KR20080069463A - Method of preparing a natural flavoring comprising pearl oyster muscle extract with enhancing flavor - Google Patents

Abstract

A method of manufacturing natural seasonings is provided to prepare natural seasonings with characteristic flavor and taste and functionality of pearl oyster muscle and utilizes pearl oyster muscle which is hitherto been discarded. Natural seasonings are prepared by the steps of: mixing pearl oyster muscle extract(Brix 20deg.), 0.4M glucose, 0.4M glycine and 0.4M cystine in a weight ratio of 4:2:1:1 and reacting at 120 to 150deg.C for 140 to 150min while maintaining a pH of 6.9 to 7.6; and additionally adding oyster concentrates thereto. The pearl oyster muscle extract is obtained by the steps of: mixing pearl oyster muscle with 2 to 3 times the weight of water; blanching at 90 to 100deg.C for 3 to 10min; adding Alcalase thereto and heating at 50 to 60deg.C for 3 to 5hr while stirring; heating at 90 to 100deg.C for 3 to 10min to deactivate the Alcalase; adding exopeptidase and hydrolyzing at 40 to 50deg.C for 3 to 5hr while stirring; heating at 90 to 100deg.C for 3 to 10min to deactivate the exopeptidase; and separating the residue, centrifuging and concentrating.

Description

Method of preparing a natural flavoring using pearl clam extract with improved flavor {Method of preparing a natural flavoring comprising pearl oyster muscle extract with enhancing flavor}

1 is an HPLC chromatogram of a standard mixture of six nucleic acid related substances.

Figure 2 is a photograph showing a stainless reactor for the sugar-amino reaction.

Figure 3 shows the central synthesis scheme for improving the flavor of pearl shell meat extract.

Figure 4 is a flow chart showing a process for producing a pearl shell meat extract using hot water, soaked water and enzyme digestion, respectively.

FIG. 5 is a photograph showing pearl shell meat extracts prepared by using hydrothermal well, heavy bath and enzymatic digestion, respectively. FIG. In this case, A represents an enzyme decomposition extract, B represents a high-moorbital extract, and C represents a hydrothermal extract.

Figure 6 is a graph showing the yield of pearl shell meat extract according to the extraction method.

7 is a graph showing the ACE inhibitory activity of pearl shells extract according to the extraction method.

8 is a graph showing the response surface analysis results for the effect of the reaction time and the pH of the reaction solution on the expression of the sweet smell of pearl shell extract, glucose, glycine-cystine sugar-amino reaction solution at 120 ℃ to be.

9 is a graph showing the response surface analysis results for the effect of the reaction time and pH of the reaction solution on the shielding expression of pearl shell meat extract-glucose-glycine-cystine sugar-amino reaction solution at 120 ℃ to be.

10 is a graph showing the response surface analysis results for the effect of the reaction time and pH of the reaction solution on the boiled beef flavor expression of pearl shell meat extract-glucose-glycine-cystine sugar-amino reaction solution at 120 ℃ to be.

11 is a flow chart showing a manufacturing process of natural seasoning using the present invention pearl clam extract.

12 is a photograph showing a granular natural seasoning using the present invention pearl clam extract.

The present invention relates to a method for preparing natural seasoning using pearl shell meat extract with improved flavor, wherein the sugar-amino reaction is carried out by adding glucose, glycine and cystine mixed solution to pearl shell meat extract prepared by two-stage enzymatic digestion. A method of producing natural seasonings containing the functional ingredients contained in pearl shellfish, while adding significantly to the shellfish's flavor and aroma while significantly reducing the amount of MSG, a chemical seasoning, by using pearl shellfish extract with improved flavor. will be.

As the standard of living in Korea improves, and consumers' awareness of food safety and functional nutrition increases, the natural extract with various functional characteristics to improve the unique taste of processed foods and give functionality to processed foods. The use of extracts is increasing day by day. This natural extract material can be found in the meat extract (meat extract) that is common in the West, but in recent years, a variety of raw materials, a variety of flavors and nutritional functional ingredients are widely used as the main raw material of the natural extract material It is becoming. Recently, the demand for flavor-based natural seasonings such as granules, powdery seasonings and concentrated extracts from natural materials with various flavors is increasing along with the safety issues of biosynthetic fermented seasonings and diversification and enhancement of taste.

Most research reports on extract material of these aquatic products are mostly general articles on general methods of extracting extracts from aquatic products. On the other hand, researches on the development and practical use of natural extract materials are mostly conducted by companies and have been the company's know-how. Therefore, there are few reports of actual research on commercialization methods. Preparation of Extracts (Park, JY et al., 1998. Korean J. Food Sci . Technol ., 20: 433 ~ 440), Studies on the Development of Seasoning Ingredients and Seasoning Materials in Fish and Shellfishes (Kim Young-Myung et al., 1988. study on material developed Korea Food research Institute research report; Koo, JK et al, 1985. Korean J. Food Sci Technol, 17:.... 283 ~ 288), manufacture of flavoring material used kelp mackerel meat processing enzymes (Lee, KH et al., 1997. Korean J. Food Sci . Technol . , 29: 77 ~ 81), Development and flavor expression of natural seasoning materials using oysters (Oh, KS et al., 1998. J. Ins Marine.Industry. , 10: 9-17; Oh, KS et al., 1998.J. Ins.Marine.Industry. , 1 0: 19 ~ 23), processing of seasoned materials using coastal shellfish (Moon, JH et al., 2002. J. Ins. Marine. Industry. , 15: 7-16).

Seafood and shellfish, such as oysters, mussels and clams, molluscs such as squids, crabs and shrimps, and invertebrate aquatic products such as crustaceans, have a slightly different taste and aroma. It has been widely used as flavor-based seasoning material in food all over the world. However, in recent years, due to the lack of resources and high prices, they have become impossible to use as seasoning raw materials. Therefore, these processed by-products are mainly used as seasoning raw materials, and efficient extraction of effective materials and development of steering technology are required. It is becoming.

Research on flavors and steering of foods, which are important food-specific characteristics that influence the quality of foods, has been actively conducted for livestock foods, mainly in Europe and the United States, and in particular certain substances included in specific foods. It has been focused on qualitative and quantitative analysis of how important it is and how much it is contained. However, in recent years, with the development of analytical equipment and technology using them, interest in flavor, which is one of the food flavor ingredients, has increased, and in order to develop new foods to satisfy the tastes of various consumers, it is necessary to improve the color and flavor of food. Development of technology for flavor enhancement is required. Meanwhile, in the case of aquatic foods, flavor enhancement techniques for masking and steering fish odor, which are the biggest obstacles to the use of aquatic products, have been studied in Korea, but in most cases, spices or herbal medicines are added. By masking off-flavor.

Process flavor for improving the flavor of food refers to the flavor produced by some form of processing technology from precursors in food. The main processing technology is heat treatment and enzymatic action, especially the flavor produced by heating Is called the reaction flavor. After World War II, research was under way to develop alternatives for meat in Europe, and as a result, reaction flavoring technology developed. The reaction, which is the basis of this reaction flavoring technology, was the Maillard reaction. to be. The Mylar reaction occurs extensively during the heating and storage of foods and produces many flavors and pigments called melanoidin through the reaction of reducing sugars with amino acids, which are first N-substituted by the reaction of reducing sugars with amino acids. Glycosylamine (N-substituted glycosylamine) is formed, followed by amadori rearrangement and a series of reactions, resulting in various types of fural, dicarbonyl, aldehyde, and hydrogen. Intermediates such as hydrogen sulfide are produced. They continue to be precursors, and meat-based fragrances such as pyrazine, pyridine, pyrrole, thiazole, thiophene and oxazole Is generated. Reaction flavoring technology is now widely used in the flavor industry to produce beef or savor, and the demand for ready-to-eat foods has increased due to changes in dietary patterns. There is an increasing demand for savor and the need to develop various reaction treatment techniques.

Beef flavor is produced by heating precursors such as amino acids, peptides, proteins, nucleic acids, fats, sugars, and thiamin, which may be Myyal reactions, splitter degradation, and myyal reaction intermediates. Reactions with hydrogen sulfide, ammonia and thiol, pyrolysis of thiamin, pyrolysis of amino acids and sugars, fatty acidization, and degradation of ribonucleotides are known to be important reactions involved in the production of beef flavor ( Nagodawithana, TW 1995. Savory flavors.Esteeekay Associate, Inc., Milwaukee: 164-224). In particular, cysteine, a sulfur-containing amino acid, produces 2-mercto-acetaldehyde, acetaldehyde, and hydrogen sulfide, which are highly reactive to cracker degradation when heated, and these compounds react as important precursors for the development of beef flavor. Do it. In Korea, a reaction flavoring described by a meat flavor development and Mai Yalta reaction vegetable protein hydrolyzate (HVP) and yeast extract is added to the precursor bars is attempted by the development of meat flavor (Yoon, SH et al., 1994. Korean J. Food Sci . Technol ., 26: 781-786).

However, the meat flavor-based flavor material developed mainly in the West has reached the level where industrial production is possible. Korea's seafood processed food also develops key flavor material products with high flavor and high added value. In the future, research on the flavor technology of aquatic foods should be carried out.

On the other hand, in recent years, the reaction surface analysis method has been used for the optimization of the reaction flavoring, the previously used experimental method is the effect of changing only one of the independent variables and all other variables fixed Although the one-factor-at-time-method for observing is mainly used, the response surface analysis method that saves time and expense and observes the interaction by factors when the result is due to the interaction of various independent variables such as food It is widely used (Moupuet, C. et al., 1992. J. Food Sci ., 57: 1395-1400).

Pearl oyster (Pinctada fucata martensii ) grows on reefs with a depth of 5 to 10 m, and is known to inhabit small scales mainly in Korea's Geojedo and southern Japan, and puts sections cut into 4-9 mm ² of holes and perforated membranes in the gonads. If you farm, you can get gemstone pearls. In the area near Tongyeong, it has succeeded in obtaining circumference by mass-producing pearl shells for the purpose of collecting precious pearls. Recently, it was selected as one of the regional development specialization projects in Korea, and its production is on the rise. As a result, pearl clam meat obtained in large quantities as a by-product when extracting columnar is restricted to use in seasoning products or canned products that require the maintenance of the shape of raw meat because there are many foreign substances and viscous substances, and it is difficult to maintain a certain form. It is only used as feed, and most of it is crushed and discarded. Pearl market using pearl shells The market size of pearl industry is about 120 billion won, while pearl shellfish meat obtained as a by-product has no proper use, and the rest is discarded except some routs.

Accordingly, the present invention has been made to solve the above problems, the pearl shell (Pearl Oyster, Pinctada) which is a by- product of the pearl culture industry fucata ) Extracts useful extracts that can be used as flavors for seasonings and instant foods using meat, and then improves the flavor of pearl shell extracts by optimizing reaction for flavor improvement. It is an object of the present invention to provide a method for producing granulated natural seasonings using the extract and preparing the extract and natural seasonings prepared using the same.

The object of the present invention is to investigate the components of pearl clam meat, and to investigate the components and sensory characteristics according to the extraction method of pearl clam meat, sugar-amino acid for flavor improvement in the most optimally selected pearl shell clam enzymatic digestion extract After selecting the optimum reactor quality of the reaction, the optimum reaction conditions were investigated through dynamic monitoring of the Mylar reaction, and pearl shell meat extract was prepared under the optimum reaction conditions investigated as above, and granular natural seasoning was prepared using the extract. And by examining its quality characteristics.

Hereinafter, the configuration of the present invention.

The present invention comprises the steps of examining the components of the pearl clam meat; Investigating the components and sensory characteristics according to the extraction method of pearl clam meat; Selecting the optimum reactor quality of the sugar-amino acid reaction for improving the flavor of the pearl shell meat extract; Investigating optimal reaction conditions through dynamic monitoring of the Mylar reaction for flavor improvement of pearl shell meat extract; Preparing pearl shell meat extract under the optimum reaction conditions; Preparing a natural seasoning using the extract; Investigate the quality characteristics of the natural seasoning.

Pearl clam meat extract in the present invention is preferably produced by the following two-stage enzymatic digestion method in terms of palatability and economics.

2 ~ 3 times of water is added to pearl clam meat and incubated at 90 ~ 100 ℃ for 3 ~ 10 minutes to inactivate autophagy. Alcalase is added and primary enzymatic digestion is performed at 50-60 ° C. for 3-5 hours with stirring. And heat treatment at 90 ~ 100 ℃ again for 3-10 minutes to inactivate the alkalase, and then add Exopeptidase such as Flavorzyme (Exopeptidase) for 3 to 5 hours while stirring at 40 ~ 50 ℃ After hydrolysis for 3 to 10 minutes at 90 ~ 100 ℃ to inactivate the exopeptidase. The residue is then separated, left to cool, centrifuged to remove the residue, and then concentrated to obtain an extract.

In the present invention, after selecting the optimum reactor quality of the sugar-amino acid reaction for flavor improvement in the pearl clam meat enzyme digestion extract, by examining the optimum reaction conditions through dynamic monitoring of the Mylar reaction, pearl clam meat extract (Brix 20 °): 0.4M Glucose: 0.4M Glycine: 0.4M Mixture of cystine 4: 2: 1: 1, keeping the pH at 6.9-7.6 while maintaining the pH at 120-150 ° C for 140-150 minutes It is possible to provide an improved pearl shell extract.

The present invention provides a natural seasoning composition comprising pearl shell meat extract with improved flavor by the sugar-amino acid reaction prepared as described above as an active ingredient.

In the present invention, the content of pearl shell meat extract with improved flavor by the sugar-amino acid reaction in the natural seasoning composition is preferably 3 to 10% by weight, but 5% by weight is the best.

Each experiment used in the present invention can be performed as follows.

1) General ingredient: The composition of the general ingredient is normal pressure (The Korean society of food science and nutrition. 2000. Handbook of experiments in food science and nutrition.Hyoil Publish Co., Seoul: 96 ~ 127) The drying method, crude protein content was measured by semimicro Kjeldahl method, crude fat content by Soxhlet method, and crude ash content by dry painting method.

2) pH, acidity and salinity: pH is homogenized and measured with a pH meter (Accumet Basic, Fisher Sci. Co., USA), and acidity is measured in 100 mL of pH sample. 0.1N NaOH solution is added dropwise to indicate the number of mL of the solution required until the pH reaches 8.3, and the salinity is measured by a salinity meter (460CP, Istek Co., Korea).

3) Volatile Basic Nitrogen, Amino Nitrogen and TBA Value: Volatile Basic Nitrogen (VBN) is measured by microdiffusion using Conway unit, and amino nitrogen (NH ₂ -N) content is formol. (Formol) Measured by titration method. TBA value (Thiobarbituric acid value) is measured by the steam distillation method of Tarladgis et al. (Tarladgis, ZG et al., 1960. J. Am. Oils Chem . Soc ., 37: 44 ~ 48)

4) Yield, Hue and Sugar: Yield of each extract is expressed as the amount of Brix 2.0 ° extract obtained from 1 kg of pearl shell meat. The hue is measured using the direct color scale (Color meter ZE-2000, Nippon Denshoku Ltd., Japan), L value (brightness), a value (redness), b value (yellowness), and ΔE for the transmission hue of the sample extract. Measure the value (color difference). The L, a and b values of the standard plate are 99.98, 0.01 and 0.01, respectively. The solid content of the sample extract is measured by Refractometers (2E, Atago, Japan).

5) Inhibitory Activity of Angiotensin-I Converting Enzyme: The inhibition of angiotensin-I converting enzyme (ACE) is measured by Horiuchi et al. (Horiuchi, M. et al., 1982. J. Chromatogr . 233: 123-130) . . In order to measure the ACE inhibition capacity, 50 μL of purified ACE (Sigma, 60 mU / mL) was added to 15 μL of the extract at a predetermined concentration, followed by preincubation (37 ° C., 5 min). To this was added 125 μL of a 5 mM solution of hippuryl-histidyl-leucine dissolved in boric acid buffer solution (pH 8.3, containing 400 mM NaCl) and reacted again (37 ° C., 30 minutes), followed by 10% Reversed-phase HPLC (LC-10AVP, Shimadzu Co., Ltd.) with 20 μL of trifluoroacetic acid (TFA), equipped with Zorbax 300SB C8 column, Hewlett Packard Co., 4.6 × 150 mm Japan) and then analyzed according to the conditions shown in Table 1. ACE inhibitory capacity is expressed as IC ₅₀ , the amount of inhibitor required to inhibit ACE activity by 50%.

HPLC conditions for investigating ACE inhibition Item Condition device HPLC (Shimadzu Co., LC-10Avp, Japan) column Zorbax 300SB C ₈ column (Hewlett Packard Co., 4.6 × 150 mm) Detector UV 228 nm Mobile phase DW / 0.1% Trifluoroacetic acid, 70% CH ₃ CN / 0.1% Trifluoroacetic acid Flow rate 1 mL / min. Sample injection amount 20 μL Column temperature 30 ℃ Sample temperature 30 ℃

6) Net ingredient

Free amino acids: The free amino acids and related compounds are dried under reduced pressure and dried on protein extracts and degreased samples with 5'-sulfosalicylic acid and ether, followed by lithium buffer (pH 2.20, 0.20 M). After the determination, the result is analyzed by an amino acid autoanalyzer (Biochrom 20, Pharmacia Biotech., England).

Nucleic Acid Related Substances: Nucleic acid related substances (ATP degradation products) are analyzed by HPLC as follows. In other words, put the 6.0 N KOH solution into the protein extract and degreasing treatment to adjust the pH to 6.5 ~ 6.8, centrifuged to give a certain amount of neutralized PCA solution, then left at 5 ℃ for 3 hours or more to remove the low-temperature crystalline material Remove This was injected into a certain amount of HPLC (Yeongin HPLC 9500 system, Yeongin Co., Korea) equipped with a Synergi 4u hydro-RP 80A column to analyze the nucleic acid-related substances, the analysis conditions are shown in Table 2 below. And HPLC chromatogram of the standard mixture of six nucleic acid-related substances obtained under these analysis conditions is shown in FIG.

HPLC Conditions for Analysis of Nucleic Acid-Related Substances Item Condition device Yeongin HPLC 9500 column Synergi 4u Hydro-RP 80A, 250 × 4.60 mm menstruum _{0.04M KH 2 PO 4 · 0.06MK 2} HPO 4 (pH 7.5) Flow rate 1.0 mL / min. Detector UV 254 nm Sample loading 20 μL

TMAO (trimethyamine oxide) and TMA (trimethyamine): 1 mL of neutral formalin solution, 10 mL of toluene and 3 mL of 50% K ₂ CO ₃ solution are mixed vigorously in 5 mL of the protein extract and degreased sample extract in the separating funnel. After shaking, transfer only the toluene layer to a test tube containing anhydrous Na ₂ SO ₄ , dehydrate it, and take it to another test tube, add 5 mL of a 0.02% toluene-picric acid solution, mix, and absorb the absorbance at 410 nm. Measure to determine the TMA content. Meanwhile, 5 mL of sample extract, 5 mL of 5% TCA solution, and 0.5 mL of 10% TiCl ₃ solution were placed in a 25 mL diaphragm flask and left for 2 hours, followed by water, and the absorbance at 410 nm was measured to determine the content of (TMAO + TMA). Here, the TMA content obtained above is subtracted to give the TMAO content.

Total creatinine: 8 mL of the protein and degreased sample extracts were taken into a test tube, 1 mL of 1.0 NH ₂ SO ₄ solution was added, and the stopper was decomposed at 121 ° C. for 30 minutes. After cooling, add a drop of m -nitrophenol, 1 mL of 0.1 N NaOH solution, and allow to stand at room temperature for 1 hour, and then measure the absorbance at 520 nm using a UV spectrophotometer (Unicam Heλios, Unicam Ltd., England). Find the content of.

Betaine: Quantified by colorimetric method using column chromatography using Dowex 50w × 20 (H ⁺ -form) cation exchange resin and reaction with ammonium reineckate salt.

Inorganic Components: The cations in the inorganic components are taken in a batch of sample extract in a batch crucible, dry ignited at 500 to 550 ° C for 5 to 6 hours, filtered through an ashless filter paper, and then fixed in a predetermined amount, followed by ICP (Inductively Coupled Plasma Atomic Emission). Spectrometer, Atomscan 25, TJA Co., USA) analyzes the contents of Na, Mg, K, Ca, Cu, Fe, Zn, S and P.

Reaction flavoring experiment for improving the flavor of pearl shell meat extract in the present invention is carried out as follows.

In other words, by the central synthesis plan, the taste of pearl clam extract is improved by using the Mylar reaction and the surface reaction analysis method using the reaction time and pH of the reaction solution as independent variables, and the reaction conditions for optimization thereof are investigated. The flavor improvement reaction solution uses an enzyme digestion extract prepared by two-stage enzymatic digestion.

Myyal reaction: After adding a certain amount of sugars such as glucose, amino acids such as glycine, cystine to the enzymatic digestion of pearl clam meat, adjust the pH, and put into a reactor (Hoke SS-DOT) as shown in FIG. The optimization condition of the Mylar reaction is investigated through the evaluation of flavor produced by heating for a certain time at a constant temperature, that is, sensory evaluation of the model reaction system.

Response surface analysis for optimization of Mylar reaction: In order to improve the flavor of pearl clam enzymatic digestion extract, the Mylar reaction is carried out by setting the reaction time and pH of the reaction solution as independent variables according to the central synthesis scheme. The experimental area is encoded in five stages of -2, -1, 0, +1, +2, and the response surface experimental plan is prepared according to the central synthesis plan using these independent variables and codes. In the present invention, the sensory properties of the reaction solution are monitored according to each independent variable in order to predict the optimum reaction condition using Response surface methodology (RSM). The reaction temperature is an important factor in the expression of boiled beef flavor and is essential for the myal reaction. However, this reaction temperature is limited to less than 15 minutes at 180 ° C. (longer temperatures at lower temperatures), which is a guideline for the manufacture of reaction flavors proposed by the Initiational Organization of Flavor Industries (IOFI). In general, when meat is heated, carcinogenic substances such as heterocyclic amines may be produced, and thus may be generated in food seasonings subjected to the myal reaction. Therefore, in the present invention, in consideration of the food hygiene safety as described above, the experiment is carried out while fixing the reaction temperature to a temperature much lower than 180 ℃ 120 ℃.

The experimental design for the reaction conditions uses a fractional factorial design and a statistical analysis system (SAS) program for response surface regression analysis. The experimental design is a factor considered as an important independent variable in the Mylar reaction, as shown in Table 3 below, reaction time (30, 60, 90, 120, 150 minutes), pH of the reaction solution (5, 6, 7, 8, 9). ) Is encoded in five stages of -2, -1, 0, +1, and +2. That is, the independent variable Xi shown in Table 3 is set to 10 sections (factor test point 4, center point 2, axis point 4) as shown in FIG. In addition, the dependent variables (Yn) affected by these independent variables, that is, sensory traits of the reaction solution (baked potato odor, Y1), masking of shellfish odor (Y2), boiled beef flavor (Boiled) meat odor, Y3), and repeated measurements three times for each condition are used for the regression analysis (panels n = 12).

Independent variable level for the central synthesis scheme for the Mylar response Xi Independent variable Level -2 -One 0 +1 +2 X ₁ Reaction time (min.) 30 60 90 120 150 X ₂ PH of reaction solution 5 6 7 8 9

In the present invention, the sensory test is performed as follows.

12 sensory panelers trained to be familiar with sensory characteristics such as shellfish extract taste, odor and color tone were used to construct several sensory characteristics of the extract's flavor and reaction variables of the Mylar response The five-stage or nine-stage rating method for the odor, odor, shielding, and boiled beef aroma is measured and evaluated (temperature: 45 to 50 ° C).

Sensory evaluation of pearl shell granulated seasoning consisted of nine panels trained to be familiar with the sensory characteristics of commercial seafood seasonings, and the sensory characteristics of flavors such as taste, odor, color tone and overall preference of this product were evaluated by five-step evaluation method (5). scale score).

The results of sensory test were ANOVA test and Duncan's multiple range test using the SPSS system (Statistical Package, SPSS Inc. USA). Test.

Hereinafter, the configuration of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to the following Examples.

Example  1: raw material Pearl shell  Ingredient survey

Pearl shell ( Pinctada) fucata ) meat was purchased from Daedeok Jinju Co., Ltd., Dosan-myeon, Tongyeong-si, Gyeongsangnam-do, South Korea. After removal, the resultant was frozen and stored in a plastic film bag and used as a raw material for extract extraction.

General ingredients, pH, salinity and the contents of volatile basic nitrogen (VBN) of the raw pearl clam meat used in the experiment are shown in Tables 4 and 5 below. The moisture content of raw pearl clam meat was 83.2%, which was somewhat higher than that of general fish and shellfish. The crude protein content was 11.3% and the crude fat content was 0.8%. The ash content was 1.4% and the carbohydrate content was 3.0%. The composition of the general components of the pearl clam meat seems to be somewhat different depending on the time of collection. On the other hand, the pH of the pearl clam meat was around 6.4, the salinity was 1.2%, and the VBN content was 16.2 mg / 100 g.

Common ingredients of pearl clam meat (g / 100 g) moisture Crude protein View minutes Geography carbohydrate content 83.2 ± 0.7 11.3 ± 0.2 1.7 ± 0.1 0.8 ± 0.2 3.0 ± 1.1

PH, Volatile Basic Nitrogen and Salt Contents in Pearl Shell Meat pH VBN (mg / 100 g) Salinity (g / 100 g) content 6.4 ± 0.1 16.2 ± 0.2 1.2 ± 0.1

Table 6 shows the results of analyzing the free amino acid composition of the raw pearl clam meat used in the experiment by an amino acid automatic analyzer. The total amount of free amino acid, the most important taste component of nitrogen-containing extract, is 508.8 mg / 100 mL, which includes taurine (150.9 mg / 100 g), urea (112.4 mg / 100 g), glutamic acid (55.6 mg / 100 g), and glycine (45.1). mg / 100 g) and alanine (27.9 mg / 100 g) were the major free amino acids, but their contents were significantly lower than those of general fish and shellfish. In general, free amino acids are known as 'taste-active components', which are the most important semi-expressing components of aquatic products (金 東 勳. 1985. Food Products Chemical., Seoul: 30-32), such as glutamic acid, glycine and alanine. It has been identified as a major taste amino acid (Hayashi, T. et al., 1981. J. Food Sci ., 46 : 479-483). As a result, the content of free amino acid or its composition is not enough to make the oyster pearl taste better than other shellfish. When used as a natural seasoning material, the use of other aquatic extracts with strong taste It was deemed necessary.

Free amino acid content of pearl clam meat (mg / 100 g) Free amino acids content Taurine 150.9 Urea 112.4 Threonine 20.4 Serine 3.9 Asparagine 19.3 Glutamic acid 55.6 α-aminoadipic acid 1.9 Proline 7.9 Glycine 45.1 Alanine 27.9 Valine 5.0 Cystine 2.3 Methionine 2.6 Isoleucine 3.4 Leucine 2.9 Tyrosine 3.4 β-alanine 6.3 Phenylalanine 2.3 Ethanolamine 4.6 Ammonium chloride 16.8 Lysine 12.7 Histidine 1.2 Arginine 10.0 Sum 508.8

The mineral composition of the pearl clam meat was measured by ICP. In pearl shells, the composition of inorganic ions such as Na, P, K, Ca and Zn was high, but its content was relatively low compared to other shellfish (Seo, SB et al., 1995. Chemical composition of marine products in korea. National Fisheries Research and Development Agency Republic of Korea ). Among the inorganic ions, Na, K, P, and Cl are known as a net-expressing ingredient that contributes to the net expression of aquatic products together with free amino acids and IMP (Hayashi, T. et al., 1978. Nippon Suisan Gakkaishi 44 : 1357-1362). On the other hand, pearl shells were somewhat unusual because they contained a relatively large amount of ingredients such as Zn (53.9 mg / 100 g) and S (33.5 mg / 100 g).

Mineral composition of pearl clam meat (mg / 100 g) Mineral content Na 524.1 ± 32.0 Mg 23.1 ± 49.0 Ca 62.8 ± 0.9 Fe 23.1 ± 5.6 Cu 0.8 ± 0.1 Zn 53.9 ± 9.3 S 33.5 ± 2.7 K 67.5 ± 4.0 P 192.4 ± 24.2

Example  2: Pearl shell Extract  Produce

Pearl clam meat extract in the present invention was prepared by three methods of hydrolysis, soaking water and enzyme digestion.

First, the hot-water extract extracts about 10 times the weight of the raw material into the raw pearl clam meat, and then trebles in hot water for 8 hours, and then separates the residue using a filter cloth made of ore. . After cooling the filtrate, the residue was removed by centrifugation, and the concentration of the solid was adjusted to Brix 2.0 DEG.

Hydro-cooked extract was extracted by pressurizing three times the weight of raw material to raw pearl clam meat and pressurized at 110 ℃ for 2 hours by using a water bath (DaeMongHong, herbal extractor DHM-23E). Next, the residue was separated using a filter cloth made of mineral wood, and after cooling, centrifuged to remove the residue, and then Brix 2.0 ° adjustment was referred to as a soothing pitch extract. At this time, the amount of water was three times the amount of the raw materials.

Enzymatic hydrolysate was prepared by two step enzyme hydrolysis. First, the mother-of-pearl shell was cut into choppers, and twice the amount of water was added to the sample weight, and the autodigestion enzyme was inactivated by boiling at 95 ° C for 5 minutes. Then, the pH of the sample solution was adjusted, and Alcalase 0.6L enzyme (Novo Nordisk Co., Denmark) was added thereto, followed by primary enzymatic degradation at 55 ° C. for 4 hours with stirring. After heat treatment at 95 ° C. for 5 minutes to deactivate the alcalase, the pH was further adjusted, and Flavorzyme (Novo Nordisk Co., Denmark) was added thereto, followed by hydrolysis for 4 hours with stirring at 45 ° C. After heat treatment at 95 ℃ for 5 minutes to inactivate the flavozyme. Next, the residue was separated by using a filter cloth, allowed to cool, centrifuged to remove the residue, and then the Brix 2.0 ° adjusted extract was used as an enzyme digestion extract. The characteristics of the proteolytic enzymes used were shown in Table 8 below. Process diagrams of the hydrothermal well, heavy bath and enzymatic pearl shell extract is as shown in Figure 4 and each extract was as shown in FIG.

Characteristics of Novosa (Denmark) Protease Used for Preparation of Enzymatic Extract enzyme Trade name Optimum reaction temperature (℃) Optimum reaction pH Manufacturing company name Primary enzymatic enzyme Alcalase 0.6L 50-60 8.0-8.5 Novo Nordisk Co., Denmark Secondary enzyme Flavorzyme 45-55 6.0-7.0 Novo Nordisk Co., Denmark

Example  3: Pearl shell  Investigation of Components and Sensory Characteristics by Extraction Method

1) General Ingredient

As a raw material of pearl clam meat, the general components of the three pearl clam extracts prepared by hydrothermal well, heavy bath and two-stage enzyme decomposition according to the process as shown in FIG. 3 were as shown in Table 9 below. The water content of the pearl clam extract with the amount of solids adjusted to Brix 2 ° was 97.9 ~ 98.0%. The crude protein content, which shows the amount of amino acid, which is the most important component of the extract, was 1.3% for enzyme digestion extract. %) And hot water treble (0.7%) showed more than 2 times higher content. Meanwhile, the content of crude ash was 0.2% in each extract, and there was no difference in content according to the extraction method.

Common Components of Pearl Clam Extract (g / 100 mL) Extract moisture Crude protein View minutes Hot water 97.9 ± 0.1 0.7 ± 0.0 0.2 ± 0.0 Heavy water 98.0 ± 0.0 0.5 ± 0.0 0.2 ± 0.0 2-stage enzyme digestion 97.5 ± 0.0 1.3 ± 0.2 0.2 ± 0.0

2) pH, salinity and acidity

The results obtained by measuring the pH, salinity and acidity of the pearl clam hot water, water bath and enzyme digestion extract are shown in Table 10 below. The pH of the pearl shell extract ranged from 6.15 to 6.30, but there was no significant difference, but the pH of the enzyme digestion extract was relatively lower than that of other extracts. This was due to the large amount of organic acid eluted from the pearl shell meat during enzymatic hydrolysis. In the case of hot water and hot water extract, the pH was slightly increased due to the effects of volatile basic nitrogen such as NH ₃ , TMA, and DMA generated by the decomposition of some proteins or free amino acids during extraction under hot water or high pressure. It was estimated.

The acidity, which can indirectly determine the content of the organic acid eluted in the extract, also showed the highest amount of enzymatic extraction (0.98 mL), so that the most of the organic acids eluted during extraction.

PH, Salinity and Acidity of Pearl Shell Extract Extract pH Salinity (g / 100 mL) PH (mL) Hot water 6.30 ± 0.2 0.17 ± 0.01 0.46 ± 0.09 Heavy water 6.29 ± 0.0 0.17 ± 0.04 0.32 ± 0.04 Enzyme digestion 6.15 ± 0.1 0.15 ± 0.02 0.98 ± 0.08

3) volatile basic nitrogen and amino nitrogen

The results of measuring the contents of volatile basic nitrogen and amino nitrogen of the pearl clam hydrolyzate, heavy bath and enzyme digestion extracts are shown in Table 11 below. The amount of volatile basic nitrogen of the pearl clam extract was 4.9 mg / 100 mL, which was higher than that of other extracts, resulting in high temperature pyrolysis resulting in ammonia and trimethylamine. Heavy water extract contains 3.1 mg / 100 mL, which produces less volatile basic nitrogen than hot water extract, while enzyme extract extracts 2.1 mg / 100 mL to lower amine substances due to deterioration of freshness or degradation of meat during extraction. It was estimated that the most effective extraction method was in terms of suppression of off-flavor production among extract extraction methods.

The amino nitrogen content, which can indirectly determine the free amino acid content of pearl shell extract, ranged from 35.0 to 74.5 mg / 100 mL. The amino nitrogen content of enzymatic digestion extract was more than twice that of other extracts. It was in extract order. Therefore, the two-stage enzymatic extraction method was also considered to be the most effective extraction method in terms of recovery of free amino acids derived from pearl shell meat.

VBN and Free Amino Acid Contents of Pearl Shell Extract Extract VBN (g / 100 mL) Free amino acid content (g / 100 mL) Hot water 4.9 ± 0.7 43.2 ± 0.06 Heavy water 3.1 ± 0.6 35.0 ± 0.28 Enzyme digestion 2.1 ± 0.6 74.5 ± 0.04

4) yield

6 shows the yields of the pearl clam hydrolysis, basal tremor and enzymatic extract obtained by adjusting the amount of solids to Brix 2.0 °. The yield of Brix 2.0 ° extract obtained from 1 kg of pearl clam is 2.65 L in hot water, 1.80 L in hot water and 6.58 L in enzymatic digestion. The two-stage enzymatic extraction method is economical to extract hot water and heavy water. It was found to be significantly more effective than

5) Inhibitory Activity of Angiotensin-I Converting Enzyme (ACE)

As a result of measuring the inhibitory ability of ACE, one of the causes of blood pressure increase in the human body, among the functional properties of protein hydrolysates such as pearl clam hydrolyzate, glutinous sonic and enzyme digestion extract, were as shown in FIG. 7. IC _50, the amount of inhibitor required to inhibit ACE activity by 50%, was 1.39 mg / mL, hydrocoated pitch 4.17 mg / mL, and hydrothermal pitch 7.95 mg / mL, which showed the best ACE inhibitory activity. The hydrothermal extract was the lowest.

Therefore, pearl shell extract was found to have a significant effect on the ACE inhibitory ability, and enzyme digestion, which produces more peptides than hot water or basin, was superior to ACE inhibition.

6) tint

Table 12 shows the results of measurement of the color tone of the pearl clam hydrolysis basin, basal bass and enzymatic digestion with a direct color difference meter. The hydrothermal and heavy bass extracts showed no significant difference in brightness (L value), redness (a value), yellowness (b value), and color difference (ΔE value, color difference) values, but enzyme digestion extract In the case of, the lightness and yellowness are slightly brighter and darker than other extracts, and the color difference value of the other extracts is different from that of other extracts.

Hue of pearl shellfish extract Extract hue L a b ΔE Hot water 24.37 ± 0.15 2.26 ± 0.78 1.96 ± 0.18 72.53 ± 0.08 Heavy water 27.79 ± 0.18 2.24 ± 0.11 1.91 ± 0.09 69.11 ± 0.07 Enzyme digestion 44.92 ± 1.45 2.59 ± 1.03 12.62 ± 0.09 56.24 ± 0.10

7) free amino acid

Table 13 shows the results of the analysis of the free amino acid composition of the pearl clam hydrolyzate, basal gland and enzymatic extract with the concentration of the solids Brix 2.0 °. The total amount of free amino acid, which is the most important taste component of nitrogen-containing extract and serves as a taste-expressing ingredient in the flavor expression of seafood food, is 660.6 mg / 100 mL of hot-water treble extract, 470.2 mg / 100 mL of hot-water treble extract, and enzyme digestion extract. Was 1,150.4 mg / 100 mL, which was much higher than other extracts. The major free amino acids of the enzymatic digestion extract were taurine, urea, asparagine, glutamic acid, valine, leucine, lysine and arginine, and other amino acids were generally evenly contained. Compared to the free amino acid composition of general fish and shellfish (Seo, SB et al., 1995. Chemical composition of marine products in Korea.National Fisheries Research and Development Agency Republic of Korea ), the taurine content is relatively low while the urea content is relatively low. It was characterized by much higher points and relatively low content of aspartic acid (aspartic acid), glutamic acid, glycine, alanine. The major free amino acids of hydrothermal and bicuspid extracts were taurine, urea, glutamic acid, glycine, and arginine. Therefore, pearl clam extract showed a difference in the production of free amino acid according to the extraction method, and the difference in the content of free amino acid significantly influenced the strength of flavor, the expression and harmony of taste, and the difference in functional characteristics. Seems mad.

Recently, it is known that protein hydrolyzate inhibits the action of angiotensin-I converting enzyme (ACE) which is one of the causes of blood pressure increase among various bioregulatory functions of food ingredients (Suzuki, T. et al., 1983. Angiotensin -Iconverting enzyme inhibiting activity in foods.Nippon Nogeikagaku Kaishi, 57: 1143-1146), it has been found that this ACE inhibitory activity is somewhat different depending on the composition and type of free amino acid and peptide (Kim, TJ et al., 1996). .. J. Kor Soc Food Sci Nutr , 25:...... 871 ~ 877; Kim, SB et al, 1993. J. Korean Fish Soc, 26: 321 ~ 329). Therefore, the free amino acids and peptides present in the pearl shell extract were expected to have a significant effect on the nutritional and functional properties in addition to the unique taste of the pearl shell extract.

Free amino acid content of pearl clam extract (mg / 100 mL) Free amino acids Extract Hot water Heavy water Enzyme digestion Taurine 82.5 62.6 134.2 Urea 299.5 194.3 138.2 Threonine 20.4 15.5 33.7 Serine 4.5 3.9 40.2 Asparagine 20.9 10.0 151.5 Glutamic acid 65.3 48.0 98.1 Proline 11.4 9.2 20.4 Glycine 41.4 34.4 36.4 Alanine 30.4 25.0 2.1 Valine 6.7 6.3 62.8 Cystine 2.1 2.0 4.1 Methionine 3.8 3.8 39.7 Isoleucine 4.6 3.7 48.3 Leucine 7.8 6.2 71.4 Tyrosine 5.5 3.9 46.3 Phenylalanine 4.0 3.3 48.1 Lysine 11.0 7.9 58.8 Histidine 2.3 1.7 21.5 Arginine 36.6 28.5 94.4 Sum 660.7 470.2 1,150.2

8) Organic Base Component

The results of the analysis of the quaternary ammonium base components, such as trimethylamine oxide (TMAO), trimethylamine (TMA), total creatinine and betaine, of pearl clam hydrolysis, hydrocoating and enzymatic extraction, are shown in Table 14. TMAO, a component involved in the cool sweetness of fresh aquatic products and controlling the osmotic pressure of aquatic organisms, was contained in a small amount of 3.0-3.9 mg / 100 mL. The content of TMA, a causative agent of, was also contained in small amounts of 0.4-0.7 mg / 100 mL. In addition, the total creatinine content, which is involved in the astringent taste of aquatic products, was also contained in a small amount of 1.3 ~ 1.6 mg / 100 mL, and there was little difference in the content according to the extraction method. Quaternary ammonium bases such as TMAO, TMA, and total creatinine play an auxiliary role in the refinement of aquatic products, but in the case of this pearl shell extract, they may play a part in the harmony of taste. The impact was not expected to be significant.

On the other hand, the content of betaine, which is the main component of fresh taste of marine invertebrates such as molluscs, cephalopods and shellfish, was 23.0 ~ 68.3 mg / 100 mL, which was much higher than that of hot water extraction or hot water extract. In comparison with the content of, it is expected that it is a tasteful ingredient which has some influence on the flavor of pearl shell extract.

Organic Base Content of Pearl Clam Extract (mg / 100 mL) Item Extract Hot water Heavy water Enzyme digestion TMAO 3.3 ± 0.1 3.0 ± 0.5 3.9 ± 0.7 TMA 0.5 ± 0.2 0.7 ± 0.4 0.4 ± 0.1 Total creatinine 1.4 ± 0.4 1.3 ± 0.5 1.6 ± 0.2 Betaine 23.0 ± 9.4 32.7 ± 7.0 68.3 ± 11.4

9) Inorganic ion

The results of analysis of the inorganic ions of the pearl clam hydrolyzate, heavy bath and enzymatic extract by ICP are shown in Table 15. The sample extract contains a large amount of Na (87.5 to 135.3 mg / 100 mL), Mg (15.2 to 35.9 mg / 100 mL), and Ca (107.6 to 135.4 mg / 100 mL) as cations, and other Fe, Zn, S, K, P, etc. were also contained in trace amounts, showing some differences in content depending on the extraction method. Compared to other aquatic extracts, the inorganic ion composition of the pearl clam extract was characterized by a high content of Ca ions, but a low content of K and P ions, and a large amount of S ions when extracted by heating such as hot water or hot water. Appeared to be. On the other hand, considering that Na, P, and Cl ions are essential ingredients for various tastes, and that inorganic ions are generally a net expression component of aquatic products along with free amino acids and IMP, Although it is not high in content, it is estimated to contribute to the net expression of pearl shell extract.

Inorganic ion content of pearl shellfish extract (mg / 100 mL) Inorganic ion Extract Hot water Heavy water Enzyme digestion Na 121.1 ± 3.7 131.9 ± 0.7 87.5 ± 2.5 Mg 15.2 ± 2.1 16.6 ± 6.0 35.9 ± 3.1 Ca 125.4 ± 23.5 135.4 ± 32.9 107.6 ± 13.8 Fe 4.1 ± 0.4 3.8 ± 2.4 5.2 ± 0.4 Cu 0.2 ± 0.0 0.2 ± 0.0 0.2 ± 0.0 Zn 3.6 ± 0.0 2.2 ± 1.7 2.4 ± 0.4 S 26.4 ± 1.1 24.5 ± 0.9 14.9 ± 0.2 K 13.5 ± 2.6 16.1 ± 4.2 4.6 ± 1.0 P 16.4 ± 3.1 7.8 ± 0.1 12.8 ± 1.8

10) Sensory Characteristics of Pearl Shell Extract

Hue, smell, taste, and synthesis of pearl clam hydroponic gong, gongmul gong and enzymatic extract

Table 16 shows the results of sensory evaluation on the red symbols. Among the various sensory elements, the hydrophilic and bicuspid extracts were slightly higher than the enzyme digestion extracts in terms of color tone, but there was no significant difference at 95% level. Rather high rating. Overall, the enzymatic digestion was slightly superior in organoleptic extract, followed by hydrothermal and bisonite extract, but there was no sensory difference between them at 95% level.

Sensory Characteristics of Pearl Clam Extract Item ¹ Extract Hot water Heavy water Enzyme digestion color 3.61 ± 0.6 ^a2 3.56 ± 0.5 ^a 3.49 ± 0.7 ^a smell 3.09 ± 0.6 ^a 2.69 ± 0.4 ^b 3.15 ± 0.6 ^a flavor 3.26 ± 0.4 ^a 3.21 ± 0.5 ^a 3.43 ± 0.2 ^b Comprehensive Preference 2.96 ± 0.5 ^a 2.94 ± 0.4 ^a 3.36 ± 0.4 ^b 1) Sensory test personnel shall be 9 (n = 9). 2) 5-step scoring method: 5, very good; 4, good; 3, normal; 2, bad; 1, very bad. 3) The same letter in each column is not significantly different (p <0.05).

Therefore, in order to manufacture an effective extract material that can be used as a natural seasoning material from pearl clam meat, it was judged that the extraction by two-stage enzymatic digestion among various extraction methods would be suitable in terms of sensuality and economics.

Example  4: Pearl shell Extract Improve flavor  Optimal Reactor Quality of Sugar-Amino Acid Reaction

Pearl shellfish extract for reaction flavoring was prepared by using an enzyme digestion extract in which the amount of solids was adjusted to Brix 20 ° in consideration of the results of the above physicochemical experiments and sensory test.

In order to select a substrate of the Mylar reaction solution to improve the flavor of pearl shell enzymatic extract, the precursors such as sugars and amino acids used in general Mylar reactions were searched. First, sensory evaluation of boiled beef odor and sweet odor generated after the myal reaction was performed by changing the combination of each amino acid using 0.1M glucose solution as the base. It was. The amino acids to be added include methionine, cystine, threonine, alanine, lysine, and sulfuric acid, which play an important role in the development of boiled beef flavor, considering the amino acid composition and taste of sample pearl clam extract, and the degree of contribution to the Mylar reaction. Glycine and the like were selected.

As a result of examining the characteristics of the fragrance generated after mixing the glucose solution with a single or multiple glucose solution as described in Table 17, the complex substrate of glucose + glycine + cystine was used to express boiled beef and soft flavor. Because it was closest and its strength was strong, this composite substrate was determined as a reaction precursor to improve the flavor of pearl shell extract.

On the other hand, in order to select a sugar suitable for this myyal reaction, 0.1 M glucose, 0.1 M ribose and 0.1 M xylose solution were mixed with 0.1 M glycine + 0.1 M cystine solution, respectively, to prepare a composite substrate, as shown in Table 17 below. As a result of the myal reaction under the condition, similar aroma was generated and there was almost no difference according to the type of sugar used in the reaction. The cheapest glucose among the three sugars was determined as a precursor of the composite substrate.

Sugar-Amino Acid Fragrance Characteristics Generated During the Mylar Reaction at 120 ° C Sugar-Amino Acid Solution ¹ Reaction time (hrs) 4 8 12 16 Boiled beef flavor A subtle fragrance Boiled beef flavor A subtle fragrance Boiled beef flavor A subtle fragrance Boiled beef flavor A subtle fragrance Methionine + Threonine Methionine + Alanine Methionine + Lysine ++ ² ++ + + Methionine + Cystine + Methionine + Glycine Threonine + alanine + Threonine + lysine + ++ ++ Threonine + cystine + + + ++ + + Threonine + glycine Alanine + Lysine ++ + Alanine + Cystine + + + ++ + Alanine + Glycine Lysine + cystine + ++ + Lysine + Glycine + + Glycine + cystine + + ++ + +++ ++ ++ ++ 1) 0.1 M glucose 10 mL + 0.1 M amino acid 5 mL + 0.1 M amino acid 5 mL 2) Odor intensity code (+: weak, +++: medium, +++++: strong)

Example  5: Pearl shell Extract Improve flavor  for Mayal  Dynamic of reaction Monitoring  Investigation of optimum reaction conditions

Pearl shellfish extract for reaction flavoring was prepared by using an enzyme digestion extract in which the amount of solids was adjusted to Brix 20 ° in consideration of the results of the above physicochemical experiments and sensory test.

To investigate the optimization condition of the mial reaction for improving the flavor of the pearl shell extract, the mial reaction was carried out using a high temperature high pressure reactor (Hake SS-DOT) as shown in FIG. 4. The composition of the reaction mixture of the pearl shell enzymatic digestion extract and the reaction precursor used was as shown in Table 18 below. The PE-Glu-GC (Pearl oyster extract-glucose-glycine-cystine) reaction system was subjected to a yalal reaction under various conditions by varying the independent variables such as reaction time and pH of the reaction solution. Based on sensory evaluation results for the expression of baked potato odor, masking of shellfish odor, and boiled meat odor, these dynamic changes are expressed in three-dimensional response surfaces. Shown in The reaction temperature is a very important independent variable in reaction flavoring, but considering the food hygiene safety, the mial reaction was performed at a fixed temperature of 120 ° C, which is much lower than the reaction limit temperature of 180 ° C. Sensory reaction of PE-Glu-GC (Pearl oyster extract-glucose-glycine-cystine) reaction system at 120 ° C. and then the three dependent variables of the reaction product: the savory flavor, the shielding of the fish and the boiled beef flavor As a result of the evaluation, it was best to adjust the pH of the reaction solution to near neutral in terms of fresh scent and patch shielding, and it was found to be affected by pH more than other reaction variables. In addition, the sensory score was high as the reaction time became longer. The expression of boiled beef flavor was much affected by reaction time and slightly less affected by pH.

Composition of the reaction solution mixed with pearl shell enzymatic digestion extract (Brix 20 °) and reaction precursor Pearl shellfish extract 0.4M glucose 0.4M glycine 0.4M cystine Composition ratio 4 2 One One

The regression equation, R ² value, and significance of the dependent variable according to the change in reaction time and the pH of the reactor mass in the myal reaction of the PE-Glu-GC reaction system are shown in Table 19 below.

Polynomials calculated by RSM for carrying out the Mayal reaction reaction Polynomial R ² valence Sweet incense Y ₁ = 6.732857 + 1.11X ₁ -0.025X ₂ -0.412857X ₁ ² -0.06X ₂ X ₁ -0.197857X ₂ ² 0.9118 0.0311 Apparel Shield Y ₂ = 6.35 + 1.1975X ₁ + 0.019167X ₂ -0.4075X ₁ ² + 0.0425X ₂ X ₁ -0.055X ₂ ² 0.9119 0.0310 Boiled beef flavor Y ₃ = 6.240714 + 0.901667X ₁ -0.01X ₂ -0.076339X ₁ ² -0.55089X ₂ ² 0.9315 0.0191

In the PE-Glu-GC reaction system, the regression equation for the expression of different flavors as the reaction time and pH of the reaction substrate were changed was Y ₁ = 6.732857 + 1.11X ₁ -0.025X ₂ -0.412857X ₁ ² -0.06X ₂ X ₁ -0.197857X ₂ ² , the regression equation for the patched shield is Y ₂ = 6.35 + 1.1975X ₁ + 0.019167X ₂ -0.4075X ₁ ² + 0.0425X ₂ X ₁ -0.055X ₂ ² , _{Y 3 = 6.240714 + 0.901667X 1 -0.01X} 2 -0.076339X 1 2 - ₂ ² was 0.55089X.

The R ₂ values for the sensory characteristics of the three dependent variables in the PE-Glu-GC reaction system were 0.9118 for the mild flavor expression, 0.9119 for the shielding of the fish, and 0.9315 for the boiled beef flavor. In addition, they were recognized for significance at levels below 5%.

The optimum reaction conditions of the expression of three dependent variables, namely fragrance, occlusion and boiled beef flavor, in the myal reaction of the PE-Glu-G-C reaction system are shown in Table 20 below. The best reaction conditions for expressing the fragrance of the mylar reaction product of PE-Glu-GC reaction system were reaction time at 120 ℃ and reaction time of 147.6 minutes, pH 7.54, and the best reaction conditions for patch shielding were reaction time of 149.9 minutes, The best reaction conditions for the expression of pH 6.98 and boiled beef flavor were 149.9 minutes and pH 6.97. Therefore, the reaction conditions for optimization of the flavor flavoring (reaction flavoring) using the PE-Glu-G-C reaction system was set to 140 ~ 150 minutes, pH 6.9 ~ 7.6 at a reaction temperature of 120 ℃.

Optimization of Reaction Conditions for Expression of Flavored, Appetized Fish, and Boiled Beef Flavor from Myal Reaction in PE-Glu-G-C Reaction System Dependent variable Temperature (℃) Time (min.) PH of reaction solution Sensory evaluation maximum ^* Sweet incense 120.0 147.6 7.54 7.97 (max) Apparel Shield 120.0 149.9 6.98 7.34 (max) Boiled beef flavor 120.0 149.9 6.97 7.71 (max) [Note] *: Maximum measured value measured by the 9-step scoring method.

On the other hand, when the reaction temperature is fixed at 120 ° C. in the myal reaction using the PE-Glu-GC reaction system, the overall effect of two reaction factors, namely the reaction time and the pH of the reaction solution on the flavor expression of the myal reaction product The results were as shown in Table 21 below. In the PE-Glu-G-C reaction system, the most important factor affecting the three dependent variables was the reaction time, followed by the pH of the reaction solution, but the pH effect of the reaction solution on the expression of boiled beef flavor was minimal.

Overall effect of reaction time and pH of reaction solution on flavor development of Mylar reaction product Response F-ratio Sweet incense Apparel Shield Boiled beef flavor time 11.30 ¹ 7.73 ² 7.38 ² pH 3.93 2.63 1.04 1) Significance of 1% or less 2) Significance of 5% or less

Optimization of the Mylar Reaction Using the PE-Glu-GC Reaction System To demonstrate the predictive reaction conditions, compare the predictions by regression equations of the dependent variables of the mial reaction under actual conditions with the actual mylar reactions under the same conditions. The results of verifying the reliability of these regression equations are shown in Table 22 below. At this time, the arbitrary conditions were set to the reaction temperature of 120 ℃, time 150 minutes, pH of the reaction solution was adjusted to 7.0. The measured values of each dependent variable were very similar to those predicted by the RSM technique, which could verify the reliability of the regression equation of the reaction flavoring using this Myyal reaction. The flavor of the meat extract could be significantly improved.

Comparison of Predicted and Actual Measures of Dependent Variables in the Optimal Mylar Response Dependent variable Prediction ¹ Found ² Sweet incense 7.48 7.1 ± 0.9 Apparel Shield 7.25 6.9 ± 0.5 Boiled beef flavor 6.70 6.4 ± 0.8 NOTE 1 Calculated using the predictive polynomial for the dependent variable. Critical values of the independent variables: reaction temperature 120 ° C., reaction time 150 minutes, pH 7.0. 2) Average value after 3 measurements (9-step scoring method).

Example  6: Pearl shell Extract  Preparation of Natural Seasoning Using

Pearl shell meat extract with improved flavor under optimum conditions was prepared based on the results of the above examples. That is, first, sample pearl clam meat was sliced with a chopper, and twice the amount of water was added to the sample weight, and the autodigestive enzyme was inactivated by ripening at 95 ° C. for 5 minutes. Then, the pH of the sample solution was adjusted, and Alcalase 0.6L enzyme (Novo Nordisk Co., Denmark) was added thereto, followed by primary enzymatic degradation at 55 ° C. for 4 hours with stirring. After heat treatment at 95 ° C. for 5 minutes to deactivate the alcalase, the pH was further adjusted, and Flavorzyme (Novo Nordisk Co., Denmark) was added thereto, followed by hydrolysis for 4 hours with stirring at 45 ° C. After heat treatment at 95 ℃ for 5 minutes to inactivate the flavozyme. Next, the residue was separated by using a filter cloth, allowed to cool, and then centrifuged to remove the residue, thereby preparing an enzyme digested pearl clam meat extract adjusted to Brix 20 °. The enzyme-degraded pearl clam meat extract: 0.4M glucose: 0.4M glycine: 0.4M by mixing the cystine 4: 2: 1: 1 to maintain a pH of 6.97 sugar-amino acid reaction for 149.9 minutes pearl shellfish improved flavor An extract was prepared.

Pearl clam extract (liquid, Brix 30 °) with improved flavor by reacting the enzymatically digested pearl clam meat extract prepared as above by spray drying to powdered pearl clam extract powder and sugar-amino acid The pearlescent granule seasoning was prepared according to the process shown in FIG. 11, and the composition and quality characteristics of the prototype were examined.

That is, first, IMP, yeast extract, kelp extract, MSG, crystalline cellulose and lactose, seasoning base powder (seasoning base powder) mixed with salt, pearl clam extract powder and lactose in a composition ratio as shown in Table 23 below Powdered materials were prepared. Next, a ribbon blender was added to the liquid flavors, such as pearly clam extract (Brix 30˚), oyster sauce (Bryx 30˚), brewed soy sauce and purified water, which improved flavor through reaction flavoring. Mixed as. The various ingredients used here were purchased from MSC Co., Ltd. and nearby markets, and were added with oyster cooked concentrate (A-class, Brix 30) to reinforce natural delicacies and to reduce the amount of MSG added to 1/5 or less than commercially available similar products. °) was purchased from Samdeok C & T Corporation and used for the experiment. The mixing ratios of the various ingredients used in this experiment were shown in Table 23.

Composition ratio of natural seasoning of the present invention using pearl clam extract Raw material properties ingredient Content (% by weight) Powder Seasoned base powder IMP 0.1 Yeast extract 1.4 Kelp Extract 0.6 MSG 5.0 Crystalline cellulose 0.6 Lactose 13.0 saline 33.3 Pearl shellfish extract powder 22.0 Lactose 14.9 Liquid Pearl shell x minutes (Brix 30˚) 5.0 Oyster Cooking Concentrate (Brix 30˚) 2.0 Brewed Soy Sauce 0.4 Purified water 1.7

The above raw materials were mixed with a ribbon mixer, molded and granulated by an extruder, dried at 60 to 65 ° C. using a fluidized bed dryer, and then cooled and sorted to obtain a pearl shell granulated seasoning. In addition, the prototype was sealed in a glass bottle and used as an analytical sample while being stored at 4 ± 1 ° C.

Example  7: Investigation of the effect of adding the oyster cooked concentrate to the natural seasoning of the present invention

General ingredients of grade A oyster cooked concentrate added to significantly reduce the amount of biosynthetic seasoning (MSG) added to existing complex seasoning foods and to reinforce the natural delicacy lacking in pearl shellfish extract. , Volatile basic nitrogen, salinity and pH are shown in Table 24, and the results of the analysis of the composition of the free amino acid, the main net expression component of the oyster cooked concentrate by the amino acid automatic analyzer, are shown in Table 25. Oyster cooked concentrate is a seasoning material widely used for reinforcing delicacy in delicacies in Japan, China, etc., water content was 63.2%, crude protein content was 9.0%, and volatile basic nitrogen was 60.2 mg / 100 g. Salinity was 9.6% and pH was 5.20. On the other hand, the free amino acid composition of the oyster cooked concentrate was 5,357.9 mg / 100 mL, taurine (1,130.9 mg / 100 mL), urea (1,438.9 mg / 100 mL), glutamic acid (403.2 mg / 100 mL), proline ( 729.1 mg / 100 mL), glycine (531.0 mg / 100 mL) and alanine (473.3 mg / 100 mL) were the major free amino acids, and β-alanine was relatively high. Therefore, it is thought that the addition of this oyster porridge concentrate can enhance the flavor of pearl shell granulated seasoning without adding a large amount of MSG by reinforcing a variety of rice components which are easily lacked in pearl shell meal.

General Components, Volatile Nitrogen, Salinity, and pH of Oyster Cooking Concentrate (g / 100 g) moisture Crude protein View minutes VBN (mg / 100 g) Salinity pH content 63.2 ± 2.6 9.0 ± 0.1 11.8 ± 0.3 60.2 ± 1.4 9.6 5.20

Free Amino Acid Composition of Oyster Cooking Concentrate (mg / 100 mL) amino acid content Phosphoserine 4.3 Taurine 1,130.9 Urea 1,438.9 Aspartic acid 65.7 Threonine 61.0 Serine 59.0 Asparagine 84.2 Glutamic acid 403.2 Sarcosine 12.0 Proline 729.1 Glycine 531.0 Alanine 473.3 α-aminoiso-n-butyric acid 5.9 Valine 20.5 Cystine 3.8 Methionine 25.9 Cystathionine 3.0 Isoleucine 10.3 Leucine 23.0 Tyrosine 15.4 β-Alanine 105.7 Phenylalanine 11.0 D, L-β-aminoisobutyric acid 7.2 γ-amino-n-butyric acid 9.3 Ethanolamine 4.6 Ornithine 21.2 Lysine 22.1 Histidine 33.1 3-Methyl-L-histidine tr Arginine 43.3 Sum 5,357.9

Example  8: Investigation of the quality characteristics of the natural seasoning of the present invention

1) General Ingredient

Pearl clam granule natural seasoning prepared in Example 6 at the compounding ratio as shown in Table 23 was the same as FIG. The present invention pearl clam granular natural seasonings and commercially available three kinds of similar complex seasoning, namely C-1 (granular clam extract powder complex seasoning, C company), C-2 (powdered beef powder extract seasoning, Company C) and C-3 (dry mussel powder complex seasoning, company D) were purchased from a nearby market and the results of the measurement of the general ingredients, volatile nitrogen, salinity and pH were as shown in Table 26 below. The moisture content of pearl shell granule seasoning was 3.8%, crude protein 12.0%, and crude ash 33.6%, similar to those of commercial products, but the contents of crude ash were slightly lower than those of commercial products.

Comparison of general ingredients, volatile basic nitrogen, salinity and pH of natural seasonings and commercial products of the present invention (g / 100g) Sample ^* moisture Crude protein View minutes Geography GPS 3.8 ± 0.2 12.0 ± 0.2 33.6 ± 0.1 0.3 ± 0.1 C-1 3.2 ± 0.3 11.1 ± 0.2 42.2 ± 0.2 0.3 ± 0.1 C-2 4.6 ± 0.2 9.8 ± 0.1 37.0 ± 0.1 1.5 ± 0.2 C-3 2.8 ± 0.2 11.6 ± 0.4 44.6 ± 0.2 1.0 ± 0.2 [GPS] * GPS: Granulated pearl clam extract seasoning C: Commercial seasoning C-1: Granular clam x-min compound seasoning food (C company) C-2: Powdered beef X-min compound seasoning food (C company) C-3: Powdered Dried Mussel Powder Seasoned Food (Company D)

2) pH, salinity, TBA value, amino nitrogen and volatile nitrogen content

The pH, salinity, TBA value, amino nitrogen and volatile basic nitrogen contents of three pearl seasoned granulated seasonings and three similar commercial seasonings were shown in Table 27. The pH of this product was 6.49, which was somewhat higher than that of commercial seasoned foods, 5.79 ~ 6.02, and the salinity was 31.4%, which was slightly lower than 33.6 ~ 40.4% of similar commercial seasoned foods.

The TBA value of lipid dispersity is 0.032 OD (531 nm), which is very low compared to 0.131 to 0.204 OD (531 nm) of commercial seasonings. It seems to be very small. On the other hand, the amino nitrogen content, which can indirectly know the content of amino acid, which is the main taste component, is 1,876.6 mg / 100 g. Was showing. Considering that the commercial complex seasoning product contains a large amount of MSG, most of the nitrogen-containing compounds contained in this product were natural free amino acids, which were judged to be good quality natural flavor materials.

The volatile basic nitrogen (VBN) content of the sample was 12.6 mg / 100 g, which was slightly higher than the commercial complex seasoned foods, from 6.9 to 10.3 mg / 100 g. It is relatively high compared to, so it can be said that it is irrelevant to freshness.

Quality Characteristics of Granulated Pearl Clam Extract Seasoning sample* pH Salinity (%) TBA (O.D. 531 nm) NH ₂ -N (mg / 100g) VBN (mg / 100g) GPS 6.49 31.4 ± 0.3 0.032 ± 0.010 1,817.6 ± 10.4 12.6 ± 0.21 C-1 6.02 39.0 ± 0.4 0.204 ± 0.009 1,491.2 ± 11.2 8.1 ± 0.14 C-2 5.79 33.6 ± 0.2 0.131 ± 0.010 842.8 ± 15.4 10.3 ± 0.30 C-3 6.02 40.4 ± 0.1 0.163 ± 0.012 1,610.2 ± 43.5 6.9 ± 0.33 [GPS] * GPS: Granulated pearl clam extract seasoning C: Commercial seasoning C-1: Granular clam x-min compound seasoning food (C company) C-2: Powdered beef X-min compound seasoning food (C company) C-3: Powdered Dried Mussel Powder Seasoned Food (Company D)

3) tint

The surface tones of three pearl clam granulant seasonings and three commercially available complex seasonings were measured with a direct color difference meter. The prototype had a lightness (L value) of 83.2, redness (a value) of 1.8, yellowness (b value) of 12.9, and ΔE value (color difference) of 18.45, similar to commercial granular clam extract powder seasoning C-1. , Compared with the other two species, the color was significantly brighter.

Hues of Granulated Pearl Clam Extract Seasoning sample* hue L a b ΔE GPS 83.2 ± 0.3 1.8 ± 0.0 12.9 ± 0.2 18.5 ± 0.6 C-1 84.5 ± 0.2 1.0 ± 0.1 11.8 ± 0.1 16.6 ± 0.5 C-2 75.0 ± 0.1 4.0 ± 0.0 19.5 ± 0.4 29.1 ± 0.3 C-3 63.9 ± 0.4 10.3 ± 0.2 23.9 ± 0.2 41.6 ± 0.5 [GPS] * GPS: Granulated pearl clam extract seasoning C: Commercial seasoning C-1: Granular clam x-min compound seasoning food (C company) C-2: Powdered beef X-min compound seasoning food (C company) C-3: Powdered Dried Mussel Powder Seasoned Food (Company D)

4) nucleotides

Table 29 shows the results of the analysis of the content of nucleotides, which is a representative taste component of C-1, the present product and granule clam extract powder combination seasoning. Six nucleotides of IMP, ATP, ADP, AMP, inosine and hypoxanthine were detected as nucleotides of the prototype, and four kinds of IMP, AMP, inosine and hypoxanthine were detected in the commercial product C-1. Among the detected nucleotides, IMP and AMP content of net taste was 88.6 mg / 100 g and 46.1 mg / 100 g, respectively, which were somewhat higher than those of commercial product C-1. Among the nucleotides, AMP and IMP, etc., are contained in fresh fish products as ATP degradation products, and have been reported to be important net expression components (Fuke, S. and Konosu, S. 1991. Taste-active compounds in some food. Physiol Behavior , 49: 863-867).

Nucleotide Content (mg / 100g) of Pearl Shellfish Extract Granular Natural Seasoning Nucleotide sample* GPS C-1 IMP 88.6 ± 0.4 59.7 ± 0.3 ATP 16.2 ± 0.9 - ADP 21.5 ± 0.5 - AMP 46.1 ± 0.4 15.1 ± 0.4 Inosine 15.5 ± 0.6 12.2 ± 0.1 Hypoxanthine 38.5 ± 0.4 66.7 ± 0.2 [GPS] * GPS: Granulated pearl clam extract seasoning C: Commercial seasoning C-1: Granular clam extract Xin complex seasoning (C company)

Considering the taste and content of these nucleotides, IMP and AMP, together with free amino acids, inorganic ions and betaines, are expected to contribute to the net expression and harmony of the prototype.

5) free amino acid

Table 30 shows the results of analysis of the contents of free amino acid, which is a representative taste component of C-1, a commercial granular clam extract powder combination seasoning product. The total content of free amino acid, which is the most important taste component of nitrogen-containing extract, which acts as taste-active components in the flavor expression of seafood, is 1,127.5 mg / 100 g, which is considerably higher than 801.3 mg / 100 g of commercial product C-1. It contained a lot. The major free amino acid of pearl shell granule seasoning was glutamic acid (313.5 mg / 100 g), followed by taurine, hydroxyproline, glycine, phosphoserine, alanine, proline, phosphoethanolamine, and urea. This contained relatively evenly. In contrast, glutamic acid accounted for more than 60% of total glutamic acid at 507.4 mg / 100 g, which was presumed to be due to the addition of large amounts of MSG for reinforcement of rice. The content of amino acids was insignificant. The difference in the composition of the free amino acids may be strong in the flavour, although the commercial product C-1 may be strong.

Free amino acid (mg / 100g) of pearl shellfish extract natural seasoning amino acid sample* GPS C-1 Phosphoserine 70.5 55.8 Taurine 181.8 28.3 Phosphoethanolamine 39.0 21.1 Urea 31.4 11.5 Hydroxyproline 90.6 65.7 Threonine 7.9 4.0 Serine 8.7 4.7 Glutamic acid 313.5 507.4 α-aminoadipic acid 15.2 - Proline 47.4 6.2 Glycine 72.2 13.3 Alanine 62.0 11.9 Citrulline 1.5 - α-aminoisobutyric acid 2.4 - Valine 8.9 4.1 Cystine 15.2 11.5 Methionine 4.7 2.1 Isoleucine 27.9 3.9 Leucine 45.7 7.4 Tyrosine 4.6 - β-alanine 9.9 2.2 Phenylalanine 6.7 3.6 Homocystathionine 2.5 - γ-aminobutyric acid 9.2 4.4 ammonia 12.2 24.9 Lysine 25.2 2.3 1-methyl-histidine 1.0 - Histidine 1.8 1.4 3-methyl-histidine 1.8 - Arginine 6.1 3.6 Sum 1,127.5 801.3 [GPS] * GPS: Granulated pearl clam extract seasoning C: Commercial seasoning C-1: Granular clam extract Xin complex seasoning (C company)

6) sensual quality

In order to compare and evaluate the sensory quality of this product and commercial complex seasonings C-2 and C-3, based on the commercial product C-1 (Sensory Rating 3.0), nine panels were mastered to familiarize themselves with the sensory characteristics of these materials. Table 31 shows the results of sensory evaluation in terms of taste, aroma, color and comprehensive evaluation. As shown in Table 31, this prototype received a much higher rating than the three commercially available products in terms of taste, aroma and comprehensive evaluation, and there was a significant difference at 95% level. Therefore, this granular seasoning material using the reaction flavoring pearl clam extract has no deterioration in sensory quality compared to similar seasoning materials on the market, and it is a natural seasoning material or instant soup with various flavors. It was concluded that the material can be sufficiently used.

Sensory evaluation results Sample ¹ division flavor incense color Overall preference GPS 4.3 ± 0.6 ^a2 4.1 ± 0.7 ^a 2.8 ± 0.3 ^a 4.2 ± 0.6 ^a C-1 3.0 ± 0.0 ^b 3.0 ± 0.0 ^b 3.0 ± 0.0 ^a 3.0 ± 0.0 ^b C-2 3.0 ± 0.4 ^b 3.3 ± 0.3 ^b 2.6 ± 0.3 ^b 2.9 ± 0.2 ^b C-3 2.0 ± 0.3 ^c 1.5 ± 0.6 ^c 2.2 ± 0.4 ^b 1.8 ± 0.5 ^c [Note] ① ¹ GPS: Granulated pearl clam extract seasoning C: Commercial seasoning C-1: Granular clam x-min complex seasoning food (C company) C-2: Powdered beef X-min compound seasoning food (C company ) C-3: Powdered dry mussel powder complex seasoning (Company D) ② ² 5 grade rating method: 5, very good; 4, good; 3, normal; 2, bad; 1, very bad. ③ The same letter in each column is not significantly different (n = 9, p <0.05).

As described above, the present invention relates to a method of preparing natural seasonings using pearl clam extract with improved flavor, and investigated the components of pearl clam meat and investigated the components and sensory characteristics according to the extraction method of pearl clam meat. Next, the most optimally selected pearlescent clam enzymatic digestion extracts were selected for the optimal reactor quality of sugar-amino acid reaction for flavor improvement, and the optimal reaction conditions through dynamic monitoring of the myal reactions were not found. Extracts with improved flavors are prepared from pearl shellfish, which are mostly discarded. By using them, the amount of MSG, a chemical seasoning, is significantly reduced, adding the unique flavor and aroma of shellfish, and the functional ingredients contained in pearl shellfish. Food industry because it has an excellent effect to provide natural seasoning It will be very useful inventions.

Claims (2)

Pearl clam extract (Brix 20 °): 0.4M Glucose: 0.4M Glycine: 0.4M Mix cystine at 4: 2: 1: 1 and maintain pH at 6.9-7.6 for 140-150 minutes at 120-150 ℃ Natural seasoning composition comprising pearl shell meat extract prepared by the reaction as an active ingredient. The natural seasoning composition according to claim 1, wherein the natural seasoning composition further comprises a oyster ripening concentrate.

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