JPH10298007A - Agent for preventing adhesion of aquatic organism - Google Patents
Agent for preventing adhesion of aquatic organismInfo
- Publication number
- JPH10298007A JPH10298007A JP9108966A JP10896697A JPH10298007A JP H10298007 A JPH10298007 A JP H10298007A JP 9108966 A JP9108966 A JP 9108966A JP 10896697 A JP10896697 A JP 10896697A JP H10298007 A JPH10298007 A JP H10298007A
- Authority
- JP
- Japan
- Prior art keywords
- aquatic organism
- carbonic anhydrase
- organism adhesion
- paint
- enzyme
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 230000003405 preventing effect Effects 0.000 title description 2
- 102000003846 Carbonic anhydrases Human genes 0.000 claims abstract description 23
- 108090000209 Carbonic anhydrases Proteins 0.000 claims abstract description 23
- 239000004480 active ingredient Substances 0.000 claims abstract description 9
- 239000003973 paint Substances 0.000 claims description 18
- 239000003112 inhibitor Substances 0.000 claims description 14
- 230000010071 organism adhesion Effects 0.000 claims description 14
- 241000243321 Cnidaria Species 0.000 claims description 3
- 230000007613 environmental effect Effects 0.000 abstract description 2
- 239000003795 chemical substances by application Substances 0.000 abstract 4
- 239000011248 coating agent Substances 0.000 abstract 2
- 238000000576 coating method Methods 0.000 abstract 2
- 230000001747 exhibiting effect Effects 0.000 abstract 1
- 230000001988 toxicity Effects 0.000 abstract 1
- 231100000419 toxicity Toxicity 0.000 abstract 1
- 230000000694 effects Effects 0.000 description 14
- 102000004190 Enzymes Human genes 0.000 description 13
- 108090000790 Enzymes Proteins 0.000 description 13
- 238000000034 method Methods 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 11
- 108090000623 proteins and genes Proteins 0.000 description 10
- 241000238586 Cirripedia Species 0.000 description 9
- 102000004169 proteins and genes Human genes 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 8
- 230000002940 repellent Effects 0.000 description 8
- 239000005871 repellent Substances 0.000 description 8
- 241000242759 Actiniaria Species 0.000 description 7
- 238000000502 dialysis Methods 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 150000001413 amino acids Chemical group 0.000 description 5
- 239000000872 buffer Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- FTOAOBMCPZCFFF-UHFFFAOYSA-N 5,5-diethylbarbituric acid Chemical compound CCC1(CC)C(=O)NC(=O)NC1=O FTOAOBMCPZCFFF-UHFFFAOYSA-N 0.000 description 4
- IKHGUXGNUITLKF-UHFFFAOYSA-N Acetaldehyde Chemical compound CC=O IKHGUXGNUITLKF-UHFFFAOYSA-N 0.000 description 4
- 241000283690 Bos taurus Species 0.000 description 4
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 4
- 235000011130 ammonium sulphate Nutrition 0.000 description 4
- 239000001569 carbon dioxide Substances 0.000 description 4
- 229910002092 carbon dioxide Inorganic materials 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 230000036961 partial effect Effects 0.000 description 4
- 239000002244 precipitate Substances 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 230000009182 swimming Effects 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 3
- 241000237536 Mytilus edulis Species 0.000 description 3
- 229920002678 cellulose Polymers 0.000 description 3
- 239000001913 cellulose Substances 0.000 description 3
- 239000000839 emulsion Substances 0.000 description 3
- 210000003743 erythrocyte Anatomy 0.000 description 3
- 235000020638 mussel Nutrition 0.000 description 3
- 229920006395 saturated elastomer Polymers 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- YBADLXQNJCMBKR-UHFFFAOYSA-M (4-nitrophenyl)acetate Chemical compound [O-]C(=O)CC1=CC=C([N+]([O-])=O)C=C1 YBADLXQNJCMBKR-UHFFFAOYSA-M 0.000 description 2
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 2
- ZPLCXHWYPWVJDL-UHFFFAOYSA-N 4-[(4-hydroxyphenyl)methyl]-1,3-oxazolidin-2-one Chemical compound C1=CC(O)=CC=C1CC1NC(=O)OC1 ZPLCXHWYPWVJDL-UHFFFAOYSA-N 0.000 description 2
- BVKZGUZCCUSVTD-UHFFFAOYSA-M Bicarbonate Chemical compound OC([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-M 0.000 description 2
- 241001474374 Blennius Species 0.000 description 2
- 108090000371 Esterases Proteins 0.000 description 2
- 239000007995 HEPES buffer Substances 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- TYMRLRRVMHJFTF-UHFFFAOYSA-N Mafenide Chemical compound NCC1=CC=C(S(N)(=O)=O)C=C1 TYMRLRRVMHJFTF-UHFFFAOYSA-N 0.000 description 2
- 240000003728 Spergula arvensis Species 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000001042 affinity chromatography Methods 0.000 description 2
- 239000011543 agarose gel Substances 0.000 description 2
- 229960002319 barbital Drugs 0.000 description 2
- 238000004166 bioassay Methods 0.000 description 2
- 150000001728 carbonyl compounds Chemical class 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 239000000498 cooling water Substances 0.000 description 2
- 239000000287 crude extract Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 210000002615 epidermis Anatomy 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- -1 hydrogen ions Chemical class 0.000 description 2
- 238000007654 immersion Methods 0.000 description 2
- 230000003100 immobilizing effect Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- VUZPPFZMUPKLLV-UHFFFAOYSA-N methane;hydrate Chemical compound C.O VUZPPFZMUPKLLV-UHFFFAOYSA-N 0.000 description 2
- 229920002401 polyacrylamide Polymers 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 230000001846 repelling effect Effects 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 239000004576 sand Substances 0.000 description 2
- FDDDEECHVMSUSB-UHFFFAOYSA-N sulfanilamide Chemical compound NC1=CC=C(S(N)(=O)=O)C=C1 FDDDEECHVMSUSB-UHFFFAOYSA-N 0.000 description 2
- 229940124530 sulfonamide Drugs 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical group C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- BTJIUGUIPKRLHP-UHFFFAOYSA-N 4-nitrophenol Chemical compound OC1=CC=C([N+]([O-])=O)C=C1 BTJIUGUIPKRLHP-UHFFFAOYSA-N 0.000 description 1
- QAUUDNIGJSLPSX-UHFFFAOYSA-N 4-nitrophenyl acetate Chemical compound CC(=O)OC1=CC=C([N+]([O-])=O)C=C1 QAUUDNIGJSLPSX-UHFFFAOYSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- HXNNRBHASOSVPG-GUBZILKMSA-N Ala-Glu-Leu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O HXNNRBHASOSVPG-GUBZILKMSA-N 0.000 description 1
- 241001083548 Anemone Species 0.000 description 1
- 241000948360 Anthopleura japonica Species 0.000 description 1
- 241001580953 Anthopleura krebsi Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 241001195836 Cypris Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- RNPGPFAVRLERPP-QEJZJMRPSA-N Gln-Trp-Asn Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(N)=O)C(O)=O RNPGPFAVRLERPP-QEJZJMRPSA-N 0.000 description 1
- RFDHKPSHTXZKLL-IHRRRGAJSA-N Glu-Gln-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCC(=O)O)N RFDHKPSHTXZKLL-IHRRRGAJSA-N 0.000 description 1
- AYBKPDHHVADEDA-YUMQZZPRSA-N Gly-His-Asn Chemical compound [H]NCC(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(N)=O)C(O)=O AYBKPDHHVADEDA-YUMQZZPRSA-N 0.000 description 1
- HFPVRZWORNJRRC-UWVGGRQHSA-N Gly-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)CN HFPVRZWORNJRRC-UWVGGRQHSA-N 0.000 description 1
- FKESCSGWBPUTPN-FOHZUACHSA-N Gly-Thr-Asn Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O FKESCSGWBPUTPN-FOHZUACHSA-N 0.000 description 1
- MJUUWJJEUOBDGW-IHRRRGAJSA-N His-Leu-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CN=CN1 MJUUWJJEUOBDGW-IHRRRGAJSA-N 0.000 description 1
- 108050004689 Inhibitor of carbonic anhydrases Proteins 0.000 description 1
- RGUXWMDNCPMQFB-YUMQZZPRSA-N Leu-Ser-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RGUXWMDNCPMQFB-YUMQZZPRSA-N 0.000 description 1
- 108010053229 Lysyl endopeptidase Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 244000300264 Spinacia oleracea Species 0.000 description 1
- 235000009337 Spinacia oleracea Nutrition 0.000 description 1
- QGXCWPNQVCYJEL-NUMRIWBASA-N Thr-Asn-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O QGXCWPNQVCYJEL-NUMRIWBASA-N 0.000 description 1
- WNQJTLATMXYSEL-OEAJRASXSA-N Thr-Phe-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O WNQJTLATMXYSEL-OEAJRASXSA-N 0.000 description 1
- QNXZCKMXHPULME-ZNSHCXBVSA-N Thr-Val-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N)O QNXZCKMXHPULME-ZNSHCXBVSA-N 0.000 description 1
- XHALUUQSNXSPLP-UFYCRDLUSA-N Tyr-Arg-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=C(O)C=C1 XHALUUQSNXSPLP-UFYCRDLUSA-N 0.000 description 1
- 229920006243 acrylic copolymer Polymers 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 238000012870 ammonium sulfate precipitation Methods 0.000 description 1
- 239000002519 antifouling agent Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- LFYJSSARVMHQJB-QIXNEVBVSA-N bakuchiol Chemical compound CC(C)=CCC[C@@](C)(C=C)\C=C\C1=CC=C(O)C=C1 LFYJSSARVMHQJB-QIXNEVBVSA-N 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000003431 cross linking reagent Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000000385 dialysis solution Substances 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 108010051307 glycyl-glycyl-proline Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- OAKJQQAXSVQMHS-UHFFFAOYSA-N hydrazine group Chemical group NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 description 1
- GPRLSGONYQIRFK-UHFFFAOYSA-N hydron Chemical compound [H+] GPRLSGONYQIRFK-UHFFFAOYSA-N 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 239000005457 ice water Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 108010012581 phenylalanylglutamate Proteins 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000011435 rock Substances 0.000 description 1
- 238000005185 salting out Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 229910021653 sulphate ion Inorganic materials 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- RSPCKAHMRANGJZ-UHFFFAOYSA-N thiohydroxylamine Chemical compound SN RSPCKAHMRANGJZ-UHFFFAOYSA-N 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
- Paints Or Removers (AREA)
Abstract
Description
【0001】[0001]
【産業上の利用分野】本発明は、水生生物付着防止剤、
特に海産汚損生物等の生物が忌避する作用を有するカル
ボニックアンヒドラーゼを有効成分とする水生生物付着
防止剤に関する。The present invention relates to an aquatic organism adhesion inhibitor,
In particular, the present invention relates to an aquatic organism adhesion inhibitor comprising, as an active ingredient, carbonic anhydrase having an action of repelling organisms such as marine fouling organisms.
【0002】[0002]
【従来の技術】船舶の船底及び漁網、火力発電所の復水
器冷却用水あるいは石油化学工業の熱交換機冷却用水の
取水路、ブイ等の海中に置かれる設備、港湾やダムの付
帯設備等ではフジツボ類、イガイ類、藻類等の水生生物
が付着、繁殖し、取水口の閉鎖等の障害が報告されてい
る。従来、これら水生生物の付着を防止するために、有
機スズ化合物を有効成分とする水生生物付着防止剤が使
用されていたが、この使用により河川もしくは海洋の環
境が汚染されたり、魚を媒体として人体への被害が発生
する恐れがあるという社会問題が生じ、有機スズ化合物
についてはその使用、製造とも法的な規制を受け、これ
に変わる水生生物付着防止剤の開発が望まれていた。2. Description of the Related Art In the bottom of ships and fishing nets, water intake channels for condenser cooling water in thermal power plants or heat exchanger cooling water in the petrochemical industry, buoys and other submarine equipment, harbors and dams, etc. It has been reported that aquatic organisms such as barnacles, mussels, and algae attach and breed, and obstacles such as closing of intakes are observed. Conventionally, in order to prevent the attachment of these aquatic organisms, an aquatic organism adhesion inhibitor containing an organotin compound as an active ingredient has been used, but this use pollutes the river or marine environment or uses fish as a medium. There has been a social problem that the human body may be damaged, and the use and production of the organotin compound are subject to legal regulations, and there has been a demand for the development of an aquatic organism adhesion inhibitor instead.
【0003】このため、最近では海洋生物等から抽出し
た生理活性物質をはじめとする様々な有機化合物を含む
安全な防止剤の開発がなされている。しかしながら前記
有機化合物では実際に人工施設等の基盤(鉄板、コンク
リート等)にこれら物質を塗布または固定化し、実際に
上記生物を忌避させるという技術は未だ開発されていな
い。そこで、環境に影響を与えず、しかもこれら問題点
を解決しうる効果的な生物忌避物質の開発が緊急の課題
となっている。[0003] For this reason, recently, safe inhibitors containing various organic compounds such as physiologically active substances extracted from marine organisms and the like have been developed. However, the technique of applying or immobilizing these substances on a base (iron plate, concrete, or the like) of an artificial facility or the like and actually repelling the above organisms has not yet been developed. Therefore, development of an effective biological repellent substance that does not affect the environment and can solve these problems has become an urgent issue.
【0004】[0004]
【発明が解決しようとする課題】本発明は、環境に悪い
影響を及ぼさず、海産汚損生物に対して有効な忌避作用
を有する酵素蛋白質、およびこの酵素蛋白質を有効成分
として含む水生生物付着防止剤を提供することを目的と
する。DISCLOSURE OF THE INVENTION The present invention relates to an enzyme protein which does not adversely affect the environment and has an effective repellent action against marine fouling organisms, and an aquatic organism adhesion inhibitor containing this enzyme protein as an active ingredient. The purpose is to provide.
【0005】[0005]
【課題を解決するための手段】本発明者らは上記目的を
達成すべく鋭意研究を行った結果、カルボニックアンヒ
ドラーゼ活性を有する蛋白質が優れた水生生物付着防止
剤効果を示すことを見いだし、本発明を完成させるにい
たった。すなわち本発明は、カルボニックアンヒドラー
ゼを有効成分として含む水生生物付着防止剤を提供する
ものである。Means for Solving the Problems As a result of intensive studies to achieve the above object, the present inventors have found that a protein having carbonic anhydrase activity exhibits an excellent aquatic organism adhesion inhibitory effect, The present invention has been completed. That is, the present invention provides an aquatic organism adhesion inhibitor containing carbonic anhydrase as an active ingredient.
【0006】上記カルボニックアンヒドラーゼとして
は、刺胞動物由来のもの等が挙げられる。また、本発明
は上記水生生物付着防止剤を含むことを特徴とする塗料
を提供するものである。Examples of the carbonic anhydrase include those derived from cnidarians. The present invention also provides a paint containing the aquatic organism adhesion inhibitor.
【0007】[0007]
【発明の実施の態様】以下本発明を詳細に説明する。本
発明の水生生物付着防止剤は、イソギンチャク類等の刺
胞動物、牛、馬等のほ乳類動物、あるいはほうれん草等
の植物、Escherichia coli等の微生物等からカルボニッ
クアンヒドラーゼ又はその含有物を採取し、これを用い
て調製されるものである。BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail. The aquatic organism antifouling agent of the present invention is obtained by collecting carbonic anhydrase or its contents from cnidarians such as sea anemones, mammals such as cattle and horses, plants such as spinach, and microorganisms such as Escherichia coli. , Prepared using this.
【0008】例えばウメボシイソギンチャク科のヨロイ
イソギンチャクからのカルボニックアンヒドラーゼを有
効成分とする生物付着防止剤の調製方法について、以下
に記載する。For example, a method for preparing a biofouling inhibitor containing carbonic anhydrase as an active ingredient from a sea anemone belonging to the family Anemonidae will be described below.
【0009】ヨロイイソギンチャクを採取し、その表皮
に付着している細砂、海藻などを取り除いたものの300g
(湿重量)を1mM EDTA、1mMDTT、0.5M NaClを含むバッ
ファー(pH7.5)1.2lに入れ、ブレンダー(トリオサイエン
ス社製)を用いて破砕し、粗抽出液を得る。[0009] 300 g of a sea anemone, which is obtained by removing fine sand and seaweed attached to its epidermis,
The (wet weight) is placed in 1.2 l of a buffer (pH 7.5) containing 1 mM EDTA, 1 mM DTT and 0.5 M NaCl, and crushed using a blender (manufactured by Trio Science) to obtain a crude extract.
【0010】次にこの抽出液を遠心分離して固形物質を
除き、上澄み液を得、この上澄み液に対し硫酸アンモニ
ウムを適量加えて塩析を行い、蛋白質のみを回収する。
回収した沈殿を以降のステップで使用する溶液に溶解
し、透析を行う。なお、透析については通常の透析方法
に従って行うことができる。Next, the extract is centrifuged to remove solid substances, and a supernatant is obtained. An appropriate amount of ammonium sulfate is added to the supernatant to carry out salting out to recover only the protein.
The collected precipitate is dissolved in a solution used in the subsequent steps, and dialysis is performed. The dialysis can be performed according to a normal dialysis method.
【0011】例えば、透析膜(セルロースチューブ:VIS
KASE社製セルロースチューブ等)に試料をいれ、試料中
に含まれる無機塩類等の低分子物質を除くのに必要な条
件を備えた十分な量の溶液、例えば0.1M NH4HCO3 5lに
入れ4℃で24 時間透析する。このようにして得られた透
析内液(蛋白質)を一般的な酵素精製に用いる方法によ
り精製する。For example, a dialysis membrane (cellulose tube: VIS
A sample is placed in a cellulose tube made by KASE Co., Ltd.) and placed in a sufficient amount of a solution having conditions necessary for removing low-molecular substances such as inorganic salts contained in the sample, for example, 0.1 M NH 4 HCO 3 5 l. Dialyze at 4 ° C for 24 hours. The inner dialysis solution (protein) thus obtained is purified by a method used for general enzyme purification.
【0012】例えば、アフィニティークロマトグラフィ
ー〔ANALYTICAL BIOCHEMISTRY 57,467-476 (1974)、THE
Journal of Biological Chemistry 270,5985-5993 (19
95)、ARCHIVES OF BIOCHEMISTORY AND BIOPHYSICS 312,
429-435 (1994)等を参照〕、あるいはイオン交換、ゲル
濾過、疎水性樹脂等のクロマトグラフィーを行うことに
より精製する。このようにして以下の記載の酵素学的性
質を有するカルボニックアンヒドラーゼ活性を有する精
製蛋白を得る。For example, affinity chromatography [ANALYTICAL BIOCHEMISTRY 57, 467-476 (1974), THE
Journal of Biological Chemistry 270,5985-5993 (19
95), ARCHIVES OF BIOCHEMISTORY AND BIOPHYSICS 312,
429-435 (1994), etc.] or by performing ion exchange, gel filtration, chromatography on a hydrophobic resin or the like. Thus, a purified protein having carbonic anhydrase activity having the following enzymatic properties is obtained.
【0013】各クロマトグラフィーで使用する機器、試
薬等については、通常市販されているものを用いること
ができる。アフィニティークロマトグラフィーに使用す
るものとしてはp-アミノメチルベンゼンスルホンアミド
・アガロース・ゲル(SIGMA社製)等が挙げられる。As the instruments, reagents and the like used in each chromatography, commercially available ones can be used. Examples of those used for affinity chromatography include p-aminomethylbenzenesulfonamide agarose gel (manufactured by SIGMA).
【0014】この場合の溶出液としては、0.4M NaN3を
含む0.1M NH4HCO3 等が用いられる。本発明で用いるカ
ルボニックアンヒドラーゼは、以下の作用及び基質特異
性を示す酵素である。[0014] as eluent in this case, like 0.1 M NH 4 HCO 3 containing 0.4 M NaN 3 is used. The carbonic anhydrase used in the present invention is an enzyme having the following action and substrate specificity.
【0015】(1)作用:二酸化炭素と水に作用し、水
素イオンと炭酸水素イオンを生ずる反応を触媒し、その
逆の反応も触媒する。 (2)基質特異性:二酸化炭素、アセトアルデヒド、そ
の他のカルボニル化合物に作用し、また、酢酸p-ニトロ
フェニルエステルのようなエステル類にも作用する。(1) Action: Acts on carbon dioxide and water to catalyze the reaction that produces hydrogen ions and hydrogen carbonate ions, and also catalyzes the reverse reaction. (2) Substrate specificity: It acts on carbon dioxide, acetaldehyde and other carbonyl compounds, and also acts on esters such as p-nitrophenyl acetate.
【0016】以上のような性質を有するカルボニックア
ンヒドラーゼを含む水生生物付着防止剤は、そのまま使
用してもよいが、例えば塗料等に混合して利用すること
ができる。上記、塗料としては水溶性エマルジョン塗
料、例えば、ヒドラジン基架橋剤を含む水溶性アクリル
共重合物塗料が好ましい。The aquatic organism adhesion inhibitor containing carbonic anhydrase having the above-mentioned properties may be used as it is, but may be used by mixing it with a paint or the like. As the above-mentioned paint, a water-soluble emulsion paint, for example, a water-soluble acrylic copolymer paint containing a hydrazine group crosslinking agent is preferable.
【0017】このような水生生物付着防止剤を含む塗料
は、陶器、プラスチック板、コンクリート等に塗布し乾
燥させて使用する。この場合、長期間(3〜6カ月)海水や
淡水中に浸漬しても剥離したり、劣化することはなく、
水生生物付着防止効果を奏する。The paint containing such an aquatic organism adhesion inhibitor is applied to pottery, a plastic plate, concrete or the like and dried before use. In this case, even if immersed in seawater or freshwater for a long time (3 to 6 months), it does not peel or deteriorate,
It has an aquatic organism adhesion preventing effect.
【0018】[0018]
【実施例】以下、実施例を示して本発明をより具体的に
説明する。ただし、本発明はこれら実施例に限定されな
い。The present invention will be described below more specifically with reference to examples. However, the present invention is not limited to these examples.
【0019】〔実施例1〕 (1)カルボニックアンヒドラーゼの精製 神奈川県城ケ島の岩礁より入手したヨロイイソギンチャ
ク(Anthopleura japonica VERRILL)について、その表皮
に付着している細砂、海藻などを取り除いたもの300g
(湿重量)を1mM EDTA、1mMDTT、0.5M NaClを含む25mM
HEPESバッファー(pH7.5)1.2lに入れ、それをブレンダー
(トリオサイエンス社製)を用いて破砕した。[Example 1] (1) Purification of carbonic anhydrase From a rock anemone (Anthopleura japonica VERRILL) obtained from a rock reef at Jogashima, Kanagawa, fine sand and seaweed attached to the epidermis were removed. 300g
(Wet weight) 25 mM containing 1 mM EDTA, 1 mM DTT, 0.5 M NaCl
Put in 1.2 L of HEPES buffer (pH 7.5) and blend it.
(Trio science).
【0020】得られるホモジナイズ懸濁液を氷冷して粗
抽出液を得た。次に該抽出液を10,000xgで20分、遠心分
離を行って(5℃)、上澄み液1.6lを採取し、これに30%
飽和になるように硫酸アンモニウムを加え硫安沈殿を行
った。The obtained homogenized suspension was cooled on ice to obtain a crude extract. Next, the extract was centrifuged at 10,000 × g for 20 minutes (5 ° C.), and 1.6 l of the supernatant was collected and 30%
Ammonium sulfate was added so as to be saturated, and ammonium sulfate precipitation was performed.
【0021】10,000xgで30分の遠心分離で沈殿を除き、
上澄み液1.6lに80%飽和になるように硫酸アンモニウム
を加えて硫安沈殿を行い、10,000xgで30分の遠心分離で
沈殿を回収した。回収した沈殿を0.1M NH4HCO3に溶解
し、セルロースチューブ(VISKASE社製)の透析膜に移
し、0.1M NH4HCO35lに対して4℃で24時間透析した。The precipitate is removed by centrifugation at 10,000 × g for 30 minutes,
Ammonium sulfate was added to 1.6 liters of the supernatant so as to be 80% saturated, and ammonium sulfate was precipitated. The precipitate was recovered by centrifugation at 10,000 × g for 30 minutes. The collected precipitate was dissolved in 0.1 M NH 4 HCO 3 , transferred to a dialysis membrane of a cellulose tube (manufactured by VISKASE), and dialyzed against 5 l of 0.1 M NH 4 HCO 3 at 4 ° C. for 24 hours.
【0022】透析した溶液1l に、0.1M NH4HCO3で平衡
化した30ml のp-アミノメチルベンゼンスルホンアミド
・アガロース・ゲル(SIGMA 社製)を加え、ときどき弱く
撹拌しながら酵素を吸着させた。これを同じ緩衝液で洗
浄し、不要な蛋白質を除いた後、ゲルをカラム(直径3cm
×20cm) に充填し0.2M NaIを含む0.1M NH4HCO3 100ml
で洗浄した後、0.4M NaN3を含む0.1M NH4HCO3 100mlで
溶出した。To 1 liter of the dialyzed solution, 30 ml of p-aminomethylbenzenesulfonamide agarose gel (manufactured by SIGMA) equilibrated with 0.1 M NH 4 HCO 3 was added, and the enzyme was adsorbed with occasional gentle stirring. . After washing with the same buffer to remove unnecessary proteins, the gel was columnized (diameter 3 cm).
0.1M NH 4 HCO 3 100ml containing the filled 0.2 M NaI in × 20 cm)
After washing with, elution was performed with 100 ml of 0.1 M NH 4 HCO 3 containing 0.4 M NaN 3 .
【0023】溶出液を透析膜に移し1mM EDTA、1mMDTT、
0.5M NaClを含む25mM HEPES緩衝液(pH7.5)3lに対し4℃
で24時間透析した。さらに、緩衝液を交換し3回透析を
行った。The eluate was transferred to a dialysis membrane, and 1 mM EDTA, 1 mM DTT,
4 ° C for 3 L of 25 mM HEPES buffer (pH 7.5) containing 0.5 M NaCl
For 24 hours. Furthermore, the buffer was exchanged and dialysis was performed three times.
【0024】アミコン社製限外濾過膜装置(分画分子量1
0000)を用いて2mlにまで濃縮した。この画分は、SDS-ポ
リアクリルアミドゲル電気泳動により、均一で単一のバ
ンドを示し、分子量が約28000のものであった。図1
は、その結果を示す図である。Ultrafiltration membrane device manufactured by Amicon (molecular weight cut off 1)
0000) to 2 ml. This fraction showed a uniform and single band by SDS-polyacrylamide gel electrophoresis and had a molecular weight of about 28,000. FIG.
Is a view showing the result.
【0025】次に、内部の部分アミノ酸配列を決定する
ために、変性させた2 n molの蛋白を含む画分にアクロ
モバクタープロテアーゼI(タカラ製)を加え、37度で一
晩処理し、多数のペプチド断片を得た。次いでこれをCA
PCELL PAK C18 SG120(資生堂製)カラムを用いて0.1%TFA
水溶液と0.1%TFAを含むアセトニトリルの0〜80%のグラ
ジエントをかけてクロマトグラフィ−を行い、ピーク部
分をそれぞれ分取した。Next, to determine the internal partial amino acid sequence, Achromobacter protease I (manufactured by Takara) was added to the denatured fraction containing 2 nmol of protein, and treated at 37 ° C. overnight. A number of peptide fragments were obtained. Then this is CA
0.1% TFA using PCELL PAK C18 SG120 (Shiseido) column
Chromatography was performed by applying a gradient of 0 to 80% of an aqueous solution and acetonitrile containing 0.1% TFA, and peak portions were respectively collected.
【0026】これらのピーク部分を473Aプロテインシー
クエンサー(アプライドバイオシステムズ製)にかけて配
列を解析した結果、配列番号1及び2に示すアミノ酸配
列を得た。These peaks were subjected to a 473A protein sequencer (manufactured by Applied Biosystems) to analyze their sequences. As a result, the amino acid sequences shown in SEQ ID NOS: 1 and 2 were obtained.
【0027】上記で得られた精製物の酵素の酵素化学的
性質を以下に示す。 1)作用:二酸化炭素と水に作用し、水素イオンと炭酸
水素イオンを生ずる反応を触媒し、その逆の反応も触媒
する。 2)基質特異性:二酸化炭素、アセトアルデヒド、その
他のカルボニル化合物に作用し、また、酢酸p-ニトロフ
ェニルエステルのようなエステル類にも作用する。The enzymatic chemical properties of the purified product obtained above are shown below. 1) Action: Acts on carbon dioxide and water to catalyze the reaction that produces hydrogen ions and bicarbonate ions, and also catalyzes the reverse reaction. 2) Substrate specificity: It acts on carbon dioxide, acetaldehyde and other carbonyl compounds, and also acts on esters such as p-nitrophenyl acetate.
【0028】3)至適pH:エステラーゼ活性を指標にし
た場合、至適pHは8〜10である。 4)分子量:約28000(SDS-ポリアクリルアミド電気泳
動) 5)部分アミノ酸配列:配列番号1および2の通りであ
る。3) Optimum pH: Optimum pH is 8 to 10 when esterase activity is used as an index. 4) Molecular weight: about 28000 (SDS-polyacrylamide electrophoresis) 5) Partial amino acid sequence: as shown in SEQ ID NOS: 1 and 2.
【0029】(2)酵素活性の測定 1)pH変化(酵素反応により生ずる水素イオン濃度の変
化)の速度を測定する方法。(2) Measurement of enzyme activity 1) A method of measuring the rate of pH change (change of hydrogen ion concentration caused by enzyme reaction).
【0030】3mlのベロナール緩衝液(0.022Mベロナール
と0.022Mナトリウム塩、pH7.95)、ブロモチモールブル
ー数滴、2.3mlの水あるいは酵素溶液を15ml容量の密閉
容器中にいれ、氷水上で15分間冷却する。これに5mlの
氷冷した飽和炭酸水を加え反応を開始する。0℃で、ブ
ロモチモールブルーの色の変化を指標に、反応液の pH
を6.3にまで下げる時間を測定する。酵素存在下での時
間をt、酵素非存在下での時間をt0としたとき、(t0-t)/
tの値を酵素の活性単位(U)とする。3 ml of veronal buffer (0.022 M veronal and 0.022 M sodium salt, pH 7.95), a few drops of bromothymol blue, 2.3 ml of water or enzyme solution are placed in a 15 ml sealed container, and placed on ice water for 15 minutes. Cool for minutes. To this, 5 ml of ice-cooled saturated carbonated water is added to start the reaction. At 0 ° C, the pH of the reaction solution is measured based on the change in color of bromothymol blue.
Measure the time to reduce to 6.3. When the time in the presence of the enzyme t, the time in the enzyme absence was t 0, (t 0 -t) /
The value of t is defined as the activity unit (U) of the enzyme.
【0031】本方法で、ヨロイイソギンチャクより精製
したカルボニックアンヒドラーゼ、及び市販の牛赤血球
由来のカルボニックアンヒドラーゼ(フナコシ製)の活
性を測定したところ、それぞれ4000U/mg、1900U/mgであ
った。According to this method, the activities of carbonic anhydrase purified from a sea anemone and a commercially available bovine erythrocyte-derived carbonic anhydrase (Funakoshi) were 4000 U / mg and 1900 U / mg, respectively. .
【0032】2)エステラーゼ活性を測定する方法 0.6mlの15mM Tris-sulfate緩衝液(pH7.6)をマイクロチ
ューブにとり、適当に希釈した酵素液15μlを加え、温
度平衡を37℃に到達させた後、0.3mlの1.5mM酢酸p-ニト
ロフェニルエステル溶液を添加し、反応を開始する。0
〜40分間反応させ、0.15mlの1NHClを加え反応を停止す
る。2) Method for measuring esterase activity: Transfer 0.6 ml of 15 mM Tris-sulfate buffer (pH 7.6) into a microtube, add 15 μl of an appropriately diluted enzyme solution, and reach a temperature equilibrium of 37 ° C. Add 0.3 ml of 1.5 mM acetic acid p-nitrophenyl ester solution to start the reaction. 0
The reaction is allowed to proceed for 4040 minutes, and the reaction is stopped by adding 0.15 ml of 1N HCl.
【0033】反応液の348nmの吸光度の変化を吸光度計
を用いて調べる。37℃で1分間あたりに1μmolのp-ニト
ロフェノールを生成する酵素量を1単位とする。The change in absorbance at 348 nm of the reaction solution is examined using an absorbance meter. One unit is the amount of the enzyme that produces 1 μmol of p-nitrophenol per minute at 37 ° C.
【0034】カルボニックアンヒドラーゼの特異的な阻
害剤であるスルファニルアミドによる阻害効果を調べる
実験では、0.6mlの15mMトリス−サルフェート(Tris-sul
fate)緩衝液(pH7.6)中に、予め所定の濃度になるように
スルファニルアミドを溶解しておき反応を行わせる。In an experiment for examining the inhibitory effect of sulfanilamide, a specific inhibitor of carbonic anhydrase, 0.6 ml of 15 mM Tris-sulphate (Tris-sul
fate) Sulfanylamide is dissolved in a buffer solution (pH 7.6) to a predetermined concentration in advance, and the reaction is performed.
【0035】上記方法でヨロイイソギンチャクより精製
したカルボニックアンヒドラーゼの比活性を測定したと
ころ、1590U/mgであった。また、100μMスルファニルア
ミド存在下では活性がほぼ消失した。The specific activity of the carbonic anhydrase purified from the sea anemone by the above method was measured and found to be 1590 U / mg. The activity almost disappeared in the presence of 100 μM sulfanilamide.
【0036】〔実施例2〕実施例1のようにして精製さ
れたカルボニックアンヒドラーゼ、及び市販の牛赤血球
由来のカルボニックアンヒドラーゼ(フナコシ製)に対
して、フジツボ類、イガイ類に対する忌避作用を指標と
したバイオアッセイを行った。その方法は特開平8-1930
97号記載の方法に従った。Example 2 Repellent action against barnacles and mussels against carbonic anhydrase purified as in Example 1 and commercially available bovine erythrocyte-derived carbonic anhydrase (Funakoshi) A bioassay was performed using as an index. The method is disclosed in JP-A-8-1930
The method described in No. 97 was followed.
【0037】径17mm、深さ15mm、容量2mlのマルチウエ
ル(培養孔24個;ファルコン社製)に各濃度に調製した分
画溶液を満たし、フジツボ類ノープリウス幼生を投入
後、経時的にその活動、生死状態を観察し、計数を行っ
た。A multiwell (17 culture holes, 24 culture holes; manufactured by Falcon) having a diameter of 17 mm, a depth of 15 mm, and a capacity of 2 ml was filled with the fractionated solution prepared at each concentration, and barnacle nauplii larvae were introduced. Activity and life and death were observed and counted.
【0038】フジツボ類ノープリウス幼生遊泳行動は、
図2に示す通り、次の5段階に分けられる。 I:活発に遊泳し、容器の底につくことはない。 II:容器の底につくことはあっても、再び遊泳する
か、動き回る。The swimming behavior of barnacle nauplii larvae is
As shown in FIG. 2, it is divided into the following five stages. I: Swim vigorously and do not stick to the bottom of the container. II: Swim or move around again, even if it hits the bottom of the container.
【0039】III:容器の底に着いたままであるが、
他の個体が接触すると動き回る。 IV:容器の底に着いたままで、付属肢がたまに動く
(他の個体が接触しても動き回らない)。 V:完全に動きが停止する(死亡個体には原生動物が発
生する)。III: While still at the bottom of the container,
It moves around when other individuals come in contact. IV: appendages occasionally move while at the bottom of the container
(It does not move around even if other individuals touch it). V: Completely stop moving (protozoa develop in dead individuals).
【0040】効果の判定は、遊泳能力を失い、マルチウ
エルの底に停止した上記III〜Vの状態(Immobilized
Condition:遊泳行動阻害・停止状態)の個体を、倒立
実体顕微鏡下で経時的に算定し、投入総個体数に対する
百分率(指数)を算出する。該指数をIMI(Immobilized In
dex)と定義し、効果判定の指標とした。結果を図3に示
した。The effect was judged by the state of the above III-V (Immobilized) in which the swimming ability was lost and stopped at the bottom of the multiwell.
Condition: swimming behavior inhibition / stop state) is calculated over time under an inverted stereomicroscope, and a percentage (index) with respect to the total number of input individuals is calculated. The index is converted to IMI (Immobilized In
dex), which was used as an index for determining the effect. The results are shown in FIG.
【0041】ヨロイイソギンチャクより精製したカルボ
ニックアンヒドラーゼ(以下、CAという)では、0.6m
g/mlの時IMI値が90%、市販の牛赤血球由来のカルボニ
ックアンヒドラーゼでは1.0mg/mlの時にIMI値が50%を
示し、忌避効果が示された。なお、対照とし牛血清アル
ブミンを同様にして測定し、結果を図3にしめした。For carbonic anhydrase (hereinafter, referred to as CA) purified from sea anemones, 0.6 m
The IMI value was 90% at g / ml, and the commercially available bovine erythrocyte-derived carbonic anhydrase had an IMI value of 50% at 1.0 mg / ml, indicating a repellent effect. As a control, bovine serum albumin was measured in the same manner, and the results are shown in FIG.
【0042】〔実施例3〕 酵素を含む塗料の調製法及
びその固定化方法 蛋白質溶液(0.5〜2.5mg/ml)に対し、等量の水溶性塗料
(水溶性エマルジョン塗料:三菱化学製)を加え混合し
た。これを素焼き板35cm2に2ml塗布し、24時間室温で放
置し、乾燥させた。Example 3 Method for Preparing Enzyme-Containing Paint and Immobilizing Method Thereof An equal amount of water-soluble paint to protein solution (0.5-2.5 mg / ml)
(Water-soluble emulsion paint: manufactured by Mitsubishi Chemical Corporation) was added and mixed. This was applied in an amount of 2 ml on an unglazed plate 35 cm 2 , allowed to stand at room temperature for 24 hours, and dried.
【0043】〔実施例4〕 塗料の野外浸漬試験 実施例3にならい、本発明の塗料(忌避物質)を素焼き板
(35cm2)に0.5〜2.5mg塗布し、神奈川県横浜市磯子区の
潮間帯位置に、14日間浸漬した。試験には、対照区とし
て、塗布しないケース(コントロール)及び本発明の蛋白
質を含まない従来の塗料を塗布したケース(エマルジョ
ン塗料)を設けた。付着状況の判定は、素焼き板に付着
したフジツボ類成体、フジツボ類、キプリス幼生、イガ
イ類の個体数を測定することで調べた。結果を図4に示
す。図4より対照区と比較して、本発明の蛋白を塗料と
混ぜて塗布した群では、フジツボ類の付着数が減少して
いた。[Example 4] Field immersion test of paint According to Example 3, a paint (repellent substance) of the present invention was applied to an unglazed plate
(35 cm 2 ) was applied at 0.5 to 2.5 mg, and immersed for 14 days in the tidal zone of Isogo Ward, Yokohama City, Kanagawa Prefecture. In the test, as a control group, a case not coated (control) and a case coated with a conventional paint not containing the protein of the present invention (emulsion paint) were provided. The state of attachment was determined by measuring the numbers of adult barnacles, barnacles, cypris larvae, and mussels attached to the unglazed plate. FIG. 4 shows the results. From FIG. 4, the number of barnacles adhered was reduced in the group where the protein of the present invention was mixed with the paint and applied compared to the control group.
【0044】[0044]
【発明の効果】本発明により、優れた生物忌避作用を有
するカルボニックアンヒドラーゼを有効成分とする塗料
及びその製造方法を提供することができる。本発明の塗
料は酵素を有効成分とするものであるから毒性が低く、
環境破壊を伴うことなく水生生物の忌避作用を奏せしめ
ることができる。According to the present invention, it is possible to provide a paint containing carbonic anhydrase having an excellent biological repellent action as an active ingredient and a method for producing the same. The paint of the present invention has a low toxicity since the enzyme is an active ingredient,
The repellent action of aquatic organisms can be exhibited without causing environmental destruction.
【0045】[0045]
配列番号 1 配列の長さ:21 配列の型:アミノ酸 配列の種類:ヨロイイソギンチャクカルボニックアンヒ
ドラーゼの部分配列 配列: Thr Asn Glu Gly Thr Asn Leu Ser Gly Gly Pro Leu Gly His Asn Tyr Arg Phe Glu Gln Phe SEQ ID NO: 1 Sequence length: 21 Sequence type: Amino acid Sequence type: Partial sequence of royal sea anemone carbonic anhydrase Sequence: Thr Asn Glu Gly Thr Asn Leu Ser Gly Gly Pro Leu Gly His Asn Tyr Arg Phe Glu Gln Phe
【0046】配列番号 2 配列の長さ:15 配列の型:アミノ酸 配列の種類:ヨロイイソギンチャクカルボニックアンヒ
ドラーゼの部分配列 配列: Thr Val Pro Ala Glu Leu His Leu Met Gln Trp Asn Thr Phe LeuSEQ ID NO: 2 Sequence length: 15 Sequence type: amino acid Sequence type: partial sequence of royal anemone carbonic anhydrase Sequence: Thr Val Pro Ala Glu Leu His Leu Met Gln Trp Asn Thr Phe Leu
【図1】SDSポリアクリルアミド電気泳動の結果を示す
図である。FIG. 1 is a view showing the results of SDS polyacrylamide electrophoresis.
【図2】フジツボノープリウス幼生の遊泳行動を示す図
である。FIG. 2 is a view showing swimming behavior of barnacle novius larvae.
【図3】フジツボノープリウス幼生を用いたバイオアッ
セイの結果を示す図である。FIG. 3 is a diagram showing the results of a bioassay using barnacle nauplii larvae.
【図4】本発明の塗料の野外浸漬試験における生物忌避
作用の結果を示す図である。FIG. 4 is a view showing the results of a biological repellent effect in a field immersion test of the paint of the present invention.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 堀内 達雄 千葉県野田市野田339番地 キッコーマン 株式会社内 (72)発明者 松崎 加奈恵 東京都大田区田園調布南23−6−204 ジ ュネス田園調布 (72)発明者 青山 郁子 神奈川県平塚市紅谷町9−15 セントアイ マンション402 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Tatsuo Horiuchi 339 Noda, Noda City, Chiba Prefecture Kikkoman Corporation (72) Inventor Kanae Matsuzaki 23-6-204 Denen Chofu Minami, Ota-ku, Tokyo Genes Denon Chofu (72 Inventor Ikuko Aoyama 9-15 Beniyacho, Hiratsuka-shi, Kanagawa 402
Claims (3)
として含む水生生物付着防止剤。1. An aquatic organism adhesion inhibitor comprising carbonic anhydrase as an active ingredient.
由来のものである請求項1記載の水生生物付着防止剤。2. The aquatic organism adhesion inhibitor according to claim 1, wherein the carbonic anhydrase is derived from a cnidarian.
剤を含むことを特徴とする塗料。3. A paint comprising the aquatic organism adhesion inhibitor according to claim 1 or 2.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP9108966A JPH10298007A (en) | 1997-04-25 | 1997-04-25 | Agent for preventing adhesion of aquatic organism |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP9108966A JPH10298007A (en) | 1997-04-25 | 1997-04-25 | Agent for preventing adhesion of aquatic organism |
Publications (1)
Publication Number | Publication Date |
---|---|
JPH10298007A true JPH10298007A (en) | 1998-11-10 |
Family
ID=14498186
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP9108966A Pending JPH10298007A (en) | 1997-04-25 | 1997-04-25 | Agent for preventing adhesion of aquatic organism |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPH10298007A (en) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2002537470A (en) * | 1996-10-29 | 2002-11-05 | ダブリュ.ピー.パワーズ カンパニー | Marine structure antifouling method and composition |
JP2004531390A (en) * | 2001-06-26 | 2004-10-14 | アクセルレイト・テクノロジー・コーポレーション | Functional surface coating |
US7501157B2 (en) | 2001-06-26 | 2009-03-10 | Accelr8 Technology Corporation | Hydroxyl functional surface coating |
-
1997
- 1997-04-25 JP JP9108966A patent/JPH10298007A/en active Pending
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2002537470A (en) * | 1996-10-29 | 2002-11-05 | ダブリュ.ピー.パワーズ カンパニー | Marine structure antifouling method and composition |
JP2004531390A (en) * | 2001-06-26 | 2004-10-14 | アクセルレイト・テクノロジー・コーポレーション | Functional surface coating |
US7501157B2 (en) | 2001-06-26 | 2009-03-10 | Accelr8 Technology Corporation | Hydroxyl functional surface coating |
US7629029B2 (en) | 2001-06-26 | 2009-12-08 | Accelr8 Technology Corporation | Functional surface coating |
US8178602B2 (en) | 2001-06-26 | 2012-05-15 | Accelr8 Technology Corporation | Functional surface coating |
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