WO2022231003A1 - Kneaded meat product with improved quality - Google Patents

Kneaded meat product with improved quality Download PDF

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Publication number
WO2022231003A1
WO2022231003A1 PCT/JP2022/019450 JP2022019450W WO2022231003A1 WO 2022231003 A1 WO2022231003 A1 WO 2022231003A1 JP 2022019450 W JP2022019450 W JP 2022019450W WO 2022231003 A1 WO2022231003 A1 WO 2022231003A1
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phospholipase
heating
weight
yield
meat
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PCT/JP2022/019450
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French (fr)
Japanese (ja)
Inventor
宗一郎 郡
洋 ▲高▼▲柳▼
広悌 薄衣
孝彰 安部
裕行 中越
哲郎 堀
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味の素株式会社
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Priority to BR112023022575A priority Critical patent/BR112023022575A2/en
Priority to JP2023517634A priority patent/JPWO2022231003A1/ja
Publication of WO2022231003A1 publication Critical patent/WO2022231003A1/en

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
    • A23L13/60—Comminuted or emulsified meat products, e.g. sausages; Reformed meat from comminuted meat product
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L35/00—Food or foodstuffs not provided for in groups A23L5/00 – A23L33/00; Preparation or treatment thereof

Definitions

  • the present invention relates to a livestock meat paste product with improved quality, specifically to a livestock meat paste product with improved yield after heating during production.
  • meat paste products after molding the dough obtained by mixing minced meat and other raw materials, it is heated (for example, baked) to make the final product, but the yield after heating (relative to the weight before heating (before baking) If the ratio of the weight after heating (after baking) is high, the meat juice is trapped inside and is considered to be a desirable quality.
  • Patent Literature 1 discloses a method for producing a processed meat food in which lipase is added to the food so that the fatty acid content per 1 mg of the food is 13 ⁇ g or more and 80 ⁇ g or less.
  • Patent Document 2 describes a method for producing a processed meat food having a salt content of 0.1% to 1% by weight, comprising adding transglutaminase and gluconate or lipase to the processed meat food;
  • a method for producing a processed meat food is disclosed, which comprises adding transglutaminase, gluconate and lipase to the processed meat food.
  • Patent Document 3 in the production of processed meat products, by using phospholipase C and / or phospholipase D, texture such as hardness and suppleness of processed meat products is not deteriorated during storage of processed meat products. It is disclosed that separation of water can be suppressed.
  • the purpose of the present invention is to provide a livestock meat paste product with improved yield after heating during production.
  • the present inventors have made intensive studies to solve the above problems, and found new findings that meat paste products such as meatballs containing choline and inositol, which are free products from phospholipids, have a high yield after heating. Found it. Based on the above findings, the present inventors have made further intensive studies and completed the present invention.
  • a livestock meat paste product containing either one or both of the following (1) and (2). (1) 65 to 9000 ppm by weight of choline; (2) 230 to 380 ppm by weight of inositol; [3] The meat paste product according to [1] or [2] above, wherein the choline and inositol are derived from phospholipids contained in raw materials of the meat paste product. [4] A method for improving the post-heating yield of a livestock meat paste product, comprising allowing phospholipase D to act on livestock meat that is a raw material (in this specification, "raw meat” is sometimes referred to as "raw material meat”).
  • Phospholipase D is Sigma-Aldrich Phospholipase D Streptomyces derived (trade name), Nagase Chemtech Denazym PMD-P1 (trade name) and Asahi Kasei Pharma PHOSPHOLIPASE D [PLDP (T-39) ] (Glycerophospholipid specific) (trade name).
  • the meat paste product of the present invention has an improved yield after heating during production, and is expected to have improved texture and taste (for example, improved juiciness).
  • FIG. 1 shows the results of calculation of the post-heating yield in the production of meatballs of Examples 1 to 3 and Comparative Example 1 in Test Example 1.
  • FIG. FIG. 2 shows the results of calculation of the post-heating yield in the production of meatballs of Examples and Comparative Examples in Test Example 2.
  • FIG. 3 shows the results of calculation of the post-heating yield during production of the meatballs of Examples and Comparative Examples in Test Example 3.
  • FIG. 4 shows the results of calculation of post-heating yields in the production of coarsely ground sausages of Examples and Comparative Examples in Test Example 4.
  • FIG. FIG. 5 shows the results of calculation of the post-heating yield in the production of meatballs of Examples and Comparative Examples in Test Example 5.
  • FIG. FIG. 1 shows the results of calculation of the post-heating yield in the production of meatballs of Examples 1 to 3 and Comparative Example 1 in Test Example 1.
  • FIG. 3 shows the results of calculation of the post-heating yield during production of the meatballs of Examples and Comparative Examples in Test Example 3.
  • FIG. 6 shows the results of calculation of the post-heating yield in the production of meatballs of Examples and Comparative Examples in Test Example 6.
  • FIG. 7 shows the results of calculation of the post-heating yield during production of the meatballs of Examples and Comparative Examples in Test Example 7.
  • FIG. 8 shows the results of calculation of post-heating yields in the production of hamburger steaks of Examples and Comparative Examples in Test Example 8.
  • FIG. 9 shows the results of calculation of post-heating yields in the production of coarsely ground sausages of Examples and Comparative Examples in Test Example 9.
  • FIG. 10 shows the results of calculation of post-heating yields in the production of chicken meatballs of Examples and Comparative Examples in Test Example 10.
  • FIG. 11 shows the results of calculation of post-heating yields during production of coarsely ground sausages of Examples and Comparative Examples in Test Example 11.
  • FIG. 11 shows the results of calculation of post-heating yields during production of coarsely ground sausages of Examples and Comparative Examples in Test Example 11.
  • the livestock meat paste product of the present invention is characterized by containing either one or both of (1) and (2) below.
  • Examples of the livestock meat paste product of the present invention include a livestock meat paste product containing the above (1) and (2).
  • Examples of the livestock meat paste product of the present invention include the following (i) to (iv).
  • the content of choline and inositol in the livestock meat paste product can be measured by the method described in Test Example 1 below.
  • the choline and inositol are preferably choline and inositol derived from phospholipids (phosphatidylcholine, phosphatidylinositol) contained in raw materials of the meat paste product.
  • the "phospholipid contained in the raw material of the meat paste product” includes not only those derived from the raw meat of the meat paste product, but also other raw materials (e.g., eggs) that may optionally be contained in the meat paste product. It also includes those derived from The raw meat is the same as those exemplified in the description below.
  • the choline and inositol may be added in the product manufacturing process (for example, the raw material mixing process) so that the choline and inositol contents in the final product are within the above ranges.
  • the meat paste product of the present invention is a food made from ground meat as a main raw material, and examples thereof include meatballs, sausages (eg, coarsely ground sausages), hamburgers, chicken meatballs, ham, and meatloaf.
  • the livestock meat paste product of the present invention contains livestock meat as a raw material.
  • the raw meat include beef, pork, horse meat, mutton, goat meat, rabbit meat, chicken, turkey meat, duck meat, goose meat, and quail meat.
  • the content of raw livestock meat is, for example, 0.1 to 99% by weight, preferably 1 to 97% by weight, more preferably 10 to 90% by weight, based on the raw materials (total weight) of the product. is.
  • the meat paste product of the present invention contains ingredients commonly used in meat paste products (e.g., proteins such as onions, bread crumbs, eggs, vegetable proteins, gluten, egg whites, gelatin, and casein) and food additives.
  • ingredients commonly used in meat paste products e.g., proteins such as onions, bread crumbs, eggs, vegetable proteins, gluten, egg whites, gelatin, and casein
  • Substances e.g.
  • Phospholipases are enzymes that have the activity of hydrolyzing phospholipids.
  • Phospholipase D activity units herein are measured and defined as follows. 0.1 mL of the enzyme solution was mixed with 0.9 mL of the substrate solution containing phosphatidylcholine, reacted at 37° C. for 30 minutes, and after stopping the reaction, 50 ⁇ L of the reaction solution was added to 1 mL of the coloring solution containing choline oxidase, peroxidase, etc. and reacted for 5 minutes. After stopping the reaction, the amount of dye produced from choline is measured. 1 U (unit) is defined as the amount of enzyme that liberates 1 ⁇ mole of choline per minute at 37° C. using phosphatidylcholine as a substrate.
  • the amount of phospholipase D added is preferably 0.001 U or more, more preferably 0.01 to 100 U, and still more preferably 0.1 to 50 U per 1 g of the raw material of the meat paste product.
  • the action time (reaction time) of phospholipase D is not particularly limited as long as it is a time during which the enzyme can act on meat, which is a substrate substance. 10 minutes or longer, 20 minutes or longer, and 30 minutes or longer. Further, for example, 72 hours or less, 48 hours or less, 24 hours or less, 12 hours or less, 6 hours or less, 3 hours or less, 2 hours or less, and 1 hour or less. A realistic action time is preferably 0 to 72 hours, more preferably 30 minutes to 72 hours.
  • the action temperature (reaction temperature) is not particularly limited as long as the enzyme maintains its activity. The enzymatic reaction can be terminated, for example, by heating at 70-75° C. for 5-10 minutes.
  • At least one enzyme selected from the group consisting of transglutaminase, lipase, and amylolytic enzyme is further added to the raw animal meat and allowed to act. is preferred.
  • the order of addition may be in any order, and all may be added simultaneously or sequentially with a time lag. is desirable.
  • at least one enzyme selected from the group consisting of transglutaminase, lipase, and amylolytic enzyme is allowed to further act on raw meat, the action time, action temperature, and method for terminating the enzymatic reaction are determined by the action time of phospholipase D described above.
  • the following enzymes (I) to (VIII) are given as the enzymes to act on the raw meat.
  • (I) Phospholipase D II) phospholipase D and transglutaminase
  • III phospholipase D and lipase
  • IV phospholipase D and amylolytic enzyme (eg ⁇ -glucosidase)
  • V phospholipase D, transglutaminase and lipase
  • VI phospholipase D, transglutaminase and amylolytic enzymes (e.g.
  • VII Phospholipase D, lipase and amylolytic enzyme (eg ⁇ -glucosidase)
  • VIII Phospholipase D, transglutaminase, lipase and amylolytic enzyme (eg ⁇ -glucosidase)
  • the transglutaminase used in the present invention is an enzyme having the activity of catalyzing an acyl transfer reaction using a glutamine residue in a protein or peptide as a donor and a lysine residue as an acceptor, and is derived from mammals, for example. , those derived from fish, those derived from microorganisms, etc. are known.
  • the origin of the transglutaminase used in the present invention is not particularly limited as long as it has the above-mentioned activity, and transglutaminase of any origin can be used, and recombinant enzymes can also be used.
  • the transglutaminase used in the present invention may be a commercial product, and as a specific example, a microorganism-derived transglutaminase commercially available from Ajinomoto Co., Inc. under the trade name of "Activa" TG can be used alone or in combination. can.
  • the enzymatic activity of transglutaminase is defined by allowing transglutaminase to act in a reaction system using benzyloxycarbonyl-L-glutamylglycine and hydroxylamine as substrates in a Tris buffer at a temperature of 37° C. and a pH of 6.0.
  • the lipase used in the present invention is an enzyme that catalyzes the reaction of hydrolyzing fatty acid esters into fatty acids and glycerol.
  • Enzymes commercially available from Amano Enzyme Co., Ltd. under the trade names of "Lipase A 'Amano'6" and “Lipase AY 'Amano'” are examples of lipases.
  • the enzymatic activity of lipase was measured by emulsifying 100 ml of olive oil and 150 ml of a 2% PVA test solution as a substrate, mixing 5 ml of the substrate, 4 ml of McIlvaine buffer (pH 7.0) and 1 ml of the enzyme solution, followed by mixing at 37° C. for 60 minutes. After the reaction is terminated, the produced fatty acid is measured by a titration method. 1 U (unit) is defined as the acid liberating activity corresponding to 1 ⁇ mol of liberated oleic acid.
  • an amylolytic enzyme is an enzyme that catalyzes the decomposition reaction and transfer reaction of sugar chains containing starch.
  • amylolytic enzymes used in the present invention include ⁇ -glucosidase, ⁇ -amylase, ⁇ -amylase, glucoamylase, isomaltase, pullulanase, sucrose- ⁇ -glucosidase, ⁇ -glucosyltransferase, cyclomaltodextrinase, Examples include isoamylase and neopullulanase, with ⁇ -glucosidase being preferred.
  • the ⁇ -glucosidase used in the present invention is an enzyme that hydrolyzes a non-reducing terminal ⁇ -1,4-glucosidic bond to produce ⁇ -glucose.
  • transglucosidase is preferred.
  • An enzyme commercially available from Amano Enzyme Co., Ltd. under the trade name of "Transglucosidase L 'Amano'" is an example of ⁇ -glucosidase.
  • the enzymatic activity of ⁇ -glucosidase was measured by adding 1 ml of 0.02 M acetate buffer (pH 5.0) to 1 ml of 1 mM ⁇ -methyl-D-glucoside, adding 0.5 ml of the enzyme solution, and precipitating at 40° C. for 60 minutes.
  • the amount of enzyme that produces 1 ⁇ g of glucose in 2.5 ml of reaction solution when acted upon is defined as 1 U (unit).
  • the amount of transglutaminase to be added is such that the enzymatic activity is, for example, 0.005 to 3.0 U, preferably 0.01 to 1.0 U, per 1 g of the raw material for the meat paste product. 0 U, more preferably 0.05 to 0.5 U, still more preferably 0.1 to 0.3 U.
  • the amount of lipase to be added is such that the enzymatic activity is, for example, 0.1 to 30.0 U, preferably 0.3 to 15.0 U, preferably 0.3 to 15.0 U, per 1 g of the raw material of the meat paste product. It is more preferably 0.5 to 10.0U, still more preferably 1.0 to 3.0U.
  • the amount of the amylolytic enzyme (eg, ⁇ -glucosidase) added is such that the enzymatic activity per 1 g of the raw material of the meat paste product is, for example, It is 0.0005 to 0.1 U, preferably 0.001 to 0.05 U, more preferably 0.002 to 0.02 U, still more preferably 0.005 to 0.01 U.
  • the meat paste product of the present invention produced by the above-described production method of the present invention has an improved yield after heating during production, and is expected to have an improved texture and taste.
  • the present invention also relates to a method for improving the post-heating yield of livestock meat paste products, which comprises allowing phospholipase D to act on raw livestock meat.
  • the phospholipase D is Sigma-Aldrich's phospholipase D actinomycete-derived (trade name), Nagase Chemtech's Denazym PMD-P1 (trade name), and Asahi Kasei Pharma's PHOSPHOLIPASE D [PLDP ( T-39)] (Glycerophospholipid specific) (trade names) are preferred.
  • At least one enzyme selected from the group consisting of transglutaminase, lipase, and amylolytic enzyme may be further acted on the raw meat.
  • the following enzymes (I) to (VIII) are listed as enzymes to act on raw meat.
  • VII Phospholipase D, lipase and amylolytic enzyme (eg ⁇ -glucosidase)
  • VIII Phospholipase D, transglutaminase, lipase and amylolytic enzyme (eg ⁇ -glucosidase)
  • examples of meat paste products examples of raw meat, amounts used, definitions, examples, addition amounts, and addition of phospholipase D, transglutaminase, lipase, and amylolytic enzyme ( ⁇ -glucosidase)
  • the method, optional ingredients, additives, etc. are described in the method for producing the livestock meat paste product of the present invention, examples of livestock meat paste products, examples of raw meat paste products, amounts used, phospholipase D, transglutaminase, lipase and It is the same as the definition, exemplification, addition amount, addition method, optionally added components, additives, etc. of the amylolytic enzyme ( ⁇ -glucosidase).
  • the post-heating yield of the meat paste product refers to the weight after heating (after baking) relative to the weight before heating (before baking) of the dough (putty) obtained by mixing the raw materials at the time of manufacturing the meat paste product. refers to the ratio (% by weight) of
  • the improved yield after heating of the meat paste product means that the yield after heating is higher than that of the meat paste product produced without the addition of enzymes.
  • Test Example 1 (Production of meatballs of Examples 1 to 3 and Comparative Example 1)
  • the beef rib lean meat and beef Kenne fat shown in Table 1 were minced to 3 mm by a meat grinder (chopper).
  • the onion was chopped into pieces of about 2 mm.
  • Each phospholipase D shown in Table 2 was added to the raw material of the composition shown in Table 1 so as to be 3.0 U per 1 g of raw material, and a standard mixer ("KitchenAid KSM5WH", manufactured by Whirlpool Corporation) was added. and mixed for 3 minutes at a setting speed of 1 to prepare a meatball dough. After degassing, it was manually molded into 30 g spheres. After molding, baked at 250 ° C.
  • cholinesterase kit-NC 289-75181, Fujifilm Wako Pure Chemical Industries, Ltd.
  • a calibration curve was prepared from the absorbance, and the content of free choline in meatballs was determined. The results are shown in Table 5-2.
  • the choline content in the meatball (estimated value: 60 ppm by weight) is the amount of choline originally contained in the raw material.
  • the choline content in the meatballs of Examples 2-1 to 2-5 and Comparative Examples 2-2 to 2-4 shown in Tables 7-1 and 7-2 is the choline originally contained in the raw materials. It is the sum of the amount (estimated value: 60 ppm by weight) and the added amount of choline. (Evaluation of yield after heating) In order to evaluate the yield after heating, the weight before baking and the weight after baking were measured at the time of meatball production.
  • Test Example 3 Inositol addition test (production of meatballs of Examples 3-1 to 3-3 and Comparative Examples 3-2 to 3-7)
  • the beef rib lean meat and beef kenne fat shown in Table 6 of Test Example 2 were minced to 3 mm by a meat grinder (chopper). The onion was chopped into pieces of about 2 mm.
  • a standard mixer (“KitchenAid KSM5WH”, manufactured by Whirlpool Corporation) was prepared using raw materials with the formulations shown in Table 6 of Test Example 2 and inositol in the amounts shown in Tables 8-1 and 8-2. was mixed for 3 minutes at a setting speed of 1 to prepare a meatball dough. After degassing, it was manually molded into 30 g spheres.
  • the inositol content in the meatball of Comparative Example 3-1 shown in Table 8-1 is the inositol content in the meatball of Comparative Example 1 of Test Example 1 produced by the same method (213 ppm by weight, Table 5- 3). Since inositol was not added in Comparative Example 3-1, the content of inositol in the meatball (estimated value: 213 ppm by weight) is the amount of inositol originally contained in the raw material.
  • the inositol content in the meatballs of Examples 3-1 to 3-3 and Comparative Examples 3-2 to 3-7 shown in Tables 8-1 and 8-2 is the inositol originally contained in the raw materials.
  • the collagen casing manufactured by Devro Co., Ltd.
  • coarsely ground sausages of Examples 4-1 to 4-7 were produced.
  • a coarsely ground sausage of Comparative Example 4-1 was obtained in the same manner as in Examples 4-1 to 4-7 except that no enzyme was added.
  • evaluation of yield after heating In order to evaluate the post-heating yield, the weight before drying and the weight after cooling (that is, after the drying-smoking-steaming-cooling process) were measured during the production of the coarsely ground sausage.
  • a standard mixer (“KitchenAid KSM5WH”, manufactured by Whirlpool Corporation) was used and mixed at a setting speed of 1 for 3 minutes to prepare a meatball dough. After degassing, it was manually molded into 30 g spheres. After molding, it was allowed to stand in a refrigerator for 1 hour. Baking at 250 ° C. for 1 minute with an impinger (FGJOA 9L, manufactured by Fujimac) and baking at 200 ° C. and RH 50% for 6 minutes in a steam convection oven (FSCC WE 61G, manufactured by Fujimac), Examples 5-1 to 5 -3 meatballs were obtained.
  • FGJOA 9L an impinger
  • FSCC WE 61G steam convection oven
  • Example 5-1 produced by adding phospholipase D
  • Examples 5-2 and 5-3 produced by adding phospholipase D and transglutaminase were Compared to Comparative Example 5-1 without enzyme addition and Comparative Example 5-2 without phospholipase D addition, the post-heating yield was higher, and an improvement in post-heating yield was observed.
  • Example 6-1 produced by adding phospholipase D and Examples 6-2 to 6-4 produced by adding phospholipase D and ⁇ -glucosidase , Compared to Comparative Example 6-1 without enzyme addition and Comparative Example 6-2 without phospholipase D addition, the yield after heating was high, and an improvement in yield after heating was observed.
  • Example 7 Enzyme addition test (combined use of phospholipase D, ⁇ -glucosidase and lipase (production of meatballs of Example 7-1) A meatball of Example 7-1 was obtained in the same manner as in Example 5-1, etc., except that the enzyme was changed to the enzyme and the amount shown in Table 14.
  • Enzymes include phospholipase D (Denazyme PMD-P1, manufactured by Nagase ChemteX Co., Ltd.), ⁇ -glucosidase (Transglucosidase L "Amano”, manufactured by Amano Enzyme Co., Ltd.) and lipase (Lipase AY "Amano", manufactured by Amano Enzyme Co., Ltd.). )It was used.
  • Production of meatballs in Comparative Example 7-1 A meatball of Comparative Example 7-1 was obtained in the same manner as in Example 7-1 except that no enzyme was added.
  • evaluation of yield after heating In order to evaluate the yield after heating, the weight before baking and the weight after baking were measured at the time of meatball production. The post-heating yield was calculated in the same manner as in Test Example 5. The results are shown in Table 14 and FIG.
  • Example 7-1 produced by adding phospholipase D, ⁇ -glucosidase and lipase had a higher yield after heating than Comparative Example 7-1 without enzyme addition. was high, and an improvement in yield was observed after heating.
  • the dough was weighed to 150 g, and after the air was removed, it was formed into an oval shape (major axis: 130 mm, minor axis: 90 mm, thickness: 12 mm).
  • the hamburger dough is baked on a griddle (FGFT60601TC, Fujimac Co., Ltd.) at 230 ° C. for 1 minute each on both sides, and after the surface is baked, baked at 250 ° C. for 4 minutes with an impinger (FGJOA9L, Fujimac Co., Ltd.).
  • FGJOA9L Fujimac Co., Ltd.
  • Example 9-1 produced by adding phospholipase D has a higher yield after heating than Comparative Example 9-1 without enzyme addition, and the yield after heating is improvement was observed.
  • Example 10-1 produced by adding phospholipase D had a higher yield after heating than Comparative Example 10-1 without enzyme addition, and a lower yield after heating. Improvement was observed.
  • Example 11-1 produced by adding phospholipase D has a high yield after heating and a high yield after heating compared to Comparative Example 11-1 without enzyme addition. improvement was observed.

Abstract

The objective of the present invention is to provide a kneaded meat product with improved post-cooking yield during production, and a method of improving post-cooking yield of a kneaded meat product.　The kneaded meat product contains one or both of (1) 65-9,000 weight ppm of choline, and (2) 230-380 weight ppm of inositol. The method of improving post-cooking yield of a kneaded meat product comprises causing phospholipase D to act on meat as a raw material.

Description

品質が向上した畜肉練り製品Livestock meat paste product with improved quality
　本発明は、品質が向上した畜肉練り製品、具体的には、製造時における加熱後歩留りが向上した畜肉練り製品に関する。 The present invention relates to a livestock meat paste product with improved quality, specifically to a livestock meat paste product with improved yield after heating during production.
　通常ソーセージやミートボールといった畜肉練り製品を製造する際には、原料挽肉と塩類を混合し、塩溶性タンパク質を溶出させる事で、乳化を促進させる。乳化を促進することで畜肉練り製品の硬さや保水性に影響を与えるが、乳化剤や卵、乳カゼインといった既存の乳化促進素材は、最終製品への表示必要性や、アレルゲンの観点から、使用することが難しかった。
　また、畜肉練り製品においては、挽肉及びその他の原材料を混合して得られた生地を成形後、加熱（例えば焼成）して最終製品とするが、加熱後歩留り（加熱前（焼成前）の重量に対する加熱後（焼成後）の重量の割合）が高いと、肉汁が内部に閉じ込められており好ましい品質と考えられる。 Normally, when producing meat paste products such as sausages and meatballs, raw minced meat and salts are mixed to elute salt-soluble proteins to promote emulsification. Promoting emulsification affects the hardness and water retention of meat paste products, but existing emulsification-promoting ingredients such as emulsifiers, eggs, and milk casein cannot be used due to the need for labeling on final products and from the perspective of allergens. was difficult.
In addition, in meat paste products, after molding the dough obtained by mixing minced meat and other raw materials, it is heated (for example, baked) to make the final product, but the yield after heating (relative to the weight before heating (before baking) If the ratio of the weight after heating (after baking) is high, the meat juice is trapped inside and is considered to be a desirable quality.
　畜肉練り製品の硬さや保水性の改善を目的として、製造時に酵素を用いる方法が検討されている。
　特許文献１には、食品にリパーゼを添加し、食品１ｍｇあたりの脂肪酸含有量を１３μｇ以上、８０μg以下とする、食肉加工食品の製造方法が開示されている。
　特許文献２には、食塩の含有量が０．１重量％～１重量％である食肉加工食品の製造方法であって、トランスグルタミナーゼとグルコン酸塩もしくはリパーゼとを食肉加工食品に添加すること、またはトランスグルタミナーゼとグルコン酸塩およびリパーゼとを食肉加工食品に添加することを含む、食肉加工食品の製造方法が開示されている。
　特許文献３には、畜肉加工製品の製造において、ホスホリパーゼＣ及び／又はホスホリパーゼＤを用いることにより、畜肉加工製品の硬さ、しなやかさ等の食感を劣化させることなく、畜肉加工製品の保存時の離水を抑制することができることが開示されている。 For the purpose of improving the hardness and water retention of livestock meat paste products, methods using enzymes during production have been investigated.
Patent Literature 1 discloses a method for producing a processed meat food in which lipase is added to the food so that the fatty acid content per 1 mg of the food is 13 µg or more and 80 µg or less.
Patent Document 2 describes a method for producing a processed meat food having a salt content of 0.1% to 1% by weight, comprising adding transglutaminase and gluconate or lipase to the processed meat food; Alternatively, a method for producing a processed meat food is disclosed, which comprises adding transglutaminase, gluconate and lipase to the processed meat food.
In Patent Document 3, in the production of processed meat products, by using phospholipase C and / or phospholipase D, texture such as hardness and suppleness of processed meat products is not deteriorated during storage of processed meat products. It is disclosed that separation of water can be suppressed.
特開２０１８－１０２２９７号公報JP 2018-102297 A 国際公開第２０２０／００４４１９号WO2020/004419 国際公開第２０１０／１４０７０８号WO2010/140708
　本発明は、製造時における加熱後歩留りが向上した畜肉練り製品の提供を目的とする。 The purpose of the present invention is to provide a livestock meat paste product with improved yield after heating during production.
　本発明者らは、上記課題の解決のために鋭意検討をしたところ、リン脂質からの遊離物であるコリン、イノシトールが含まれるミートボール等の畜肉練り製品が、加熱後歩留りが高いという新知見を見出した。
　上記知見に基づいて、本発明者らは、さらに鋭意検討をして、本発明を完成させた。 The present inventors have made intensive studies to solve the above problems, and found new findings that meat paste products such as meatballs containing choline and inositol, which are free products from phospholipids, have a high yield after heating. Found it.
Based on the above findings, the present inventors have made further intensive studies and completed the present invention.
　すなわち、本発明は、下記を提供する。
［１］下記（１）、（２）のいずれか一方又は両方を含有する、畜肉練り製品。
（１）６５～９０００重量ｐｐｍのコリン
（２）２３０～３８０重量ｐｐｍのイノシトール
［２］前記（１）及び（２）を含有する、上記［１］記載の畜肉練り製品。
［３］前記コリン及びイノシトールが、畜肉練り製品の原材料に含まれるリン脂質に由来する、上記［１］又は［２］に記載の畜肉練り製品。
［４］原材料である畜肉（本明細書において「原材料である畜肉」を「原料畜肉」と称する場合がある。）にホスホリパーゼＤを作用させることを含む、畜肉練り製品の加熱後歩留り向上方法。
［５］ホスホリパーゼＤが、シグマ　アルドリッチ（Ｓｉｇｍａ－Ａｌｄｒｉｃｈ）製ホスホリパーゼＤ　放線菌由来（商品名）、ナガセケムテック製デナチームＰＭＤ－Ｐ１（商品名）及び旭化成ファーマ製ＰＨＯＳＰＨＯＬＩＰＡＳＥ　Ｄ［ＰＬＤＰ（Ｔ－３９）］（Ｇｌｙｃｅｒｏｐｈｏｓｐｈｏｌｉｐｉｄ　ｓｐｅｃｉｆｉｃ）（商品名）からなる群から選択される、上記［４］記載の方法。
［６］トランスグルタミナーゼ、リパーゼ、及び澱粉分解酵素からなる群から選択される少なくとも１つの酵素をさらに作用させることを含む、上記［４］または［５］に記載の方法。
［７］原材料である畜肉に、シグマ　アルドリッチ製ホスホリパーゼＤ　放線菌由来（商品名）、ナガセケムテック製デナチームＰＭＤ－Ｐ１（商品名）及び旭化成ファーマ製ＰＨＯＳＰＨＯＬＩＰＡＳＥ　Ｄ［ＰＬＤＰ（Ｔ－３９）］（Ｇｌｙｃｅｒｏｐｈｏｓｐｈｏｌｉｐｉｄ　ｓｐｅｃｉｆｉｃ）（商品名）からなる群から選択されるホスホリパーゼＤを作用させることを含む、上記［１］～［３］のいずれかに記載の畜肉練り製品の製造方法。
［８］トランスグルタミナーゼ、リパーゼ、及び澱粉分解酵素からなる群から選択される少なくとも１つの酵素をさらに作用させることを含む、上記［７］記載の方法。
［９］原材料である畜肉に、シグマ　アルドリッチ製ホスホリパーゼＤ　放線菌由来（商品名）、ナガセケムテック製デナチームＰＭＤ－Ｐ１（商品名）及び旭化成ファーマ製ＰＨＯＳＰＨＯＬＩＰＡＳＥ　Ｄ［ＰＬＤＰ（Ｔ－３９）］（Ｇｌｙｃｅｒｏｐｈｏｓｐｈｏｌｉｐｉｄ　ｓｐｅｃｉｆｉｃ）（商品名）からなる群から選択されるホスホリパーゼＤを作用させることを含む、畜肉練り製品の製造方法。
［１０］トランスグルタミナーゼ、リパーゼ、及び澱粉分解酵素からなる群から選択される少なくとも１つの酵素をさらに作用させることを含む、上記［９］記載の方法。 That is, the present invention provides the following.
[1] A livestock meat paste product containing either one or both of the following (1) and (2).
(1) 65 to 9000 ppm by weight of choline; (2) 230 to 380 ppm by weight of inositol;
[3] The meat paste product according to [1] or [2] above, wherein the choline and inositol are derived from phospholipids contained in raw materials of the meat paste product.
[4] A method for improving the post-heating yield of a livestock meat paste product, comprising allowing phospholipase D to act on livestock meat that is a raw material (in this specification, "raw meat" is sometimes referred to as "raw material meat").
[5] Phospholipase D is Sigma-Aldrich Phospholipase D Streptomyces derived (trade name), Nagase Chemtech Denazym PMD-P1 (trade name) and Asahi Kasei Pharma PHOSPHOLIPASE D [PLDP (T-39) ] (Glycerophospholipid specific) (trade name).
[6] The method according to [4] or [5] above, which further comprises allowing at least one enzyme selected from the group consisting of transglutaminase, lipase, and amylolytic enzyme to act.
[7] For livestock meat, which is a raw material, Phospholipase D derived from actinomycetes (trade name) manufactured by Sigma-Aldrich, Denathym PMD-P1 (trade name) manufactured by Nagase Chemtech, and PHOSPHOLIPASE D [PLDP (T-39)] (Glycerophospholipid) manufactured by Asahi Kasei Pharma specific) (trade name).
[8] The method according to [7] above, which further comprises allowing at least one enzyme selected from the group consisting of transglutaminase, lipase, and amylolytic enzyme to act.
[9] For livestock meat, which is a raw material, Phospholipase D derived from actinomycetes (trade name) manufactured by Sigma-Aldrich, Denathym PMD-P1 (trade name) manufactured by Nagase Chemtech, and PHOSPHOLIPASE D [PLDP (T-39)] (Glycerophospholipid) manufactured by Asahi Kasei Pharma specific) (trade name).
[10] The method according to [9] above, which further comprises allowing at least one enzyme selected from the group consisting of transglutaminase, lipase, and amylolytic enzyme to act.
　本発明の畜肉練り製品は、製造時における加熱後歩留りが向上しており、ひいては食感及び呈味の向上（例えば、ジューシーさの向上）が期待できる畜肉練り製品である。 The meat paste product of the present invention has an improved yield after heating during production, and is expected to have improved texture and taste (for example, improved juiciness).
図１は、試験例１において、実施例１～３及び比較例１のミートボールの製造時の加熱後歩留りを算出した結果を示す。FIG. 1 shows the results of calculation of the post-heating yield in the production of meatballs of Examples 1 to 3 and Comparative Example 1 in Test Example 1. FIG. 図２は、試験例２において、実施例及び比較例のミートボールの製造時の加熱後歩留りを算出した結果を示す。FIG. 2 shows the results of calculation of the post-heating yield in the production of meatballs of Examples and Comparative Examples in Test Example 2. As shown in FIG. 図３は、試験例３において、実施例及び比較例のミートボールの製造時の加熱後歩留りを算出した結果を示す。FIG. 3 shows the results of calculation of the post-heating yield during production of the meatballs of Examples and Comparative Examples in Test Example 3. FIG. 図４は、試験例４において、実施例及び比較例の粗挽きソーセージの製造時の加熱後歩留りを算出した結果を示す。FIG. 4 shows the results of calculation of post-heating yields in the production of coarsely ground sausages of Examples and Comparative Examples in Test Example 4. FIG. 図５は、試験例５において、実施例及び比較例のミートボールの製造時の加熱後歩留りを算出した結果を示す。FIG. 5 shows the results of calculation of the post-heating yield in the production of meatballs of Examples and Comparative Examples in Test Example 5. As shown in FIG. 図６は、試験例６において、実施例及び比較例のミートボールの製造時の加熱後歩留りを算出した結果を示す。FIG. 6 shows the results of calculation of the post-heating yield in the production of meatballs of Examples and Comparative Examples in Test Example 6. As shown in FIG. 図７は、試験例７において、実施例及び比較例のミートボールの製造時の加熱後歩留りを算出した結果を示す。FIG. 7 shows the results of calculation of the post-heating yield during production of the meatballs of Examples and Comparative Examples in Test Example 7. FIG. 図８は、試験例８において、実施例及び比較例のハンバーグの製造時の加熱後歩留りを算出した結果を示す。FIG. 8 shows the results of calculation of post-heating yields in the production of hamburger steaks of Examples and Comparative Examples in Test Example 8. FIG. 図９は、試験例９において、実施例及び比較例の粗挽きソーセージの製造時の加熱後歩留りを算出した結果を示す。FIG. 9 shows the results of calculation of post-heating yields in the production of coarsely ground sausages of Examples and Comparative Examples in Test Example 9. FIG. 図１０は、試験例１０において、実施例及び比較例の鶏つみれの製造時の加熱後歩留りを算出した結果を示す。FIG. 10 shows the results of calculation of post-heating yields in the production of chicken meatballs of Examples and Comparative Examples in Test Example 10. FIG. 図１１は、試験例１１において、実施例及び比較例の粗挽きソーセージの製造時の加熱後歩留りを算出した結果を示す。FIG. 11 shows the results of calculation of post-heating yields during production of coarsely ground sausages of Examples and Comparative Examples in Test Example 11. FIG.
　以下に、本発明を詳細に説明する。
　本発明の畜肉練り製品は、以下（１）、（２）のいずれか一方又は両方を含有することを特徴とする。
（１）６５～９０００重量ｐｐｍ（好ましくは８０～７０００重量ｐｐｍ、より好ましくは１００～５０００重量ｐｐｍ、さらに好ましくは２６０～２０６０重量ｐｐｍ）のコリン
（２）２３０～３８０重量ｐｐｍ（好ましくは２３５～３６３重量ｐｐｍ、より好ましくは２４０～３２０重量ｐｐｍ、さらに好ましくは２４３～２７３重量ｐｐｍ）のイノシトール The present invention will be described in detail below.
The livestock meat paste product of the present invention is characterized by containing either one or both of (1) and (2) below.
(1) 65 to 9000 ppm by weight (preferably 80 to 7000 ppm by weight, more preferably 100 to 5000 ppm by weight, more preferably 260 to 2060 ppm by weight) of choline; 363 ppm by weight, more preferably 240 to 320 ppm by weight, more preferably 243 to 273 ppm by weight) of inositol
　本発明の畜肉練り製品としては、例えば、上記（１）及び（２）を含有する畜肉練り製品が挙げられる。
　本発明の畜肉練り製品としては、例えば、以下（i）～（iv）が挙げられる。
（i）（１）６５～９０００重量ｐｐｍのコリン及び（２）２３０～３８０重量ｐｐｍのイノシトールを含有する、畜肉練り製品。
（ii）（１）８０～７０００重量ｐｐｍのコリン及び（２）２３５～３６３重量ｐｐｍのイノシトールを含有する、畜肉練り製品。
（iii）（１）１００～５０００重量ｐｐｍのコリン及び（２）２４０～３２０重量ｐｐｍのイノシトールを含有する、畜肉練り製品。
（iv）（１）２６０～２０６０重量ｐｐｍのコリン及び（２）２４３～２７３重量ｐｐｍのイノシトールを含有する、畜肉練り製品。 Examples of the livestock meat paste product of the present invention include a livestock meat paste product containing the above (1) and (2).
Examples of the livestock meat paste product of the present invention include the following (i) to (iv).
(i) A meat paste containing (1) 65-9000 ppm by weight choline and (2) 230-380 ppm by weight inositol.
(ii) A meat paste containing (1) 80-7000 ppm by weight choline and (2) 235-363 ppm by weight inositol.
(iii) A livestock meat paste product containing (1) 100 to 5000 ppm by weight of choline and (2) 240 to 320 ppm by weight of inositol.
(iv) A meat paste containing (1) 260-2060 ppm by weight choline and (2) 243-273 ppm by weight inositol.
　本発明において、コリン、イノシトールの畜肉練り製品中の含有量は、後述の試験例１に記載の方法によって、測定することができる。 In the present invention, the content of choline and inositol in the livestock meat paste product can be measured by the method described in Test Example 1 below.
　本発明の畜肉練り製品において、前記コリン、イノシトールは、畜肉練り製品の原材料に含まれるリン脂質（ホスファチジルコリン、ホスファチジルイノシトール）に由来するコリン、イノシトールであることが好ましい。本発明において、「畜肉練り製品の原材料に含まれるリン脂質」には、畜肉練り製品の原料畜肉に由来するものだけでなく、畜肉練り製品に任意に含有していてもよいその他の原材料（例えば、卵）に由来するものも含まれる。該原料畜肉は、後述の説明で例示するものと同様である。
　本発明の畜肉練り製品において、前記コリン、イノシトールは、最終製品中のコリン、イノシトール含有量が上記した範囲となるように、製品の製造工程（例えば、原材料の混合工程）で添加してもよい。 In the meat paste product of the present invention, the choline and inositol are preferably choline and inositol derived from phospholipids (phosphatidylcholine, phosphatidylinositol) contained in raw materials of the meat paste product. In the present invention, the "phospholipid contained in the raw material of the meat paste product" includes not only those derived from the raw meat of the meat paste product, but also other raw materials (e.g., eggs) that may optionally be contained in the meat paste product. It also includes those derived from The raw meat is the same as those exemplified in the description below.
In the livestock meat paste product of the present invention, the choline and inositol may be added in the product manufacturing process (for example, the raw material mixing process) so that the choline and inositol contents in the final product are within the above ranges.
　本発明の畜肉練り製品は、挽肉を主原材料にして作られる食品であり、例えば、ミートボール、ソーセージ（例えば、粗挽きソーセージ）、ハンバーグ、鶏つみれ、ハム、ミートローフが挙げられる。 The meat paste product of the present invention is a food made from ground meat as a main raw material, and examples thereof include meatballs, sausages (eg, coarsely ground sausages), hamburgers, chicken meatballs, ham, and meatloaf.
　本発明の畜肉練り製品は、原材料として畜肉を含有する。該原料畜肉としては、例えば、牛肉、豚肉、馬肉、めん羊肉、山羊肉、家兎肉、鶏肉、七面鳥肉、カモ肉、ガチョウ肉、鶉肉が挙げられる。
　本発明の畜肉練り製品において、原料畜肉の含有量は、製品の原材料（合計重量）に対して、例えば０．１～９９重量％、好ましくは１～９７重量％、より好ましくは１０～９０重量％である。 The livestock meat paste product of the present invention contains livestock meat as a raw material. Examples of the raw meat include beef, pork, horse meat, mutton, goat meat, rabbit meat, chicken, turkey meat, duck meat, goose meat, and quail meat.
In the livestock meat paste product of the present invention, the content of raw livestock meat is, for example, 0.1 to 99% by weight, preferably 1 to 97% by weight, more preferably 10 to 90% by weight, based on the raw materials (total weight) of the product. is.
　本発明の畜肉練り製品は、上記した成分の他に、畜肉練り製品において一般的に使用される食材（例えば、玉ねぎ、パン粉、卵、植物蛋白、グルテン、卵白、ゼラチン、カゼイン等の蛋白質）や食品添加物（例えば、食塩、澱粉、加工澱粉、小麦粉、畜肉エキス等の調味料、香辛料、発色剤、酸化防止剤、デキストリン等の賦形剤、蛋白加水分解物、蛋白部分分解物、乳化剤、クエン酸塩、重合リン酸塩等のキレート剤、グルタチオン、システイン等の還元剤、アルギン酸、かんすい、色素、酸味料、香料等）を原材料として含有していてもよい。 In addition to the above ingredients, the meat paste product of the present invention contains ingredients commonly used in meat paste products (e.g., proteins such as onions, bread crumbs, eggs, vegetable proteins, gluten, egg whites, gelatin, and casein) and food additives. Substances (e.g. salt, starch, modified starch, wheat flour, seasonings such as meat extracts, spices, coloring agents, antioxidants, excipients such as dextrin, protein hydrolysates, protein partial hydrolysates, emulsifiers, citric acid Salts, chelating agents such as polymerized phosphates, reducing agents such as glutathione and cysteine, alginic acid, lye salt, pigments, acidulants, fragrances, etc.) may be contained as raw materials.
　本発明の畜肉練り製品は、原料畜肉に、シグマ　アルドリッチ製ホスホリパーゼＤ　放線菌由来（商品名）、ナガセケムテック製デナチームＰＭＤ－Ｐ１（商品名）及び旭化成ファーマ製ＰＨＯＳＰＨＯＬＩＰＡＳＥ　Ｄ［ＰＬＤＰ（Ｔ－３９）］（Ｇｌｙｃｅｒｏｐｈｏｓｐｈｏｌｉｐｉｄ　ｓｐｅｃｉｆｉｃ）（商品名）からなる群から選択されるホスホリパーゼＤを作用させることで製造することができる。該方法によって、原料畜肉に由来するコリンを上記範囲で含む、本発明の畜肉練り製品を製造することができる。 In the livestock meat paste product of the present invention, Sigma-Aldrich's Phospholipase D Actinomycete-derived (trade name), Nagase Chemtech's Denathym PMD-P1 (trade name), and Asahi Kasei Pharma's PHOSPHOLIPASE D [PLDP (T-39)] are added to raw meat. (Glycerophospholipid specific) (trade name). By this method, the meat paste product of the present invention containing the choline derived from the raw material meat within the above range can be produced.
　ホスホリパーゼは、リン脂質を加水分解する活性を有する酵素である。
　本明細書においてホスホリパーゼＤの活性単位は、次のように測定され、かつ、定義される。
　ホスファチジルコリンを含む基質溶液０．９ｍＬに酵素溶液０．１ｍＬを混和し、３７℃にて３０分間反応させ、反応停止後、コリンオキシダーゼ、ペルオキシダーゼ等を含む発色溶液１ｍＬに反応液５０μＬを加え５分反応させ、反応停止後、コリンより生成した色素量を測定する。ホスファチジルコリンを基質として３７℃、１分間に１μモルのコリンを遊離する酵素量を１Ｕ（ユニット）と定義する。 Phospholipases are enzymes that have the activity of hydrolyzing phospholipids.
Phospholipase D activity units herein are measured and defined as follows.
0.1 mL of the enzyme solution was mixed with 0.9 mL of the substrate solution containing phosphatidylcholine, reacted at 37° C. for 30 minutes, and after stopping the reaction, 50 μL of the reaction solution was added to 1 mL of the coloring solution containing choline oxidase, peroxidase, etc. and reacted for 5 minutes. After stopping the reaction, the amount of dye produced from choline is measured. 1 U (unit) is defined as the amount of enzyme that liberates 1 μmole of choline per minute at 37° C. using phosphatidylcholine as a substrate.
　本発明において、ホスホリパーゼＤの添加量は、畜肉練り製品の原材料１ｇに対して酵素活性が、好ましくは０．００１Ｕ以上、より好ましくは０．０１～１００Ｕ、さらに好ましくは０．１～５０Ｕである。 In the present invention, the amount of phospholipase D added is preferably 0.001 U or more, more preferably 0.01 to 100 U, and still more preferably 0.1 to 50 U per 1 g of the raw material of the meat paste product.
　ホスホリパーゼＤの作用時間（反応時間）は、酵素が基質物質である肉に作用することが可能な時間であれば特に限定されないが、例えば、０分以上、１分以上、３分以上、５分以上、１０分以上、２０分以上、３０分以上が挙げられる。また、例えば、７２時間以下、４８時間以下、２４時間以下、１２時間以下、６時間以下、３時間以下、２時間以下、１時間以下が挙げられる。現実的な作用時間としては０～７２時間が好ましく、３０分～７２時間がより好ましい。また、作用温度（反応温度）に関しても酵素が活性を保つ範囲であれば特に限定されないが、現実的な温度としては０～６０℃で作用させることが好ましい。酵素反応は、例えば、７０～７５℃で、５～１０分間加熱により終了させることができる。 The action time (reaction time) of phospholipase D is not particularly limited as long as it is a time during which the enzyme can act on meat, which is a substrate substance. 10 minutes or longer, 20 minutes or longer, and 30 minutes or longer. Further, for example, 72 hours or less, 48 hours or less, 24 hours or less, 12 hours or less, 6 hours or less, 3 hours or less, 2 hours or less, and 1 hour or less. A realistic action time is preferably 0 to 72 hours, more preferably 30 minutes to 72 hours. Also, the action temperature (reaction temperature) is not particularly limited as long as the enzyme maintains its activity. The enzymatic reaction can be terminated, for example, by heating at 70-75° C. for 5-10 minutes.
　上記した本発明の製造方法において、上記したホスホリパーゼＤに加えて、トランスグルタミナーゼ、リパーゼ、及び澱粉分解酵素からなる群から選択される少なくとも１つの酵素を、さらに原料畜肉に添加して、作用させることが好ましい。
　複数の酵素を添加する場合の添加順序は、いかなる順序でもよく、全てを同時に添加しても、時間差をつけて順に添加しても構わないが、簡便性の観点から、全てを同時に添加するのが望ましい。
　トランスグルタミナーゼ、リパーゼ、及び澱粉分解酵素からなる群から選択される少なくとも１つの酵素をさらに原料畜肉に作用させる場合の、作用時間、作用温度、酵素反応の終了方法は、上記したホスホリパーゼＤの作用時間、作用温度、酵素反応の終了方法と同じである。
　本発明の製造方法において、原料畜肉に作用させるための酵素としては、以下（I）～（VIII）が挙げられる。
（I）ホスホリパーゼＤ
（II）ホスホリパーゼＤ及びトランスグルタミナーゼ
（III）ホスホリパーゼＤ及びリパーゼ
（IV）ホスホリパーゼＤ及び澱粉分解酵素（例えば、α－グルコシダーゼ）
（V）ホスホリパーゼＤ、トランスグルタミナーゼ及びリパーゼ
（VI）ホスホリパーゼＤ、トランスグルタミナーゼ及び澱粉分解酵素（例えば、α－グルコシダーゼ）
（VII）ホスホリパーゼＤ、リパーゼ及び澱粉分解酵素（例えば、α－グルコシダーゼ）
（VIII）ホスホリパーゼＤ、トランスグルタミナーゼ、リパーゼ及び澱粉分解酵素（例えば、α－グルコシダーゼ） In the above-described production method of the present invention, in addition to the above-described phospholipase D, at least one enzyme selected from the group consisting of transglutaminase, lipase, and amylolytic enzyme is further added to the raw animal meat and allowed to act. is preferred.
When a plurality of enzymes are added, the order of addition may be in any order, and all may be added simultaneously or sequentially with a time lag. is desirable.
When at least one enzyme selected from the group consisting of transglutaminase, lipase, and amylolytic enzyme is allowed to further act on raw meat, the action time, action temperature, and method for terminating the enzymatic reaction are determined by the action time of phospholipase D described above. , the working temperature, and the termination method of the enzymatic reaction.
In the production method of the present invention, the following enzymes (I) to (VIII) are given as the enzymes to act on the raw meat.
(I) Phospholipase D
(II) phospholipase D and transglutaminase (III) phospholipase D and lipase (IV) phospholipase D and amylolytic enzyme (eg α-glucosidase)
(V) phospholipase D, transglutaminase and lipase (VI) phospholipase D, transglutaminase and amylolytic enzymes (e.g. α-glucosidase)
(VII) Phospholipase D, lipase and amylolytic enzyme (eg α-glucosidase)
(VIII) Phospholipase D, transglutaminase, lipase and amylolytic enzyme (eg α-glucosidase)
　本発明に用いられるトランスグルタミナーゼは、タンパク質やペプチド中のグルタミン残基を供与体とし、リジン残基を受容体とするアシル転移反応を触媒する活性を有する酵素であり、例えば、哺乳動物由来のもの、魚類由来のもの、微生物由来のもの等、種々の起源のものが知られている。本発明において用いられるトランスグルタミナーゼは、上述の活性を有すればその起源は特に制限されず、いかなる起源のトランスグルタミナーゼであっても使用でき、また組み換え酵素を使用してもよい。本発明において用いられるトランスグルタミナーゼは市販品であってもよく、具体例としては、味の素株式会社より「アクティバ」ＴＧの商品名で市販されている微生物由来のトランスグルタミナーゼを単独または組み合わせて用いることができる。
　本明細書において、トランスグルタミナーゼの酵素活性は、温度３７℃、ｐＨ６．０のトリス緩衝液中、ベンジルオキシカルボニル－Ｌ－グルタミルグリシンおよびヒドロキシルアミンを基質とする反応系で、トランスグルタミナーゼを作用せしめ、生成したヒドロキサム酸をトリクロロ酢酸存在下で鉄錯体を形成させた後、５２５ｎｍにおける吸光度を測定し、ヒドロキサム酸量を検量線により求め、１分間に１μモルのヒドロキサム酸を生成せしめる酵素量を１ユニット（１Ｕ）と定義する（特開昭６４－２７４７１号公報参照）。 The transglutaminase used in the present invention is an enzyme having the activity of catalyzing an acyl transfer reaction using a glutamine residue in a protein or peptide as a donor and a lysine residue as an acceptor, and is derived from mammals, for example. , those derived from fish, those derived from microorganisms, etc. are known. The origin of the transglutaminase used in the present invention is not particularly limited as long as it has the above-mentioned activity, and transglutaminase of any origin can be used, and recombinant enzymes can also be used. The transglutaminase used in the present invention may be a commercial product, and as a specific example, a microorganism-derived transglutaminase commercially available from Ajinomoto Co., Inc. under the trade name of "Activa" TG can be used alone or in combination. can.
As used herein, the enzymatic activity of transglutaminase is defined by allowing transglutaminase to act in a reaction system using benzyloxycarbonyl-L-glutamylglycine and hydroxylamine as substrates in a Tris buffer at a temperature of 37° C. and a pH of 6.0. After forming an iron complex with the produced hydroxamic acid in the presence of trichloroacetic acid, the absorbance at 525 nm is measured, the amount of hydroxamic acid is obtained from a calibration curve, and the amount of enzyme that produces 1 μmole of hydroxamic acid per minute is 1 unit. (1U) (see JP-A-64-27471).
　本発明に用いられるリパーゼは、脂肪酸エステルを脂肪酸とグリセリンとに加水分解する反応の触媒となる酵素である。尚、「リパーゼＡ「アマノ」６」、「リパーゼＡＹ「アマノ」」という商品名で天野エンザイム株式会社より市販されている酵素が、リパーゼの一例である。
　本明細書において、リパーゼの酵素活性は、オリーブ油１００ｍｌと２％ＰＶＡ試液１５０ｍｌを乳化させ基質とし、基質５ｍｌ、マッキルベイン緩衝液(ｐＨ７．０)４ｍｌ及び酵素液１ｍｌを混和し、３７℃にて６０分間反応させ、反応停止後、生成した脂肪酸を滴定法で測定する。遊離したオレイン酸１μｍｏｌに相当する酸を遊離させる活性を１Ｕ（ユニット）と定義する。 The lipase used in the present invention is an enzyme that catalyzes the reaction of hydrolyzing fatty acid esters into fatty acids and glycerol. Enzymes commercially available from Amano Enzyme Co., Ltd. under the trade names of "Lipase A 'Amano'6" and "Lipase AY 'Amano'" are examples of lipases.
In the present specification, the enzymatic activity of lipase was measured by emulsifying 100 ml of olive oil and 150 ml of a 2% PVA test solution as a substrate, mixing 5 ml of the substrate, 4 ml of McIlvaine buffer (pH 7.0) and 1 ml of the enzyme solution, followed by mixing at 37° C. for 60 minutes. After the reaction is terminated, the produced fatty acid is measured by a titration method. 1 U (unit) is defined as the acid liberating activity corresponding to 1 μmol of liberated oleic acid.
　本発明において、澱粉分解酵素とは、澱粉を含む糖鎖の分解反応、転移反応の触媒となる酵素である。本発明に用いられる澱粉分解酵素としては、例えば、α－グルコシダーゼ、α－アミラーゼ、β－アミラーゼ、グルコアミラーゼ、イソマルターゼ、プルラナーゼ、スクロース－α－グルコシダーゼ、α－グルコシルトランスフェラーゼ、シクロマルトデキストリナーゼ、イソアミラーゼ、ネオプルラナーゼなどが挙げられ、α－グルコシダーゼが好ましい。 In the present invention, an amylolytic enzyme is an enzyme that catalyzes the decomposition reaction and transfer reaction of sugar chains containing starch. Examples of amylolytic enzymes used in the present invention include α-glucosidase, α-amylase, β-amylase, glucoamylase, isomaltase, pullulanase, sucrose-α-glucosidase, α-glucosyltransferase, cyclomaltodextrinase, Examples include isoamylase and neopullulanase, with α-glucosidase being preferred.
　本発明に用いられるα－グルコシダーゼは、非還元末端α－１，４－グルコシド結合を加水分解し、α－グルコースを生成する酵素である。α－グルコシダーゼのうち、トランスグルコシダーゼが好ましい。尚、「トランスグルコシダーゼＬ「アマノ」」という商品名で天野エンザイム株式会社より市販されている酵素が、α－グルコシダーゼの一例である。
　本明細書において、α－グルコシダーゼの酵素活性は、１ｍＭ　α－メチル－Ｄ－グルコシド１ｍｌに０．０２Ｍ酢酸バッファー（ｐＨ５．０）１ｍｌを加え、酵素溶液０．５ｍｌ添加して、４０℃６０分間作用させたときに、反応液２．５ｍｌ中に１μｇのブドウ糖を生成する酵素量を１Ｕ（ユニット）と定義する。 The α-glucosidase used in the present invention is an enzyme that hydrolyzes a non-reducing terminal α-1,4-glucosidic bond to produce α-glucose. Among α-glucosidases, transglucosidase is preferred. An enzyme commercially available from Amano Enzyme Co., Ltd. under the trade name of "Transglucosidase L 'Amano'" is an example of α-glucosidase.
As used herein, the enzymatic activity of α-glucosidase was measured by adding 1 ml of 0.02 M acetate buffer (pH 5.0) to 1 ml of 1 mM α-methyl-D-glucoside, adding 0.5 ml of the enzyme solution, and precipitating at 40° C. for 60 minutes. The amount of enzyme that produces 1 μg of glucose in 2.5 ml of reaction solution when acted upon is defined as 1 U (unit).
　本発明の製造方法において、トランスグルタミナーゼを使用する場合、トランスグルタミナーゼの添加量は、畜肉練り製品の原材料1ｇに対して酵素活性が、例えば０．００５～３．０Ｕ、好ましくは０．０１～１．０Ｕ、より好ましくは０．０５～０．５Ｕ、さらに好ましくは０．１～０．３Ｕである。 When transglutaminase is used in the production method of the present invention, the amount of transglutaminase to be added is such that the enzymatic activity is, for example, 0.005 to 3.0 U, preferably 0.01 to 1.0 U, per 1 g of the raw material for the meat paste product. 0 U, more preferably 0.05 to 0.5 U, still more preferably 0.1 to 0.3 U.
　本発明の製造方法において、リパーゼを使用する場合、リパーゼの添加量は、畜肉練り製品の原材料1ｇに対して酵素活性が、例えば０．１～３０．０Ｕ、好ましくは０．３～１５．０Ｕ、より好ましくは０．５～１０．０Ｕ、さらに好ましくは１．０～３．０Ｕである。 In the production method of the present invention, when lipase is used, the amount of lipase to be added is such that the enzymatic activity is, for example, 0.1 to 30.0 U, preferably 0.3 to 15.0 U, preferably 0.3 to 15.0 U, per 1 g of the raw material of the meat paste product. It is more preferably 0.5 to 10.0U, still more preferably 1.0 to 3.0U.
　本発明の製造方法において、澱粉分解酵素（例えば、α－グルコシダーゼ）を使用する場合、澱粉分解酵素（例えば、α－グルコシダーゼ）の添加量は、畜肉練り製品の原材料1ｇに対して酵素活性が、例えば０．０００５～０．１Ｕ、好ましくは０．００１～０．０５Ｕ、より好ましくは０．００２～０．０２Ｕ、さらに好ましくは０．００５～０．０１Ｕである。 In the production method of the present invention, when an amylolytic enzyme (eg, α-glucosidase) is used, the amount of the amylolytic enzyme (eg, α-glucosidase) added is such that the enzymatic activity per 1 g of the raw material of the meat paste product is, for example, It is 0.0005 to 0.1 U, preferably 0.001 to 0.05 U, more preferably 0.002 to 0.02 U, still more preferably 0.005 to 0.01 U.
　上記した本発明の製造方法により製造される本発明の畜肉練り製品は、製造時における加熱後歩留りが向上しており、ひいては食感及び呈味の向上が期待できる畜肉練り製品である。 The meat paste product of the present invention produced by the above-described production method of the present invention has an improved yield after heating during production, and is expected to have an improved texture and taste.
　本発明はまた、原料畜肉にホスホリパーゼＤを作用させることを含む、畜肉練り製品の加熱後歩留り向上方法に関する。
　本発明の、該加熱後歩留り向上方法において、ホスホリパーゼＤは、シグマ　アルドリッチ製ホスホリパーゼＤ　放線菌由来（商品名）、ナガセケムテック製デナチームＰＭＤ－Ｐ１（商品名）及び旭化成ファーマ製ＰＨＯＳＰＨＯＬＩＰＡＳＥ　Ｄ［ＰＬＤＰ（Ｔ－３９）］（Ｇｌｙｃｅｒｏｐｈｏｓｐｈｏｌｉｐｉｄ　ｓｐｅｃｉｆｉｃ）（商品名）からなる群から選択されるホスホリパーゼＤが好ましい。 The present invention also relates to a method for improving the post-heating yield of livestock meat paste products, which comprises allowing phospholipase D to act on raw livestock meat.
In the method for improving the post-heating yield of the present invention, the phospholipase D is Sigma-Aldrich's phospholipase D actinomycete-derived (trade name), Nagase Chemtech's Denazym PMD-P1 (trade name), and Asahi Kasei Pharma's PHOSPHOLIPASE D [PLDP ( T-39)] (Glycerophospholipid specific) (trade names) are preferred.
　本発明の、該加熱後歩留り向上方法において、上記したホスホリパーゼＤに加えて、トランスグルタミナーゼ、リパーゼ、及び澱粉分解酵素からなる群から選択される少なくとも１つの酵素をさらに、原料畜肉に作用させることが好ましい。
　本発明の、該加熱後歩留り向上方法において、原料畜肉に作用させるための酵素としては、以下（I）～（VIII）が挙げられる。
（I）ホスホリパーゼＤ
（II）ホスホリパーゼＤ及びトランスグルタミナーゼ
（III）ホスホリパーゼＤ及びリパーゼ
（IV）ホスホリパーゼＤ及び澱粉分解酵素（例えば、α－グルコシダーゼ）
（V）ホスホリパーゼＤ、トランスグルタミナーゼ及びリパーゼ
（VI）ホスホリパーゼＤ、トランスグルタミナーゼ及び澱粉分解酵素（例えば、α－グルコシダーゼ）
（VII）ホスホリパーゼＤ、リパーゼ及び澱粉分解酵素（例えば、α－グルコシダーゼ）
（VIII）ホスホリパーゼＤ、トランスグルタミナーゼ、リパーゼ及び澱粉分解酵素（例えば、α－グルコシダーゼ） In the method for improving the post-heating yield of the present invention, in addition to the above-described phospholipase D, at least one enzyme selected from the group consisting of transglutaminase, lipase, and amylolytic enzyme may be further acted on the raw meat. preferable.
In the method for improving yield after heating according to the present invention, the following enzymes (I) to (VIII) are listed as enzymes to act on raw meat.
(I) Phospholipase D
(II) phospholipase D and transglutaminase (III) phospholipase D and lipase (IV) phospholipase D and amylolytic enzyme (eg α-glucosidase)
(V) phospholipase D, transglutaminase and lipase (VI) phospholipase D, transglutaminase and amylolytic enzymes (e.g. α-glucosidase)
(VII) Phospholipase D, lipase and amylolytic enzyme (eg α-glucosidase)
(VIII) Phospholipase D, transglutaminase, lipase and amylolytic enzyme (eg α-glucosidase)
　本発明の、該加熱後歩留り向上方法において、畜肉練り製品の例示、原料畜肉の例示、使用量、ホスホリパーゼＤ、トランスグルタミナーゼ、リパーゼ及び澱粉分解酵素（α－グルコシダーゼ）の定義、例示、添加量、添加方法、任意に添加してもよい成分、添加剤等は、上記本発明の畜肉練り製品の製造方法において説明した、畜肉練り製品の例示、原料畜肉の例示、使用量、ホスホリパーゼＤ、トランスグルタミナーゼ、リパーゼ及び澱粉分解酵素（α－グルコシダーゼ）の定義、例示、添加量、添加方法、任意に添加してもよい成分、添加剤等と同様である。 In the method for improving yield after heating of the present invention, examples of meat paste products, examples of raw meat, amounts used, definitions, examples, addition amounts, and addition of phospholipase D, transglutaminase, lipase, and amylolytic enzyme (α-glucosidase) The method, optional ingredients, additives, etc. are described in the method for producing the livestock meat paste product of the present invention, examples of livestock meat paste products, examples of raw meat paste products, amounts used, phospholipase D, transglutaminase, lipase and It is the same as the definition, exemplification, addition amount, addition method, optionally added components, additives, etc. of the amylolytic enzyme (α-glucosidase).
　本明細書において、畜肉練り製品の加熱後歩留りとは、畜肉練り製品の製造時において、原材料を混合して得た生地（パテ）の加熱前（焼成前）の重量に対する加熱後（焼成後）の重量の割合（重量％）をいう。
　本発明において、畜肉練り製品の加熱後歩留り向上とは、酵素無添加で製造した畜肉練り製品と比較して、加熱後歩留まりが高いことをいう。 In this specification, the post-heating yield of the meat paste product refers to the weight after heating (after baking) relative to the weight before heating (before baking) of the dough (putty) obtained by mixing the raw materials at the time of manufacturing the meat paste product. refers to the ratio (% by weight) of
In the present invention, the improved yield after heating of the meat paste product means that the yield after heating is higher than that of the meat paste product produced without the addition of enzymes.
　以下、実施例、試験例に基づいて本発明をより詳細に説明するが、本発明はこれらに限定されるものではない。 The present invention will be described in more detail below based on examples and test examples, but the present invention is not limited to these.
［試験例１］
（実施例１～３、比較例１のミートボールの製造）
　表１に示した牛バラ赤身、牛ケンネ脂を肉挽き機（チョッパー）により３ｍｍのミンチとした。玉ねぎは約２ｍｍのみじん切りとした。
　表１に示した配合の原材料に、表２に示した各ホスホリパーゼＤを原材料１ｇあたり３．０Ｕとなるよう添加し、スタンダードミキサー（「キッチンエイド（ＫｉｔｃｈｅｎＡｉｄ）ＫＳＭ５ＷＨ」、ワールプール・コーポレーション製）を用い、設定速度１にて３分間混合し、ミートボール生地を調製した。空気抜きした後に３０ｇの球形に手で成型した。成型後、インピンジャー（ＦＧＪＯＡ９Ｌ、フジマック製）にて２５０℃で１分間焼成及びスチームコンベクションオーブン（ＦＳＣＣ　ＷＥ　６１Ｇ、フジマック製）にて２００℃、ＲＨ５０％で６分間焼成し、実施例１～３のミートボールを得た。
　また、ホスホリパーゼＤを添加しない以外は同様の方法で、比較例１のミートボールを得た。
　得られた実施例１～３、比較例１のミートボールについて、後述の（１）加熱後歩留りの評価、及び(２)ミートボール中遊離コリン及び遊離イノシトール測定を行った。加熱後歩留りの評価のために、ミートボール製造時において、上記焼成前の重量と焼成後の重量を測定した。
　表２に示す各ホスホリパーゼＤのメーカー、酵素活性を表３に示す。 [Test Example 1]
(Production of meatballs of Examples 1 to 3 and Comparative Example 1)
The beef rib lean meat and beef Kenne fat shown in Table 1 were minced to 3 mm by a meat grinder (chopper). The onion was chopped into pieces of about 2 mm.
Each phospholipase D shown in Table 2 was added to the raw material of the composition shown in Table 1 so as to be 3.0 U per 1 g of raw material, and a standard mixer ("KitchenAid KSM5WH", manufactured by Whirlpool Corporation) was added. and mixed for 3 minutes at a setting speed of 1 to prepare a meatball dough. After degassing, it was manually molded into 30 g spheres. After molding, baked at 250 ° C. for 1 minute with an impinger (FGJOA9L, manufactured by Fujimac) and baked at 200 ° C. for 6 minutes at RH 50% in a steam convection oven (FSCC WE 61G, manufactured by Fujimac). Got meatballs.
Moreover, meatballs of Comparative Example 1 were obtained in the same manner except that phospholipase D was not added.
The meatballs obtained in Examples 1 to 3 and Comparative Example 1 were subjected to (1) evaluation of post-heating yield and (2) measurement of free choline and free inositol in meatballs, which will be described later. In order to evaluate the yield after heating, the weight before baking and the weight after baking were measured at the time of meatball production.
Table 3 shows the manufacturer and enzymatic activity of each phospholipase D shown in Table 2.
Figure JPOXMLDOC01-appb-T000001
Figure JPOXMLDOC01-appb-T000001
Figure JPOXMLDOC01-appb-T000002
Figure JPOXMLDOC01-appb-T000002
Figure JPOXMLDOC01-appb-T000003
Figure JPOXMLDOC01-appb-T000003
（１）加熱後歩留りの評価
　一般的に、挽き肉加工食品においては、加熱後歩留りが高いと、肉汁が内部に閉じ込められており、好ましい品質であると考えられる。
　実施例１～３、比較例１のミートボールの製造時における焼成前の重量と焼成後の重量から、下記の式に従って、加熱後歩留りを算出した。
（式）
加熱後歩留り（重量％）＝（焼成後の重量／焼成前の重量）×１００
　結果を表４及び図１に示す。 (1) Evaluation of post-heating yield In general, when the minced meat processed food has a high post-heating yield, meat juice is trapped inside, and it is considered to be of desirable quality.
The post-heating yield was calculated according to the following formula from the weight before baking and the weight after baking when the meatballs of Examples 1 to 3 and Comparative Example 1 were produced.
(formula)
Yield after heating (% by weight) = (weight after firing/weight before firing) x 100
The results are shown in Table 4 and FIG.
Figure JPOXMLDOC01-appb-T000004
Figure JPOXMLDOC01-appb-T000004
　表４の結果より、ホスホリパーゼＤを添加して製造した実施例１～３のミートボールは、酵素無添加の比較例１に対して、加熱後歩留りが高く、加熱後歩留りの向上が認められた。なかでも、実施例１及び３において、顕著な加熱後歩留りの向上が認められた。 From the results of Table 4, the meatballs of Examples 1 to 3 produced by adding phospholipase D had a higher yield after heating than Comparative Example 1 without enzyme addition, and an improvement in yield after heating was observed. . Among them, in Examples 1 and 3, a remarkable improvement in post-heating yield was observed.
（２）ミートボール中遊離コリン及び遊離イノシトール測定
（２－１）サンプルの調製
　実施例１～３、比較例１のミートボールについて、アルミパウチに入れて外側から手でつぶし、凍結乾燥機を用いて凍結乾燥した。凍結乾燥による減量割合を表５－１に示す。乾燥後、麺棒でたたいて粉砕し、ミートボール粉末を得た。得られたミートボール粉末を、下記の（２－２）ミートボール中遊離コリン測定、（２－３）ミートボール中遊離イノシトール測定のサンプルとして用いた。 (2) Measurement of free choline and free inositol in meatballs (2-1) Preparation of samples The meatballs of Examples 1 to 3 and Comparative Example 1 were placed in an aluminum pouch, crushed by hand from the outside, and then using a freeze dryer. and lyophilized. Table 5-1 shows the weight loss rate due to freeze-drying. After drying, it was pulverized by hitting with a rolling pin to obtain meatball powder. The obtained meatball powder was used as a sample for the following (2-2) measurement of free choline in meatballs and (2-3) measurement of free inositol in meatballs.
Figure JPOXMLDOC01-appb-T000005
Figure JPOXMLDOC01-appb-T000005
（２－２）ミートボール中遊離コリン測定
（測定前処理（抽出）方法）
（i）０．５ｇのミートボール粉末に３．０ｍｌの水を加えた。
（ii）氷冷しながら１００００ｒｐｍで２分間、ポリトロンで粉砕した。
（iii）１．５ｍｌチューブ２本に全量を均等に分注し、１００００ｘｇ，１０度，１分で遠心分離した。
（iv）上清を２．５ｍｌシリンジに移し、０．４５μｍクロマトディスクを通してサンプル液を得た。
（遊離コリン測定方法）
　コリンオキシダーゼ、ペルオキシダーゼ等を含む発色溶液（コリンエステラーゼキット－ＮＣ、２８９－７５１８１、富士フイルム和光純薬株式会社）１ｍＬにサンプル液５０μＬを加え５分反応させ、赤色キノン色素を生成した。その後、反応停止液（コリンエステラーゼキット－ＮＣ、２８９－７５１８１、富士フイルム和光純薬株式会社）にて反応停止後、１０分間室温にて放置した。その後、波長５０５ｎｍにおける吸光度を測定した。基準液（コリンエステラーゼキット－ＮＣ、２８９－７５１８１、富士フイルム和光純薬株式会社）を用いて、０、０．７５、１、１．２５、１．５及び１．７５ｍＭコリン標準液を作成し、上記と同様の操作にて吸光度を測定した。吸光度より検量線を作成し、ミートボール中遊離コリン含有量を求めた。結果を表５－２に示す。 (2-2) Measurement of free choline in meatballs (measurement pretreatment (extraction) method)
(i) 3.0 ml of water was added to 0.5 g of meatball powder.
(ii) pulverized with a polytron at 10,000 rpm for 2 minutes while cooling on ice;
(iii) The total amount was evenly dispensed into two 1.5 ml tubes and centrifuged at 10000 xg, 10°C for 1 minute.
(iv) The supernatant was transferred to a 2.5 ml syringe and passed through a 0.45 μm chromatodisk to obtain a sample solution.
(Method for measuring free choline)
50 μL of the sample solution was added to 1 mL of a coloring solution containing choline oxidase, peroxidase, etc. (Cholinesterase Kit-NC, 289-75181, Fuji Film Wako Pure Chemical Industries, Ltd.) and allowed to react for 5 minutes to generate a red quinone dye. After that, the reaction was terminated with a reaction termination solution (cholinesterase kit-NC, 289-75181, Fuji Film Wako Pure Chemical Industries, Ltd.) and allowed to stand at room temperature for 10 minutes. After that, absorbance at a wavelength of 505 nm was measured. 0, 0.75, 1, 1.25, 1.5 and 1.75 mM choline standard solutions were prepared using a standard solution (cholinesterase kit-NC, 289-75181, Fujifilm Wako Pure Chemical Industries, Ltd.), Absorbance was measured in the same manner as above. A calibration curve was prepared from the absorbance, and the content of free choline in meatballs was determined. The results are shown in Table 5-2.
Figure JPOXMLDOC01-appb-T000006
Figure JPOXMLDOC01-appb-T000006
（２－３）ミートボール中遊離イノシトール測定
（測定前処理（抽出）方法）
（i）１～２ｇのミートボール粉末に６０ｍｌの水を加えた。
（ii）３０分振とう後、１ｍｏｌ／Ｌ水酸化ナトリウム溶液及び１ｍｏｌ／Ｌ塩酸を用いてｐＨ５．０に調整した。
（iii）水で１００ｍＬに定容し、ろ過し、サンプル液を得た。
（遊離イノシトール測定方法）
　Ｓａｃｃｈａｒｏｍｙｃｅｓ　ｃｅｒｅｖｉｓｉａｅ　ＡＴＣＣ　９０８０をブドウ糖ペプトン培地「ニッスイ」（日水製薬株式会社）を用い３０℃±１℃、２０時間±３時間で前培養し、接種菌液を調製した。滅菌したイノシトール分析培地に標準溶液又はサンプル液を塗布し、３０℃±１℃、１９時間±３時間で培養し、波長６００ｎｍで測定した。標準溶液（イノシトール０．５ｍｇ／ｍｌ、２５％エタノール溶液）を用いて検量線を作成し、ミートボール中遊離イノシトール含有量を求めた。結果を表５－３に示す。 (2-3) Measurement of free inositol in meatballs (measurement pretreatment (extraction) method)
(i) 60 ml of water was added to 1-2 g of meatball powder.
(ii) After shaking for 30 minutes, the pH was adjusted to 5.0 using 1 mol/L sodium hydroxide solution and 1 mol/L hydrochloric acid.
(iii) The volume was adjusted to 100 mL with water and filtered to obtain a sample liquid.
(Method for measuring free inositol)
Saccharomyces cerevisiae ATCC 9080 was pre-cultured at 30° C.±1° C. for 20 hours±3 hours using a glucose peptone medium "Nissui" (Nissui Pharmaceutical Co., Ltd.) to prepare an inoculum solution. A standard solution or a sample solution was applied to a sterilized inositol analysis medium, cultured at 30° C.±1° C. for 19 hours±3 hours, and measured at a wavelength of 600 nm. A calibration curve was prepared using a standard solution (0.5 mg/ml inositol, 25% ethanol solution) to determine the content of free inositol in meatballs. The results are shown in Table 5-3.
Figure JPOXMLDOC01-appb-T000007
Figure JPOXMLDOC01-appb-T000007
［試験例２］コリン添加試験
（実施例２－１～実施例２－５、比較例２－２～比較例２－４のミートボールの製造）
　表６に示した牛バラ赤身、牛ケンネ脂を肉挽き機（チョッパー）により３ｍｍのミンチとした。玉ねぎは約２ｍｍのみじん切りとした。
　表６に示した配合の原材料および表７－１、７－２に記載の添加量のコリンを使用し、スタンダードミキサー（「キッチンエイド（ＫｉｔｃｈｅｎＡｉｄ）ＫＳＭ５ＷＨ」、ワールプール・コーポレーション製）を用い、設定速度１にて３分間混合し、ミートボール生地を調製した。空気抜きした後に３０ｇの球形に手で成型した。成型後、1時間冷蔵庫で静置した。インピンジャー（ＦＧＪＯＡ９Ｌ、フジマック製）にて２５０℃で１分間焼成及びスチームコンベクションオーブン（ＦＳＣＣ　ＷＥ　６１Ｇ、フジマック製）にて２００℃、ＲＨ５０％で６分間焼成し、実施例２－１～実施例２－５、比較例２－２～比較例２－４のミートボールを得た。
（比較例２－１のミートボールの製造）
　コリンを添加しない以外は実施例２－１等と同じ方法で、比較例２－１のミートボールを得た。
（ミートボール中コリン含有量）
　表７－１に示す、比較例２－１のミートボール中コリン含有量は、同様の方法で製造した試験例１の比較例１のミートボール中のコリン含有量（６０重量ｐｐｍ、表５－２参照）に基づく推定値である。比較例２－１はコリンを添加していないので、ミートボール中コリン含有量（推定値：６０重量ｐｐｍ）は、原材料中に元々含まれるコリン量である。
　表７－１、７－２に示す、実施例２－１～実施例２－５、比較例２－２～比較例２－４のミートボール中コリン含有量は、原材料中に元々含まれるコリン量（推定値：６０重量ｐｐｍ）とコリン添加量との合計である。
（加熱後歩留りの評価）
　加熱後歩留りの評価のために、ミートボール製造時において、上記焼成前の重量と焼成後の重量を測定した。製造時における焼成前の重量と焼成後の重量から、下記の式に従って、加熱後歩留りを算出した。
（式）
加熱後歩留り（重量％）＝（焼成後の重量／焼成前の重量）×１００
　結果を表７－１、７－２及び図２に示す。 [Test Example 2] Choline addition test (production of meatballs of Examples 2-1 to 2-5 and Comparative Examples 2-2 to 2-4)
The beef rib lean meat and beef Kenne fat shown in Table 6 were minced to 3 mm by a meat grinder (chopper). The onion was chopped into pieces of about 2 mm.
A standard mixer (“KitchenAid KSM5WH”, manufactured by Whirlpool Corporation) was used using raw materials with the formulations shown in Table 6 and choline in the amounts shown in Tables 7-1 and 7-2. Mix for 3 minutes at speed 1 to prepare a meatball dough. After degassing, it was manually molded into 30 g spheres. After molding, it was allowed to stand in a refrigerator for 1 hour. Baking at 250 ° C. for 1 minute with an impinger (FGJOA 9L, manufactured by Fujimac) and baking at 200 ° C. and RH 50% for 6 minutes in a steam convection oven (FSCC WE 61G, manufactured by Fujimac), Examples 2-1 to 2 -5, meatballs of Comparative Examples 2-2 to 2-4 were obtained.
(Production of meatballs of Comparative Example 2-1)
Meatballs of Comparative Example 2-1 were obtained in the same manner as in Example 2-1, etc., except that choline was not added.
(choline content in meatball)
The choline content in the meatball of Comparative Example 2-1 shown in Table 7-1 is the choline content in the meatball of Comparative Example 1 of Test Example 1 produced by the same method (60 ppm by weight, Table 5-1). 2). Since choline was not added in Comparative Example 2-1, the choline content in the meatball (estimated value: 60 ppm by weight) is the amount of choline originally contained in the raw material.
The choline content in the meatballs of Examples 2-1 to 2-5 and Comparative Examples 2-2 to 2-4 shown in Tables 7-1 and 7-2 is the choline originally contained in the raw materials. It is the sum of the amount (estimated value: 60 ppm by weight) and the added amount of choline.
(Evaluation of yield after heating)
In order to evaluate the yield after heating, the weight before baking and the weight after baking were measured at the time of meatball production. From the weight before firing and the weight after firing at the time of production, the post-heating yield was calculated according to the following formula.
(formula)
Yield after heating (% by weight) = (weight after firing/weight before firing) x 100
The results are shown in Tables 7-1, 7-2 and FIG.
Figure JPOXMLDOC01-appb-T000008
Figure JPOXMLDOC01-appb-T000008
Figure JPOXMLDOC01-appb-T000009
Figure JPOXMLDOC01-appb-T000009
Figure JPOXMLDOC01-appb-T000010
Figure JPOXMLDOC01-appb-T000010
　表７－１、７－２、図２の結果より、コリン含有量を２６０～２０６０重量ｐｐｍの範囲に調整した実施例２－２、２－３、２－４、２－５のミートボールにおいて、加熱後歩留りが特に高いことが示された。 From the results in Tables 7-1, 7-2, and FIG. 2, the meatballs of Examples 2-2, 2-3, 2-4, and 2-5 in which the choline content was adjusted to the range of 260 to 2060 ppm by weight , the yield after heating was shown to be particularly high.
［試験例３］イノシトール添加試験
（実施例３－１～実施例３－３、比較例３－２～比較例３－７のミートボールの製造）
　試験例２の表６に示した牛バラ赤身、牛ケンネ脂を肉挽き機（チョッパー）により３ｍｍのミンチとした。玉ねぎは約２ｍｍのみじん切りとした。
　試験例２の表６に示した配合の原材料および表８－１、８－２に記載の添加量のイノシトールを使用し、スタンダードミキサー（「キッチンエイド（ＫｉｔｃｈｅｎＡｉｄ）ＫＳＭ５ＷＨ」、ワールプール・コーポレーション製）を用い設定速度１にて３分間混合し、ミートボール生地を調製した。空気抜きした後に３０ｇの球形に手で成型した。成型後、1時間冷蔵庫で静置した。インピンジャー（ＦＧＪＯＡ９Ｌ、フジマック製）にて２５０℃で１分間焼成及びスチームコンベクションオーブン（ＦＳＣＣ　ＷＥ　６１Ｇ、フジマック製）にて２００℃、ＲＨ５０％で６分間焼成し、実施例３－１～実施例３－３、比較例３－２～比較例３－７のミートボールを得た。
（比較例３－１のミートボールの製造）
　イノシトールを添加しない以外は実施例３－１等と同じ方法で、比較例３－１のミートボールを得た。
（ミートボール中イノシトール含有量）
　表８－１に示す、比較例３－１のミートボール中イノシトール含有量は、同様の方法で製造した試験例１の比較例１のミートボール中のイノシトール含有量（２１３重量ｐｐｍ、表５－３参照）に基づく推定値である。比較例３－１はイノシトールを添加していないので、ミートボール中イノシトール含有量（推定値：２１３重量ｐｐｍ）は、原材料中に元々含まれるイノシトール量である。
　表８－１、８－２に示す、実施例３－１～実施例３－３、比較例３－２～比較例３－７のミートボール中イノシトール含有量は、原材料中に元々含まれるイノシトール量（推定値：２１３重量ｐｐｍ）とイノシトール添加量との合計である。
（加熱後歩留りの評価）
　加熱後歩留りの評価のために、ミートボール製造時において、上記焼成前の重量と焼成後の重量を測定した。製造時における焼成前の重量と焼成後の重量から、下記の式に従って、加熱後歩留りを算出した。
（式）
加熱後歩留り（重量％）＝（焼成後の重量／焼成前の重量）×１００
　結果を表８－１、８－２及び図３に示す。 [Test Example 3] Inositol addition test (production of meatballs of Examples 3-1 to 3-3 and Comparative Examples 3-2 to 3-7)
The beef rib lean meat and beef kenne fat shown in Table 6 of Test Example 2 were minced to 3 mm by a meat grinder (chopper). The onion was chopped into pieces of about 2 mm.
A standard mixer (“KitchenAid KSM5WH”, manufactured by Whirlpool Corporation) was prepared using raw materials with the formulations shown in Table 6 of Test Example 2 and inositol in the amounts shown in Tables 8-1 and 8-2. was mixed for 3 minutes at a setting speed of 1 to prepare a meatball dough. After degassing, it was manually molded into 30 g spheres. After molding, it was allowed to stand in a refrigerator for 1 hour. Baking at 250 ° C. for 1 minute with an impinger (FGJOA 9L, manufactured by Fujimac) and baking at 200 ° C. and RH 50% for 6 minutes in a steam convection oven (FSCC WE 61G, manufactured by Fujimac), Examples 3-1 to 3 -3, meatballs of Comparative Examples 3-2 to 3-7 were obtained.
(Production of meatballs in Comparative Example 3-1)
Meatballs of Comparative Example 3-1 were obtained in the same manner as in Example 3-1, etc., except that inositol was not added.
(Inositol content in meatballs)
The inositol content in the meatball of Comparative Example 3-1 shown in Table 8-1 is the inositol content in the meatball of Comparative Example 1 of Test Example 1 produced by the same method (213 ppm by weight, Table 5- 3). Since inositol was not added in Comparative Example 3-1, the content of inositol in the meatball (estimated value: 213 ppm by weight) is the amount of inositol originally contained in the raw material.
The inositol content in the meatballs of Examples 3-1 to 3-3 and Comparative Examples 3-2 to 3-7 shown in Tables 8-1 and 8-2 is the inositol originally contained in the raw materials. It is the sum of the amount (estimated value: 213 ppm by weight) and the added amount of inositol.
(Evaluation of yield after heating)
In order to evaluate the yield after heating, the weight before baking and the weight after baking were measured at the time of meatball production. From the weight before firing and the weight after firing at the time of production, the post-heating yield was calculated according to the following formula.
(formula)
Yield after heating (% by weight) = (weight after firing/weight before firing) x 100
The results are shown in Tables 8-1, 8-2 and FIG.
Figure JPOXMLDOC01-appb-T000011
Figure JPOXMLDOC01-appb-T000011
Figure JPOXMLDOC01-appb-T000012
Figure JPOXMLDOC01-appb-T000012
　表８－１、８－２、図３の結果より、イノシトール含有量を２４３～２７３重量ｐｐｍの範囲に調整した実施例３－１、３－２のミートボールにおいて、加熱後歩留りが特に高いことが示された。 From the results in Tables 8-1, 8-2, and FIG. 3, the meatballs of Examples 3-1 and 3-2, in which the inositol content was adjusted to the range of 243 to 273 ppm by weight, had a particularly high yield after heating. It has been shown.
［試験例４］酵素添加試験（ホスホリパーゼＤとリパーゼとの併用）
（実施例４－１～実施例４－７の粗挽きソーセージの製造）
　表９中の群Ａの原材料を肉挽き機（チョッパー）により５ｍｍのミンチとし、ニーダー（「卓上ニーダーＰＮＶ－５」、株式会社入江商会製）にて、前記ミンチに表９中の群Ｂの原材料および水を添加し、更に表１０に記載のユニット量となるようにホスホリパーゼＤ（デナチーム　ＰＭＤ－Ｐ１、ナガセケムテックス株式会社製）およびリパーゼ（リパーゼＡＹ「アマノ」、天野エンザイム株式会社製）を加えて１０分間混合した後、冷蔵庫にて４時間静置し塩漬処理をした。
　次いで、表９中の群Ｃの原材料を添加してスタンダードミキサー（「キッチンエイド（ＫｉｔｃｈｅｎＡｉｄ）　ＫＳＭ５ＷＨ」、ワールプール・コーポレーション製）にて２速で１分間混合した後、真空包装した。
　次に、コラーゲンケーシング（デブロ（Ｄｅｖｒｏ）株式会社製）に充填し、６０℃で３０分間乾燥した後、燻煙（６０℃、１０分）および蒸煮（７５℃、３０分）を行い、一晩冷却して、実施例４－１～実施例４－７の粗挽きソーセージを製造した。
（比較例４－１の粗挽きソーセージの製造）
　酵素を添加しない以外は実施例４－１～実施例４－７と同じ方法で、比較例４－１の粗挽きソーセージを得た。
（加熱後歩留りの評価）
　加熱後歩留りの評価のために、粗挽きソーセージ製造時において、上記乾燥前の重量と、冷却後（すなわち、乾燥－燻煙－蒸煮－冷却工程の後）の重量を測定した。製造時における乾燥前の重量と冷却後の重量から、下記の式に従って、加熱後歩留りを算出した。
（式）
加熱後歩留り（重量％）＝（冷却後の重量／乾燥前の重量）×１００
　結果を表１０及び図４に示す。 [Test Example 4] Enzyme addition test (combination of phospholipase D and lipase)
(Production of coarsely ground sausages of Examples 4-1 to 4-7)
The raw materials of Group A in Table 9 are minced to 5 mm by a meat grinder (chopper), and the minced meat of Group B in Table 9 is minced with a kneader ("Desktop Kneader PNV-5", manufactured by Irie Shokai Co., Ltd.). Raw materials and water were added, and phospholipase D (Denazyme PMD-P1, manufactured by Nagase ChemteX Corporation) and lipase (Lipase AY "Amano", manufactured by Amano Enzyme Co., Ltd.) were added so that the unit amounts shown in Table 10 were obtained. After the mixture was added and mixed for 10 minutes, it was allowed to stand in a refrigerator for 4 hours for pickling.
Next, the raw materials of group C in Table 9 were added and mixed in a standard mixer (“KitchenAid KSM5WH”, manufactured by Whirlpool Corporation) at 2nd speed for 1 minute, followed by vacuum packaging.
Next, the collagen casing (manufactured by Devro Co., Ltd.) was filled and dried at 60°C for 30 minutes, followed by smoking (60°C, 10 minutes) and steaming (75°C, 30 minutes) overnight. After cooling, coarsely ground sausages of Examples 4-1 to 4-7 were produced.
(Production of coarsely ground sausage of Comparative Example 4-1)
A coarsely ground sausage of Comparative Example 4-1 was obtained in the same manner as in Examples 4-1 to 4-7 except that no enzyme was added.
(Evaluation of yield after heating)
In order to evaluate the post-heating yield, the weight before drying and the weight after cooling (that is, after the drying-smoking-steaming-cooling process) were measured during the production of the coarsely ground sausage. From the weight before drying and the weight after cooling at the time of production, the post-heating yield was calculated according to the following formula.
(formula)
Yield after heating (% by weight) = (weight after cooling/weight before drying) x 100
The results are shown in Table 10 and FIG.
Figure JPOXMLDOC01-appb-T000013
Figure JPOXMLDOC01-appb-T000013
Figure JPOXMLDOC01-appb-T000014
Figure JPOXMLDOC01-appb-T000014
　表１０、図４の結果より、ホスホリパーゼＤ及びリパーゼを添加して製造した実施例４－１～実施例４－７の粗挽きソーセージは、酵素無添加の比較例４－１に対して、加熱後歩留りが高く、加熱後歩留りの向上が認められた。 From the results in Table 10 and FIG. 4, the coarsely ground sausages of Examples 4-1 to 4-7 produced by adding phospholipase D and lipase were heated compared to Comparative Example 4-1 without enzyme addition. The post-yield was high, and the improvement of the post-heating yield was recognized.
［試験例５］酵素添加試験（ホスホリパーゼＤとトランスグルタミナーゼとの併用）
（実施例５－１～実施例５－３のミートボールの製造）
　表１１に示した牛バラ赤身、牛ケンネ脂を肉挽き機（チョッパー）により３ｍｍのミンチとした。玉ねぎは約２ｍｍのみじん切りとした。
　表１１に示した配合の原材料および表１２に記載のユニット量となるようにホスホリパーゼＤ（デナチーム　ＰＭＤ－Ｐ１、ナガセケムテックス株式会社製）およびトランスグルタミナーゼ（「アクティバ」ＴＧ、味の素株式会社製）を使用し、スタンダードミキサー（「キッチンエイド（ＫｉｔｃｈｅｎＡｉｄ）ＫＳＭ５ＷＨ」、ワールプール・コーポレーション製）を用い設定速度１にて３分間混合し、ミートボール生地を調製した。空気抜きした後に３０ｇの球形に手で成型した。成型後、1時間冷蔵庫で静置した。インピンジャー（ＦＧＪＯＡ９Ｌ、フジマック製）にて２５０℃で１分間焼成及びスチームコンベクションオーブン（ＦＳＣＣ　ＷＥ　６１Ｇ、フジマック製）にて２００℃、ＲＨ５０％で６分間焼成し、実施例５－１～実施例５－３のミートボールを得た。
（比較例５－１、５－２のミートボールの製造）
　酵素を添加しない以外は実施例５－１等と同じ方法で、比較例５－１のミートボールを得た。ホスホリパーゼＤを添加しない以外は実施例５－１等と同じ方法で、比較例５－２のミートボールを得た。
（加熱後歩留りの評価）
　加熱後歩留りの評価のために、ミートボール製造時において、上記焼成前の重量と焼成後の重量を測定した。製造時における焼成前の重量と焼成後の重量から、下記の式に従って、加熱後歩留りを算出した。
（式）
加熱後歩留り（重量％）＝（焼成後の重量／焼成前の重量）×１００
　結果を表１２及び図５に示す。 [Test Example 5] Enzyme addition test (combination of phospholipase D and transglutaminase)
(Production of meatballs of Examples 5-1 to 5-3)
The beef rib lean meat and beef kenne fat shown in Table 11 were minced to 3 mm by a meat grinder (chopper). The onion was chopped into pieces of about 2 mm.
Phospholipase D (Denazyme PMD-P1, manufactured by Nagase ChemteX Corporation) and transglutaminase ("Activa" TG, manufactured by Ajinomoto Co., Inc.) are added to the raw materials of the formulation shown in Table 11 and the unit amounts shown in Table 12. A standard mixer (“KitchenAid KSM5WH”, manufactured by Whirlpool Corporation) was used and mixed at a setting speed of 1 for 3 minutes to prepare a meatball dough. After degassing, it was manually molded into 30 g spheres. After molding, it was allowed to stand in a refrigerator for 1 hour. Baking at 250 ° C. for 1 minute with an impinger (FGJOA 9L, manufactured by Fujimac) and baking at 200 ° C. and RH 50% for 6 minutes in a steam convection oven (FSCC WE 61G, manufactured by Fujimac), Examples 5-1 to 5 -3 meatballs were obtained.
(Production of meatballs of Comparative Examples 5-1 and 5-2)
Meatballs of Comparative Example 5-1 were obtained in the same manner as in Example 5-1, etc., except that no enzyme was added. A meatball of Comparative Example 5-2 was obtained in the same manner as in Example 5-1, etc., except that phospholipase D was not added.
(Evaluation of yield after heating)
In order to evaluate the yield after heating, the weight before baking and the weight after baking were measured at the time of meatball production. From the weight before firing and the weight after firing at the time of production, the post-heating yield was calculated according to the following formula.
(formula)
Yield after heating (% by weight) = (weight after firing/weight before firing) x 100
The results are shown in Table 12 and FIG.
Figure JPOXMLDOC01-appb-T000015
Figure JPOXMLDOC01-appb-T000015
Figure JPOXMLDOC01-appb-T000016
Figure JPOXMLDOC01-appb-T000016
　表１２、図５の結果より、ホスホリパーゼＤを添加して製造した実施例５－１、ホスホリパーゼＤ及びトランスグルタミナーゼを添加して製造した実施例５－２及び実施例５－３のミートボールは、酵素無添加の比較例５－１及びホスホリパーゼＤ無添加の比較例５－２に対して、加熱後歩留りが高く、加熱後歩留りの向上が認められた。 From the results of Table 12 and FIG. 5, the meatballs of Example 5-1 produced by adding phospholipase D, and Examples 5-2 and 5-3 produced by adding phospholipase D and transglutaminase were Compared to Comparative Example 5-1 without enzyme addition and Comparative Example 5-2 without phospholipase D addition, the post-heating yield was higher, and an improvement in post-heating yield was observed.
［試験例６］酵素添加試験（ホスホリパーゼＤとαグルコシダーゼとの併用）
（実施例６－１～実施例６－４のミートボールの製造）
　酵素を表１３に示す酵素及び配合量に変更した以外は、実施例５－１等と同じ方法で、実施例６－１～実施例６－４のミートボールを得た。酵素はホスホリパーゼＤ（デナチーム　ＰＭＤ－Ｐ１、ナガセケムテックス株式会社製）およびα―グルコシダーゼ（トランスグルコシダーゼＬ「アマノ」」、天野エンザイム株式会社製）を使用した。
（比較例６－１、６－２のミートボールの製造）
　酵素を添加しない以外は実施例６－１等と同じ方法で、比較例６－１のミートボールを得た。ホスホリパーゼＤを添加しない以外は実施例６－１等と同じ方法で、比較例６－２のミートボールを得た。
（加熱後歩留りの評価）
　加熱後歩留りの評価のために、ミートボール製造時において、上記焼成前の重量と焼成後の重量を測定した。試験例５と同じ方法で、加熱後歩留りを算出した。結果を表１３及び図６に示す。 [Test Example 6] Enzyme addition test (combination of phospholipase D and α-glucosidase)
(Production of meatballs of Examples 6-1 to 6-4)
Meatballs of Examples 6-1 to 6-4 were obtained in the same manner as in Example 5-1, etc., except that the enzyme was changed to the enzyme and the blending amount shown in Table 13. Phospholipase D (Denazyme PMD-P1, manufactured by Nagase ChemteX Corporation) and α-glucosidase (Transglucosidase L "Amano", manufactured by Amano Enzyme Co., Ltd.) were used as enzymes.
(Production of meatballs of Comparative Examples 6-1 and 6-2)
A meatball of Comparative Example 6-1 was obtained in the same manner as in Example 6-1, etc., except that no enzyme was added. A meatball of Comparative Example 6-2 was obtained in the same manner as in Example 6-1, etc., except that phospholipase D was not added.
(Evaluation of yield after heating)
In order to evaluate the yield after heating, the weight before baking and the weight after baking were measured at the time of meatball production. The post-heating yield was calculated in the same manner as in Test Example 5. The results are shown in Table 13 and FIG.
Figure JPOXMLDOC01-appb-T000017
Figure JPOXMLDOC01-appb-T000017
　表１３、図６の結果より、ホスホリパーゼＤを添加して製造した実施例６－１、ホスホリパーゼＤ及びα－グルコシダーゼを添加して製造した実施例６－２～実施例６－４のミートボールは、酵素無添加の比較例６－１及びホスホリパーゼＤ無添加の比較例６－２に対して、加熱後歩留りが高く、加熱後歩留りの向上が認められた。 From the results of Table 13 and FIG. 6, the meatballs of Example 6-1 produced by adding phospholipase D and Examples 6-2 to 6-4 produced by adding phospholipase D and α-glucosidase , Compared to Comparative Example 6-1 without enzyme addition and Comparative Example 6-2 without phospholipase D addition, the yield after heating was high, and an improvement in yield after heating was observed.
［試験例７］酵素添加試験（ホスホリパーゼＤとα－グルコシダーゼとリパーゼとの併用
（実施例７－１のミートボールの製造） 
　酵素を表１４に示す酵素及び配合量に変更した以外は、実施例５－１等と同じ方法で、実施例７－１のミートボールを得た。酵素はホスホリパーゼＤ（デナチーム　ＰＭＤ－Ｐ１、ナガセケムテックス株式会社製）、α―グルコシダーゼ（トランスグルコシダーゼＬ「アマノ」」、天野エンザイム株式会社製）およびリパーゼ（リパーゼＡＹ「アマノ」、天野エンザイム株式会社製）を使用した。
（比較例７－１のミートボールの製造）
　酵素を添加しない以外は実施例７－１と同じ方法で、比較例７－１のミートボールを得た。
（加熱後歩留りの評価）
　加熱後歩留りの評価のために、ミートボール製造時において、上記焼成前の重量と焼成後の重量を測定した。試験例５と同じ方法で、加熱後歩留りを算出した。結果を表１４及び図７に示す。 [Test Example 7] Enzyme addition test (combined use of phospholipase D, α-glucosidase and lipase (production of meatballs of Example 7-1)
A meatball of Example 7-1 was obtained in the same manner as in Example 5-1, etc., except that the enzyme was changed to the enzyme and the amount shown in Table 14. Enzymes include phospholipase D (Denazyme PMD-P1, manufactured by Nagase ChemteX Co., Ltd.), α-glucosidase (Transglucosidase L "Amano", manufactured by Amano Enzyme Co., Ltd.) and lipase (Lipase AY "Amano", manufactured by Amano Enzyme Co., Ltd.). )It was used.
(Production of meatballs in Comparative Example 7-1)
A meatball of Comparative Example 7-1 was obtained in the same manner as in Example 7-1 except that no enzyme was added.
(Evaluation of yield after heating)
In order to evaluate the yield after heating, the weight before baking and the weight after baking were measured at the time of meatball production. The post-heating yield was calculated in the same manner as in Test Example 5. The results are shown in Table 14 and FIG.
Figure JPOXMLDOC01-appb-T000018
Figure JPOXMLDOC01-appb-T000018
　表１４、図７の結果より、ホスホリパーゼＤとα－グルコシダーゼとリパーゼとを添加して製造した実施例７－１のミートボールは、酵素無添加の比較例７－１に対して、加熱後歩留りが高く、加熱後歩留りの向上が認められた。 From the results of Table 14 and FIG. 7, the meatballs of Example 7-1 produced by adding phospholipase D, α-glucosidase and lipase had a higher yield after heating than Comparative Example 7-1 without enzyme addition. was high, and an improvement in yield was observed after heating.
［試験例８］ホスホリパーゼＤ添加試験（ハンバーグ）
（実施例８－１、８－２のハンバーグの製造）
　表１５に示した肉原料（牛バラ、豚ウデ）を肉挽き機（チョッパー）により３ｍｍのミンチとした。
　表１５に示した配合の各原料に、ホスホリパーゼＤ（デナチーム　ＰＭＤ－Ｐ１、ナガセケムテックス株式会社製）を対原材料１ｇあたり０．４Ｕもしくは４．０Ｕとなるよう添加し、スタンダードミキサー（「キッチンエイド（ＫｉｔｃｈｅｎＡｉｄ）ＫＳＭ５ＷＨ」、ワールプール・コーポレーション製）を用い、設定速度１にて３分間混合し、ハンバーグ生地を調製した。
　生地を１５０ｇに計量し、空気抜きした後に小判形（長径１３０ｍｍ、短径９０ｍｍ、厚さ１２ｍｍ）に成型した。成型後にハンバーグ生地を２３０℃のグリドル（ＦＧＦＴ６０６０１ＴＣ、フジマック社製）にて両面１分間ずつ焼成し、表面に焼き目を付けた後、インピンジャー（ＦＧＪＯＡ９Ｌ、フジマック製）にて２５０℃で４分間焼成し、実施例８－１、８－２のハンバーグを得た。
（比較例８－１のハンバーグの製造）
　酵素を添加しない以外は実施例８－１、８－２と同じ方法で、比較例８－１のハンバーグを得た。
（加熱後歩留りの評価）
　加熱後歩留りの評価のために、ハンバーグ製造時において、上記焼成前の重量と焼成後の重量を測定した。製造時における焼成前の重量と焼成後の重量から、下記の式に従って、加熱後歩留りを算出した。
（式）
加熱後歩留り（重量％）＝（焼成後の重量／焼成前の重量）×１００
　結果を表１６及び図８に示す。 [Test Example 8] Phospholipase D addition test (hamburger)
(Production of hamburgers of Examples 8-1 and 8-2)
The meat raw materials (beef ribs, pork ribs) shown in Table 15 were minced to 3 mm by a meat grinder (chopper).
Phospholipase D (Denazyme PMD-P1, manufactured by Nagase ChemteX Corporation) was added to each raw material of the formulation shown in Table 15 so as to be 0.4 U or 4.0 U per 1 g of the raw material, and a standard mixer ("KitchenAid (KitchenAid) KSM5WH", manufactured by Whirlpool Corporation) was mixed at a setting speed of 1 for 3 minutes to prepare a hamburger dough.
The dough was weighed to 150 g, and after the air was removed, it was formed into an oval shape (major axis: 130 mm, minor axis: 90 mm, thickness: 12 mm). After molding, the hamburger dough is baked on a griddle (FGFT60601TC, Fujimac Co., Ltd.) at 230 ° C. for 1 minute each on both sides, and after the surface is baked, baked at 250 ° C. for 4 minutes with an impinger (FGJOA9L, Fujimac Co., Ltd.). Then, hamburgers of Examples 8-1 and 8-2 were obtained.
(Production of hamburger in Comparative Example 8-1)
A hamburger of Comparative Example 8-1 was obtained in the same manner as in Examples 8-1 and 8-2 except that no enzyme was added.
(Evaluation of yield after heating)
In order to evaluate the yield after heating, the weight before baking and the weight after baking were measured at the time of hamburger production. From the weight before firing and the weight after firing at the time of production, the post-heating yield was calculated according to the following formula.
(formula)
Yield after heating (% by weight) = (weight after firing/weight before firing) x 100
The results are shown in Table 16 and FIG.
Figure JPOXMLDOC01-appb-T000019
Figure JPOXMLDOC01-appb-T000019
Figure JPOXMLDOC01-appb-T000020
Figure JPOXMLDOC01-appb-T000020
　表１６、図８の結果より、ホスホリパーゼＤを添加して製造した実施例８－１、８－２のハンバーグは、酵素無添加の比較例８－１に対して、加熱後歩留りが高く、加熱後歩留りの向上が認められた。 From the results in Table 16 and FIG. 8, the hamburger steaks of Examples 8-1 and 8-2 produced by adding phospholipase D had a high yield after heating compared to Comparative Example 8-1 without enzyme addition, and heating An improvement in post-yield was observed.
［試験例９］ホスホリパーゼＤ添加試験（ソーセージ）
（実施例９－１の粗挽きソーセージの製造）
　表１７中の群Ａの原材料を肉挽き機（チョッパー）により５ｍｍのミンチとし、ニーダー（「卓上ニーダーＰＮＶ－５」、株式会社入江商会製）にて、前記ミンチに表１７中の群Ｂの原材料および水を添加し、更に表１８に記載の配合量でホスホリパーゼＤ（デナチーム　ＰＭＤ－Ｐ１、ナガセケムテックス株式会社製）を加えて１０分間混合した後、冷蔵庫にて４時間静置し塩漬処理をした。
　次いで、表１７中の群Ｃの原材料を添加してスタンダードミキサー（「キッチンエイド（ＫｉｔｃｈｅｎＡｉｄ）　ＫＳＭ５ＷＨ」、ワールプール・コーポレーション製）にて２速で１分間混合した後、真空包装した。
　次に、コラーゲンケーシング（デブロ（Ｄｅｖｒｏ）株式会社製）に充填し、６０℃で３０分間乾燥した後、燻煙（６０℃、１０分）および蒸煮（７５℃、３０分）を行い、一晩冷却して、実施例９－１の粗挽きソーセージを製造した。
（比較例９－１の粗挽きソーセージの製造）
　酵素を添加しない以外は実施例９－１と同じ方法で、比較例９－１の粗挽きソーセージを得た。
（加熱後歩留りの評価）
　加熱後歩留りの評価のために、粗挽きソーセージ製造時において、上記乾燥前の重量と、冷却後（すなわち、乾燥－燻煙－蒸煮－冷却工程の後）の重量を測定した。製造時における乾燥前の重量と冷却後の重量から、下記の式に従って、加熱後歩留りを算出した。
（式）
加熱後歩留り（重量％）＝（冷却後の重量／乾燥前の重量）×１００
　結果を表１８及び図９に示す。 [Test Example 9] Phospholipase D addition test (sausage)
(Production of coarsely ground sausage of Example 9-1)
The raw materials of Group A in Table 17 were minced to 5 mm by a meat grinder (chopper), and the minced meat of Group B in Table 17 was minced with a kneader ("Desktop Kneader PNV-5", manufactured by Irie Shokai Co., Ltd.). Raw materials and water are added, and phospholipase D (Denazyme PMD-P1, manufactured by Nagase ChemteX Corporation) is added in the amount shown in Table 18 and mixed for 10 minutes, then left for 4 hours in a refrigerator and salted. processed.
Next, the raw materials of group C in Table 17 were added and mixed in a standard mixer (“KitchenAid KSM5WH”, manufactured by Whirlpool Corporation) at 2nd speed for 1 minute, followed by vacuum packaging.
Next, the collagen casing (manufactured by Devro Co., Ltd.) was filled and dried at 60°C for 30 minutes, followed by smoking (60°C, 10 minutes) and steaming (75°C, 30 minutes) overnight. After cooling, coarsely ground sausages of Example 9-1 were produced.
(Production of coarsely ground sausage of Comparative Example 9-1)
A coarsely ground sausage of Comparative Example 9-1 was obtained in the same manner as in Example 9-1 except that no enzyme was added.
(Evaluation of yield after heating)
In order to evaluate the post-heating yield, the weight before drying and the weight after cooling (that is, after the drying-smoking-steaming-cooling process) were measured during the production of the coarsely ground sausage. From the weight before drying and the weight after cooling at the time of production, the post-heating yield was calculated according to the following formula.
(formula)
Yield after heating (% by weight) = (weight after cooling/weight before drying) x 100
The results are shown in Table 18 and FIG.
Figure JPOXMLDOC01-appb-T000021
Figure JPOXMLDOC01-appb-T000021
Figure JPOXMLDOC01-appb-T000022
Figure JPOXMLDOC01-appb-T000022
　表１８、図９の結果より、ホスホリパーゼＤを添加して製造した実施例９－１の粗挽きソーセージは、酵素無添加の比較例９－１に対して、加熱後歩留りが高く、加熱後歩留りの向上が認められた。 From the results in Table 18 and FIG. 9, the coarsely ground sausage of Example 9-1 produced by adding phospholipase D has a higher yield after heating than Comparative Example 9-1 without enzyme addition, and the yield after heating is improvement was observed.
［試験例１０］ホスホリパーゼＤ添加試験（鶏つみれ）
（実施例１０－１の鶏つみれの製造）
　表１９中の肉原料（鶏ムネ肉、豚脂肪）を肉挽き機（チョッパー）により５ｍｍのミンチとし、スタンダードミキサー（「キッチンエイド（ＫｉｔｃｈｅｎＡｉｄ）　ＫＳＭ５ＷＨ」、ワールプール・コーポレーション製）にて、前記ミンチに表１９中の各原料を添加し、更に表２０に記載の配合量でホスホリパーゼＤ（デナチーム　ＰＭＤ－Ｐ１、ナガセケムテックス株式会社製）を加えて２速３分３０秒間混合した。
　生地を１５ｇに計量し、冷蔵庫にて２時間静置した。成型後にスチームコンベクションオーブン（ＦＳＣＣ　ＷＥ　６１Ｇ、フジマック製）にて１００℃、ＲＨ１００％で１０分間焼成し、実施例１０－１の鶏つみれを得た。
（比較例１０－１の鶏つみれの製造）
　酵素を添加しない以外は実施例１０－１と同じ方法で、比較例１０－１の鶏つみれを得た。
（加熱後歩留りの評価）
　加熱後歩留りの評価のために、鶏つみれ製造時において、上記焼成前の重量と焼成後の重量を測定した。製造時における焼成前の重量と焼成後の重量から、下記の式に従って、加熱後歩留りを算出した。
（式）
加熱後歩留り（重量％）＝（焼成後の重量／焼成前の重量）×１００
　結果を表２０及び図１０に示す。 [Test Example 10] Phospholipase D addition test (chicken fishball)
(Production of chicken fishballs in Example 10-1)
The meat raw materials (chicken breast meat, pork fat) in Table 19 were minced to 5 mm by a meat grinder (chopper), and the minced meat was obtained with a standard mixer ("KitchenAid KSM5WH", manufactured by Whirlpool Corporation). In addition, each raw material in Table 19 was added to the mixture, and phospholipase D (Denazyme PMD-P1, manufactured by Nagase ChemteX Corporation) was added in the amount shown in Table 20 and mixed at 2nd speed for 3 minutes and 30 seconds.
The dough was weighed to 15 g and allowed to stand in the refrigerator for 2 hours. After molding, it was baked in a steam convection oven (FSCC WE 61G, manufactured by Fujimac) at 100° C. and RH 100% for 10 minutes to obtain chicken meatballs of Example 10-1.
(Production of chicken fishballs in Comparative Example 10-1)
Chicken meatballs of Comparative Example 10-1 were obtained in the same manner as in Example 10-1 except that no enzyme was added.
(Evaluation of yield after heating)
In order to evaluate the post-heating yield, the weight before baking and the weight after baking were measured during the production of chicken fishballs. From the weight before firing and the weight after firing at the time of production, the post-heating yield was calculated according to the following formula.
(formula)
Yield after heating (% by weight) = (weight after firing/weight before firing) x 100
The results are shown in Table 20 and FIG.
Figure JPOXMLDOC01-appb-T000023
Figure JPOXMLDOC01-appb-T000023
Figure JPOXMLDOC01-appb-T000024
Figure JPOXMLDOC01-appb-T000024
　表２０、図１０の結果より、ホスホリパーゼＤを添加して製造した実施例１０－１の鶏つみれは、酵素無添加の比較例１０－１に対して、加熱後歩留りが高く、加熱後歩留りの向上が認められた。 From the results in Table 20 and FIG. 10, the chicken meatballs of Example 10-1 produced by adding phospholipase D had a higher yield after heating than Comparative Example 10-1 without enzyme addition, and a lower yield after heating. Improvement was observed.
［試験例１１］ホスホリパーゼＤ添加試験（高脂肪ソーセージ）
（実施例１１－１の粗挽きソーセージの製造）
　表２１中の群Ａの原材料を肉挽き機（チョッパー）により５ｍｍのミンチとし、ニーダー（「卓上ニーダーＰＮＶ－５」、株式会社入江商会製）にて、前記ミンチに表２１中の群Ｂの原材料および水を添加し、更に表２２に記載の配合量でホスホリパーゼＤ（デナチーム　ＰＭＤ－Ｐ１、ナガセケムテックス株式会社製）を加えて１０分間混合した後、冷蔵庫にて４時間静置し塩漬処理をした。
　次いで、表２１中の群Ｃの原材料を添加してスタンダードミキサー（「キッチンエイド（ＫｉｔｃｈｅｎＡｉｄ）　ＫＳＭ５ＷＨ」、ワールプール・コーポレーション製）にて２速で１分間混合した後、真空包装した。
　次に、コラーゲンケーシング（デブロ（Ｄｅｖｒｏ）株式会社製）に充填し、６０℃で３０分間乾燥した後、燻煙（６０℃、１０分）および蒸煮（７５℃、３０分）を行い、一晩冷却して、実施例１１―１の粗挽きソーセージを製造した。
（比較例１１－１の粗挽きソーセージの製造）
　酵素を添加しない以外は実施例１１－１と同じ方法で、比較例１１－１の粗挽きソーセージを得た。
（加熱後歩留りの評価）
　加熱後歩留りの評価のために、粗挽きソーセージ製造時において、上記乾燥前の重量と、冷却後（すなわち、乾燥－燻煙－蒸煮－冷却工程の後）の重量を測定した。製造時における乾燥前の重量と冷却後の重量から、下記の式に従って、加熱後歩留りを算出した。
（式）
加熱後歩留り（重量％）＝（冷却後の重量／乾燥前の重量）×１００
　結果を表２２及び図１１に示す。 [Test Example 11] Phospholipase D addition test (high-fat sausage)
(Production of coarsely ground sausage of Example 11-1)
The raw materials of Group A in Table 21 were minced to 5 mm by a meat grinder (chopper), and the minced meat of Group B in Table 21 was minced with a kneader ("Desktop Kneader PNV-5", manufactured by Irie Shokai Co., Ltd.). Raw materials and water are added, and phospholipase D (Denazyme PMD-P1, manufactured by Nagase ChemteX Corporation) is added in the amount shown in Table 22 and mixed for 10 minutes, then left for 4 hours in a refrigerator and salted. processed.
Next, the raw materials of Group C in Table 21 were added and mixed in a standard mixer (“KitchenAid KSM5WH”, manufactured by Whirlpool Corporation) at 2nd speed for 1 minute, and then vacuum-packaged.
Next, the collagen casing (manufactured by Devro Co., Ltd.) was filled and dried at 60°C for 30 minutes, followed by smoking (60°C, 10 minutes) and steaming (75°C, 30 minutes) overnight. After cooling, coarsely ground sausages of Example 11-1 were produced.
(Production of coarsely ground sausage of Comparative Example 11-1)
A coarsely ground sausage of Comparative Example 11-1 was obtained in the same manner as in Example 11-1 except that no enzyme was added.
(Evaluation of yield after heating)
In order to evaluate the post-heating yield, the weight before drying and the weight after cooling (that is, after the drying-smoking-steaming-cooling process) were measured during the production of the coarsely ground sausage. From the weight before drying and the weight after cooling at the time of production, the post-heating yield was calculated according to the following formula.
(formula)
Yield after heating (% by weight) = (weight after cooling/weight before drying) x 100
The results are shown in Table 22 and FIG.
Figure JPOXMLDOC01-appb-T000025
Figure JPOXMLDOC01-appb-T000025
Figure JPOXMLDOC01-appb-T000026
Figure JPOXMLDOC01-appb-T000026
　表２２、図１１の結果より、ホスホリパーゼＤを添加して製造した実施例１１－１の粗挽きソーセージは、酵素無添加の比較例１１－１に対して、加熱後歩留りが高く、加熱後歩留りの向上が認められた。 From the results of Table 22 and FIG. 11, the coarsely ground sausage of Example 11-1 produced by adding phospholipase D has a high yield after heating and a high yield after heating compared to Comparative Example 11-1 without enzyme addition. improvement was observed.
　本発明によれば、加熱後歩留りが向上した畜肉練り製品を提供することができる。 According to the present invention, it is possible to provide a meat paste product with improved yield after heating.
　本出願は、日本で出願された特願２０２１－０７７２５７を基礎としており、その内容は本明細書にすべて包含されるものである。 This application is based on Japanese Patent Application No. 2021-077257 filed in Japan, the contents of which are all incorporated herein.

Claims (6)

  1. 　下記（１）、（２）のいずれか一方又は両方を含有する、畜肉練り製品。
    （１）６５～９０００重量ｐｐｍのコリン
    （２）２３０～３８０重量ｐｐｍのイノシトール     A livestock meat paste product containing either one or both of the following (1) and (2).
    (1) 65-9000 ppm by weight choline (2) 230-380 ppm by weight inositol
  2. 　前記（１）及び（２）を含有する、請求項１記載の畜肉練り製品。 The meat paste product according to claim 1, which contains the above (1) and (2).
  3. 　前記コリン及びイノシトールが、畜肉練り製品の原材料に含まれるリン脂質に由来する、請求項１又は２に記載の畜肉練り製品。 The meat paste product according to claim 1 or 2, wherein the choline and inositol are derived from phospholipids contained in raw materials of the meat paste product.
  4. 　原材料である畜肉にホスホリパーゼＤを作用させることを含む、畜肉練り製品の加熱後歩留り向上方法。 A method for improving the post-heating yield of livestock meat paste products, which includes allowing phospholipase D to act on livestock meat, which is a raw material.
  5. 　ホスホリパーゼＤが、シグマ　アルドリッチ製ホスホリパーゼＤ　放線菌由来（商品名）、ナガセケムテック製デナチームＰＭＤ－Ｐ１（商品名）及び旭化成ファーマ製ＰＨＯＳＰＨＯＬＩＰＡＳＥ　Ｄ［ＰＬＤＰ（Ｔ－３９）］（Ｇｌｙｃｅｒｏｐｈｏｓｐｈｏｌｉｐｉｄ　ｓｐｅｃｉｆｉｃ）（商品名）からなる群から選択される、請求項４記載の方法。 Phospholipase D is Sigma-Aldrich's Phospholipase D Actinomycete-derived (trade name), Nagase Chemtech's Denazym PMD-P1 (trade name), and Asahi Kasei Pharma's PHOSPHOLIPASE D [PLDP (T-39)] (Glycerophospholipid specific) (trade name) 5. The method of claim 4, wherein the method is selected from the group consisting of:
  6. 　トランスグルタミナーゼ、リパーゼ、及び澱粉分解酵素からなる群から選択される少なくとも１つの酵素をさらに作用させることを含む、請求項４または５に記載の方法。 The method according to claim 4 or 5, further comprising allowing at least one enzyme selected from the group consisting of transglutaminase, lipase, and amylolytic enzyme to act.
PCT/JP2022/019450 2021-04-30 2022-04-28 Kneaded meat product with improved quality WO2022231003A1 (en)

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH11346719A (en) * 1998-06-03 1999-12-21 Izu Rice Center:Kk Preparation of lye using phyllostachys pubescens and modification of meat using the same
WO2006075771A1 (en) * 2005-01-13 2006-07-20 Ajinomoto Co., Inc. Processed meat product or fish paste product and process for producing the same
WO2010140708A1 (en) * 2009-06-05 2010-12-09 味の素株式会社 Enzyme preparation for reforming processed meat products and manufacturing method for processed meat products
WO2013172447A1 (en) * 2012-05-17 2013-11-21 ナガセケムテックス株式会社 Enzyme preparation for modifying food material
CN107593874A (en) * 2017-08-24 2018-01-19 安徽靖童科技农业发展有限公司 A kind of meat product processing stabilizers
JP2020145977A (en) * 2019-03-14 2020-09-17 奥野製薬工業株式会社 Texture-improving agent and texture-improving method of meat product

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH11346719A (en) * 1998-06-03 1999-12-21 Izu Rice Center:Kk Preparation of lye using phyllostachys pubescens and modification of meat using the same
WO2006075771A1 (en) * 2005-01-13 2006-07-20 Ajinomoto Co., Inc. Processed meat product or fish paste product and process for producing the same
WO2010140708A1 (en) * 2009-06-05 2010-12-09 味の素株式会社 Enzyme preparation for reforming processed meat products and manufacturing method for processed meat products
WO2013172447A1 (en) * 2012-05-17 2013-11-21 ナガセケムテックス株式会社 Enzyme preparation for modifying food material
CN107593874A (en) * 2017-08-24 2018-01-19 安徽靖童科技农业发展有限公司 A kind of meat product processing stabilizers
JP2020145977A (en) * 2019-03-14 2020-09-17 奥野製薬工業株式会社 Texture-improving agent and texture-improving method of meat product

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