JPH0829084B2 - Method for producing enzyme for degrading fish protein - Google Patents
Method for producing enzyme for degrading fish proteinInfo
- Publication number
- JPH0829084B2 JPH0829084B2 JP61296626A JP29662686A JPH0829084B2 JP H0829084 B2 JPH0829084 B2 JP H0829084B2 JP 61296626 A JP61296626 A JP 61296626A JP 29662686 A JP29662686 A JP 29662686A JP H0829084 B2 JPH0829084 B2 JP H0829084B2
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- present
- fish protein
- protease
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Description
【発明の詳細な説明】 （１）産業上の利用分野 本発明は、新規微生物（アスペルギルス・ソエーS 29
7）を利用する魚類蛋白質分解用酵素の製造法に関する
ものである。DETAILED DESCRIPTION OF THE INVENTION (1) Field of Industrial Application The present invention is directed to a novel microorganism (Aspergillus soe S 29
The present invention relates to a method for producing an enzyme for proteolytic decomposition of fish using 7).
カツオやマグロ等の缶詰製造時に副生する煮汁は年間
約10万トンあり、この煮汁は蛋白質を多量に含んでいる
ため、プロテアーゼ処理、その他種々加工されて、微生
物培地成分や調味料の素材として利用されている。There are about 100,000 tons of broth produced as a by-product during the production of canned bonito, tuna, etc. Since this broth contains a large amount of protein, it is processed with protease and various other processes and used as a material for microbial medium components and seasonings. It's being used.
しかしながら、魚特有の異臭を消し、旨味・風味の豊
かな加工品を工業的有利に製造することは非常に困難で
あるため、実際に利用されている煮汁が約60％に止ま
り、残りの40％は廃棄されているのが現状である。However, since it is very difficult to industrially produce processed products with rich umami and flavor by eliminating the offensive odor peculiar to fish, the actual broth used is only about 60%, and the remaining 40 % Is currently abandoned.
本発明は、これら魚類蛋白質を処理するに適した酵素
を製造する方法に関するものであり、本発明の酵素は従
来市販されているいづれの酵素と比較してもはるかに高
い処理効果を示すものである。The present invention relates to a method for producing an enzyme suitable for treating these fish proteins, and the enzyme of the present invention exhibits a much higher treatment effect than any of the commercially available enzymes. is there.
（２）従来の技術 魚類蛋白質を蛋白質源として酵素で加水分解し飲料、
調味料および治療食等の素材を得る例は古くから見られ
る（東海水研報第43巻、87−90頁、1965年；東海水研報
第73巻、103−112頁、1973年）。新しくは、市販アルカ
リ性プロテアーゼをそれらの活性至適pH域であるpH7.5
−9.0で魚類蛋白質に作用させ、加水分解の程度を15−2
5％の範囲で停止することにより、不快味が少なくしか
も魚臭の少ない加水分解物およびその製造法が発表され
ている（特開昭61−28370号）。(2) Conventional technology Beverages obtained by hydrolyzing fish proteins with enzymes as protein sources
Examples of obtaining ingredients such as seasonings and therapeutic foods have long been seen (Tokai Suiho Kenkyu 43, 87-90, 1965; Tokai Suiho 73, 103-112, 1973). Newly, commercially available alkaline proteases are added to their activity optimum pH range of pH 7.5.
It acts on fish protein at -9.0, and the degree of hydrolysis is 15-2.
A hydrolyzate having less unpleasant taste and less fishy odor and a method for producing the same have been announced by stopping in the range of 5% (JP-A-61-28370).
また、藤巻正生等はストレプトマイセス・グリセウス
の産生する酵素、アクチナーゼが魚類蛋白質の分解に適
していることを報告している。（アグリカルチュラル・
アンド・バイオロジカル・ケミストリー第37巻、2891−
2898頁、1973年） しかしながら、これら従来の方法は、かならずしも満
足出来るものではない。Also, Masaaki Fujimaki et al. Reported that actinase, an enzyme produced by Streptomyces griseus, is suitable for degrading fish proteins. (Agricultural
And Biological Chemistry Vol. 37, 2891-
(Page 2898, 1973) However, these conventional methods are not always satisfactory.
（３）発明が解決しようとする問題点 本発明はカツオやマグロ等の缶詰製造時あるいはサバ
やイワシの加工時に多量副生する煮汁等に含まれる魚類
蛋白質を処理し、魚特有の異臭を消し、旨味や風味の優
れた加工食品にするために使用する酵素が市販品ではか
なずしも満足出来ないので、この目的に合致する新規な
秀れた酵素を創製することを目的とするものである。(3) Problems to be Solved by the Invention The present invention treats fish proteins contained in broth and the like that are by-produced in large amounts during canning production of skipjack, tuna, etc. or during processing of mackerel and sardines to eliminate fish-specific off-odor. Since the enzyme used to make processed food with excellent umami and flavor is not commercially available, it is aimed to create a new excellent enzyme that meets this purpose. is there.
本発明者等は、独自に分離した種々のプロテアーゼ産
生菌を培養して得た酵素と市販のプロテアーゼについ
て、魚類蛋白質分解効果を比較し、本発明者等が分離し
た菌株アスペルギルス・ソエーS 297の産生するプロテ
アーゼが最も秀れていることを見出し、本発明を完成し
た。The inventors of the present invention, comparing the enzyme obtained by culturing various protease-producing bacteria independently isolated and commercially available protease, comparing the proteolytic effect of fish, the strain of Aspergillus soe S 297 isolated by the present inventors The inventors have found that the protease produced is the most excellent and completed the present invention.
（４）発明の構成 本発明は、「アスペルギルス・ソエー（Aspergillus
sojae）S297（微工研菌寄第9073号）又はその変異株を
培養し培地中にプロテアーゼを生成蓄積させ、これを採
取することを特徴とする魚類蛋白質の分解用酵素の製造
法。」に関するものである。(4) Structure of the Invention The present invention relates to “Aspergillus
A method for producing an enzyme for degrading fish protein, which comprises culturing Sjae) S297 (Microtechnology Research Institute No. 9073) or a mutant strain thereof to produce and accumulate a protease in a medium, and collecting the protease. It is about ".
本発明に係る微生物、アスペルギルス・ソエーS 297
は本発明者等が市販のショウユ用種麹から分離した新規
菌株であり、後述する通り、魚類蛋白質分解用酵素とし
て非常にすぐれたプロテアーゼを産生し、分生子柄が極
めて短かく、豊富な分生子頭を速やかに形成する点で他
のアスペルギルス・ソエーと区別される。Microorganisms according to the invention, Aspergillus soe S 297
Is a novel strain isolated by the present inventors from commercially available seed koji for Shoyu, and as described below, produces a very excellent protease as an enzyme for proteolysis of fish, and the conidia stalk is extremely short and rich in content. Distinguished from other Aspergillus soe by the rapid formation of the natal head.
以下、アスペルギルス・ソエーS 297の菌学的性状を
ツアペック寒天平板上、25℃で生育させて観察した結果
に基いて説明する。Hereinafter, the mycological properties of Aspergillus soe S 297 will be described based on the results of observation on Tuapeck agar plates grown at 25 ° C.
（生育） ツアペック寒天平板上では速く広がりコロニー直径は
7cm前後に発達する、基底菌糸層は白色で密、分生子柄
は極めて短かく、中心部はほとんど羊毛状に盛り上がる
ことがなく、表面は平たんである。分生子頭と分生子の
形成は特に良好で、集落は若い時期が輝黄色〜黄色、後
に黄緑色〜緑色、成熟すると黄褐色、古くなると黄緑色
を若干含む褐色となる。集落表面はクリーム色〜明淡褐
色。(Growth) It spreads quickly on Tuapec agar plates and the colony diameter is
Approximately 7 cm, basal hyphae are white and dense, conidia peduncle is very short, center does not rise like wool, and surface is flat. The formation of conidia and conidia is particularly good, and the colony is bright yellow to yellow in the young period, yellowish green to green later, yellowish brown when mature, and brownish with some yellowish green when old. The surface of the village is cream to light brown.
（形態） 分生子頭：直径350〜400μｍ、若いと放射状、成熟す
るとゆるい放射状、黄緑色〜褐色。(Morphology) Conidial head: Diameter 350-400 μm, radial when young, loose radial when mature, yellow-green to brown.
分生子柄：長さ200〜500μｍ、直径は頂のう下部で10
〜13μｍ、基底菌糸近くが４〜７μｍ、壁表面は平滑で
あり、頂のうの下部附近だけが粗面。Conidia peduncle: length 200-500 μm, diameter 10 at the bottom of the apex
~ 13μm, 4 ~ 7μm near the basal hypha, the wall surface is smooth, only the lower part of the apex is rough.
頂のう：直径20〜22μｍ、亜球形。 The apex: 20-22 μm in diameter, subspherical.
フィアライド：頂のうのほぼ前面より形成され、長さ
（７〜９μｍ）×幅（３〜４μｍ） 分生子：直径５〜6.3μｍ、球形〜亜球形、表面が刺
状（coarsely echinulate）。Phialide: Formed almost from the anterior surface of the apex, length (7 to 9 μm) × width (3 to 4 μm) Conidia: diameter 5 to 6.3 μm, spherical to subspherical shape, coarsely echinulate.
菌核及びメトラ：形成されない。 Sclerotium and metola: Not formed.
その他:p−アニスアルデヒド含有ツアペック寒天平板
上でピンク色素の形成能無し。コウジ酸を産生する。ア
フラトキシンB1,B2,G1,G2を産生しない。Others: No ability to form pink dye on Tuapec agar plates containing p-anisaldehyde. Produces kojic acid. Does not produce the aflatoxins B 1 , B 2 , G 1 and G 2 .
以上の菌学的性状をケイ・ビー・ラパー・アンド・デ
ィー・アイ・フェンネル（K.B.Raper ＆ D.I.Fennell）
著、ザ・ジーナス・アスペルギルス（The Genus Asperg
illus）、375−403頁、1965年；エス・ウシジマ（S.Ush
ijima）等、アグリカルチュラル・アンド・バイオロジ
カル・ケミスロリー（Agricultural and Biological Ch
emistry）第46巻、2365−2367頁、1982年；エッチ・ム
ラカミ（H.Murakami）等、ジャーナル・オブ・ジェネラ
ル・アンド・アプライド・ミクロバイオロジー（Journa
l of General and Applied Microbiology）第28巻、55
−60頁、1982年の文献に基づいて検討し、本発明のS 29
7株はアスペルギルス・ソエー（Aspergillus sojae）に
属する菌株であることを確認した。The above bacteriological properties are described by KB Raper & DIFennell
By The Genus Asperg
illus), pages 375-403, 1965; S. Ush
ijima) etc., Agricultural and Biological Ch
emistry) Vol. 46, pp. 2365-2367, 1982; H. Murakami et al., Journal of General and Applied Microbiology (Journa).
l of General and Applied Microbiology) 28, 55
-60 pages, reviewed based on the literature of 1982, S 29 of the present invention
It was confirmed that 7 strains belong to Aspergillus sojae.
本発明に利用出来る微生物は、工業技術院微生物工業
技術研究所に微工研菌寄第9073号として寄託されている
アスペルギルス・ソエーS 297菌株のみに止まらず、そ
の自然的又は人工的変異株も当然包含される。Microorganisms that can be used in the present invention are not limited to Aspergillus soe S 297 strain that has been deposited in the Institute of Microbial Science and Technology of the Institute of Industrial Science and Technology as Micromachine Research Institute No. 9073, as well as its natural or artificial mutants. Naturally included.
本発明に於ける培養は、液体培養でもよいが、一般的
には、好気的固体培養の方が好ましい。The culture in the present invention may be liquid culture, but in general, aerobic solid culture is preferable.
通常、本発明の菌株S 297は小麦▲麩▼等の固体培地
に接種し、40〜55時間好気的に培養するとプロテアーゼ
を培地（麹）を分泌する。培養後、麹を水に懸濁するこ
とによりプロテアーゼは容易に水層中い移行し、不溶物
を除去することにより粗酵素液を得ることが出来る。Usually, the strain S297 of the present invention secretes a protease (medium) when inoculated into a solid medium such as wheat bran and aerobically cultured for 40 to 55 hours. After culturing, by suspending koji in water, the protease easily migrates into the aqueous layer, and insoluble matter can be removed to obtain a crude enzyme solution.
この粗酵素液から、公知の酵素精製法により、精製プ
ロテアーゼを得ることが出来る。From this crude enzyme solution, a purified protease can be obtained by a known enzyme purification method.
以下、実施例により本発明の実施態様を詳細に説明す
る。Hereinafter, embodiments of the present invention will be described in detail with reference to Examples.
実施例 ▲麩▼4.8kg、KH2PO4320g、Ｌ−グルタミン酸ナトリ
ウム160g、水8lからなる固体培地を121℃、１気圧下20
分間滅菌した後、アスペルギルス・ソエーS 297の分生
子を植菌し、30℃で２日間通風培養し、麹6.9kgを得
た。これを水60l中に懸濁させた充分攪拌後遠心分離（9
000rpm）し、上清液55l（酵素力価220U/ml）を得た。次
いで、この液をホロファイバーシステム（小松川化工機
製）により分離膜PM1000を通過させた後、分離膜PM5で
濃縮し、精製回収した。Example ▲ A solid medium consisting of 4.8 kg of wheat, 320 g of KH 2 PO 4 , 160 g of sodium L-glutamate, and 8 liters of water was added at 121 ° C. under 1 atmosphere of 20.
After sterilizing for a minute, conidia of Aspergillus soe S297 were inoculated and aerobically cultured at 30 ° C. for 2 days to obtain 6.9 kg of koji. This was suspended in 60 liters of water and stirred thoroughly, followed by centrifugation (9
000 rpm) to obtain 55 l of supernatant (enzyme titer 220 U / ml). Next, this liquid was passed through a separation membrane PM1000 by a hollow fiber system (manufactured by Komatsugawa Kakoki Co., Ltd.), then concentrated with a separation membrane PM5, and purified and recovered.
得られた濃縮液は4lで、280nmでの吸光度が26であっ
た。The resulting concentrated liquid had a volume of 41 and an absorbance at 280 nm of 26.
この液に安定剤として420gのデキストリン（日本資糧
工業社製NSD1318）を加えた後、凍結乾燥を行い、422g
の酵素粉末を得た。After adding 420 g of dextrin (NSD1318 manufactured by Nippon Shigyo Co., Ltd.) as a stabilizer to this solution, lyophilization was performed to obtain 422 g.
The enzyme powder of was obtained.
酵素力価は24000U/gであり、回収率は83.7％であっ
た。The enzyme titer was 24000 U / g, and the recovery rate was 83.7%.
この酵素粉末は本発明酵素の標品として以後の試験に
供した。This enzyme powder was used in the subsequent tests as a standard of the enzyme of the present invention.
この標品酵素は、単一のプロテアーゼからなるもので
はなく、多種類のプロテアーゼの混合物から構成されて
いる。このことを明らかにするため、その中に含まれる
比較的強い活性を示したプロテアーゼ６種について、酵
素活性を測定した結果を第１表に示す。This standard enzyme does not consist of a single protease, but is composed of a mixture of many kinds of proteases. In order to clarify this, Table 1 shows the results of measuring the enzyme activity of 6 kinds of proteases contained therein, which showed a relatively strong activity.
＊文献 :Agricultural and Biological Chemistry 37,3685
（1973）． :Method in Enzymology,G.E.Perlmann and L. Loran
d,Academic Press,18,397（1970）． :Biochemica et Biophysica Acta 397,443（1975）． ：和光純薬工業（株）製LAPC−Test Wako使用の活性
測定法。 * Reference: Agricultural and Biological Chemistry 37 , 3685
(1973). : Method in Enzymology, GE Perlmann and L. Loran
d, Academic Press, 18 , 397 (1970). : Biochemica et Biophysica Acta 397 , 443 (1975). : Activity measurement method using LAPC-Test Wako manufactured by Wako Pure Chemical Industries, Ltd.
本発明酵素は第１表に示すように多種類の酵素を含ん
でおり、それら個々の酵素を分離して使用することも出
来るが、通常、混合物のままで使用して好適な結果を得
ることが出来る。The enzyme of the present invention contains various kinds of enzymes as shown in Table 1, and the individual enzymes can be separated and used, but usually, the mixture is used as it is to obtain suitable results. Can be done.
本発明酵素により魚類蛋白質を分解処理する場合、酵
素は不活性担体に固定化して使用することも出来る。When a fish protein is decomposed by the enzyme of the present invention, the enzyme may be immobilized on an inert carrier before use.
この分解処理は50〜60℃で行なわれる場合が多いが、
本発明酵素は、市販酵素に比較して耐熱性が秀れている
ので実際の使用場面で好適な処理効果を得ることが出来
る。This decomposition process is often performed at 50-60 ° C,
Since the enzyme of the present invention has excellent heat resistance as compared with the commercially available enzyme, it is possible to obtain a suitable treatment effect in actual use.
次に市販の代表的プロテアーゼと本発明酵素の耐熱性
試験結果を第２表に示す。Next, Table 2 shows the results of heat resistance test of typical commercially available proteases and the enzyme of the present invention.
＊残存活性率は、各供試酵素について、その中に含まれ
る表記３種の酵素活性を次の方法により測定して求め
た。 * The residual activity rate was determined by measuring the enzyme activity of the three kinds of notation contained in each of the test enzymes by the following method.
（イ）中性及びアルカリ性プロテイナーゼの活性は20mM
リン酸バッファー（pH7.0）中で酵素を60℃、10分間処
理して残存する活性をアンソン−荻原改変法で測定し
た。(A) The activity of neutral and alkaline proteinases is 20 mM
The enzyme was treated in a phosphate buffer (pH 7.0) at 60 ° C. for 10 minutes, and the remaining activity was measured by the Anson-Ogihara modification method.
（ロ）カルボキシペプチダーゼの活性は20mMリン酸化バ
ッファー（pH6.0）中で酵素を50℃、10分間処理して残
存する活性を中田等の方法〔Agricultural and Biologi
cal Chemistry,36,1343（1972）〕に準じ基質カルボベ
ンゾキシ−グルタミン−チロシンを使用して測定した。(B) The activity of carboxypeptidase was treated with the enzyme in 20 mM phosphorylation buffer (pH 6.0) at 50 ° C for 10 minutes and the remaining activity was determined by the method of Nakata et al. [Agricultural and Biologi
cal Chemistry, 36 , 1343 (1972)], using the substrate carbobenzoxy-glutamine-tyrosine.
（ハ）アミノペプチターゼの活性は20mMリン酸バッファ
ー（pH7.0）中で酵素を60℃、10分間処理して残存する
活性をLAP C−Test Wako（和光純薬製）を用いて測定し
た。(C) The activity of aminopeptidase was treated with the enzyme in 20 mM phosphate buffer (pH 7.0) at 60 ° C for 10 minutes, and the remaining activity was measured using LAP C-Test Wako (manufactured by Wako Pure Chemical Industries). .
第２表の成績から明らかなように、本発明酵素は、魚
類蛋白質の分解に直接関与すると思われる中性及びアル
カリ性プロテイナーゼとアルボキシペプチダーゼの耐熱
性が市販酵素に比較して著しく優れていた。As is clear from the results shown in Table 2, the enzyme of the present invention was remarkably superior to the commercially available enzyme in heat resistance of neutral and alkaline proteinases and arboxypeptidase which are considered to be directly involved in the degradation of fish proteins.
５ 作用効果 本発明酵素の作用効果を説明するため、以下に試験例
を示す。5 Effects In order to explain the effects of the enzyme of the present invention, test examples are shown below.
試験例１ 本発明酵素及び市販酵素で処理して得られる魚類蛋白
質分解産物を味に関する官能検査。Test Example 1 Sensory test for taste of a fish protein degradation product obtained by treating with the enzyme of the present invention and a commercially available enzyme.
試験方法 カルチベーター350（マグロ缶詰製造時に副生する煮
汁エキスの商品名、焼津水産化学工業株式会社製、水分
40％）を基質とし、供試プロテアーゼを20単位/mlの割
合になるように添加し、50℃、３時間加水分解処理し
た。処理液を５分間煮沸した後、水で２倍稀釈したもの
について、60℃で専門家８名により、旨味が強く苦味が
少ないことを総合的に評価して順位法（日科技連官能検
査委員会編、官能検査ハンドブック）により官能検査し
た。Test method cultivator 350 (trade name of broth extract by-produced during canned tuna production, manufactured by Yaizu Suisan Chemical Co., Ltd., moisture
40%) as a substrate, the protease to be tested was added at a rate of 20 units / ml, and hydrolyzed at 50 ° C. for 3 hours. After the treatment solution was boiled for 5 minutes and then diluted twice with water, 8 experts at 60 ° C comprehensively evaluated that it had a strong umami and little bitterness, and ranked it by the rank method (Nikki Giren Sensory Inspection Committee). Sensory test was conducted according to the sensory test handbook edited by the Society.
供試酵素 試験結果 本試験の結果は第３表に示す。Test enzyme Test results The results of this test are shown in Table 3.
第３表に試験結果から明らかなように、本発明酵素で
処理した煮汁エキスは最も良い評点を得ており、いづれ
の市販酵素を用いて処理した煮汁エキスよりも官能的に
優れたものを与えることが判明した。 As is clear from the test results in Table 3, the broth extract treated with the enzyme of the present invention has the highest score, and gives a sensually superior broth extract treated with any of the commercially available enzymes. It has been found.
試験例２ 本発明酵素及び公知のアスペルギルス・ソエー菌産生
のプロテアーゼで処理して得られる魚類蛋白質分解産物
の味に関する官能検査。 Test Example 2 Sensory test for taste of fish protein degradation products obtained by treatment with the enzyme of the present invention and a known protease produced by Aspergillus soe.
試験方法 魚類蛋白質分解産物の調製は試験例１と同様にして行
い、官能検査は専門家９名により供試酵素とプロテアー
ゼＰ（試験例１で、市販酵素の中で最も官能的に好まし
い分解物を与えた酵素）との間で味を比較する２点嗜好
試験法（官能検査ハンドブック参照）で行った。Test method The fish protein degradation product was prepared in the same manner as in Test Example 1, and a sensory test was conducted by 9 experts to test enzyme and Protease P (in Test Example 1, the most functionally preferable degradation product among commercially available enzymes. It was conducted by a two-point preference test method (see sensory test handbook) in which the taste was compared with that of the enzyme that gave the product.
供試酵素 本発明酵素と、比較的耐熱性の良いプロテアーゼを産
生することの知られている食糧研究所保存のアスペルギ
ルス・ソエーに属する菌株８種（保存番号:R1921II,R19
21III,R6110II,R1312V,R6423II,S−12−3,0−14−9II,0
−20−２）を選び前記実施例と同様にしてそれぞれの菌
を培養し供試酵素を調製した。Test enzyme Eight strains belonging to Aspergillus soe preserved by the Food Research Institute known to produce the enzyme of the present invention and a protease having relatively good thermostability (preservation number: R1921II, R19
21III, R6110II, R1312V, R6423II, S-12-3, 0-14-9II, 0
-20-2) was selected and each bacterium was cultured in the same manner as in the above-mentioned Example to prepare the test enzyme.
試験結果 本試験の結果は第５表に示す。Test results The results of this test are shown in Table 5.
第５表の成績から明らかなように、アスペルギルス・
ソエーに属する菌株の産生するプロテアーゼは、市販酵
素の中から最も好ましい結果を得たプロテアーゼＰに比
較して遜色無いものが多かったが、本発明酵素はそれら
の中で抜群の成績を得た。 As you can see from the results in Table 5, Aspergillus
Many of the proteases produced by the strains belonging to Soe were comparable to Protease P, which had the most favorable results, among the commercially available enzymes, but the enzyme of the present invention obtained outstanding results among them.
試験例３ 本発明酵素及び市販酵素の魚類蛋白質分解活性試験 試験方法 カルチベーター350（前記、マグロ煮汁エキス）をア
ンバーライトIR−120B（Ｈ＋形）カラムに通液し、その
中に含まれている遊離のアミノ酸を吸着除去し、A260nm
/A280nm比が1.5以下の画分を集め、pH6.0に調製した後
凍結乾燥して基質として用いる魚蛋白質を調製した。Test Example 3 Fish Proteolytic Activity Test of Enzymes of the Present Invention and Commercial Enzymes Test Method Calciter 350 (above, tuna broth extract) was passed through an Amberlite IR-120B (H + form) column and contained therein. Adsorbs and removes free amino acids present, A260nm
Fractions having a / A280 nm ratio of 1.5 or less were collected, adjusted to pH 6.0, and then lyophilized to prepare a fish protein used as a substrate.
この魚蛋白質5.6％（w/v）水溶液に供試酵素を100U/m
l（アンソン−萩原改変法で求めた力価）の割合で添加
し、50℃、10分間加水分解反応を行い、生成したα−ア
ミノ基をTNBS法（奥山典五，笠井久隆著、蛋白質・核散
・酵素第18巻、1153−1159頁、1973年）に従って測定
し、魚蛋白質分解率を次の式から算出した。To this 5.6% (w / v) fish protein solution, 100 U / m of the test enzyme was added.
It was added at a ratio of 1 (Anson-Hagiwara modification method) and hydrolyzed at 50 ° C for 10 minutes, and the α-amino group produced was subjected to the TNBS method (Okuyama Norigo, Kasai Hisataka, Protein. (Nansan and Enzyme, Vol. 18, 1153-1159, 1973), and the fish protein degradation rate was calculated from the following formula.
（但し、式中〔 〕は〔 〕内の物のα−アミノ基の分
析値を示す。） 試験結果 本試験の結果は第６表に示す。 (However, [] in the formula represents the analysis value of the α-amino group of the substance in [].) Test Results The results of this test are shown in Table 6.
第６表の成績が示すように、本発明酵素は市販の魚蛋
白分解に適した酵素及び代表的植物起源及び動物起源の
プロテアーゼに比較してはるかに高い魚蛋白質分解率を
示した。 As shown by the results in Table 6, the enzyme of the present invention showed a much higher fish protein degradation rate than commercially available enzymes suitable for fish protein degradation and typical plant- and animal-derived proteases.
なお、本明細書に於いて酵素単位は、特記しない限
り、中台らによりアグリカルチュラル・アンド・バイオ
ロジカル・ケミストリー第37巻、2685−2694頁、1973年
に報告されたプロテイナーゼ活性測定法（アンソン−荻
原改変法）により、ミルクカゼイン（メルク社製）を供
試酵素で30℃、pH7.0で分解する条件下、１分間当り１
μmoleのチロシンを遊離させる酵素量で示した。The enzyme unit in the present specification, unless otherwise specified, is a proteinase activity assay method (Annson method reported by Agricultural and Biological Chemistry Vol. 37, 2685-2694, 1973 by Nakadai et al. -Ogihara modification method), under the condition that milk casein (manufactured by Merck & Co., Inc.) is decomposed with a test enzyme at 30 ° C and pH 7.0, 1 per minute.
The amount is shown as the amount of enzyme that liberates tyrosine of μmole.
フロントページの続き (72)発明者 又平 芳春 静岡県島田市東町1510−２ 審査官 冨永 みどり (56)参考文献 特開 昭57−174087（ＪＰ，Ａ) 特開 昭61−242596（ＪＰ，Ａ) 特開 昭62−248485（ＪＰ，Ａ)Front Page Continuation (72) Inventor Yoshiharu Matahira 1510-2 Higashimachi, Shimada City, Shizuoka Prefecture Midori Tominaga (56) References JP 57-174087 (JP, A) JP 61-242596 (JP, A) ) JP-A-62-248485 (JP, A)
Claims (1)
jae）S297（微工研菌寄第9073号）又は、その変異株を
培養することを特徴とする煮汁に含まれる魚類蛋白質の
分解用酵素の製造法。1. Aspergillus soe
jae) A method for producing an enzyme for degrading a fish protein contained in broth, which comprises culturing S297 (Microtechnology Research Institute No. 9073) or a mutant strain thereof.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61296626A JPH0829084B2 (en) | 1986-12-15 | 1986-12-15 | Method for producing enzyme for degrading fish protein |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61296626A JPH0829084B2 (en) | 1986-12-15 | 1986-12-15 | Method for producing enzyme for degrading fish protein |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS63148985A JPS63148985A (en) | 1988-06-21 |
| JPH0829084B2 true JPH0829084B2 (en) | 1996-03-27 |
Family
ID=17835980
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP61296626A Expired - Fee Related JPH0829084B2 (en) | 1986-12-15 | 1986-12-15 | Method for producing enzyme for degrading fish protein |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0829084B2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DK175501B1 (en) | 2002-12-02 | 2004-11-15 | Green Earth | Plant and process for continuous hydrolysis of a proteinaceous animal or vegetable feedstock |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS57174087A (en) * | 1981-04-21 | 1982-10-26 | Higeta Shoyu Kk | Breeding method of koji mold |
| JPS61242596A (en) * | 1985-04-22 | 1986-10-28 | Wako Pure Chem Ind Ltd | Determination of body fluid component of living body |
| JPS62248485A (en) * | 1985-12-27 | 1987-10-29 | Japanese Res & Dev Assoc Bio Reactor Syst Food Ind | Production of protease |
-
1986
- 1986-12-15 JP JP61296626A patent/JPH0829084B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JPS63148985A (en) | 1988-06-21 |
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