for the development of microalgae with excellent lipid content.

In particular, in the case of microalgae used for biodiesel production, there is a lack of research on intracellular metabolic characteristics, so target gene selection to increase lipid productivity is necessary. In other words, there is a lack of prior research on which genes should be amplified to develop recombinant strains with increased lipid content.

The present inventor completed the present invention after confirming that when a glutathione peroxidase-like gene derived from Pyropia yezoensis was introduced into microalgae and overexpressed, the lipid content in the recombinant microorganism increased.

An object of the present invention is to provide a polynucleotide encoding a glutathione peroxidase-like protein.

Another object of the present invention is to provide a recombinant vector containing a polynucleotide encoding a glutathione peroxidase-like protein.

Another object of the present invention is to provide a microorganism with improved lipid productivity transformed with a recombinant vector containing a polynucleotide encoding a glutathione peroxidase-like protein.

Another object of the present invention is to provide a method for producing a microorganism with improved lipid productivity, which includes a transformation step of transforming the microorganism with a recombinant vector containing a polynucleotide encoding a glutathione peroxidase-like protein.

Another object of the present invention is to provide a method for producing lipids comprising the following steps:

A culturing step of culturing a microorganism transformed with a recombinant vector containing a polynucleotide encoding a glutathione peroxidase-like protein; and

Extraction step to extract lipids.

The present invention relates to a microorganism with improved lipid productivity into which a glutathione peroxidase-like gene has been introduced and a lipid production method using the same. Chlamydomonas reinhardtii into which a glutathione peroxidase-like gene has been introduced according to the present invention. reinhardtii ) strain showed improved lipid productivity.

The present inventor discovered a glutathione peroxidase-like gene among genes that are specifically highly expressed from changes in expression of the Pyropia yezoensis gene depending on the environment, and transformed this gene into the microalgae Chlamydomonas rheinhardti. Lipid content was assessed.

As a result, it was confirmed that recombinant Chlamydomonas reinhardtii transformed with a glutathione peroxidase-like gene showed excellent growth in a culture environment for lipid production and synthesized a higher content of lipids compared to the wild type. This was used to produce lipids. It is expected that it will be possible to mass-produce metabolites such as , starch, etc.

Hereinafter, the present invention will be described in more detail.

One aspect of the invention relates to a polynucleotide encoding a glutathione peroxidase-like protein.

In the present invention, the glutathione peroxidase-like protein may be a peptide containing the amino acid sequence of SEQ ID NO: 4, for example, may be composed of the amino acid sequence of SEQ ID NO: 4, but is not limited thereto.

In the present invention, the polynucleotide encoding a glutathione peroxidase-like protein may be a polynucleotide containing the base sequence of SEQ ID NO: 3 or a base sequence substantially identical thereto, for example, the base sequence of SEQ ID NO: 3 It may consist of, but is not limited to this.

In the present invention, 'substantial identity' means aligning the nucleotide sequence encoding the glutathione peroxidase-like protein and any other nucleotide sequence to correspond as much as possible, and analyzing the sequence to determine whether any other nucleotide sequence is a glutathione peroxidase-like protein. It means having at least 70%, at least 80%, at least 90%, or at least 98% sequence homology with the nucleotide sequence encoding.

In the present invention, the polynucleotide encoding a glutathione peroxidase-like protein may be derived from radioactive seaweed, but is not limited thereto.

Another aspect of the present invention relates to a recombinant vector comprising a polynucleotide encoding a glutathione peroxidase-like protein.

In the present invention, the glutathione peroxidase-like protein may be a peptide containing the amino acid sequence of SEQ ID NO: 4, for example, may be composed of the amino acid sequence of SEQ ID NO: 4, but is not limited thereto.

In the present invention, the polynucleotide encoding a glutathione peroxidase-like protein may be a polynucleotide containing the base sequence of SEQ ID NO: 3 or a base sequence substantially identical thereto, for example, the base sequence of SEQ ID NO: 3 It may consist of, but is not limited to this.

In the present invention, 'substantial identity' means aligning the nucleotide sequence encoding the glutathione peroxidase-like protein and any other nucleotide sequence to correspond as much as possible, and analyzing the sequence to determine whether any other nucleotide sequence is a glutathione peroxidase-like protein. It means having at least 70%, at least 80%, at least 90%, or at least 98% sequence homology with the nucleotide sequence encoding.

The term 'vector' in this specification refers to a means for expressing a target gene in a host cell, and recombinant vectors for transformation of host cells include both cloning vectors and expression vectors. Examples include plasmid vectors, cosmid vectors and viral vectors such as bacteriophage vectors, adenovirus vectors, retroviral vectors and adeno-associated viral vectors. In addition, vectors that can be used as recombinant vectors include plasmids often used in the art (e.g., pSC101, pGV1106, pACYC177, ColE1, pKT230, pME290, pBR322, pUC8/9, pUC6, pBD9, pHC79, pIJ61, pLAFR1, pHV14 , pGEX series, pET series, and pUC19, etc.), phages (e.g., λgt4λB, λ-Charon, λΔz1, and M13, etc.), or viruses (e.g., SV40, etc.), but are not limited thereto.

Another aspect of the present invention relates to a microorganism with improved lipid productivity transformed with a recombinant vector containing a polynucleotide encoding a glutathione peroxidase-like protein.

In the present invention, the glutathione peroxidase-like protein may be a peptide containing the amino acid sequence of SEQ ID NO: 4, for example, may be composed of the amino acid sequence of SEQ ID NO: 4, but is not limited thereto.

In the present invention, the polynucleotide encoding a glutathione peroxidase-like protein may be a polynucleotide containing the base sequence of SEQ ID NO: 3 or a base sequence substantially identical thereto, for example, the base sequence of SEQ ID NO: 3 It may consist of, but is not limited to this.

In the present invention, 'substantial identity' means aligning the nucleotide sequence encoding the glutathione peroxidase-like protein and any other nucleotide sequence to correspond as much as possible, and analyzing the sequence to determine whether any other nucleotide sequence is a glutathione peroxidase-like protein. It means having at least 70%, at least 80%, at least 90%, or at least 98% sequence homology with the nucleotide sequence encoding.

In the present invention, lipid is a general term for biomolecules soluble in non-polar solvents. Lipids include fatty acids, glycerol lipids, sterol lipids, phospholipids, and lipids. There may be sphingolipid, prenol lipid, etc., but it is not limited thereto.

In the present invention, the polynucleotide encoding a glutathione peroxidase-like protein may be derived from radioactive seaweed, but is not limited thereto.

In the present invention, the microorganism may be microalgae, for example, one or more of the group consisting of Chlamydomonas reinhardtii, Schizochytrium mangrovei , and Botryococcus braunii . However, it is not limited to this.

Another aspect of the present invention relates to a method for producing a microorganism with improved lipid productivity, which includes a transformation step of transforming the microorganism with a recombinant vector containing a polynucleotide encoding a glutathione peroxidase-like protein.

In one embodiment of the present invention, a transformant with improved lipid production ability can be created by inserting an expression vector containing a gene encoding a target protein into a host cell.

In the present invention, the glutathione peroxidase-like protein may be a peptide containing the amino acid sequence of SEQ ID NO: 4, for example, may be composed of the amino acid sequence of SEQ ID NO: 4, but is not limited thereto.

In the present invention, the polynucleotide encoding a glutathione peroxidase-like protein may be a polynucleotide containing the base sequence of SEQ ID NO: 3 or a base sequence substantially identical thereto, for example, the base sequence of SEQ ID NO: 3 It may consist of, but is not limited to this.

In the present invention, 'substantial identity' means aligning the nucleotide sequence encoding the glutathione peroxidase-like protein and any other nucleotide sequence to correspond as much as possible, and analyzing the sequence to determine whether any other nucleotide sequence is a glutathione peroxidase-like protein. It means having at least 70%, at least 80%, at least 90%, or at least 98% sequence homology with the nucleotide sequence encoding.

In the present invention, the polynucleotide encoding a glutathione peroxidase-like protein may be derived from radioactive seaweed, but is not limited thereto.

In the present invention, the host cell is a cell capable of stably and continuously cloning or expressing the inserted expression vector, and any host cell known in the art can be used, and the transformant is created by introducing the recombinant vector into an appropriate host cell. It may have been obtained.

In the present invention, the microorganism may be any one or more of the group consisting of microalgae, for example, Chlamydomonas reinhardtii, Schizochytrium mangrovei , and Botryococcus braunii . It is not limited.

In the present invention, a glutathione peroxidase-like gene or a recombinant vector containing it can be transported (introduced) into a host cell using a transport method widely known in the art. For example, if the host cell is a prokaryotic cell, the CaCl ₂ method or electroporation method can be used as a transport method for the recombinant vector, and if the host cell is a eukaryotic cell, microinjection method, calcium phosphate precipitation method, or electroporation method can be used. , liposome-mediated transfection, and gene bombardment can be used, but are not limited to these.

In the present invention, selection of transformed host cells can be easily performed using a phenotype expressed by a selection marker, according to methods widely known in the art. For example, if the selection marker is a specific antibiotic resistance gene, the transformant can be easily selected by culturing the transformant in a medium containing the specific antibiotic.

Another aspect of the invention relates to a method for producing lipids comprising the following steps:

A culturing step of culturing a microorganism transformed with a recombinant vector containing a polynucleotide encoding a glutathione peroxidase-like protein; and

Extraction step to extract lipids.

In the present invention, the glutathione peroxidase-like protein may be a peptide containing the amino acid sequence of SEQ ID NO: 4, for example, may be composed of the amino acid sequence of SEQ ID NO: 4, but is not limited thereto.

In the present invention, the polynucleotide encoding a glutathione peroxidase-like protein may be a polynucleotide containing the base sequence of SEQ ID NO: 3 or a base sequence substantially identical thereto, for example, the base sequence of SEQ ID NO: 3 It may consist of, but is not limited to this.

In the present invention, 'substantial identity' means aligning the nucleotide sequence encoding the glutathione peroxidase-like protein and any other nucleotide sequence to correspond as much as possible, and analyzing the sequence to determine whether any other nucleotide sequence is a glutathione peroxidase-like protein. It means having at least 70%, at least 80%, at least 90%, or at least 98% sequence homology with the nucleotide sequence encoding.

In the present invention, the extracted lipid may be extracted in the form of an ester through a transesterification process of fatty acids.

In the present invention, the polynucleotide encoding a glutathione peroxidase-like protein may be derived from radioactive seaweed, but is not limited thereto.

In the present invention, the microorganism with improved lipid productivity may be any one or more of the group consisting of microalgae, for example, Chlamydomonas reinhardtii, Schizochytrium mangrovei , and Botryococcus braunii . However, it is not limited to this.

In one embodiment of the present invention, the recombinant Chlamydomonas reinhardtii has an increased lipid content compared to the wild-type control in which a polynucleotide encoding a glutathione peroxidase-like protein is not introduced, resulting in a significant increase in biodiesel production. It showed an effective effect, and this can be used for biodiesel production in the future.

In the present invention, biodiesel may be produced by converting fatty acids into methyl ester or ethyl ester form through a transesterification process, but is not limited thereto.

The present invention relates to Chlamydomonas reinhardtii transformed with a recombinant vector containing an isolated polynucleotide encoding a glutathione peroxidase-like gene derived from Pyropia yezoensi, and the same. Regarding the lipid production method used, Chlamydomonas reinhardtii produced according to the present invention shows excellent growth in a culture environment for lipid production, and is expected to be able to mass produce metabolites such as lipids and starch through this. .

Figure 1 is a graph showing the lipid content of recombinant Chlamydomonas reinhardtii into which a glutathione peroxidase-like gene has been introduced according to an embodiment of the present invention compared to the wild type.

Hereinafter, the present invention will be described in more detail through the following examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Example 1: RNA extraction and cDNA synthesis from spiny-patterned seaweed

White seaweed ( Pyropia yezoensi ) was used after receiving it from the Seaweed Research Center of the National Institute of Fisheries Science.

Specifically, MGM (Modified Grund Media) medium was composed of 3.72 g of C10H14O6N2Na _2· 2H2O (Na ₂ EDTA), 0.28 g of FeSO _4· 7H ₂ O, 10.74 g of Na ₂ HPO _4· 12H ₂ O, 42.5 g of NaNO ₃ , and MnCl _{2 ·} It was prepared using 1 liter (ℓ) of sterilized seawater containing 0.019197 g of 7H ₂ O and 1 mg of vitamin B12 (Cyanocobalamin).

Next, put the spiny-patterned seaweed ^into the MGM medium, add the MGM medium containing the spiny-patterned seaweed to a plant culture dish (100 The culture was maintained for 1 to 10 weeks at ² s ^-1 , 12:12 Light:Dark cycle, and temperature of 12°C.

To obtain RNA from cultured spiny seaweed, an RNA purification kit [QIAGEN RNeasy ^® Mini kit] was used.

Specifically, 100 mg of spiny seaweed fronds were frozen in liquid nitrogen and finely ground in a sterilized mortar and pestle.

100 mg of powdered tissue was placed in a 1.5 mL micro centrifuge tube, added with 450 uL RLT buffer, stirred, then transferred to a QIAshredder spin column and centrifuged at 15,000 rpm for 2 minutes.

Next, a 200 uL volume of ethanol was mixed with the separated buffer, placed on an RNeasy spin column, and centrifuged at >8,000 g for 15 seconds.

The buffer that fell off was removed and the column was washed with 700 uL RW1 buffer for 15 seconds at >8,000 g.

Next, the spilled buffer was removed and washed twice with 500 uL RPE buffer for 15, 2 minutes at >8,000 g.

The column was transferred to a new 1.5 mL micro centrifuge tube, 30 to 50 uL of RNA removal water (RNase-free water) was added, and each RNA sample was obtained after centrifugation at >8,000 g for 1 minute.

The spectrophotometer [Biodrop, Biochrom, German] was set to a wavelength range of 230 nm and 280 nm, and 2 uL of RNA removal water was added to the spot to set the base. 2 uL of the previously obtained RNA sample was added to the spot, and the RNA quantification value was confirmed through absorbance at 260 nm.

cDNA was synthesized for the quantified 2 uL RNA sample using a cDNA synthesis kit [Transcriptor Fiest Strand cDNA Synthesis Kit, Roche, Germany].

Specifically, each quantified 2 uL RNA sample was mixed with 1 uL of oligo-dT and DEPC-treated water (DEPC-water) to make a total of 13 uL and incubated at 65°C for 10 minutes.

After treatment, 4 uL of RT reaction (5x), 1 uL of reverse transcriptase, 0.5 uL of RNase inhibitor, and 2 uL of DNTP were added and incubated at 50 ℃ for 60 minutes and 85 ℃ for 5 minutes to produce each cDNA. was synthesized.

Afterwards, the glutathione peroxidase-like gene was PCR amplified from the synthesized cDNA using the primers shown in Table 1 below.

Specifically, for each cycle, denaturation of cDNA was performed at 94°C for 5 minutes, and primer binding step (annealing) was performed at 65°C for 1 minute. The DNA synthesis step (extension) was performed at 72°C for 2 minutes using Pfu DNA Polymerase.

PCR was performed for a total of 30 cycles, and in the last cycle, the DNA synthesis step was performed at 72°C for 10 minutes.

The base sequence and amino acid sequence of the amplified glutathione peroxidase-like gene are shown in Table 1 below.

sequence number designation Sequence (5'->3') One forward primer ATGAAAAAAGTGATAGGTATAGG 2 reverse primer TCAGTAATTCTTTTGGCAAAAACT 3 glutathione peroxidase (DNA) ATGAAAAAAGTGATAGGTATAGCCATAGGATTAGTAGTAGTTGCAGGTATAGCAATAGCATTTACATTACCCAATAAAGATGCAGACCCTGCCCGAAAAGTTGGTGAGTTGCAAGCACAATCTTTTTATGATTTGAATATTTTAAGTTTAGATGGAGATGACTCTTTAAAACTTTGCTAATTTTAAAGATAAAAAAGTACTGTGTGTTAATGTTGCCAGTAAATGCGGGTATACCTACCAATACGAAGATTTGCAAAAACTACAC GAGAAGTATAAAAGAGAAGTTGGTAATAATAGGTTTTCCTTGCAATCAGTTTTTAGGGCAAGAACCGGCGATGCTGAAGAAATTGAGAGTTTTGCCAAAAGAATTACTGA 4 glutathione peroxidase (protein) MKKVIGIAIGLVVVAGIAIAFTLPNKDADPARKVGELQAQSFYDLNILSLDGDDSLNFANFKDKKVLCVNVASKCGYTYQYEDLQKLHEKYKEKLVIIGFPCNQFLGQEPGDAEEIESFCQKNY

Example 2: Obtaining Chlamydomonas transformants

It was confirmed whether the amplified glutathione peroxidase-like gene affects lipid production in Chlamydomonas reinhardtii .

Specifically, the glutathione peroxidase-like gene was PCR amplified using the primers in Table 2 below, and then the gene was amplified using the vector pChlamy_3/D-TOPO of the recombinant protein expression kit [GeneArt ^® Chlamydomonas TOPO ^® Engineering Kits (A14264)]. It was expressed.

At this time, the PCR amplification was carried out in the same manner as the PCR amplification process in Example 1 described above, except that the temperature of the primer binding step was set to 68°C.

sequence number designation Sequence (5'->3') 5 forward primer CACCATGAAAAAAGTGATAGGTATAGG 2 reverse primer TCAGTAATTCTTTTGGCAAAAACT

Since the vector pChlamy_1/D-TOPO has a hygromycin resistance gene, wild type Chlamydomonas Rheinhardti C137 was transformed using Gene Pulser (Bio-Rad, US) and cultured for 2 to 3 weeks. Afterwards, transformants were obtained by selection in a medium supplemented with 15 mg/L hygromycin.

Example 3: Comparison of lipid content of recombinant Chlamydomonas

Recombinant Chlamydomonas reinhardtii, which expressed a glutathione peroxidase-like gene derived from Pyropia yezoensi, and wild-type Chlamydomonas reinhardtii were grown in MBBM (Modified Bold's Basal medium). After placing it in the medium, it was cultured for 7 days by maintaining a light intensity of 20 umol photon m ^-2 s ^-1 , a 12:12 Light:Dark cycle, and a temperature of 20°C.

The lipid content of the cultivated strains was analyzed, and the lipid content analysis was performed in the form of fatty acid methyl esters (FA methyl esters; FAMEs) according to known techniques.

Specifically, 5 mg of freeze-dried strain cells were suspended in 1 mL 2 M NaOH-CH ₃ OH solution, shaken (110 rpm) for 1 hour at room temperature (RT), and incubated at 75°C for 15 minutes. ) was done.

After cooling, 1 mL 4 M HCl-CH ₃ OH solution was added to the mixture, the pH was adjusted to less than 2.0 with HCl, and the mixture was incubated at 75°C for 15 minutes.

Afterwards, FAME was extracted with 1 mL of hexane, shaken by hand for 30 seconds, and then centrifuged at 3500 g for 2 minutes.

Finally, the amount of FAME in the mixed solution was measured, and the results are shown in Figure 1 and Table 3.

Experimental group (recombinant strain) Control (wild type strain) lipid content
(mg/L) 70.24 51.21

As can be seen in Figure 1 and Table 3, a lipid content of 70.24 mg/L was obtained in recombinant Chlamydomonas reinhardt expressing a glutathione peroxidase-like gene derived from spiny seaweed, and the lipid amount measured in wild-type Chlamydomonas reinhardt It was confirmed that it had a 37.1% higher lipid content compared to 51.21 mg/L.

<110> INDUSTRY FOUNDATION OF CHONNAM NATIONAL UNIVERSITY <120> Microorganism with improved lipid productivity into which glutathione peroxidase presumed gene is introduced, and method for manufacturing lipids using the same <130> PN220072 <160> 5 <170> KoPatentIn 3.0 <210> 1 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 1 atgaaaaaag tgataggtat agg 23 <210> 2 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 2 tcagtaattc ttttggcaaa aact 24 <210> 3 <211> 375 <212> DNA <213> Unknown <220> <223> glutathione peroxidase (DNA) <400> 3 atgaaaaaag tgataggtat agccatagga ttagtagtag ttgcaggtat agcaatagca 60 tttacattac ccaataaaga tgcagaccct gcccgaaaag ttggtgagtt gcaagcacaa 120 tctttttatg atttgaatat tttaagttta gatggagatg actctttaaa ctttgctaat 180 tttaaagata aaaaagtact gtgtgttaat gttgccagta aatgcgggta tacctaccaa 240 tacgaagatt tgcaaaaact acacgagaag tataaagaga agttggtaat aataggtttt 300 ccttgcaatc agtttttagg gcaagaaccg ggcgatgctg aagaaattga gagtttttgc 360 caaaagaatt actga 375 <210> 4 <211> 124 <212> PRT <213> Unknown <220> <223> glutathione peroxidase (protein) <400> 4 Met Lys Lys Val Ile Gly Ile Ala Ile Gly Leu Val Val Val Ala Gly 1 5 10 15 Ile Ala Ile Ala Phe Thr Leu Pro Asn Lys Asp Ala Asp Pro Ala Arg 20 25 30 Lys Val Gly Glu Leu Gln Ala Gln Ser Phe Tyr Asp Leu Asn Ile Leu 35 40 45 Ser Leu Asp Gly Asp Asp Ser Leu Asn Phe Ala Asn Phe Lys Asp Lys 50 55 60 Lys Val Leu Cys Val Asn Val Ala Ser Lys Cys Gly Tyr Thr Tyr Gln 65 70 75 80 Tyr Glu Asp Leu Gln Lys Leu His Glu Lys Tyr Lys Glu Lys Leu Val 85 90 95 Ile Ile Gly Phe Pro Cys Asn Gln Phe Leu Gly Gln Glu Pro Gly Asp 100 105 110 Ala Glu Glu Ile Glu Ser Phe Cys Gln Lys Asn Tyr 115 120 <210> 5 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 5 caccatgaaa aaagtgatag gtatagg 27

Claims (11)

A polynucleotide encoding a glutathione peroxidase-like protein. The polynucleotide of claim 1, wherein the polynucleotide includes the base sequence of SEQ ID NO: 3 or a base sequence substantially identical thereto. A recombinant vector containing a polynucleotide encoding a glutathione peroxidase-like protein. The recombinant vector according to claim 3, wherein the polynucleotide includes the base sequence of SEQ ID NO: 3 or a base sequence substantially identical thereto. A microorganism transformed with a recombinant vector containing a polynucleotide encoding a glutathione peroxidase-like protein. The microorganism according to claim 5, wherein the polynucleotide includes the base sequence of SEQ ID NO: 3 or a base sequence substantially identical thereto. The microorganism according to claim 5, wherein the microorganism is Chlamydomonas reinhardtii . Method for producing lipids, comprising the following steps:
A culturing step of culturing a microorganism transformed with a recombinant vector containing a polynucleotide encoding a glutathione peroxidase-like protein; and
Extraction step to extract lipids. The method of claim 8, wherein the lipid is extracted in the form of an ester through a transesterification process of fatty acid. The method of claim 8, wherein the polynucleotide includes the base sequence of SEQ ID NO: 3 or a base sequence substantially identical thereto. The method of claim 8, wherein the microorganism is Chlamydomonas reinhardtii .