JPH01291799A - Production of chitosan-oligosaccharide and use thereof - Google Patents
Production of chitosan-oligosaccharide and use thereofInfo
- Publication number
- JPH01291799A JPH01291799A JP12185288A JP12185288A JPH01291799A JP H01291799 A JPH01291799 A JP H01291799A JP 12185288 A JP12185288 A JP 12185288A JP 12185288 A JP12185288 A JP 12185288A JP H01291799 A JPH01291799 A JP H01291799A
- Authority
- JP
- Japan
- Prior art keywords
- chitosan
- solution
- added
- chitosan oligosaccharide
- 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
- RQFQJYYMBWVMQG-IXDPLRRUSA-N chitotriose Chemical compound O[C@@H]1[C@@H](N)[C@H](O)O[C@H](CO)[C@H]1O[C@H]1[C@H](N)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)N)[C@@H](CO)O1 RQFQJYYMBWVMQG-IXDPLRRUSA-N 0.000 title claims abstract description 48
- 238000004519 manufacturing process Methods 0.000 title claims description 5
- 229920001661 Chitosan Polymers 0.000 claims abstract description 52
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 14
- 108010059820 Polygalacturonase Proteins 0.000 claims abstract description 13
- 108010093305 exopolygalacturonase Proteins 0.000 claims abstract description 13
- 239000005452 food preservative Substances 0.000 claims abstract description 8
- 235000019249 food preservative Nutrition 0.000 claims abstract description 8
- 108090000790 Enzymes Proteins 0.000 claims description 30
- 102000004190 Enzymes Human genes 0.000 claims description 30
- 238000000034 method Methods 0.000 claims description 16
- 229920001542 oligosaccharide Polymers 0.000 claims description 9
- 150000002482 oligosaccharides Chemical class 0.000 claims description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 abstract description 60
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 29
- 238000000354 decomposition reaction Methods 0.000 abstract description 21
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 abstract description 15
- 230000002335 preservative effect Effects 0.000 abstract description 10
- 230000002255 enzymatic effect Effects 0.000 abstract description 9
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 abstract description 8
- 235000013305 food Nutrition 0.000 abstract description 7
- 239000003755 preservative agent Substances 0.000 abstract description 7
- 239000002244 precipitate Substances 0.000 abstract description 5
- 241000228245 Aspergillus niger Species 0.000 abstract description 3
- 238000005406 washing Methods 0.000 abstract description 3
- 238000001035 drying Methods 0.000 abstract description 2
- 239000003960 organic solvent Substances 0.000 abstract description 2
- 238000000926 separation method Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 49
- 229940088598 enzyme Drugs 0.000 description 29
- 239000000047 product Substances 0.000 description 29
- 239000000052 vinegar Substances 0.000 description 16
- 235000021419 vinegar Nutrition 0.000 description 16
- 235000019640 taste Nutrition 0.000 description 15
- 239000000203 mixture Substances 0.000 description 14
- 239000007788 liquid Substances 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 9
- 239000002253 acid Substances 0.000 description 7
- 238000003860 storage Methods 0.000 description 7
- 230000000844 anti-bacterial effect Effects 0.000 description 6
- 238000009835 boiling Methods 0.000 description 6
- 150000003839 salts Chemical class 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 235000019606 astringent taste Nutrition 0.000 description 5
- 108010089807 chitosanase Proteins 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000011156 evaluation Methods 0.000 description 5
- 235000011194 food seasoning agent Nutrition 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 240000007594 Oryza sativa Species 0.000 description 4
- 235000007164 Oryza sativa Nutrition 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 238000006911 enzymatic reaction Methods 0.000 description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
- 238000005554 pickling Methods 0.000 description 4
- 235000009566 rice Nutrition 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 3
- 235000010149 Brassica rapa subsp chinensis Nutrition 0.000 description 3
- 235000000536 Brassica rapa subsp pekinensis Nutrition 0.000 description 3
- 241000499436 Brassica rapa subsp. pekinensis Species 0.000 description 3
- 244000061456 Solanum tuberosum Species 0.000 description 3
- 235000002595 Solanum tuberosum Nutrition 0.000 description 3
- 150000007513 acids Chemical class 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 235000013601 eggs Nutrition 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 238000005187 foaming Methods 0.000 description 3
- 235000012149 noodles Nutrition 0.000 description 3
- 235000012015 potatoes Nutrition 0.000 description 3
- 230000035484 reaction time Effects 0.000 description 3
- 238000005096 rolling process Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 238000005063 solubilization Methods 0.000 description 3
- 230000007928 solubilization Effects 0.000 description 3
- 229920002101 Chitin Polymers 0.000 description 2
- 241000238424 Crustacea Species 0.000 description 2
- 241000238557 Decapoda Species 0.000 description 2
- 239000004278 EU approved seasoning Substances 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 241000209140 Triticum Species 0.000 description 2
- 235000021307 Triticum Nutrition 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 210000002421 cell wall Anatomy 0.000 description 2
- 238000007598 dipping method Methods 0.000 description 2
- 235000019634 flavors Nutrition 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000006864 oxidative decomposition reaction Methods 0.000 description 2
- 235000015277 pork Nutrition 0.000 description 2
- 239000004576 sand Substances 0.000 description 2
- 235000013580 sausages Nutrition 0.000 description 2
- 235000013555 soy sauce Nutrition 0.000 description 2
- 238000005507 spraying Methods 0.000 description 2
- 238000010025 steaming Methods 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- 239000012085 test solution Substances 0.000 description 2
- 239000007218 ym medium Substances 0.000 description 2
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- 240000002234 Allium sativum Species 0.000 description 1
- 241000194110 Bacillus sp. (in: Bacteria) Species 0.000 description 1
- 102000005575 Cellulases Human genes 0.000 description 1
- 108010084185 Cellulases Proteins 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 240000008067 Cucumis sativus Species 0.000 description 1
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 240000008415 Lactuca sativa Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- YBHQCJILTOVLHD-YVMONPNESA-N Mirin Chemical compound S1C(N)=NC(=O)\C1=C\C1=CC=C(O)C=C1 YBHQCJILTOVLHD-YVMONPNESA-N 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-N Nitrous acid Chemical compound ON=O IOVCWXUNBOPUCH-UHFFFAOYSA-N 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 241000228143 Penicillium Species 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 241000952054 Rhizopus sp. Species 0.000 description 1
- 244000166071 Shorea robusta Species 0.000 description 1
- 235000015076 Shorea robusta Nutrition 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 241001136496 Talaromyces islandicus Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 230000003627 anti-cholesterol Effects 0.000 description 1
- 230000000843 anti-fungal effect Effects 0.000 description 1
- 210000000436 anus Anatomy 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 230000034303 cell budding Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 210000000991 chicken egg Anatomy 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 238000010227 cup method (microbiological evaluation) Methods 0.000 description 1
- 230000000850 deacetylating effect Effects 0.000 description 1
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 210000003278 egg shell Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007071 enzymatic hydrolysis Effects 0.000 description 1
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 235000004611 garlic Nutrition 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 125000001475 halogen functional group Chemical group 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 229960001438 immunostimulant agent Drugs 0.000 description 1
- 239000003022 immunostimulating agent Substances 0.000 description 1
- 230000003308 immunostimulating effect Effects 0.000 description 1
- 238000004898 kneading Methods 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 235000013923 monosodium glutamate Nutrition 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 235000021573 pickled cucumbers Nutrition 0.000 description 1
- 229920001592 potato starch Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 235000012045 salad Nutrition 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 238000009938 salting Methods 0.000 description 1
- 229920006298 saran Polymers 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 235000015170 shellfish Nutrition 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 229940073490 sodium glutamate Drugs 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 235000020138 yakult Nutrition 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Food Preservation Except Freezing, Refrigeration, And Drying (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明はキトサンを酵素分解してキトサンオリゴ糖を製
造する方法および得られたキトサンオリゴ糖からなる食
品保存料に関する。DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a method for producing chitosan oligosaccharide by enzymatically decomposing chitosan, and a food preservative comprising the obtained chitosan oligosaccharide.
〔従来の技術および発明が解決しようとする課題〕キト
サンは、エビやカニなど甲殻類や微生物の細胞壁などに
含有されるキチンを脱アセチル化したものであり、最近
では水処理剤やキレート刑など工業用；抗コレステロー
ル作用や人工皮膚、免疫賦活剤など医薬用；発芽促進効
果、抗カビ性など農業用；保湿効果による化粧品用など
のほか食品分野での利用など様々な用途に用いられはじ
めている。[Prior art and problems to be solved by the invention] Chitosan is a deacetylated chitin contained in the cell walls of crustaceans such as shrimp and crabs and microorganisms, and has recently been used as a water treatment agent and chelating agent. It has started to be used for various purposes such as industrial use; medicinal use such as anti-cholesterol effects, artificial skin, and immunostimulant; agricultural use such as germination promoting effect and anti-fungal effect; cosmetic use due to its moisturizing effect, and use in the food field. .
しかし、キトサンは高分子物質であり、溶液とした場合
、非常に粘性を示す。例えばキトサン２〜３％（ｗ／ｖ
）溶液ではゲル状を呈し、キトサンを高濃度に溶解する
ことができない。そこで、キトサンの低分子化の方法と
して、これまでに塩酸による加水分解法（Ｓ、 Ｔ、
Ｈｏｒｏｗｉｔｚら、ジャーナル・オブ・アメリカン・
ケミカル・ソサイエティ。However, chitosan is a polymeric substance, and when it is made into a solution, it is extremely viscous. For example, chitosan 2-3% (w/v
) In solution, it appears gel-like and chitosan cannot be dissolved in high concentrations. Therefore, as a method for reducing the molecular weight of chitosan, hydrolysis using hydrochloric acid (S, T,
Horowitz et al., Journal of American
Chemical Society.
７９巻、５０４６〜５０４９頁（１９５７））、亜硝酸
による酸化分解法（Ｆ、 Ｙａｋｕら、セルロース・ケ
ミストリー・アンド・テクノロジー、１１巻。79, pp. 5046-5049 (1957)), oxidative decomposition method using nitrous acid (F, Yaku et al., Cellulose Chemistry and Technology, Vol. 11).
４２１〜４３０頁（１９７７））および塩素による酸化
分解法（平野茂博ら２日本農芸化学会 昭和５９年度大
会講演要旨集、３３０頁）などの化学的分解法が知られ
ているが、使用する薬品類の安全性の問題や取扱い上の
繁雑さ、さらには多大なエネルギーが必要であることな
ど難点がある。421-430 (1977)) and the oxidative decomposition method using chlorine (Shigehiro Hirano et al. 2 Japanese Society of Agricultural Chemistry, Abstracts of the 1980 Conference, p. 330), but the chemicals used There are some drawbacks, such as safety issues, complicated handling, and the need for a large amount of energy.
また、キトサン分解酵素であるキトサナーゼに関する報
告としては、バチルス・エスピー（Ｂａｃｉｌｌｕｓｓ
ｐ、）の生産するキトサナーゼ、ペニシリウム・イスラ
ンデイカム（Ｐｅｎ ｉｃ ｉ Ｉ Ｉ ｉｕｍ １ｓ
ｌａｎｄｉｃｕｌ１１）の生産するキトサナーゼ、スト
レプトマイセス（Ｓｔｒｅ ｔｏｗ ｃｅｓ）爲の生産
するキトサナーゼなどが知られている。In addition, reports regarding chitosanase, a chitosan-degrading enzyme, include Bacillus sp.
Chitosanase produced by Penicillium islandicum (Penicillium p.)
Chitosanase produced by S. randicul 11) and chitosanase produced by Streptomyces (Stret tow ces) are known.
しかし、これらキトサナーゼは活性が低いなど生産性の
問題や、高度の精製技術が必要であるなどの問題があり
、市販には到っていない。一般市販酵素を用いたキトサ
ン分解法として、パパイン。However, these chitosanases have problems with productivity such as low activity and require advanced purification techniques, and are not commercially available. Papain is a chitosan decomposition method using a commercially available enzyme.
セルラーゼおよび酸性プロテアーゼの使用（特開昭６３
−６３３８８）があるが、キトサンに対する各酵素の使
用量が多く、コスト的に難点があり、十分に満足できる
方法は未だに得られていない。Use of cellulases and acidic proteases (JP-A-63)
-63388), but the amount of each enzyme used relative to chitosan is large, and there are problems in terms of cost, and a fully satisfactory method has not yet been obtained.
そこで、本発明者らは上記課題を解決すべく鋭意研究を
重ねた結果、キトサン溶液をペクチナーゼを主成分とす
る酵素剤で酵素分解することによりキトサンオリゴ塘を
製造することができ、しかも得られたキトサンオリゴ糖
を用いて渋味のない食品保存料が得られることを見出し
て本発明を完成するに至った。Therefore, as a result of intensive research to solve the above problems, the present inventors were able to produce chitosan oligotang by enzymatically decomposing a chitosan solution with an enzyme agent whose main component is pectinase. The present inventors have discovered that a food preservative without astringency can be obtained using chitosan oligosaccharide, and have completed the present invention.
すなわち、本発明はキトサン溶液をペクチナーゼを主成
分とする酵素剤で酵素分解し、キトサンオリゴ糖を製造
することを特徴とするキトサンオリゴ糖の製造方法およ
び該キトサンオリゴ糖からなる食品保存料を提供するも
のである。That is, the present invention provides a method for producing chitosan oligosaccharides, which comprises enzymatically decomposing a chitosan solution with an enzyme agent containing pectinase as a main component to produce chitosan oligosaccharides, and a food preservative made from the chitosan oligosaccharides. It is something to do.
本発明で用いるキトサンは、甲殻類、昆虫類。The chitosan used in the present invention is derived from crustaceans and insects.
貝類などの下等動物の外皮骨格組織の成分および菌類の
細胞壁成分として知られているキチンを原料とし、これ
をアルカリ処理などの常法により脱アセチル化して得ら
れるものである。Chitin, which is known as a component of the integumentary skeletal tissue of lower animals such as shellfish and a component of the cell wall of fungi, is used as a raw material and is obtained by deacetylating it using conventional methods such as alkali treatment.
キトサンは酸性下でのみ溶解するため、キトサン溶液を
調製するにあたり酸を使用するが、その際に用いる酸と
しては有機酸および無機酸のいずれでもよいが、食品に
添加することを考慮し、酢酸、乳酸、クエン酸、リンゴ
酸など通常食用に供される酸を用いることが望ましい。Chitosan dissolves only under acidic conditions, so an acid is used to prepare the chitosan solution.The acid used at this time can be either an organic acid or an inorganic acid, but in consideration of adding it to food, acetic acid is used. It is desirable to use acids that are commonly used for food, such as lactic acid, citric acid, and malic acid.
特に、得られたキトサンオリゴ糖を食品保存料として用
いることを考えた場合、酸自身に抗菌力の強さが知られ
ている酢酸が望ましい。酢酸を含有する食品としては食
酢が良く知られており、香味の面、抗菌性の面から最も
好ましい。また、その他の保存効果を有する保存料や調
味料を併用してもよい。具体的には酢酸ソーダ、特許第
９５９２２２号等に記載の食酢含有保存料や市販食酢な
どが利用できる。In particular, when considering the use of the obtained chitosan oligosaccharide as a food preservative, acetic acid is desirable because the acid itself is known to have strong antibacterial activity. Vinegar is well known as a food containing acetic acid, and is most preferred from the standpoint of flavor and antibacterial properties. In addition, other preservatives and seasonings having a preservative effect may be used in combination. Specifically, sodium acetate, the vinegar-containing preservative described in Japanese Patent No. 959222, commercially available vinegar, and the like can be used.
次に、本発明で用いるペクチナーゼ（ペクチン分解酵素
）を主成分とする酵素剤としては一般に市販されている
酵素剤で十分であるが、至適ｐＨが３〜５であり、酢酸
によるキトサンの可溶化および酵素分解時の最適ｐＨの
調整の条件がそのまま適用できるものが好適に利用でき
る。また、ペクチナーゼは一般にアスペルギルス・ニガ
ー（ＭＰ旧庄■Ｕ頂１肛）やリゾープス属（−）などに
より生産されたものが市販されており、いずれを用いて
もよいが、粘性低下の点から前者の方が好ましい。Next, as the enzyme agent containing pectinase (pectin-degrading enzyme) as the main component used in the present invention, generally commercially available enzyme agents are sufficient, but the optimum pH is 3 to 5, and chitosan can be degraded by acetic acid. It is preferable to use one that allows the conditions for adjusting the optimum pH during solubilization and enzymatic decomposition to be applied as is. In addition, pectinases produced by Aspergillus niger (MP Kyu-sho ■U top 1 anus) and Rhizopus sp. is preferable.
キトサン溶液のペクチナーゼを主成分とする酵素剤によ
る酵素分解の条件は、キトサン１重量部に対し酢酸０．
１重量部以上、好ましくは０．３〜６重量部を含むｐ［
＋３〜５のキトサン溶液にペクチナーゼを主成分とする
酵素剤０．００１重量部以上、好ましくは０．００５〜
０．０５重量部添加し、４０〜６０°Ｃで１時間以上の
酵素分解を行い、キトサン分解液とする。次いで、得ら
れたキトサン分解液をアセトン、イソプロパツールなど
の有機溶媒や水酸化ナトリウムなどによりｐＨ６，５以
上に調整することにより沈澱を生成させ、濾過。The conditions for enzymatic decomposition of a chitosan solution using an enzyme agent whose main component is pectinase are as follows: 1 part by weight of chitosan and 0.00% acetic acid.
p [ containing 1 part by weight or more, preferably 0.3 to 6 parts by weight
0.001 part by weight or more of an enzyme agent containing pectinase as a main component, preferably 0.005 to 5 parts by weight, to a chitosan solution of +3 to 5.
Add 0.05 parts by weight and perform enzymatic decomposition at 40 to 60°C for 1 hour or more to obtain a chitosan decomposition solution. Next, the resulting chitosan decomposition solution is adjusted to pH 6.5 or higher using an organic solvent such as acetone, isopropanol, or sodium hydroxide to form a precipitate, and then filtered.
遠心分離などにより沈澱物を分取し、水洗、乾燥するこ
とによりキトサンオリゴ糖を得ることができる。Chitosan oligosaccharide can be obtained by separating the precipitate by centrifugation, washing with water, and drying.
このようにして得られたキトサンオリゴ糖を食品保存料
として使用する場合、キトサン分解液を加熱して酵素を
失活せしめたもの、該分解液から分離、精製したキトサ
ンオリゴ糖あるいは該キトサンオリゴ糖を酸に溶解せし
めたものなどのいずれでもよい。添加量は、キトサンオ
リゴ糖として０．００５〜０．５％（ｗ／ｗ）、好まし
くは０．０１〜０．２％（ｗ／ｗ）である。添加量が少
ないと、抗菌効果が期待できず、添加量が多いと、渋味
が発現するので注意する必要がある。添加するキトサン
オリゴ糖の形状は液状または粉末状、顆粒状などの固体
状で使用でき、添加方法は溶解、混練、噴霧。When using the chitosan oligosaccharide thus obtained as a food preservative, the chitosan oligosaccharide obtained by heating the chitosan decomposition solution to inactivate the enzyme, the chitosan oligosaccharide separated and purified from the decomposition solution, or the chitosan oligosaccharide separated from the decomposition solution and purified. Any one obtained by dissolving in an acid may be used. The amount added is 0.005 to 0.5% (w/w), preferably 0.01 to 0.2% (w/w) as chitosan oligosaccharide. If the amount added is small, no antibacterial effect can be expected, and if the amount added is too large, an astringent taste will develop, so care must be taken. The chitosan oligosaccharide to be added can be used in liquid, powder, or solid forms such as granules, and can be added by dissolving, kneading, or spraying.
散布、浸漬など任意であり、いずれも食品に添加しやす
い形状、方法を適宜選択すればよい。Spraying, dipping, etc. are optional, and the shape and method that are easy to add to food may be selected as appropriate.
本発明は、ペクチナーゼを主成分とする酵素剤によるキ
トサンの酵素加水分解法によってキトサンオリゴ糖を得
る方法であるが、キトサンを可？容化する段階から分解
の終了段階まで酢酸または食酢などを使用することによ
り、（ア）酵素分解段階でも可溶化工程の酢酸または食
酢などが残存するので、そのままもしくは筒車な調製工
程で分解することができる。（イ）酢酸酸性下での酵素
活性を有する酵素剤を使用するので、充分に酵素分解を
行うことができる。（つ）ペクチナーゼを主成分とする
酵素剤によって加水分解された分解液中に防腐力の強い
酢酸または食酢などが存在するので、分解終了後のキト
サンオリゴ糖液の長期間の保存が可能であること（１）
さらに、分解終了後のキトサンオリゴ糖の防腐力と酢酸
または食酢の防腐力が相乗的に増強された顕著な防腐力
を有する保存料を得ることができる等の種々の効果を得
ることができる。The present invention is a method for obtaining chitosan oligosaccharides by enzymatic hydrolysis of chitosan using an enzyme agent containing pectinase as a main component. By using acetic acid or vinegar from the solubilization stage to the final stage of decomposition, (a) the acetic acid or vinegar from the solubilization process remains even in the enzymatic decomposition stage, so it can be decomposed as it is or in an hourly preparation process. be able to. (a) Since an enzyme agent having enzymatic activity under acetic acid is used, sufficient enzymatic decomposition can be carried out. (1) Since acetic acid or vinegar, which have strong preservative properties, are present in the decomposition solution that is hydrolyzed by an enzyme agent whose main component is pectinase, it is possible to preserve the chitosan oligosaccharide solution for a long period of time after completion of decomposition. Thing (1)
Furthermore, various effects can be obtained, such as the ability to obtain a preservative with remarkable preservative power in which the preservative power of chitosan oligosaccharide and the preservative power of acetic acid or vinegar are synergistically enhanced after completion of decomposition.
以下に実施例をあげて本発明を説明するが、本発明はそ
れのみに限定されることはない。The present invention will be explained below with reference to Examples, but the present invention is not limited thereto.
実施例１
キトサン２．０ｇを１％酢酸水溶液１００ｄに溶解した
０次いで、第１表に示した各酵素を０．０４ｇ加え、４
５°Ｃ恒温水槽中でゆっくり攪拌しながら６時間反応せ
しめた。反応終了後、沸とう水中に５分間保ち、酵素を
失活せしめ、キトサンオリゴ糖液を得た。得られたキト
サンオリゴ糖液をジャーレス（ｓｈａｌｅｓ）変法（Ａ
ｇｒｉｃｕｌｔｕｒａｌ ＢｉｏｌｏｇｉｃａｌＣｈｅ
ｍｉｓｔｒｙ、 ３５巻、１１５４〜１１５６頁、１９
７１）により測定したところ、４２０ｒ＋ｍで吸収が認
められ、キトサンオリゴ糖が生成していることが確認さ
れた。また、得られた各キトサンオリゴ糖液について分
解程度、抗菌活性および渋味を測定した。Example 1 2.0 g of chitosan was dissolved in 100 d of 1% acetic acid aqueous solution. Next, 0.04 g of each enzyme shown in Table 1 was added.
The reaction was allowed to proceed for 6 hours with slow stirring in a 5°C constant temperature water bath. After the reaction was completed, the mixture was kept in boiling water for 5 minutes to inactivate the enzyme and obtain a chitosan oligosaccharide solution. The obtained chitosan oligosaccharide solution was subjected to a modified Shales method (A
gricultural Biological Che
mistry, vol. 35, pp. 1154-1156, 19
71), absorption was observed at 420 r+m, confirming that chitosan oligosaccharides were produced. Furthermore, the degree of decomposition, antibacterial activity, and astringency of each obtained chitosan oligosaccharide solution were measured.
この結果を第１表に示す。なお、分解程度は東京計器Ｂ
型粘度計でＢＬアダプタを用い、２０°Ｃにおける粘度
を測定した。抗菌活性は、被験菌としてサツカロマイセ
ス−セレビシェ（Ｓａｃｃｈａｒｏｍ ｃｅｓｃｅｒｅ
ｖｉｓｉａｅ）ＩＦＯＯ２０３をＹＭ培地（酵母エキス
３ｇ、ＩＥ芽芽生キス３ｇペプトン５ｇ、グルコースＬ
ｏｇを蒸留水で１２にしたもの、ｐＨ５，５）に３０°
Ｃで２４時間前培養した液２ｄを、常法に従い殺菌処理
したＹＭ寒天培地（上記ＹＭ培地＋寒天１．５％（ｉｖ
／ｖ）） １００ ｍｌに加えプレートを作成し、常法
に従い適宜希釈したキトサンオリゴ糖液３０μ２をカッ
プ法により３０″Ｃで２４時間後のハロー形成の有無を
調べ、ハロー形成するキトサンオリゴ糖の最小濃度をＭ
ＩＣとした。渋味は、キトサンオリゴ糖液を水で１０倍
希釈し、官能評価で渋味の強弱を判定した。また、対照
は酵素を添加しなかったこと以外は上記と同様の操作を
行ったものとした。The results are shown in Table 1. The degree of disassembly is Tokyo Keiki B.
The viscosity at 20°C was measured using a BL adapter with a type viscometer. The antibacterial activity was determined using Saccharomyces cerevisiae as the test bacterium.
visiae) IFOO203 in YM medium (yeast extract 3g, IE budding kiss 3g peptone 5g, glucose L
og to 12 with distilled water, pH 5,5) at 30°
2d of the solution pre-cultured for 24 hours in C was mixed with YM agar medium (the above YM medium + agar 1.5% (iv.
/v)) A plate was prepared by adding 100 ml of the chitosan oligosaccharide solution, and 30μ2 of the chitosan oligosaccharide solution was appropriately diluted according to a conventional method.The presence or absence of halo formation was examined after 24 hours at 30"C using the cup method. The minimum concentration is M
It was made into an IC. For the astringency, the chitosan oligosaccharide solution was diluted 10 times with water, and the strength of the astringency was determined by sensory evaluation. In addition, as a control, the same operation as above was performed except that no enzyme was added.
第１表より明らかなように、ペクチナーゼ、特にアスペ
ルギルス・ニガーにより生産されたペクチナーゼを主成
分とする酵素剤を用いて得られたキトサンオリゴ糖がす
ぐれていることがわかった。As is clear from Table 1, it was found that the chitosan oligosaccharide obtained using an enzyme agent whose main component is pectinase, especially pectinase produced by Aspergillus niger, is superior.
次に、キトサンを溶解する酸について検討した。Next, we investigated acids that dissolve chitosan.
キトサン２．０ｇを第２表に示した酸の１％水溶液に溶
解後、水酸化ナトリウムにてｐＨ４，０に調整した。そ
こにスミチームＡＰ２ （新日本化学工業■製）０．０
５ｇを加え、４５°Ｃ恒温水槽中でゆっくり攪拌しなが
ら６時間反応せしめた。反応終了後、沸とう水中に５分
間保ち、酵素を失活せしめ、キトサンオリゴ糖液を得た
。得られた各キトサンオリゴ糖液について分解程度、抗
菌活性を前記と同様の方法により測定した。この結果を
第２表に示す。After dissolving 2.0 g of chitosan in a 1% aqueous solution of the acids shown in Table 2, the pH was adjusted to 4.0 with sodium hydroxide. There Sumiteam AP2 (manufactured by Shin Nippon Chemical Industry ■) 0.0
5 g was added thereto, and the reaction was allowed to proceed for 6 hours with slow stirring in a constant temperature water bath at 45°C. After the reaction was completed, the mixture was kept in boiling water for 5 minutes to inactivate the enzyme and obtain a chitosan oligosaccharide solution. The degree of decomposition and antibacterial activity of each obtained chitosan oligosaccharide solution were measured by the same method as described above. The results are shown in Table 2.
第２表
第２表から明らかなように、キトサンを可溶化し、酵素
分解する場合、酢酸が好ましいことがわかった。As is clear from Table 2, acetic acid was found to be preferable when solubilizing chitosan and enzymatically decomposing it.
次に、酵素の添加量について検討した。キトサン２．Ｏ
ｇを１％酢酸水溶液１００ｍ１に溶解し、スミチームＡ
Ｐ２（新日本化学工業■製）をキトサンに対して０．２
５〜４％（ｗ／ｗ）加え、５０°Ｃ恒温水槽中でゆっく
り攪拌しながら６時間反応せしめた。反応終了後、沸と
う水中に５分間保ち、酵素を失活せしめ、キトサンオリ
ゴ糖液を得た。Next, we examined the amount of enzyme added. Chitosan 2. O
Dissolve g in 100 ml of 1% acetic acid aqueous solution,
P2 (manufactured by Shin Nippon Chemical Co., Ltd.) at 0.2 for chitosan.
5 to 4% (w/w) was added, and the mixture was reacted for 6 hours with slow stirring in a constant temperature water bath at 50°C. After the reaction was completed, the mixture was kept in boiling water for 5 minutes to inactivate the enzyme and obtain a chitosan oligosaccharide solution.
得られたキトサンオリゴ糖液の粘度を前記と同様の方法
により測定し、結果を第１図に示した。第１図より明ら
かなように、酵素無添加時の粘度１４６０ｃｐが酵素添
加により低下し、酵素添加量は基質に対して約１％（ｗ
／ｗ）ではほぼ一定となり、キトサンが分解しているこ
とがわかった。The viscosity of the obtained chitosan oligosaccharide solution was measured by the same method as described above, and the results are shown in FIG. As is clear from Figure 1, the viscosity of 1460 cp when no enzyme is added is reduced by the addition of enzyme, and the amount of enzyme added is approximately 1% (w) of the substrate.
/w) remained almost constant, indicating that chitosan was decomposed.
次に、酵素反応時間について検討した。キトサン２．０
ｇを１％酢酸水溶液１００ ｍｌに溶解し、スミチーム
ＡＰ２（新日本化学工業■製）をキトサンに対して２％
（ｗ／ｗ）加え、５０℃恒温水槽中でゆっくり攪拌しな
がら１〜１８時間反応せしめた。反応終了後、沸とう水
中に５分間保ち、酵素を失活せしめ、キトサンオリゴ糖
液を得た。得られたキトサンオリゴ糖液の粘度を前記と
同様の方法により測定し、結果を第２図に示した。Next, we examined the enzyme reaction time. chitosan 2.0
Dissolve g in 100 ml of 1% acetic acid aqueous solution, and add Sumiteem AP2 (manufactured by Shin Nippon Chemical Co., Ltd.) to 2% chitosan.
(w/w) and reacted for 1 to 18 hours with slow stirring in a constant temperature water bath at 50°C. After the reaction was completed, the mixture was kept in boiling water for 5 minutes to inactivate the enzyme and obtain a chitosan oligosaccharide solution. The viscosity of the obtained chitosan oligosaccharide solution was measured by the same method as described above, and the results are shown in FIG.
第２図より明らかなように、酵素反応は６時間程度でキ
トサンの分解がほぼ一定となることがわかった。As is clear from FIG. 2, it was found that the decomposition of chitosan became almost constant in the enzymatic reaction after about 6 hours.
実施例２
キトサン５．０ｇを市販食酢（酢酸酸度４．２％（ｗ／
ｖ）） １００ ｔｔｔｉに溶解した。次いで、スミチ
ームＡＰ２（新日本化学工業■製） ０．１ ｇを加え
、５０°Ｃ恒温水槽中でゆっくり攪拌しながら６時間反
応せしめた。反応終了後、沸とう水中に５分間保ち、酵
素を失活せしめ、キトサンオリゴ糖液（Ａ）を得た。Example 2 5.0 g of chitosan was added to commercially available vinegar (acetic acid acidity 4.2% (w/
v)) Dissolved in 100 ttti. Next, 0.1 g of Sumiteam AP2 (manufactured by Shin Nihon Kagaku Kogyo ■) was added, and the mixture was reacted for 6 hours with slow stirring in a constant temperature water bath at 50°C. After the reaction was completed, the mixture was kept in boiling water for 5 minutes to inactivate the enzyme and obtain a chitosan oligosaccharide solution (A).
適当な大きさに切断した白菜１０ｋｇに１０％食塩水１
０ｋｇを添加し、５ｋｇの重石をして１８時間塩漬けを
行なった後、白菜を取り出して軽く水切りした。この塩
漬白菜３００ｇに、食塩５ｋｇ、グルタミン酸ソーダ０
．５ｋｇおよびキトサンオリゴ糖液（Ａ）を第３表に示
す量加えたものに水を加えて１００ｆとした調味液３０
０ｄを漬液として添加し、これをビニール袋に入れて密
封し試作品を得た。また、対照としてキトサンオリゴ糖
液（Ａ）を添加しないこと以外は上記と同様の組成の調
味液を用いた対照品を作った。10 kg of Chinese cabbage cut into appropriate size and 1 part of 10% salt solution
After adding 0 kg and salting for 18 hours using a 5 kg weight, the Chinese cabbage was taken out and lightly drained. 300g of this salted Chinese cabbage, 5kg of salt, 0 of sodium glutamate
．． 5 kg and chitosan oligosaccharide solution (A) in the amount shown in Table 3, and then added water to make 100 f Seasoning liquid 30
0d was added as a dipping solution, which was then placed in a plastic bag and sealed to obtain a prototype. In addition, as a control, a control product was prepared using a seasoning liquid having the same composition as above except that the chitosan oligosaccharide liquid (A) was not added.
得られた試作品および対照品それぞれ２０検体を室温（
１８〜２６°Ｃ）で７日間保存し、漬液の濁り具合から
防腐効果を判定した。また、保存１８目のものを１０人
のパネラ−に試食してもらい食味評価を行なった。この
結果を第３表に示す。20 samples each of the obtained prototype and control products were kept at room temperature (
The preservative effect was determined from the degree of turbidity of the soaking solution. In addition, 10 panelists tasted the 18th preserved product and evaluated its taste. The results are shown in Table 3.
第３表
第３表より明らかなように、防腐効果があり、かつ食味
に問題ないキトサンオリゴ糖量（固型分として）は、原
料野菜に対し０．００５％〜０．５％（Ｗ／Ｗ）が望ま
しいことが判った。Table 3 As is clear from Table 3, the amount of chitosan oligosaccharide (as solid content), which has a preservative effect and does not cause any taste problems, is 0.005% to 0.5% (W/ W) was found to be desirable.
実施例３
鶏卵の全卵液２２５ｇを溶きほぐし、だし汁５０ｄを入
れ、次いで醤油５ｄ、塩１ｇ、砂糖１４ｇおよび実施例
２で得たキトサンオリゴ糖液（Ａ）３ＩＩ１．に水を加
えて４０戚とした試験液を加えて混合した０次に、卵焼
なべに薄（油をひいて熱し、上記の卵焼き原料の１／４
程度を流し入れ、底の部分が固まってきたところで手前
に巻き込んだ。Example 3 Beat 225 g of whole chicken egg liquid, add 50 d of stock, then add 5 d of soy sauce, 1 g of salt, 14 g of sugar, and the chitosan oligosaccharide solution (A) 3II1 obtained in Example 2. Add water to make a 40% test solution and mix. Next, heat a thin layer of oil in an omelet pan and add 1/4 of the above omelet ingredients.
Pour in the amount, and when the bottom part has hardened, roll it toward you.
巻いたものをなべの向う側に移し、再びなべに油をひき
、先と同様に原料を流し込み、底の部分が固まってきた
ところで初め（＝焼いた部分を芯にして手前に向って巻
き込んだ。同様にして次々に巻き込みながら焼いた後、
好みの大きさに切って卵焼きの製品を得た。また、対照
としてキトサンオリゴ糖液（Ａ）を添加しないこと以外
は上記と同様にして卵焼きを作った。Transfer the rolled item to the other side of the pot, reapply oil to the pot, pour in the ingredients as before, and when the bottom part has hardened, start rolling (= using the baked part as the core and rolling it towards you. After baking in the same way, rolling in one after another,
Cut it into the desired size to obtain a fried egg product. In addition, as a control, fried eggs were prepared in the same manner as above except that the chitosan oligosaccharide solution (A) was not added.
得られた製品および対照品について、保存試験および食
味評価を行った。この結果を第１表に示す。なお、保存
試験は製品および対照品をトレイバックに入れサランラ
ップで包装し、２０°Ｃで保存した場合の腐敗の程度を
示す。なお、表中の−は腐敗せず２士は腐敗した。＋＋
は腐敗が著しいを示す。食味評価は２０人のパネラ−に
できたての製品を試食してもらい、食味評価を行なった
。Storage tests and taste evaluations were conducted on the obtained products and control products. The results are shown in Table 1. The storage test shows the degree of spoilage when the product and control product are placed in a tray bag, wrapped in Saran wrap, and stored at 20°C. In addition, - in the table was not corrupted, and 2 players were corrupted. ++
indicates significant corruption. Taste evaluation was conducted by having 20 panelists taste the freshly made products.
この結果を第４表に示す。The results are shown in Table 4.
第４表
第４表から明らかなように、キトサンオリゴ糖液を添加
することにより、卵焼きの保存性が向上し、食味の面か
らも良好であることがわかった。As is clear from Table 4, it was found that the addition of the chitosan oligosaccharide solution improved the preservability of the fried eggs and also provided good taste.
また、一部のパネラ−により製品の方が対照品より黒ず
みが少なく色調が良好であるとの評価が得られた。Additionally, some panelists rated the product as having less darkening and better color tone than the control product.
実施例５
キトサン５．０ｇを２％（−／い）乳酸液１００ ｍｌ
に溶解した９次いで、マセロチーム ２Ｓ（Ｉ＄３ヤク
ルト本社製）０．２ｇを加え、４５°Ｃ恒温水槽中で６
時間ゆっくり攪拌しながら反応せしめた。反応終了後、
沸とう水中に５分間保ち、酵素を失活せしめ、キトサン
オリゴ糖液（Ｂ）を得た。得られたキトサンオリゴ糖液
（Ｂ）を用いて下記の組成のきゅうりの浅漬けを作り、
保存試験を行った。Example 5 5.0 g of chitosan and 100 ml of 2% (-/I) lactic acid solution
Next, 0.2 g of Macerozyme 2S (I$3 manufactured by Yakult Honsha Co., Ltd.) dissolved in
The reaction was allowed to proceed with slow stirring for several hours. After the reaction is complete,
The enzyme was inactivated by keeping it in boiling water for 5 minutes to obtain a chitosan oligosaccharide solution (B). Using the obtained chitosan oligosaccharide solution (B), make pickled cucumbers with the following composition,
A storage test was conducted.
なお、きゅうりと漬液の固体：液体＝ｌ：１とした。Note that the solid:liquid ratio of cucumber and pickling liquid was 1:1.
この結果を第５表に示す。なお、表中の−は漬液の混濁
なし、＋は漬液の混濁あり、十＋は漬液はげしく混濁を
示す。The results are shown in Table 5. In the table, - indicates no turbidity of the pickling solution, + indicates turbidity of the pickling solution, and 10+ indicates severe turbidity of the pickling solution.
第５表
表より明らかなように、キトサンオリゴ糖の添加により
保存効果が著しく向上した。As is clear from Table 5, the preservation effect was significantly improved by the addition of chitosan oligosaccharide.
実施例５
糖類５０％（ｗ／ｗ）、呈味剤フレーバー香味液２０％
（ｗ／ｗ）に下記に示す試験液を加え、水で加水して１
００％としたみりん風調味料を作った。Example 5 Sugars 50% (w/w), flavoring agent flavor liquid 20%
Add the test solution shown below to (w/w) and add water to 1
I made a mirin-style seasoning with 00%.
ｈ −民」 ？
Ａ 実施例２で得られたキトサンオリゴ糖液（Ａ）
ＬＸ（ｖ／ｗ）Ｂ 醸造酢（酢酸酸度５％のもの）
２χ（ｖ／ｗ）Ｃ水
これらの製品をそれぞれ小袋に密封し、３０°Ｃで保存
して酵母による発泡の有無を調べた。また、食味評価を
実施例１と同様にして実施した。この結果を第６表に示
す。h-min”? A Chitosan oligosaccharide solution obtained in Example 2 (A)
LX (v/w) B Brewed vinegar (acetic acid acidity 5%)
2χ (v/w) C water Each of these products was sealed in a sachet and stored at 30°C to examine the presence or absence of foaming caused by yeast. In addition, taste evaluation was conducted in the same manner as in Example 1. The results are shown in Table 6.
第６表
（＝）・・・発泡認められず
（＋）・・・発泡あり
第６表より明らかなように、製品Ａが保存性および食味
がよいことがわかった。Table 6 (=): Foaming not observed (+): Foaming present As is clear from Table 6, product A was found to have good preservability and good taste.
実施例６
皮をむき切断したジャガイモ４００ ｇ、豚肉１００ｇ
をサラダ油１２ａｔで炒め、ここに醤油３７ｍ１．酒１
５ｄ、 ミリン８ｄ、砂ｔ７！１３．５ｇ、食塩２．
５ｇ、実施例２で得られたキトサンオリゴ糖液（Ａ）１
０ｍ、醸造酢（酢酸酸度１０％のもの）５成および水１
７５ｄを加え、汁気がなくなるまで煮込み、肉ジャガ（
製品Ｂｌ）５５０ｇを得た。Example 6 400 g of peeled and cut potatoes, 100 g of pork
Stir-fry with 12 at of salad oil and add 37 ml of soy sauce. Alcohol 1
5d, Mirin 8d, sand T7!13.5g, salt 2.
5g, chitosan oligosaccharide solution (A) obtained in Example 2 1
0m, 5 parts of brewed vinegar (acetic acid acidity 10%) and 1 part of water
Add 75d and simmer until the liquid evaporates.
550 g of product Bl) was obtained.
また、対照としてキトサンオリゴ糖液（Ａ）を添加せず
、醸造酢（酢酸酸度１０％のもの）１５ｉＩｌおよび水
１１５ｍ１を用いたこと以外は上記と同様にして肉ジャ
ガ（製品Ｂ２）を作り、さらにキトサンオリゴ糖液（Ａ
）と醸造酢を添加せず、水１９０ｄを用いたこと以外は
上記と同様にして肉ジャガ（対照品）を作った。In addition, as a control, meat potatoes (product B2) were made in the same manner as above, except that 15 μl of brewed vinegar (10% acetic acid acidity) and 115 ml of water were used without adding the chitosan oligosaccharide solution (A). Furthermore, chitosan oligosaccharide solution (A
) and brewed vinegar were not added and meat potatoes (control product) were made in the same manner as above except that 190 d of water was used.
得られた製品および対照品をパック詰めし、２５°Ｃで
所定期間保存試験を行い、−船主菌数を測定した。−船
主菌数の測定は、常法に従い毎日標準寒天培地を用いた
混釈法で測定し、１０’個／ｇに達した時を腐敗日と判
断した。また、できたての製品をパネラ−に試食させ食
味評価を行った。The obtained products and control products were packed and subjected to a storage test at 25°C for a predetermined period of time, and the number of bacteria was measured. - The shipowner's bacterial count was measured every day by the pouring method using a standard agar medium according to the usual method, and when it reached 10' bacteria/g, it was judged as the date of spoilage. In addition, panelists were asked to sample the freshly made products and evaluate their taste.
この結果を第７表に示す。The results are shown in Table 7.
第７表
第７表より明らかなように、製品Ｂｌは微生物による腐
敗を抑制し、かつ食味も良好であった。As is clear from Table 7, product B1 suppressed spoilage caused by microorganisms and had good taste.
実施例７
タラすり身１０００ｇに食塩３０ｇを加え、よく播潰し
、次いで馬鈴薯澱粉６０ｇ、砂Ｗ１０ｇ。Example 7 30 g of salt was added to 1000 g of ground cod, and the mixture was thoroughly crushed, followed by 60 g of potato starch and 10 g of sand W.
実施例２で得られたキトサンオリゴ糖液（Ａ）２５戚と
、醸造酢（酢酸酸度１０％のもの）１１に水酸化ナトリ
ウム３３．３ｇを加えて混合して得た食酢液１５ｍ！お
よび水１６０ｄを加え、さらによく播潰し板付した後、
蒸化に並べ４０分間蒸煮した。蒸煮終了後、蒸化から取
り出してかまぼこ（製品ＣＩ）を得た。また、対照とし
て食酢液を添加せず、水１７５成を用いたこと以外は上
記と同様にしてかまぼこ（製品Ｃ２）を作り、キトサン
オリゴ糖液（Ａ）と食酢液を添加せず、水２００ ｍｌ
を用いたこと以外は上記と同様にしてかまぼこ（対照品
）を作った。15 m of vinegar solution obtained by mixing 25 relatives of the chitosan oligosaccharide solution (A) obtained in Example 2 with 11 brewed vinegar (10% acetic acidity) and 33.3 g of sodium hydroxide! After adding 160 d of water and thoroughly crushing and attaching a plate,
The mixture was placed in a steamer and steamed for 40 minutes. After the steaming was completed, the mixture was removed from the steaming process to obtain kamaboko (product CI). In addition, as a control, kamaboko (product C2) was made in the same manner as above except that no vinegar solution was added and 175% of water was used. ml
Kamaboko (control product) was made in the same manner as above except that .
得られた製品および対照品を２０°Ｃで所定期間保存し
、実施例２と同様にして保存試験および食味評価を行っ
た。この結果を第８表に示す。The obtained products and control products were stored at 20°C for a predetermined period of time, and a storage test and taste evaluation were conducted in the same manner as in Example 2. The results are shown in Table 8.
第８表
表より明らかなように、製品ＣＩおよび製品Ｃ２は対照
品に比べて保存性１食味ともに優れており、特に製品Ｃ
１が優れていることが判った。As is clear from Table 8, product CI and product C2 are superior to the control product in terms of shelf life and taste, especially product C.
1 was found to be superior.
実施例８
精白米１．５ｋｇを水洗したのち、実施例２で得られた
キトサンオリゴ糖液（Ａ）３０ｍｆ、水２２２０ｍｋを
加えて市販炊飯器にて炊飯したところ、保存性に優れ、
食味の良好な米飯を得ることができた。Example 8 After washing 1.5 kg of polished rice with water, 30 mf of the chitosan oligosaccharide solution (A) obtained in Example 2 and 2220 mk of water were added and the rice was cooked in a commercially available rice cooker.
It was possible to obtain cooked rice with good taste.
実施例９
実施例２で得られたキトサンオリゴ糖液（Ａ）をアセト
ン中に滴下し、沈澱物を得た。この沈澱物を分別し、水
洗、乾燥させてキトサンオリゴ糖を得た。Example 9 The chitosan oligosaccharide solution (A) obtained in Example 2 was dropped into acetone to obtain a precipitate. This precipitate was separated, washed with water, and dried to obtain chitosan oligosaccharide.
小麦わ）４ｋｇに対し食塩１５０ｇ、上記で得られたキ
トサンオリゴＦ’５ｇおよび水１５００ ｍＡを加え、
よく混ねてめん打ちを行いうどんを作ったところ、保存
性に優れ、食味の良好なうどんを得ることができた。Add 150 g of salt, 5 g of chitosan oligo F' obtained above and 1500 mA of water to 4 kg of wheat (wheat flour),
When udon noodles were made by mixing the mixture well and making noodles, it was possible to obtain udon noodles with excellent storage stability and good taste.
実施例１０
豚の可食部分をひき肉にしたもの３００ｋｇにデンプン
１５ｋｇ、塩３ｋｇ、すりおろしニンニク５ｋｇ。Example 10 300 kg of minced pork edible parts, 15 kg of starch, 3 kg of salt, and 5 kg of grated garlic.
化学調味料および香辛料を適量加え、さらに実施例２で
得られたキトサンオリゴ糖ｉ （Ａ） ８ ｆｆｉと醸
造酢（酢酸酸度１０′／、のちの）ＩＮに卵殻３０．０
ｇ。Add appropriate amounts of chemical seasonings and spices, and add eggshell 30.0 to chitosan oligosaccharide i (A) 8 ffi obtained in Example 2 and brewed vinegar (acetic acid acidity 10'/, later) IN.
g.
重炭酸ナトリウム１４．０ｇおよび水酸化ナトリウム６
．６７ｇを加えて混合した液４Ｎをすべて加えてよく混
合し、常法に従いケーシング充填後、加熱、冷却してソ
ーセージを製造したところ、食味の良い保存性に優れた
ソーセージを製造することができた。14.0 g of sodium bicarbonate and 6 g of sodium hydroxide
．． 67g was added and all of the mixed liquid 4N was added and mixed well, and sausages were manufactured by filling casings according to the conventional method, heating and cooling, and it was possible to manufacture sausages with good taste and excellent storage stability. .
本発明によれば、キトサンオリゴ糖を容易、かつ安価に
製造することができる。また、本発明により得られるキ
トサンオリゴ糖は食品用保存料等として有用である。According to the present invention, chitosan oligosaccharides can be produced easily and at low cost. Furthermore, the chitosan oligosaccharides obtained by the present invention are useful as food preservatives and the like.
第１図は実施例１における酵素添加量とキトサン分解程
度（粘度）の関係を示し、第２図は実施例１における酵
素反応時間とキトサン分解程度（粘度）の関係を示す。
第）図
酵素添加量（キトサン１こ対する％）
酵素反応時間（時間）FIG. 1 shows the relationship between the amount of enzyme added and the degree of chitosan decomposition (viscosity) in Example 1, and FIG. 2 shows the relationship between the enzyme reaction time and the degree of chitosan decomposition (viscosity) in Example 1. Figure) Enzyme addition amount (% for 1 chitosan) Enzyme reaction time (hours)
Claims (2)
剤で酵素分解し、キトサンオリゴ糖を製造することを特
徴とするキトサンオリゴ糖の製造方法。(1) A method for producing chitosan oligosaccharides, which comprises producing chitosan oligosaccharides by enzymatically decomposing a chitosan solution with an enzyme agent containing pectinase as a main component.
糖からなる食品保存料。(2) A food preservative comprising chitosan oligosaccharide produced by the method according to claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP12185288A JPH01291799A (en) | 1988-05-20 | 1988-05-20 | Production of chitosan-oligosaccharide and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP12185288A JPH01291799A (en) | 1988-05-20 | 1988-05-20 | Production of chitosan-oligosaccharide and use thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH01291799A true JPH01291799A (en) | 1989-11-24 |
Family
ID=14821526
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP12185288A Pending JPH01291799A (en) | 1988-05-20 | 1988-05-20 | Production of chitosan-oligosaccharide and use thereof |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH01291799A (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04108396A (en) * | 1990-08-28 | 1992-04-09 | Higeta Shoyu Kk | Production of high polymerization degree chito-oligosaccharide |
| US5762992A (en) * | 1991-08-01 | 1998-06-09 | San-Ei Chemical Industries, Ltd. | Method of preventing a decrease in sweetness of thaumatin |
| JP2001333700A (en) * | 2000-05-26 | 2001-12-04 | Sanin Kensetsu Kogyo Kk | Pet food |
| KR100334132B1 (en) * | 1999-08-31 | 2002-04-26 | 김길환 | Method for Producing of Seasoned Chicken Meat Containing Chitosan Oligosaccharides |
| JP2002238466A (en) * | 2001-01-31 | 2002-08-27 | Iji Biosystem:Kk | Feed additive |
| JP2002315551A (en) * | 2001-04-19 | 2002-10-29 | Yoshinaga Nakai | Preservative for food and method for producing the same |
| WO2004101804A1 (en) * | 2003-05-16 | 2004-11-25 | Obschestvo S Ogranichennoy Otvetstvennost'yu 'invest-Farm' | Method for producing oligosaccharide chitosan and oligosaccharide chitosan |
| JP2013079217A (en) * | 2011-10-05 | 2013-05-02 | Koyo Chemical Kk | Oligoglucosamine that has reduced browning and method of manufacturing the oligoglucosamine |
| WO2014061705A1 (en) * | 2012-10-16 | 2014-04-24 | 味の素株式会社 | Food or drink with enhanced salty taste and/or umami and manufacturing method therefor, salty taste and/or umami-enhancing composition for food or drink, and method for enhancing salty taste and/or umami of food or drink |
| CN108949862A (en) * | 2018-08-24 | 2018-12-07 | 广东药科大学 | A kind of chitosan oligosaccharide and preparation method thereof of complex enzyme preparation |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5512222B2 (en) * | 1976-12-21 | 1980-03-31 |
-
1988
- 1988-05-20 JP JP12185288A patent/JPH01291799A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5512222B2 (en) * | 1976-12-21 | 1980-03-31 |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04108396A (en) * | 1990-08-28 | 1992-04-09 | Higeta Shoyu Kk | Production of high polymerization degree chito-oligosaccharide |
| US5762992A (en) * | 1991-08-01 | 1998-06-09 | San-Ei Chemical Industries, Ltd. | Method of preventing a decrease in sweetness of thaumatin |
| KR100334132B1 (en) * | 1999-08-31 | 2002-04-26 | 김길환 | Method for Producing of Seasoned Chicken Meat Containing Chitosan Oligosaccharides |
| JP2001333700A (en) * | 2000-05-26 | 2001-12-04 | Sanin Kensetsu Kogyo Kk | Pet food |
| JP2002238466A (en) * | 2001-01-31 | 2002-08-27 | Iji Biosystem:Kk | Feed additive |
| JP2002315551A (en) * | 2001-04-19 | 2002-10-29 | Yoshinaga Nakai | Preservative for food and method for producing the same |
| WO2004101804A1 (en) * | 2003-05-16 | 2004-11-25 | Obschestvo S Ogranichennoy Otvetstvennost'yu 'invest-Farm' | Method for producing oligosaccharide chitosan and oligosaccharide chitosan |
| JP2013079217A (en) * | 2011-10-05 | 2013-05-02 | Koyo Chemical Kk | Oligoglucosamine that has reduced browning and method of manufacturing the oligoglucosamine |
| WO2014061705A1 (en) * | 2012-10-16 | 2014-04-24 | 味の素株式会社 | Food or drink with enhanced salty taste and/or umami and manufacturing method therefor, salty taste and/or umami-enhancing composition for food or drink, and method for enhancing salty taste and/or umami of food or drink |
| CN108949862A (en) * | 2018-08-24 | 2018-12-07 | 广东药科大学 | A kind of chitosan oligosaccharide and preparation method thereof of complex enzyme preparation |
| CN108949862B (en) * | 2018-08-24 | 2021-02-26 | 广东药科大学 | Chitosan oligosaccharide prepared by complex enzyme and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| DE3781840T2 (en) | PASTA PROTEIN MATERIAL OR PROTEIN FOOD FROM CRUSTIES. | |
| JP5424181B2 (en) | Process for producing food for persons with difficulty in chewing / swallowing and food for persons having difficulty in chewing / swallowing | |
| KR20100075991A (en) | Food material suitable for patients with disturbance of mastication/swallowing | |
| DE2114066A1 (en) | New polysaccharide, process for its manufacture and use in the manufacture of food | |
| JP7109900B2 (en) | Flavor improving agent and method for improving flavor for vegetable protein-containing composition | |
| PL121126B1 (en) | Method of manufacture of dry sausages | |
| JP2006149384A (en) | Method for producing processed food | |
| KR101699823B1 (en) | Method for manufacturing kimchi | |
| JPH01291799A (en) | Production of chitosan-oligosaccharide and use thereof | |
| JPH0698738A (en) | Preservative for food | |
| CN107981253B (en) | Salted fish salt and preparation process thereof | |
| KR101425386B1 (en) | Sweet potato mayonnaise using enzymatically modified and method for the mayonnaise | |
| KR100515973B1 (en) | Food preservative composition and food comprising the same | |
| KR101105317B1 (en) | Method for manufacturing smoked-chicken breast containing chitosan and pomegranate | |
| US4788070A (en) | Process for acidifying ground meats | |
| JP2002171933A (en) | Kimchi essence health food | |
| JPH02238856A (en) | Seasoning and condiment containing miraculin as effective component and food and drink containing same | |
| JPH07147951A (en) | Production of food preservative and highly preservable food | |
| JP4658272B2 (en) | Method for producing egg white powder with high water retention | |
| JP2000175669A (en) | Quality improvement of pressurized and heatsterilized food | |
| JPH06153882A (en) | Preserving agent for food | |
| JPH1066521A (en) | Additive for beverage and food product | |
| JP2001327266A (en) | Method for producing meat-suspended dispersion | |
| KR20020082731A (en) | A manufacturing method of seasoning chitosan cuttlefish | |
| JP2849834B2 (en) | Frozen food, chilled food, livestock meat, fish meat paste product containing chitin or chitosan, and method for producing the same |