CN106148363A - Thallus Laminariae (Thallus Eckloniae) vanadium ion dependent form haloperoxidase gene and encoding proteins thereof - Google Patents
Thallus Laminariae (Thallus Eckloniae) vanadium ion dependent form haloperoxidase gene and encoding proteins thereof Download PDFInfo
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- CN106148363A CN106148363A CN201610565700.4A CN201610565700A CN106148363A CN 106148363 A CN106148363 A CN 106148363A CN 201610565700 A CN201610565700 A CN 201610565700A CN 106148363 A CN106148363 A CN 106148363A
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- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 48
- 230000001419 dependent effect Effects 0.000 title claims abstract description 9
- 229910001456 vanadium ion Inorganic materials 0.000 title claims abstract description 5
- 102000004169 proteins and genes Human genes 0.000 title abstract description 17
- 230000000694 effects Effects 0.000 claims abstract description 14
- 229910052740 iodine Inorganic materials 0.000 claims abstract description 12
- 239000011630 iodine Substances 0.000 claims abstract description 12
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 claims abstract description 11
- 108010073997 Bromide peroxidase Proteins 0.000 claims abstract description 9
- 102000003992 Peroxidases Human genes 0.000 claims abstract description 8
- 108040007629 peroxidase activity proteins Proteins 0.000 claims abstract description 8
- 125000002346 iodo group Chemical group I* 0.000 claims abstract description 6
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 6
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract description 5
- 108090000790 Enzymes Proteins 0.000 claims description 10
- 239000002773 nucleotide Substances 0.000 claims description 6
- 230000006872 improvement Effects 0.000 claims description 4
- 230000002255 enzymatic effect Effects 0.000 claims description 2
- 125000001309 chloro group Chemical group Cl* 0.000 claims 1
- 235000018102 proteins Nutrition 0.000 abstract description 16
- 108010035722 Chloride peroxidase Proteins 0.000 abstract description 8
- 238000005516 engineering process Methods 0.000 abstract description 6
- 241000195493 Cryptophyta Species 0.000 abstract description 3
- 238000001952 enzyme assay Methods 0.000 abstract description 3
- 238000012163 sequencing technique Methods 0.000 abstract description 3
- 241001466453 Laminaria Species 0.000 abstract description 2
- 230000009465 prokaryotic expression Effects 0.000 abstract description 2
- 102000004190 Enzymes Human genes 0.000 description 9
- 239000000047 product Substances 0.000 description 7
- 241000894006 Bacteria Species 0.000 description 6
- 238000001514 detection method Methods 0.000 description 5
- 229910052736 halogen Inorganic materials 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 241000199919 Phaeophyceae Species 0.000 description 4
- 150000002367 halogens Chemical class 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 229910052720 vanadium Inorganic materials 0.000 description 4
- LEONUFNNVUYDNQ-UHFFFAOYSA-N vanadium atom Chemical group [V] LEONUFNNVUYDNQ-UHFFFAOYSA-N 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 3
- 241000015177 Saccharina japonica Species 0.000 description 3
- 238000000246 agarose gel electrophoresis Methods 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 230000014509 gene expression Effects 0.000 description 3
- 125000005843 halogen group Chemical group 0.000 description 3
- 230000026030 halogenation Effects 0.000 description 3
- 238000005658 halogenation reaction Methods 0.000 description 3
- VOBIHUAWDXUCPH-UHFFFAOYSA-N 2-chloro-5,5-dimethylcyclohexane-1,3-dione Chemical compound CC1(C)CC(=O)C(Cl)C(=O)C1 VOBIHUAWDXUCPH-UHFFFAOYSA-N 0.000 description 2
- 102000013563 Acid Phosphatase Human genes 0.000 description 2
- 108010051457 Acid Phosphatase Proteins 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 102000004316 Oxidoreductases Human genes 0.000 description 2
- 108090000854 Oxidoreductases Proteins 0.000 description 2
- 239000011543 agarose gel Substances 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 125000001246 bromo group Chemical group Br* 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000010369 molecular cloning Methods 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- GZCWLCBFPRFLKL-UHFFFAOYSA-N 1-prop-2-ynoxypropan-2-ol Chemical compound CC(O)COCC#C GZCWLCBFPRFLKL-UHFFFAOYSA-N 0.000 description 1
- OPIFSICVWOWJMJ-AEOCFKNESA-N 5-bromo-4-chloro-3-indolyl beta-D-galactoside Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1OC1=CNC2=CC=C(Br)C(Cl)=C12 OPIFSICVWOWJMJ-AEOCFKNESA-N 0.000 description 1
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 1
- 241000195649 Chlorella <Chlorellales> Species 0.000 description 1
- 241001062009 Indigofera Species 0.000 description 1
- 241000199900 Laminariales Species 0.000 description 1
- 206010025421 Macule Diseases 0.000 description 1
- 229910020700 Na3VO4 Inorganic materials 0.000 description 1
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 241000206572 Rhodophyta Species 0.000 description 1
- 108091081024 Start codon Proteins 0.000 description 1
- 108020005038 Terminator Codon Proteins 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 1
- 229910052794 bromium Inorganic materials 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 description 1
- 101150093249 hpo gene Proteins 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 125000002883 imidazolyl group Chemical group 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- IHIXIJGXTJIKRB-UHFFFAOYSA-N trisodium vanadate Chemical compound [Na+].[Na+].[Na+].[O-][V]([O-])([O-])=O IHIXIJGXTJIKRB-UHFFFAOYSA-N 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0065—Oxidoreductases (1.) acting on hydrogen peroxide as acceptor (1.11)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
- C12Y111/01—Peroxidases (1.11.1)
- C12Y111/01806—Iodide peroxidase (vanadium-containing) (1.11.1.B6)
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
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- Biotechnology (AREA)
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- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention relates to gene engineering technology field, specifically one main laminaria vanadium ion dependent form haloperoxidase gene, the aminoacid sequence of this gene nucleotide series and coded protein is respectively SEQ ID NO:1 37 and SEQ ID NO:38 74.The present invention obtains gene order by transcript profile sequencing technologies binding sequence analytical technology, build prokaryotic expression carrier, by recombiant protein is carried out Enzyme assay it was confirmed it has the highest iodo peroxidase activity, without bromoperoxidase and chloro peroxidase activity.This gene has important using value for improving the content of iodine in the algae such as Thallus Laminariae (Thallus Eckloniae).
Description
Technical field
The present invention relates to the codase gene of a class Thallus Laminariae (Thallus Eckloniae) haloperoxidase.Particularly to Thallus Laminariae (Thallus Eckloniae) (Saccharina
Japonica) nucleotide sequence of haloperoxidase gene, and encoding proteins and during Thallus Laminariae (Thallus Eckloniae) halogenation and
Application in improvement Important Economic composition character.
Background technology
Having been found that multiple halogenide in ocean, wherein most has higher medicinal and using value.In ocean,
The Producer scope that can produce halogenide is extremely extensive, of a great variety, various informative.Wherein marine algae has stronger absorption
The ability of halogen, its internal halogenide is abundanter.Thallus Laminariae (Thallus Eckloniae) is sea-plant (algae) kind of whole world Main Cultivation, its
Content of iodine is high, in the Thallus Laminariae (Thallus Eckloniae) of 1g dry weight can the iodine of isolated 10mg, account for the 1% of self dry weight.Thallus Laminariae (Thallus Eckloniae) is many in China
Based on cultured population, it is a kind of extremely important large-scale economical alga, forms because the chemical products such as iodine, Algin can be produced
For the raw material of industry being widely cultivated.Meanwhile, Thallus Laminariae (Thallus Eckloniae) amount of iodine is the highest, is used to the edible vegetable that supplementing iodine is best, nutriture value
It is worth the abundantest.Along with the exploitation of the medicine effect of polysaccharose substance in Thallus Laminariae (Thallus Eckloniae) in recent years, its medical value is more people institute
Pay attention to, promoted the further development of laminaria culture industry.Owing to the I content in Thallus Laminariae (Thallus Eckloniae) is high, excite numerous studies person
Concern to it, some scientists guess, the reason of Thallus Laminariae (Thallus Eckloniae) enrichment I, may be namely due to the existence of its iodine oxidase
Particularity, therefore becomes focus to its halogenation mechanism and the oxidasic research of halogen.
Biological halogenation process depends on the effect of halo enzyme, and halo enzyme mainly includes haloperoxidase and Huang
Element dependent form halo enzyme, wherein most study is vanadium dependent form haloperoxidase (vHPO), according to its oxidation halogen from
The difference of sub-ability, can be classified as chloro peroxidase (chloroperoxidase), bromoperoxidase
And iodo peroxidase (iodoperoxidase) (bromoperoxidase).These three enzyme all has on gene structure
Identical conservative metal ion binding site, the i.e. imidazole ring of a histidine residues, this position in order to combine vanadium ion, its ammonia
Base acid sequence and acid phosphatase family (acid phosphatases) have the homology of height.At present, vanadium dependent form halogen
Oxidase is all found in red algae, chlorella and Brown algae；But vanadium dependent form iodine peroxidase is only found, especially in Brown algae
It is some more high monoids, such as Laminariales.
Clone products synthesis related gene also verifies its function, discloses the relation between gene and product, and auxiliary carries out product
Plant improvement and become one of International Agriculture breeding field effective way improving economic sector content；Meanwhile, genetic engineering is utilized
Technology production active substance and economic product have become the core content of modern biotechnology industry development.At present, vanadium in Brown algae
The clone of dependent form haloperoxidase (vHPO) gene extremely lacks, and its function is confirmed not yet.
Summary of the invention
For technical problem present in currently available technology, the present invention provides a class haloperoxidase gene, its
Codified one haloperoxidase albumen, described albumen has the highest iodo peroxidase
(iodoperoxidase) activity, without bromoperoxidase (bromoperoxidase) and chloro peroxidase
(chloroperoxidase) activity, described gene and encoding proteins thereof can be used for improving the content of iodine in frond.
An object of the present invention is to provide a class haloperoxidase gene, and described gene is all from Thallus Laminariae (Thallus Eckloniae)
The haloperoxidase gene separated in (Saccharina japonica), the nucleotide sequence of described gene such as SEQ
Shown in ID NO:1-37.
The two of the purpose of the present invention are to provide the albumen of gene code described in a class, and described albumen is by SEQ ID NO:1-
Nucleotide sequence coded shown in 37, its aminoacid sequence is as shown in SEQ ID NO:38-74；It has the highest iodo mistake
Peroxidase activity, without bromo and chloro peroxidase activity.
In the present invention, the encoding amino acid sequence of haloperoxidase gene and correspondence thereof is listed as follows:
The three of the object of the invention are the application in terms of described gene and encoding proteins content of iodine in improving frond thereof.
The four of the object of the invention are described gene and encoding proteins application in improvement economic sector character thereof.
The method that the present invention is checked order by transcript profile, obtains a series of haloperoxidase from Thallus Laminariae (Thallus Eckloniae) transcript profile
Gene, obtains full length sequence by gene synthetic, and the albumen being experimentally confirmed its coding has the highest iodine
For peroxidase activity, without bromoperoxidase and chloro peroxidase activity.Research Thallus Laminariae (Thallus Eckloniae) haloperoxidase
Gene contributes to the deep mechanism understanding that in the Brown algaes such as Thallus Laminariae (Thallus Eckloniae), the halogen such as bromine, iodine accumulates, and also educates for genetic engineering and molecule simultaneously
Plant and genetic resources is provided.The present invention obtains haloperoxidase gene first from Thallus Laminariae (Thallus Eckloniae) (Saccharina japonica),
And by prokaryotic expression carrier, gene carried out expression of recombinant proteins and carry out Enzyme assay it was confirmed it has the highest
Iodo peroxidase activity, without bromo and chloro peroxidase activity.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is expanded on further.Should be understood that these examples be merely to illustrate the present invention and
It is not used in restriction the scope of the present invention.The experimental technique of unreceipted specific experiment condition in the following example, generally according to routine
Condition, Molecular Cloning: A Laboratory guide (Sambrook J, et al.2008.Molecular Cloning:A Laboratory
Manual, 3rd Ed.) described in condition, or according to the condition proposed by manufacturer.
The preparation of embodiment 1:SjvIPO1 sequential coding albumen
By Shanghai Xu Guan biotechnology Development Co., Ltd synthesize SEQ ID NO:1 full length sequence, the purpose fragment of recovery,
Overnight it is connected to cloning vehicle pMD19-T in 16 DEG C of metal baths, and is transformed into competent escherichia coli cell E.coli Top10
In, coat on the LB solid medium containing 100mg/mL Amp, 37 DEG C of incubated overnight, sieve through IPTG/X-gal indigo plant white macula
After choosing, 4-10 positive colony of picking checks order.By sequencing result by sequence alignment, it is separated to Thallus Laminariae (Thallus Eckloniae) HPO gene, name
For SjvIPO1.SjvIPO1CDS sequence is 1905bp, and its nucleotide sequence, as shown in SEQ ID NO:1, encodes 634
Aminoacid, with ATG as start codon, TAA is termination codon.
Thallus Laminariae (Thallus Eckloniae) SjvIPO1PCR product, after the agarose gel electrophoresis detection of 1%, cuts purpose bar under uviol lamp
Band, agarose gel reclaims, and reclaims product SjvIPO1 and pET32a plasmid carries out EcoRV and NotI double digestion, at 37 DEG C of metals
After bath 3-4h, with 1% agarose gel electrophoresis detection, use agarose gel to reclaim test kit and reclaim.By purpose fragment
SjvIPO1 and plasmid pET32a is attached, and 16 DEG C overnight, the named pET32a-SjvIPO1 of recombiant plasmid built.
By pET32a-SjvIPO1 recombinant plasmid transformed E. coli expression strains BL21, picking BL21 positive colony, shake
Bacterium also preserves bacterial strain.PCR detects recon.PCR primer detects in 1% agarose gel electrophoresis, automatic gel image analysis instrument
Imaging.Picking electrophoresis detection inserts the cloning and sequencing that band is correct, detects the presence of sudden change, and whether frameshit frame changes.
The BL21 bacterial strain of purpose fragment will be successfully connected, with according to 1:1000 ratio (10mL LB fluid medium+
10uL AP+10uL bacterium solution) shake bacterium cultivation, surveyed OD value every one hour, until OD600nm value reaches 0.6 (3-4h).With
The IPTG concentration induction bacterium solution of 0.1mM, inductive condition is 25 DEG C, 160rpm, induces 6-8h.Induction takes 2mL bacterium solution 4 DEG C after terminating
12000rpm5min is centrifuged, and abandons supernatant, is inverted in absorbent paper by pipe；Precipitation adds 1/2 volume and shifts to an earlier date 1 × PBS that pre-cooling is good
(PH=7.4) in (i.e. 2mL centrifugal after precipitation in add 1mL 1 × PBS), fully mix.Ultrasonic method crushes bacterium solution, collects broken
Supernatant after broken, with the membrane filtration of 0.45 μm, after Ni column purification, the protein sample of collection is put in bag filter and is dialysed, with
Remove unwanted ion, it is thus achieved that restructuring SjvIPO1 albumen, its aminoacid sequence as shown in SEQ ID NO:38, total length
634aa；Use the expression of SDS-PAGE, Western-Blot detection recombiant protein.
Embodiment 2
According to the method for embodiment 1, to SjvIPO2-SjvIPO37 (nucleotide sequence is as shown in SEQ ID NO:2-37)
Encoding proteins is prepared, through order-checking show prepared protein sequence as shown in SEQ ID NO:39-74, specific as follows:
The functional verification of embodiment 3:SjvIPO1-37 encoding proteins
The detection of SjvIPO1 protein active uses monochloro 1,1-Dimethyl-3,5-diketocyclohexane (MCD) standard method.Enzyme activity assay reaction is as follows: will
SjvIPO1 albumen PBS is diluted to 0.1mM (100 microlitres are diluted to 500 microlitres), adds 56 microlitre 10mM Na3VO4, low speed mixes
Even process is overnight.50mM MES (pH 6.50), 200mM KBr (or KCl, KI), 0.1mM MCD
And appropriate Na (monochlorodimedone)3VO4The restructuring SjvIPO1 albumen processed, adds 1mM H2O2Initial action.
By above-mentioned removing H2O2System mixing after under the conditions of relevant temperature, hatch initial action after 2min, with corresponding buffer be
Blank, use high performance liquid chromatography at 278nm, measure the content of MCD in liquid phase, each reaction arrange 4 parallel
Sample.After testing, I is utilized-It is 5895U/mg that enzyme is lived, and optimal reactive temperature is 25 DEG C, and optimum pH is 6.5.To Cl-And Br-Live without enzyme
Property.This enzyme is high temperature enzyme, basic protein.
And the albumen that SjvIPO1 sequence is similar is detected, but its enzymatic activity is all far smaller than the present invention and prepares
SjvIPO recombiant protein, enzyme activity value contrast as follows:
Comparative example 1 is protein sequence disclosed in CAF04025.1 for GenBank accession number
Comparative example 2 is protein sequence disclosed in CAQ51446.1 for GenBank accession number
Although the present invention describes specific example, but has any will be apparent to practitioners skilled in the art,
The present invention can be made various changes and change the most under the premise without departing from the spirit and scope of the present invention.Therefore, appended right
Require to cover all these variation within the scope of the present invention.
Claims (4)
1. one kind encodes the gene of vanadium ion dependent form haloperoxidase in Thallus Laminariae (Thallus Eckloniae), it is characterised in that the core of described gene
Nucleotide sequence is as shown in SEQ ID NO:1-37.
2. one kind by the albumen of gene code described in claim 1, it is characterised in that the aminoacid sequence of described albumen such as SEQ
Shown in ID NO:38-74, it has the highest iodo peroxidase activity, without bromoperoxidase and chloro peroxidating
Thing enzymatic activity.
3. the application in terms of the content of iodine in improving frond of the enzyme gene described in claim 1.
4. the application in improvement economic sector character of the albumen described in claim 2.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108913703A (en) * | 2018-07-31 | 2018-11-30 | 中国海洋大学 | A kind of bromoperoxidase and its encoding gene |
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| CN101037675A (en) * | 2006-03-15 | 2007-09-19 | 中国科学院大连化学物理研究所 | Isolation and extraction method for bromine peroxide enzyme |
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Non-Patent Citations (1)
| Title |
|---|
| 吕辉: "海带（Saccharina japonica）钒依赖型溴过氧化酶基因（vBPO）的分子系统学研究、原核表达与酶活检测", 《中国优秀硕士学位论文全文数据库 基础科学辑》 * |
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