JPH0427997B2 - - Google Patents
Info
- Publication number
- JPH0427997B2 JPH0427997B2 JP58110124A JP11012483A JPH0427997B2 JP H0427997 B2 JPH0427997 B2 JP H0427997B2 JP 58110124 A JP58110124 A JP 58110124A JP 11012483 A JP11012483 A JP 11012483A JP H0427997 B2 JPH0427997 B2 JP H0427997B2
- Authority
- JP
- Japan
- Prior art keywords
- amylase
- substance
- wai
- serine
- amino acid
- 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.)
- Expired - Lifetime
Links
- 239000000126 substance Substances 0.000 claims description 20
- 241000209140 Triticum Species 0.000 claims description 19
- 235000021307 Triticum Nutrition 0.000 claims description 18
- 108010065511 Amylases Proteins 0.000 claims description 16
- 239000004382 Amylase Substances 0.000 claims description 15
- 102000013142 Amylases Human genes 0.000 claims description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 15
- 235000019418 amylase Nutrition 0.000 claims description 15
- 239000003392 amylase inhibitor Substances 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 13
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 230000005764 inhibitory process Effects 0.000 claims description 9
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 7
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 6
- 210000003079 salivary gland Anatomy 0.000 claims description 6
- 102000004190 Enzymes Human genes 0.000 claims description 5
- 108090000790 Enzymes Proteins 0.000 claims description 5
- 229920005654 Sephadex Polymers 0.000 claims description 5
- 239000012507 Sephadex™ Substances 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 4
- 125000001429 N-terminal alpha-amino-acid group Chemical group 0.000 claims description 4
- 238000004458 analytical method Methods 0.000 claims description 4
- 238000001962 electrophoresis Methods 0.000 claims description 4
- 150000001413 amino acids Chemical class 0.000 claims description 3
- 235000003704 aspartic acid Nutrition 0.000 claims description 3
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims description 3
- 229920002401 polyacrylamide Polymers 0.000 claims description 3
- 125000001433 C-terminal amino-acid group Chemical group 0.000 claims description 2
- 102000005367 Carboxypeptidases Human genes 0.000 claims description 2
- 108010006303 Carboxypeptidases Proteins 0.000 claims description 2
- 102000004142 Trypsin Human genes 0.000 claims description 2
- 108090000631 Trypsin Proteins 0.000 claims description 2
- 238000010521 absorption reaction Methods 0.000 claims description 2
- 235000001014 amino acid Nutrition 0.000 claims description 2
- 238000003277 amino acid sequence analysis Methods 0.000 claims description 2
- 230000029087 digestion Effects 0.000 claims description 2
- 238000002523 gelfiltration Methods 0.000 claims description 2
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 2
- 239000012266 salt solution Substances 0.000 claims description 2
- 239000012588 trypsin Substances 0.000 claims description 2
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims 3
- 239000004220 glutamic acid Substances 0.000 claims 3
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims 2
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims 2
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 claims 2
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 claims 2
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 claims 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims 2
- 239000004472 Lysine Substances 0.000 claims 2
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 claims 2
- 235000004279 alanine Nutrition 0.000 claims 2
- 235000013922 glutamic acid Nutrition 0.000 claims 2
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 claims 2
- 229960000310 isoleucine Drugs 0.000 claims 2
- 229930182817 methionine Natural products 0.000 claims 2
- 239000004474 valine Substances 0.000 claims 2
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 claims 1
- 239000004475 Arginine Substances 0.000 claims 1
- 239000004471 Glycine Substances 0.000 claims 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 claims 1
- LEVWYRKDKASIDU-IMJSIDKUSA-N L-cystine Chemical compound [O-]C(=O)[C@@H]([NH3+])CSSC[C@H]([NH3+])C([O-])=O LEVWYRKDKASIDU-IMJSIDKUSA-N 0.000 claims 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 claims 1
- JZKXXXDKRQWDET-QMMMGPOBSA-N L-m-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC(O)=C1 JZKXXXDKRQWDET-QMMMGPOBSA-N 0.000 claims 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 claims 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 claims 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 claims 1
- 239000004473 Threonine Substances 0.000 claims 1
- 229940024606 amino acid Drugs 0.000 claims 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims 1
- 235000018417 cysteine Nutrition 0.000 claims 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims 1
- 229960003067 cystine Drugs 0.000 claims 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 claims 1
- 102000004139 alpha-Amylases Human genes 0.000 description 16
- 108090000637 alpha-Amylases Proteins 0.000 description 16
- 229940024171 alpha-amylase Drugs 0.000 description 16
- 101710171801 Alpha-amylase inhibitor Proteins 0.000 description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 12
- 235000013312 flour Nutrition 0.000 description 12
- 239000006228 supernatant Substances 0.000 description 8
- 108010044467 Isoenzymes Proteins 0.000 description 7
- 239000003112 inhibitor Substances 0.000 description 7
- 239000002244 precipitate Substances 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 238000005119 centrifugation Methods 0.000 description 6
- 239000011780 sodium chloride Substances 0.000 description 6
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 5
- 238000004587 chromatography analysis Methods 0.000 description 5
- 238000000502 dialysis Methods 0.000 description 5
- 239000008213 purified water Substances 0.000 description 5
- 229920002684 Sepharose Polymers 0.000 description 4
- 229940088598 enzyme Drugs 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 108010029785 Pancreatic alpha-Amylases Proteins 0.000 description 3
- 102000001746 Pancreatic alpha-Amylases Human genes 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000011033 desalting Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000011002 quantification Methods 0.000 description 3
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 2
- 241000238413 Octopus Species 0.000 description 2
- 244000098338 Triticum aestivum Species 0.000 description 2
- 244000098345 Triticum durum Species 0.000 description 2
- 235000007264 Triticum durum Nutrition 0.000 description 2
- 239000008351 acetate buffer Substances 0.000 description 2
- 239000003729 cation exchange resin Substances 0.000 description 2
- 238000010908 decantation Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 238000005227 gel permeation chromatography Methods 0.000 description 2
- 238000004255 ion exchange chromatography Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 210000000496 pancreas Anatomy 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 210000003296 saliva Anatomy 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- PFRGGOIBYLYVKM-UHFFFAOYSA-N 15alpha-hydroxylup-20(29)-en-3-one Natural products CC(=C)C1CCC2(C)CC(O)C3(C)C(CCC4C5(C)CCC(=O)C(C)(C)C5CCC34C)C12 PFRGGOIBYLYVKM-UHFFFAOYSA-N 0.000 description 1
- 229940122816 Amylase inhibitor Drugs 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000693011 Homo sapiens Pancreatic alpha-amylase Proteins 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010033645 Pancreatitis Diseases 0.000 description 1
- 206010034038 Parotitis Diseases 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- SOKRNBGSNZXYIO-UHFFFAOYSA-N Resinone Natural products CC(=C)C1CCC2(C)C(O)CC3(C)C(CCC4C5(C)CCC(=O)C(C)(C)C5CCC34C)C12 SOKRNBGSNZXYIO-UHFFFAOYSA-N 0.000 description 1
- MIJWOJAXARLEHA-WDSKDSINSA-N Ser-Gly-Glu Chemical compound OC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O MIJWOJAXARLEHA-WDSKDSINSA-N 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 229940039227 diagnostic agent Drugs 0.000 description 1
- 239000000032 diagnostic agent Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000013213 extrapolation Methods 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 238000009738 saturating Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Plant Substances (AREA)
- Peptides Or Proteins (AREA)
Description
【発明の詳細な説明】
本発明は小麦種子ないしは小麦粉からの新規な
α−アミラーゼ阻害物質WAI−65およびその製
法に関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel α-amylase inhibitor WAI-65 from wheat seeds or wheat flour and a method for producing the same.
α−アミラーゼは各種生物に広く存在する加水
分解酵素でヒトに関して云えば主として唾液腺お
よび膵臓から由来し、その動態は各種疾病特に膵
炎、耳下腺炎、肝疾患、ある種の癌等の病態によ
り健康時と比較して大きく変動することが知られ
ている。従つてこれらの病態を正しく診断するた
めには、総アミラーゼ活性定量だけでなく、唾液
腺および膵臓由来のα−アミラーゼアイソザイム
の正確な逐次的動向を把握することが望まれてい
る。 α-Amylase is a hydrolytic enzyme that is widely present in various organisms, and in humans, it is mainly derived from the salivary glands and pancreas, and its dynamics vary depending on the pathological condition of various diseases, especially pancreatitis, parotitis, liver disease, and certain cancers. It is known that there are large fluctuations compared to when healthy. Therefore, in order to correctly diagnose these pathological conditions, it is desirable to not only quantify total amylase activity but also to understand the accurate sequential trends of α-amylase isozymes derived from salivary glands and pancreas.
従来臨床検査の場でヒトのα−アミラーゼアイ
ソザイムの分別定量には電気泳動法が用いられて
いるが、この方法は非常に煩瑣であり迅速に検体
を処理する方法としては難点が多く、更に結果の
判定には熟練を要し、容易な方法とは云い難い。
この問題点の解決として小麦粉由来のアミラーゼ
阻害物質を用いての分析方法がオドンネル氏らに
よつて研究されてきたが、いまだ満足するもので
なく実用的でなかつた〔J.Clin.Chem.23，560〜
566（1977）参照〕。 Conventionally, electrophoresis has been used to separate and quantify human α-amylase isozyme in clinical testing, but this method is extremely cumbersome and has many drawbacks as a method for processing specimens quickly. Judgment requires skill and is far from an easy method.
As a solution to this problem, an analytical method using an amylase inhibitor derived from wheat flour has been studied by Mr. O'Donnell et al., but it is still unsatisfactory and not practical [J.Clin.Chem. 23 ,560~
566 (1977)].
これらの欠点を補うα−アミラーゼの分別定量
用試薬として本発明者らは既に小麦粉を給源にし
てヒト唾液中のα−アミラーゼを特異的に阻害す
るα−アミラーゼ阻害物質WAI−53を発見した
（特開昭57−140727号公報参照）。WAI−53はヒ
ト唾液および膵臓のα−アミラーゼに対する単位
重量当りの阻害活性化（単位当りの各酵素を50％
阻害するために必要なWAI−53の量比Ｐ／Ｓ）
が250であり、オドンネル氏らのα−アミラーゼ
阻害剤の阻害活性比が高々100であるのと比較す
ると、α−アミラーゼアイソザイム分別定量の観
点から前者の優位性は疑う余地がない。本発明者
らは小麦粉中のα−アミラーゼを更に詳細に研究
する過程でWAI−53を含めた従来の既知α−ア
ミラーゼ阻害物質とは物理化学的および酵素化学
的に全く異なる新規なα−アミラーゼ阻害物質
WAI−65を発見し、この物質が生体中のα−ア
ミラーゼアイソザイムの分別定量用臨床検査薬と
なり得る優れた特長を有することを見出した。 As a reagent for the fractional determination of α-amylase that compensates for these shortcomings, the present inventors have already discovered WAI-53, an α-amylase inhibitor that specifically inhibits α-amylase in human saliva using wheat flour as a source ( (Refer to Japanese Patent Application Laid-Open No. 140727/1983). WAI-53 inhibits and activates human salivary and pancreatic α-amylase (50% of each enzyme per unit weight).
Amount ratio of WAI-53 required for inhibition (P/S)
is 250, and when compared with the inhibitory activity ratio of the α-amylase inhibitor of O'Donnell et al., which is at most 100, there is no doubt that the former is superior from the viewpoint of differential quantification of α-amylase isozymes. In the process of studying α-amylase in wheat flour in more detail, the present inventors discovered a new α-amylase that is completely different from the conventional known α-amylase inhibitors including WAI-53 in terms of physicochemical and enzymatic chemistry. inhibitor
We discovered WAI-65 and found that this substance has excellent features that can be used as a clinical test drug for fractionating and quantifying α-amylase isozyme in living organisms.
本発明によれば、小麦種子または小麦粉中の水
溶性区分からの抽出液を少なくとも含水有機溶媒
による分別沈殿、加熱処理、ゲル過クロマト処
理、イオン交換クロマト処理からなる一連の工程
で精製することにより新規且つ有用なα−アミラ
ーゼ阻害物質WAI−65が提供されるものである。 According to the present invention, an extract from a water-soluble fraction of wheat seeds or wheat flour is purified through a series of steps consisting of at least fractional precipitation with a water-containing organic solvent, heat treatment, gel permeation chromatography, and ion exchange chromatography. A novel and useful α-amylase inhibitor WAI-65 is provided.
本発明による新規なα−アミラーゼ阻害物質を
うるための原料となる小麦種子または小麦粉は硬
質小麦、内地小麦、軟質小麦、デユラム小麦ない
しはそれらより由来する強力小麦粉、中力小麦
粉、薄力小麦粉の種類および等級を問わない。含
有量の差異こそあれすべての小麦の胚乳部分には
所望のα−アミラーゼ阻害物質が含まれている。 The wheat seeds or wheat flour that are the raw materials for obtaining the novel α-amylase inhibitor of the present invention are hard wheat, inland wheat, soft wheat, durum wheat, or types of strong wheat flour, medium wheat flour, and soft wheat flour derived therefrom. and any grade. The endosperm of all wheat plants contains the desired α-amylase inhibitor, although the content may vary.
本発明のα−アミラーゼ阻害物質WAI−65の
取得方法としては次の工程によつて行なえる。 The α-amylase inhibitor WAI-65 of the present invention can be obtained by the following steps.
原料としての小麦または小麦粉を原料に対し３
〜10倍量の水好ましくは精製水で１〜５時間室温
において撹拌してWAI−65物質の抽出を行なう。
所定の時間撹拌した後、例えば遠心分離、傾瀉、
過等の適宜な操作で上澄みを採取する。この上
澄みは真空または常圧下での加熱により処理れさ
る。通常約50〜70℃において10分ないし１時間の
加熱が行なわれる。70℃で30分の加熱が好まし
い。所望によつてはこの加熱処理された液を再び
遠心分離、傾瀉または過等の操作に付して上澄
みを採取する。 Wheat or wheat flour as a raw material
The WAI-65 substance is extracted by stirring with ~10 times the amount of water, preferably purified water, for 1 to 5 hours at room temperature.
After stirring for a predetermined time, for example, centrifugation, decantation,
Collect the supernatant using an appropriate procedure such as filtering. This supernatant is treated by heating under vacuum or normal pressure. Heating is usually carried out at about 50-70°C for 10 minutes to 1 hour. Heating at 70°C for 30 minutes is preferred. If desired, the heat-treated liquid is again subjected to operations such as centrifugation, decantation, or filtration to collect the supernatant.
ここで加熱処理された抽出液（またはその上澄
み）を水性アセトン、エタノールまたはメタノー
ルのような含水有機溶媒〔濃度40〜70％（ｖ／
ｖ）〕で処理して生成する沈殿物を除去し、得ら
れた残留液に更に上記溶媒を加えて溶媒濃度90％
（ｖ／ｖ）とし且つその混合物を低温（０〜10℃）
に放置して沈殿を形成させ、そして得られた沈殿
を採取しそしてこれを精製水に溶解させる。次い
で得られた溶液を１〜20mM NaClを含むトリス
塩酸緩衝液（PH9.0〜9.5）を用いて充填させたセ
フアデツクスＧ−75（商標名）を用いてゲル過
カラムクロマトを行なう。活性区分を集めて透析
処理により脱塩し透析内液を濃縮した後、CM−
セフアロースCL−6B（商標名）やCM−セフアデ
ツクスＣ−25（商標名）などを用いたカチオン系
イオン交換クロマトを数回実施して所望の物質の
純度を高める。 Here, the heat-treated extract (or its supernatant) is mixed with a water-containing organic solvent such as aqueous acetone, ethanol or methanol [concentration 40-70% (v/
v)] to remove the generated precipitate, and add the above solvent to the resulting residual liquid to make the solvent concentration 90%.
(v/v) and the mixture at low temperature (0-10℃)
to form a precipitate, and the resulting precipitate is collected and dissolved in purified water. The resulting solution is then subjected to gel permeation column chromatography using Sephadex G-75 (trade name) filled with Tris-HCl buffer (PH 9.0 to 9.5) containing 1 to 20 mM NaCl. After collecting the active fraction and desalting it by dialysis treatment and concentrating the dialysate fluid, CM-
Cationic ion exchange chromatography using Sepharose CL-6B (trade name) or CM-Sephadex C-25 (trade name) is performed several times to increase the purity of the desired substance.
CM−セフアロースCL−6Bによる液体クロマ
ト処理は最も好便な方法で、通常30mM酢酸緩衝
液（PH4.5）で樹脂を飽和させ、活性物質を含む
溶液を充填し、ひきつづいて０〜0.5モルのNaCl
を含む同じ緩衝液で濃度勾配クロマト処理を実施
する。 Liquid chromatography with CM-Sepharose CL-6B is the most convenient method, usually by saturating the resin with 30 mM acetate buffer (PH 4.5) and loading the solution containing the active substance, followed by 0-0.5 molar NaCl
Concentration gradient chromatography is performed with the same buffer containing:
通常セフアデツクスＧ−75ゲルクロマト処理の
段階までの過程でWAI−53およびWAI−65を分
離することはできないが、上述したカチオン交換
樹脂を用いたクロマト処理で能率よく分別でき
る。WAI−65の純度を充分高めるために１回以
上のカチオン交換樹脂によるクロマト処理を実施
する必要があるが通常はこの操作を２回繰り返せ
ば高純度のWAI−65フラクシヨンが得られる。 Normally, WAI-53 and WAI-65 cannot be separated up to the stage of Sephadex G-75 gel chromatography, but they can be efficiently separated by chromatography using the above-mentioned cation exchange resin. In order to sufficiently increase the purity of WAI-65, it is necessary to carry out chromatography treatment using a cation exchange resin one or more times, but normally a highly pure WAI-65 fraction can be obtained by repeating this operation twice.
得られた0.65フラクシヨンを含む区分を集め、
透析処理で脱塩後凍結乾燥を行えば目的とする
WAI−65の白色無定形凍結乾燥物が得られる。 Collect the segments containing the obtained 0.65 fraction,
The target can be achieved by desalting through dialysis treatment and freeze-drying.
A white amorphous freeze-dried product of WAI-65 is obtained.
このようにして得られたWAI−65物質の物理
化学的な性質は次のとおりである。 The physicochemical properties of the WAI-65 substance thus obtained are as follows.
(1) 水または希薄塩溶液に可溶、メタノール、エ
タノール、アセトン、クロロホルムおよびヘキ
サンに不溶。(1) Soluble in water or dilute salt solutions, insoluble in methanol, ethanol, acetone, chloroform and hexane.
(2) 紫外部吸収 １％水溶液（室温）
λnax 278nm
Ｅ278nm
１％１cm＝7.19
(3) 分子量セフアデツクスＧ−75を用いたゲル
過法により24000
(4) Daris氏等の方法〔「Annals New York
Academy of Science」121，404（1964）参照〕
によるポリアクリルアミド電気泳動における泳
動度は0.65で単一なバンドを示す。(2) Ultraviolet absorption 1% aqueous solution (room temperature) λ nax 278nm E278nm 1% 1cm = 7.19 (3) Molecular weight 24000 by gel filtration method using Cephadex G-75 (4) Daris et al.'s method [Annals New York
Academy of Science” 121 , 404 (1964)]
The mobility in polyacrylamide electrophoresis is 0.65, showing a single band.
(5) Oath氏等の方法〔「Cereal Chemistry」50，
190〜７（1973）参照〕によるSDSポリアクリル
アミド電気泳動は分子量12500の位置に単一な
バンドを与える。これは本物質が二つの同一な
サブユニツトからなることを示す。(5) Oath et al.'s method ["Cereal Chemistry" 50 ,
190-7 (1973)] gives a single band at a molecular weight of 12,500. This indicates that the substance consists of two identical subunits.
(6) エドマン法によるＮ末端アミノ酸配列解析の
結果によればこの物質はSer−Gly−Gluなるア
ミノ酸配列をなす。(6) According to the results of N-terminal amino acid sequence analysis using the Edman method, this substance has the amino acid sequence Ser-Gly-Glu.
(7) カルボキシペプチダーゼを用いたＣ末端アミ
ノ酸解析ではこの物質はSerを与える。(7) This substance gives Ser in C-terminal amino acid analysis using carboxypeptidase.
(8) トリプシン消化による得られるペプチドは11
個でそれらのＮ末端は次のとおりである。 Ｎ−末端アミノ酸
ペプチドフラグメント
数
AspまたはCysH ３
Glu ２
LysまたはIleu ２
Ser、Ala、Val、Met 各１
(9) アミノ酸組成はWAI−65が二つの同じサブ
ユニツトよりなり、一つのサブユニツトにアス
パラギン酸が８残基含まれるとすると次のとお
りである。(8) The number of peptides obtained by trypsin digestion is 11.
Their N-termini are: Number of N-terminal amino acid peptide fragments Asp or CysH 3 Glu 2 Lys or Ileu 2 Ser, Ala, Val, Met 1 each (9) As for the amino acid composition, WAI-65 consists of two identical subunits, with aspartic acid in one subunit. Assuming that 8 residues are included, they are as follows.
Cys 10.4 Glu 18.1 Val 5.9
Asp 8.0 Pro 11.5 Met 4.7
Thr 7.3 Gly 7.1 Ileu 4.7
Ser 6.3 Ala 6.4 Leu 10.3
Tyr 4.9 Lys 2.3 Arg 7.1
Phe 2.1 His 1.0 Trp
(10) ヒト唾液腺および膵臓アミラーゼの本発明の
物質との阻害曲線において単位あたりの各酵素
を50％阻害する本発明の物質の所要量比は約
（１：500〜１：600）である（添付図面参照）。Cys 10.4 Glu 18.1 Val 5.9 Asp 8.0 Pro 11.5 Met 4.7 Thr 7.3 Gly 7.1 Ileu 4.7 Ser 6.3 Ala 6.4 Leu 10.3 Tyr 4.9 Lys 2.3 Arg 7.1 Phe 2.1 His 1.0 Trp (10) Combination of human salivary gland and pancreatic amylase with the substance of the present invention The required ratio of amounts of the substances of the invention to inhibit 50% of each enzyme per unit in the inhibition curve is approximately (1:500 to 1:600) (see attached drawings).
本発明のα−アミラーゼ阻害物質（WAI−65）
はヒトの唾液腺α−アミラーゼに対して極めて特
異的に阻害作用を示し、一方ヒトの膵臓α−アミ
ラーゼに対する阻害作用は極めて微弱である。更
に本発明の物質は広域な酵素濃度範囲で唾液腺型
α−アミラーゼおよび膵臓型α−アミラーゼアイ
ソザイムに対する阻害比が大きな値をとりうるの
でヒトの体液をはじめとした各種臨床検体のα−
アミラーゼアイソザイムの分別定量のための優れ
た手段となりうるものである。 α-amylase inhibitor of the present invention (WAI-65)
shows a very specific inhibitory effect on human salivary gland α-amylase, while its inhibitory effect on human pancreatic α-amylase is extremely weak. Furthermore, the substance of the present invention can have a large inhibition ratio against salivary gland α-amylase and pancreatic α-amylase isoenzymes over a wide range of enzyme concentrations, and therefore can inhibit α-amylase from various clinical samples including human body fluids.
This can be an excellent means for differentially quantifying amylase isozymes.
既にオドンネル氏等は、小麦粉中にヒトの唾液
腺型α−アミラーゼを膵臓型のそれよりも強く阻
害するα−アミラーゼ阻害物質を報告し、この物
質を利用した臨床検体中のα−アミラーゼアイソ
ザイムの分別定量の可能性を示している。
〔Clinical Chemistry23，560〜566（1977）〕。しか
しながらオドンネル氏等の報告したα−アミラー
ゼ阻害物質はその電気泳動の易動度において、本
物質が0.65であるのに対して0.20であり、著るし
く相違している。更にまた特開昭57−140727号公
報で開示されているα−アミラーゼ阻害物質
WAI−53（電気泳動度0.53）とも相異することは
明らかである。 O'Donnell et al. have already reported an α-amylase inhibitor in wheat flour that inhibits human salivary gland-type α-amylase more strongly than pancreatic-type α-amylase, and has used this substance to separate α-amylase isozymes in clinical samples. It shows the possibility of quantification.
[Clinical Chemistry 23 , 560-566 (1977)]. However, the electrophoretic mobility of the α-amylase inhibitor reported by Mr. O'Donnell et al. is 0.20, compared to 0.65 for the present substance, which is a significant difference. Furthermore, the α-amylase inhibitor disclosed in JP-A No. 57-140727
It is clear that it is different from WAI-53 (electrophoretic mobility 0.53).
添付図面にWAI−65阻害物質のヒト唾液およ
び膵臓のα−アミラーゼに対する阻害曲線を示
す。図面における各α−アミラーゼの阻害率は次
のようにして測定した。すなわち、小麦抽出物よ
り精製したWAI−65をタンパク質量として０〜
10mg含有する10mMトリス塩酸（PH8.0）、10mM
NaClの溶液１mlより10mlを採り、ヒトスイ由来
又はヒトダ液由来アミラーゼ0.015IU（国際単位）
を含有する10mMトリス塩酸（PH8.0）、10mM
NaClの溶液１mlより10μを採り、ヒトスイ由来
又はヒトダ液由来アミラーゼ0.015IU（国際単位）
を含有する10ｍＭトリス塩酸（PH8.0）、10ｍ
MNaClの溶液30μと混じ、室温で25分間保温し
た。この混液10μをアミラーゼDS試薬（米国ベ
ツクマン社製）の基質溶液250μに加え、37℃、
５分間反応した。残存するアミラーゼ活性は、ア
ボツト・オートアナライザーABA−100を用いて
測定した。 The accompanying drawing shows inhibition curves of WAI-65 inhibitors against human salivary and pancreatic α-amylase. The inhibition rate of each α-amylase in the drawing was measured as follows. In other words, WAI-65 purified from wheat extract has a protein content of 0 to 0.
Contains 10mg of 10mM Tris-HCl (PH8.0), 10mM
Take 10ml from 1ml of NaCl solution and add 0.015IU (international unit) of amylase derived from human water or human octopus fluid.
10mM Tris-HCl (PH8.0) containing 10mM
Take 10 μ from 1 ml of NaCl solution and 0.015 IU (international unit) of amylase derived from human water or human octopus fluid.
10mM Tris-HCl (PH8.0) containing
Mixed with 30μ of MNaCl solution and incubated at room temperature for 25 minutes. Add 10μ of this mixture to 250μ of amylase DS reagent (manufactured by Beckman, USA) substrate solution, and heat at 37°C.
It reacted for 5 minutes. Residual amylase activity was measured using Abbott Autoanalyzer ABA-100.
そして図面では、ダ液及びスイ液アミラーゼが
横軸に示すWAI−65の各々の量（μｇ）によつ
て阻害される割合を百分率で示してある。この図
面において等単位量のダ液アミラーゼとスイ液ア
ミラーゼを50％阻害するに要するWAI−65の量
はそれぞれ3.1μｇおよび1500〜1900μｇである。
なおこれらの数値はそれぞれの曲線（スイ液アミ
ラーゼに於いては曲線の延長線）が50％阻害レベ
ルと交わる点から算出することができる。従つ
て、各アミラーゼを50％阻害するWAI−65の所
要量の比は3.1：1500〜3.1：1900≒１：500〜
１：600となる。 In the drawing, the percentage inhibition of Da and Sox amylase by each amount (μg) of WAI-65 shown on the horizontal axis is shown. In this figure, the amounts of WAI-65 required to inhibit 50% of equal unit amounts of dairy liquor amylase and water liquor amylase are 3.1 μg and 1500 to 1900 μg, respectively.
Note that these values can be calculated from the point where each curve (in the case of water syrup amylase, the extension line of the curve) intersects with the 50% inhibition level. Therefore, the ratio of the amount of WAI-65 required to inhibit each amylase by 50% is 3.1:1500~3.1:1900≒1:500~
It will be 1:600.
また、図面より計算されるＰ／Ｓ値（単位当り
のアミラーゼアイソザイムの酵素活性を50％阻害
する阻害剤の所要量比）は外挿法によれば548で
あつた。一方WAI−53阻害物質は250であり、従
つてこのことからみても本発明のWAI−65阻害
物質はWAI−53阻害物質とは相異するものであ
り、WAI−65阻害物質はヒトα−アミラーゼの
分別定量に適した生化学試薬で優れた臨床検査薬
となり得るものと期待される。 Further, the P/S value (required amount ratio of inhibitor to inhibit 50% of amylase isozyme enzyme activity per unit) calculated from the drawing was 548 by extrapolation. On the other hand, the WAI-53 inhibitor is 250, and therefore, from this point of view, the WAI-65 inhibitor of the present invention is different from the WAI-53 inhibitor, and the WAI-65 inhibitor is human α- It is a biochemical reagent suitable for the differential quantification of amylase and is expected to be an excellent clinical diagnostic agent.
以下に本発明のα−アミラーゼ阻害物質WAI
−65物質の製造法を実施例により詳述する。 The α-amylase inhibitor WAI of the present invention is described below.
The method for producing the -65 substance will be explained in detail with reference to Examples.
実施例
小麦粉25Kgに精製水100を加えて１時間ゆる
やかに撹拌し、遠心分離法により上清73.4を得
た。得られた上清液を凍結乾燥して1.13Kgの凍結
乾燥物を得た。これに精製水を加えて液量を13.5
となし、この水溶液を70℃で30分間加熱処理
し、生ずる沈殿を遠心分離で除いて上清13.0を
得た。得られた上清液に４℃の温度下に99％エタ
ノールを加えてエタノール濃度を60％に保ち、生
ずる沈殿を遠心分離で除いた。ひき続いて更に99
％エタノールを加えてエタノール濃度が90％にな
るようにし、１昼夜４℃で放置して沈殿を形成さ
せた。生成した沈殿を遠心分離で集め、精製水に
溶解させてWAI−65を含む水溶液1.8を得た。Example 100 kg of purified water was added to 25 kg of wheat flour, gently stirred for 1 hour, and 73.4 kg of supernatant was obtained by centrifugation. The obtained supernatant liquid was freeze-dried to obtain 1.13 kg of freeze-dried product. Add purified water to this to make the liquid volume 13.5
This aqueous solution was heated at 70° C. for 30 minutes, and the resulting precipitate was removed by centrifugation to obtain a supernatant 13.0. 99% ethanol was added to the obtained supernatant at a temperature of 4°C to maintain the ethanol concentration at 60%, and the resulting precipitate was removed by centrifugation. Then another 99
% ethanol was added to make the ethanol concentration 90%, and the mixture was allowed to stand at 4°C for one day and night to form a precipitate. The generated precipitate was collected by centrifugation and dissolved in purified water to obtain an aqueous solution 1.8 containing WAI-65.
次に2mM塩化ナトリウムを含む10mMトリス
塩酸緩衝液（PH9.2）を溶出液としてセフアデツ
クスＧ−75ゲル過クロマト処理を行ない、
WAI−65を含む画分を集めた。得られたWAI−
53区分を水溶液の形で透析膜にて脱塩後、CM−
セフアロースCL−6Bを担体とし20〜0.5Mの塩化
ナトリウムを含む30mM酢酸緩衝液（PH4.5）を
用いて濃度勾配クロマトグラフイーを実施した。
溶出されたWAI−53区分を集め、透析処理を行
つて脱塩し、凍結乾燥を行つてWAI−65の粗乾
固物0.377ｇを得た。 Next, Sephadex G-75 gel perchromatography was performed using 10mM Tris-HCl buffer (PH9.2) containing 2mM sodium chloride as the eluent.
Fractions containing WAI-65 were collected. Obtained WAI−
After desalting 53 sections in the form of an aqueous solution using a dialysis membrane, CM-
Concentration gradient chromatography was performed using Sepharose CL-6B as a carrier and 30 mM acetate buffer (PH4.5) containing 20 to 0.5 M sodium chloride.
The eluted WAI-53 fraction was collected, subjected to dialysis treatment to desalt, and freeze-dried to obtain 0.377 g of a crude dried product of WAI-65.
WAI−65の純度を更に高めるためにCM−セフ
アロースCL−6B11（商標名）クロマト処理を同
じ条件で繰り返して得られたWAI−65を含む画
分を集め、透析処理を行つて脱塩し、凍結乾燥し
てWAI−65の乾燥物56mgを得た。 In order to further increase the purity of WAI-65, the fractions containing WAI-65 obtained by repeating CM-Sepharose CL-6B11 (trade name) chromatography under the same conditions were collected and desalted by dialysis treatment. Freeze-drying yielded 56 mg of dried WAI-65.
添付図面はWAI−65物質のヒト唾液および膵
臓アミラーゼに対する阻害曲線を示す図である。
The accompanying drawing shows inhibition curves of WAI-65 substance against human saliva and pancreatic amylase.
Claims (1)
タノール、アセトン、クロロホルムおよびヘキ
サンに不溶であること、 (2) 紫外部吸収 １％水溶液（室温） λnax 278nm Ｅ278nm １％１cm 7.19 (3) 分子量 セフアデツクスＧ−75を用いたゲル
過法により24000 (4) Davis氏等の方法によるポリアクリルアミド
電気泳動における泳動度は0.65で単一なバンド
を示す、 (5) Oath氏らの方法によるSDSポリアクリルア
ミド電気泳動は分子量12500の位置に単一なバ
ンドを与え、これは本物質が二つの同一なサブ
ユニツトからなることを示す、 (6) エドマン法によるＮ末端アミノ酸配列解析の
結果によればこの物質はセリン−グリシン−グ
ルタミン酸なるＮ末端アミノ酸配列を示す、 (7) カルボキシペプチターゼを用いたＣ末端アミ
ノ酸解析ではこの物質はセリンを与える、 (8) トリプシン消化による得られるペプチドは11
個でそれらのＮ末端は次のとおりである Ｎ−末端アミノ酸 ペプチドフラグメント数 アスパラギン酸またはシステイン ３ グルタミン酸 ２ リシンまたはイソロイシン ２ セリン、アラニン、バリン、メチオニン 各１ ９ アミノ酸組成はWAI−65が二つの同じサブ
ユニツトよりなり、一つのサブユニツトにアス
パラギン酸が８残基含まれるとすると次のとお
りである シスチン 10.4 グルタミン酸 18.1 バリン 5.9 アスパラギン酸 8.0 プロリン 11.5 メチオニン 4.7 スレオニン 7.3 グリシン 7.1 イソロイシン 4.7 セリン 6.3 アラニン 6.4 ロイシン 10.3 チロシン 4.9 リジン 2.3 アルギニン 7.1 フエニルアラニン 2.1 ヒスチジン 1.0 (10) ヒト唾液腺および膵臓アミラーゼに対する本
物質の阻害曲線において単位あたりの各酵素を
50％阻害する本物質の所要量比は１：500〜
１：600である を有することを特徴とする、小麦由来の新規なα
−アミラーゼ阻害物質。[Claims] 1 Physical and chemical properties (1) Soluble in water or dilute salt solution, insoluble in methanol, ethanol, acetone, chloroform and hexane, (2) Ultraviolet absorption 1% aqueous solution (room temperature) λ nax 278nm E278nm 1%1cm 7.19 (3) Molecular weight 24000 by gel filtration using Sephadex G-75 (4) The mobility in polyacrylamide electrophoresis by the method of Davis et al. is 0.65 and shows a single band. 5) SDS polyacrylamide electrophoresis using the method of Oath et al. gave a single band at a molecular weight of 12,500, indicating that this substance consists of two identical subunits. (6) N-terminal analysis using the Edman method According to the results of amino acid sequence analysis, this substance has an N-terminal amino acid sequence of serine-glycine-glutamic acid. (7) C-terminal amino acid analysis using carboxypeptidase reveals that this substance yields serine. (8) Trypsin digestion The resulting peptide is 11
Their N -terminal amino acids are as follows: Aspartic acid or cysteine 3 Glutamic acid 2 Lysine or isoleucine 2 Serine, alanine, valine, methionine 1 each 9 Amino acid composition WAI-65 has the same two Assuming that each subunit contains 8 aspartic acid residues, the following are cystine 10.4 glutamic acid 18.1 valine 5.9 aspartic acid 8.0 proline 11.5 methionine 4.7 threonine 7.3 glycine 7.1 isoleucine 4.7 serine 6.3 alanine 6.4 leucine 10.3 tyrosine 4.9 Lysine 2.3 Arginine 7.1 Phenylalanine 2.1 Histidine 1.0 (10) In the inhibition curve of this substance against human salivary gland and pancreatic amylase, each enzyme per unit
The required amount ratio of this substance to inhibit 50% is 1:500 ~
A novel α derived from wheat characterized by having a ratio of 1:600.
-Amylase inhibitors.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP58110124A JPS604132A (en) | 1983-06-21 | 1983-06-21 | Novel alpha-amylase inhibitory substance |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP58110124A JPS604132A (en) | 1983-06-21 | 1983-06-21 | Novel alpha-amylase inhibitory substance |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS604132A JPS604132A (en) | 1985-01-10 |
| JPH0427997B2 true JPH0427997B2 (en) | 1992-05-13 |
Family
ID=14527631
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP58110124A Granted JPS604132A (en) | 1983-06-21 | 1983-06-21 | Novel alpha-amylase inhibitory substance |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS604132A (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2620695B1 (en) * | 1987-09-21 | 1990-01-05 | Rhone Poulenc Chimie | LIQUID-LIQUID EXTRACTION GALLIUM RECOVERY PROCESS |
| JP2757404B2 (en) * | 1988-12-09 | 1998-05-25 | 日清製粉株式会社 | Method for obtaining α-amylase inhibitor-containing substance from wheat |
| JP2757405B2 (en) * | 1988-12-09 | 1998-05-25 | 日清製粉株式会社 | Diet preparation containing α-amylase inhibitor obtained from wheat |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5885899A (en) * | 1981-11-16 | 1983-05-23 | Fujirebio Inc | Amylase inhibitor and its preparation |
-
1983
- 1983-06-21 JP JP58110124A patent/JPS604132A/en active Granted
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5885899A (en) * | 1981-11-16 | 1983-05-23 | Fujirebio Inc | Amylase inhibitor and its preparation |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS604132A (en) | 1985-01-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Oppenheim et al. | The primary structure and functional characterization of the neutral histidine-rich polypeptide from human parotid secretion. | |
| CA2050300C (en) | Tnf-inhibiting proteins and the preparation thereof | |
| Walker et al. | Primary structure and subunit stoichiometry of F1-ATPase from bovine mitochondria | |
| US5849560A (en) | Proteases causing degradation of amyloid β-protein precursor | |
| US4760130A (en) | Serine protease inhibitors and methods for isolation of same | |
| EP0546084A1 (en) | PROTEASES CAUSING ABNORMAL DEGRADATION OF AMYLOID $g(b)-PROTEIN PRECURSOR | |
| JPH01206992A (en) | Basic protein, its derivative, fragment and use thereof | |
| AU591474B2 (en) | Serine protease inhibitors and methods for isolation of same | |
| David et al. | Calpain II induced insolubilization of lens β-crystallin polypeptides may induce cataract | |
| Laine et al. | Primary structure and microheterogeneities of rat chloroleukemia histone H2A (histone ALK, IIbl, or F2a2) | |
| US4806626A (en) | α-amylase inhibitor | |
| JPH0427997B2 (en) | ||
| Biedermann et al. | The amino acid sequence of proteinase A inhibitor 3 from baker's yeast | |
| Maruyama et al. | Purification and amino acid analysis of plasma acid stable trypsin inhibitor | |
| JPS5885899A (en) | Amylase inhibitor and its preparation | |
| Škrtić et al. | Methionine-preferring broad specificity aminopeptidase from chicken egg-white | |
| JPH10298199A (en) | New peptide and angiotensin i converting enzyme inhibitor containing the peptide | |
| JPH06220088A (en) | Tripeptide inhibiting angiotensin i converting enzyme, its production and food containing the tripeptide | |
| Chakrabarti et al. | Purification and characterization of a low molecular weight basic protein from marine turtle egg white | |
| JPS62501291A (en) | Purified serine protease inhibitor and pharmaceutical composition containing same | |
| JPH0543718B2 (en) | ||
| JP2714430B2 (en) | Novel peptide having Na, K-ATPase inhibitory action | |
| KR0122430B1 (en) | Method for assay of aminopeptidase by use of synthetic oligopeptide | |
| JPH03112999A (en) | Physiologically active protein of momordica charantia | |
| JPH0291100A (en) | Protein inhibiting human phospholipase a2 |