CN111887417A

CN111887417A - Method for preparing black bean-flavor rib-flavor spice

Info

Publication number: CN111887417A
Application number: CN202010707051.3A
Authority: CN
Inventors: 汪薇; 任文彬; 白卫东; 曾晓房; 刘功良; 刘旋斌; 杜婉怡
Original assignee: Zhongkai University of Agriculture and Engineering
Current assignee: Zhongkai University of Agriculture and Engineering
Priority date: 2020-07-21
Filing date: 2020-07-21
Publication date: 2020-11-06

Abstract

The invention provides a preparation method of black bean-flavor rib-flavor spice, which comprises the following steps: s1: enzymolysis: grinding pork, adding water, stirring, mixing, adjusting the pH value to 6.0-8.0, adding protease for enzymolysis, and obtaining an enzymolysis solution of the pork; s2: maillard reaction: adding a Maillard reaction substrate into the enzymatic hydrolysate in the step (1), and reacting for 30-90 min; s3: blending: adding salt, ginger powder, onion powder, anise, 5' -flavour nucleotide disodium and soy sauce into the material prepared in the step (2) for blending to obtain the black bean-flavor rib flavor spice. The invention provides a preparation method, and solves the problems of weak overall aroma intensity, insufficient and obvious characteristic aroma and insufficient mouthfeel of the conventional seasoned soy bean spare rib flavor. The preparation method combines protein enzymolysis and Maillard reaction, so that the protein is fully hydrolyzed, the content of free amino acid in the base material is improved, and the content of fragrance precursor substances produced in the Maillard reaction is increased.

Description

Method for preparing black bean-flavor rib-flavor spice

Technical Field

The invention belongs to the technical field of food processing, and particularly relates to a method for preparing a black bean-flavor rib flavor spice.

Background

Meat flavor has been developed into an important component of food flavor in the last forty years, and the production capacity and sales volume of the meat flavor are gradually increased, so that the development of the food industry is promoted. Meanwhile, the expectation of broad consumers on obtaining the meat flavor with strong, vivid, seasoning and nutrition is also higher and higher.

At present, the pork essence is commonly used in the aspects of cooked meat products, leisure food, fast food, sauce marinated products and the like. However, the public often has certain contradiction psychology to the essence and spice synthesized by chemistry, and people are afraid of the fact that the essence and spice is harmful to human bodies and wants to obtain certain pork characteristic flavor. The essence and flavor prepared by the Maillard reaction are popular, and the reason is that the method is simple and easy and has higher safety. Since the development of the modern industry, the maturation process of the method is rapid, and the production of meat flavor and fragrance by using the Maillard reaction is an indispensable way in the industry.

The enzyme is used as a biocatalyst with high specificity and high efficiency, the catalytic reaction conditions are mild, and the enzymatic hydrolysis of protein is different from the acid hydrolysis and the alkaline hydrolysis of protein, and the enzyme does not destroy amino acid. The enzymolysis protein can not generate racemization phenomenon, and can not damage amino acid, and the obtained product has stable physicochemical property. The pork substrate is subjected to enzymolysis before the Maillard reaction, so that high molecular substances in the pork substrate can be effectively decomposed, favorable substrates are improved for the Maillard reaction, and the flavor of the pork substrate is enhanced.

Meat essence is one of salty essences with the most extensive application in the market, and the defect of the characteristic flavor is always a difficult problem in essence research. At present, a lot of manufacturers mainly use various reducing sugars and amino acid reactions as the basis to prepare spices, and the produced meat essence has the problems of weak overall aroma intensity, insufficient obvious characteristic aroma, unsaturated mouthfeel and short aroma retention time.

Disclosure of Invention

The invention aims to provide a preparation method of a black bean-flavored sparerib-flavored spice, which comprises the following steps:

s1: enzymolysis: grinding pork, adding water, stirring, mixing, adjusting the pH value to 6.0-8.0, adding protease for enzymolysis, and obtaining an enzymolysis solution of the pork;

s2: maillard reaction: adding a Maillard reaction substrate into the enzymatic hydrolysate in the step (1), and reacting for 30-90 min.

S3: blending: adding salt, ginger powder, onion powder, anise, 5' -flavour nucleotide disodium and soy sauce into the material prepared in the step (2) for blending.

Preferably, the protease is selected from one or more of the following enzyme classes: papain, compound protease, hydrolytic protease, flavourzyme and compound protease. More preferably, the protease is a mixed enzyme of papain and flavourzyme. The invention screens out the enzyme suitable for pork enzymolysis through a large number of experiments, then realizes the best enzymolysis effect through pairwise compounding, and finally screens out the papain and the flavourzyme to carry out compounding to obtain the best enzymolysis effect.

Preferably, the addition amount of the protease is 1-3% of the mass of the pork; more preferably, the protease is added in an amount of 2% by mass of the pork meat. In the enzymolysis process, the addition amount of the enzyme is crucial to the enzymolysis reaction, so that the addition amount of the enzyme is optimized and screened by the inventor, in the subsequent embodiment, the addition amount of the enzyme is 1-3%, the pork can be degraded more fully, and especially, the effect of adding 2% of protease is optimal.

Preferably, the enzymolysis temperature is 40-65 ℃. More preferably, the enzymatic hydrolysis temperature is 55 ℃. Because the enzyme is a protein and is sensitive to temperature, the temperature of the enzymolysis reaction is optimized by the inventor, and the papain and the flavourzyme can better play a degradation role within the range of 40-65 ℃, particularly the best enzymolysis effect is achieved at 55 ℃.

Preferably, the enzymolysis time is 1-4 h; more preferably, the enzymolysis time is 2.5 h. The enzymolysis reaction time is also a relatively important parameter, so that the reaction time is optimized by the inventor, and in the subsequent embodiment, when the enzymolysis time is 1-4 hours, the papain and the flavourzyme can better perform the degradation effect, and particularly, the optimum effect is achieved when the enzymolysis time is 2.5 hours.

Preferably, the enzymatic hydrolysis pH is 7.0. The enzyme is also a protein, and the pH value influences the function of the enzyme, so the inventor optimizes the enzymolysis pH value and finds that the papain and the flavourzyme can better perform the degradation effect when the pH value is 7.0.

Preferably, the Maillard reaction substrate comprises xylose, glucose, L-cysteine, L-proline, DL-methionine, glycine; more preferably, the Maillard reaction substrate comprises xylose, glucose, L-cysteine, glycine. In the Maillard reaction, the type of amino acid can influence the reaction, thereby influencing the fragrance of the spice, therefore, the inventor screens the type of the amino acid, and finds that the spice obtained by compounding L-cysteine and glycine has better fragrance.

Preferably, the addition amount of the xylose is 2-4% of the mass of the pork; more preferably, the addition amount of the xylose is 2.5 percent of the mass of the pork. Xylose can influence the Maillard reaction activity, so the inventor optimizes the addition amount of xylose, and finds that the reaction activity is higher when the addition amount of xylose is 2-4% of the pork mass, and the activity is highest particularly when the addition amount of xylose is 2.5%.

Preferably, the mass ratio of the xylose to the glucose is 1-3: 1-3; more preferably, the mass ratio of xylose to glucose is 2: 1. the compounding ratio of xylose to glucose can influence the whole system of the Maillard reaction, so that the mixture ratio of xylose to glucose is optimized by the inventor, and the mass ratio of xylose to glucose is found to be 1-3: 1-3, the flavor after Maillard reaction is better, especially the mass ratio of xylose to glucose is 2: at 1, the highest score was obtained for the perfume.

Preferably, the addition amount of the L-cysteine is 0.1-0.4% of the mass of the pork; more preferably, the addition amount of the L-cysteine is 0.2 percent of the mass of the pork. L-cysteine plays a leading role in the synthesis of some important meat flavor substances in the Maillard reaction, and a proper amount of cysteine can generate soft meat flavor. Therefore, the inventor optimizes the addition amount of the L-cysteine, and finds that the obtained flavor has a high score when the addition amount of the L-cysteine is 0.1-0.4% of the pork quality, and particularly the score is the highest when the addition amount of the L-cysteine is 0.2%.

Preferably, the Maillard reaction time is 30-90 min; more preferably, the maillard reaction time is 60 min. The reaction time is an important factor influencing the Maillard reaction, so that the Maillard reaction time is optimized by the inventor, the score of the perfume obtained within 30-90 min is found to be higher, and particularly the score is the highest when the reaction time is 60 min.

Preferably, the Maillard reaction temperature is 90-110 ℃; more preferably, the maillard reaction temperature is 100 ℃. The Maillard reaction temperature greatly influences the reaction route, so that the inventor optimizes the Maillard reaction temperature, and finds that the obtained perfume has higher score when the temperature is 90-110 ℃, and particularly has the highest score when the temperature is 100 ℃.

Preferably, the Maillard reaction substrate further comprises VB1, vegetable protein hydrolysate and yeast extract.

The invention has the beneficial effects that: the invention provides a preparation method of a black bean-flavored spare rib flavor, and solves the problems of weak overall aroma intensity, insufficient and obvious characteristic aroma, unsaturated taste and short aroma retention time of the conventional black bean-flavored spare rib flavor. The preparation method combines the protein enzymolysis technology with the Maillard reaction, so that the protein is fully hydrolyzed, the content of free amino acid in the base material is improved, and the content of fragrance-producing precursor substances in the Maillard reaction is increased. Meanwhile, the reaction time can be greatly shortened, the scorched flavor phenomenon caused by long-time high-temperature reaction is avoided, the nutritive value and the flavor of the product are improved, and the taste is more full.

Detailed Description

In order to show technical solutions, purposes and advantages of the present invention more concisely and clearly, the technical solutions of the present invention are described in detail below with reference to specific embodiments.

Experimental materials, reagents and instruments

Shanghai bright ham pork, soy sauce, onion powder, star anise powder, ginger powder and salt are purchased from Tianmao supermarket.

TABLE 1 test materials and reagents

Sodium hydroxide, ethanol, potassium hydrogen phthalate, phenolphthalein indicator and the like are all in analytical purity on the market.

Main experimental equipment

TABLE 2 Main test Equipment

Experimental methods

1. Pork enzymolysis method

Cleaning fresh pork hind leg meat, mincing, treating with boiling water for 5min, weighing the treated pork, adding water according to a certain water-meat ratio to prepare a mixed solution, adjusting pH, adding different protease combinations, performing enzymolysis under specific conditions, inactivating enzyme at 90 ℃, and taking an enzymolysis solution for later use after 15 min.

2. Maillard reaction method

Adding appropriate amount of xylose, glucose, L-cysteine, glycine, VB1, vegetable protein hydrolysate, yeast extract, salt, rhizoma Zingiberis recens powder, Bulbus Allii Cepae powder, fructus Anisi Stellati, 5' -flavor nucleotide disodium, and soy sauce into the enzymolysis solution, performing Maillard reaction under specific conditions, cooling to 35-45 deg.C, and performing sensory evaluation on the final product.

3. Measurement of free amino acid Nitrogen

The measurement was carried out by the first acidimetric method for amino acid nitrogen in GB5009.235-2016 foods [12 ].

4. Sensory evaluation

In order to make the experimentally prepared black bean-flavor spare rib essence more consistent with the target, a sensory evaluation group (10 persons) is established, products with the optimal enzymolysis conditions and Maillard reaction conditions are respectively selected from the single-factor experiment, and the products are subjected to sensory evaluation strictly according to the aroma grading standard.

Table 3 shows the reference standard for grading, which is mainly used for grading from three aspects of spare rib flavor intensity, fermented soybean flavor intensity and flavor harmony, wherein the sum of the three is the result of grading, and the score is 10.

TABLE 3 Scent criteria for fragrance

Example 1 Effect of the type of protease on the fragrance-producing Effect

Respectively adding 10g of minced and heat-treated pork serving as a substrate into 5 triangular bottles with the volume ratio of 1:1, and adjusting the pH to 7.0. 2% of papain and compound protease, papain and hydrolase, papain and flavourzyme, flavourzyme and hydrolase, flavourzyme and compound protease are respectively added into 5 triangular flasks, the triangular flasks are placed in a shaking incubator (55 ℃, 120r/min), and after reaction for 2 hours, the triangular flasks are placed in a constant-temperature water bath kettle (90 ℃, 15min) for enzyme deactivation, and enzymatic hydrolysate is obtained. Adding Maillard reaction substrates, namely 3% of xylose, 1.5% of glucose, 0.5% of VB1, 0.1% of L-cysteine, 0.5% of glycine, 0.5% of plant protein hydrolysate, 0.5% of yeast extract, 4% of salt, 2.5% of ginger powder, 1% of onion powder, 0.1% of anise, 0.2% of 5' -flavor nucleotide disodium and 3.5% of soy sauce into the enzymatic hydrolysate, reacting for 1h at 100 ℃, and cooling to 35-45 ℃ to be tested.

As shown in the following table, the types and amounts of polypeptides and amino acids obtained by the enzymatic hydrolysis of the same substrate by different proteases differ from each other because the action sites of the proteases differ from each other. Therefore, the selection of the protease has an important influence on the enzymatic products, thereby greatly influencing the formation of the aroma after the Maillard reaction. Meanwhile, the protease compound ratio is good in single protease enzymolysis effect, the hydrolysis degree is higher, enzyme cutting sites are increased after compounding, pork enzymolysis is facilitated, the hydrolysis degree of pork and the types of amino acids are influenced by compounding different proteases, and the flavor of the essence is greatly influenced. Therefore, the inventors screened the proteases in table 4 by a large number of tests to obtain better enzymolysis effect.

TABLE 4 influence of the type of protease on the fragrance-producing Effect

As can be seen from Table 4, when papain and flavourzyme are compounded, the content of the obtained free amino acid nitrogen is the highest and is 0.46g/mL, and the hydrolysis effect of pork is the best. The amino compounds are mainly derived from enzymolysis liquid, and the content of free amino acid can obviously and directly influence the aroma production effect of the Maillard reaction. Experiments show that the final product of the enzymolysis liquid obtained by compounding papain and flavourzyme through Maillard reaction has the highest fragrance score of 8.9, and at the moment, the pork ribs are rich in flavor, the fermented soybeans are prominent in flavor, the harmony is strong, and no peculiar smell exists.

The papain and the hydrolase are compounded, and the content of the obtained free amino acid nitrogen is the lowest and is 0.28 g/mL. It is possible that the degree of enzymatic hydrolysis is limited because both papain and hydrolase are endonucleases. And the final product obtained by taking the pork rib as the substrate and carrying out Maillard reaction has the lowest fragrance score of 4.5 points, and the mellow fragrance and the fermented soybean fragrance of the pork ribs are not outstanding and have poor harmony.

The method is characterized in that papain and compound protease compounding, flavor protease and compound protease compounding and flavor protease and hydrolysis protease compounding are used, the content of free amino acid nitrogen is sequentially reduced, the obtained free amino acid nitrogen serves as the fragrance score of a final product of a substrate after Maillard reaction, the papain and compound protease compounding is 7.4 minutes, the flavor protease and compound protease compounding is 8.4 minutes, and the flavor protease and hydrolysis protease compounding is 7.0 minutes.

In conclusion, the best scheme of the invention is to use papain and flavourzyme to carry out enzymolysis on pork by compounding.

EXAMPLE 2 Effect of protease addition amount on fragrance-producing Effect

Respectively adding 10g of minced and heat-treated pork serving as a substrate into 5 triangular bottles with the volume ratio of 1:1, and adjusting the pH to 7.0. Adding papain and flavourzyme accounting for 1%, 1.5%, 2%, 2.5% and 3% of the pork mass into 5 triangular bottles respectively, placing the bottles in a shaking incubator (55 ℃, 120r/min), reacting for 2h, and then placing the triangular bottles in a constant-temperature water bath kettle (90 ℃, 15min) for enzyme deactivation. Adding Maillard reaction substrates, namely 3% of xylose, 1.5% of glucose, 0.5% of VB1, 0.1% of L-cysteine, 0.5% of glycine, 0.5% of plant protein hydrolysate, 0.5% of yeast extract, 4% of salt, 2.5% of ginger powder, 1% of onion powder, 0.1% of anise, 0.2% of 5' -flavor nucleotide disodium and 3.5% of soy sauce into the enzymatic hydrolysate, reacting for 1h at 100 ℃, and cooling to 35-45 ℃ to be tested.

The amount of protease added has an effect on the flavour of the final base. If less enzyme is added during the preparation, the generated flavor substances are relatively less, and the flavor of the produced black bean-flavor spareribs is relatively light; however, if too much enzyme is added during the preparation process, a large amount of flavor substances are generated, or pork is excessively hydrolyzed, or other undesirable phenomena occur, resulting in an undesirable flavor, and therefore, the amount of protease to be added is desirably controlled.

TABLE 5 influence of the amount of protease added on the fragrance-producing effect

As can be seen from Table 5, as the total enzyme amount increases, the content of free amino acid nitrogen is continuously increased, the enzyme reaction is more and more thorough, and the aroma score is changed. When the addition amount of the protease is less than 2% of the pork mass, the content of free amino acid nitrogen is rapidly increased along with the increase of the addition amount of the protease. When the addition amount of the protease is more than 2 percent of the pork mass, the content of the free amino acid nitrogen is slowly increased along with the increase of the addition amount of the protease. When the addition amount of the protease is equal to 2% of the pork quality, the fragrance score of the Maillard reaction product prepared from the enzymatic hydrolysate is the highest and is 8.8, and the spare rib flavor and the fermented soybean flavor are strong and have good harmony. Meanwhile, the experimental result shows that when the content of free amino acid nitrogen in the enzymolysis liquid is 0.46-0.49 g/mL, the aroma production effect ratio is better than that of the level below or above.

The reason for the phenomenon is presumably that the enzymolysis effect is stronger and stronger along with the increase of the total enzyme amount, the enzyme is more fully contacted with the substrate, various characteristic flavor substances are generated, and the flavor of the black bean-flavor spare rib base material is strong; and when the protease in the enzymolysis liquid reaches a certain amount, the concentration of the substrate reaches saturation, the enzymolysis rate changes slowly along with the increase of the enzyme adding amount, although the content of free amino acid nitrogen still increases in a small range, the aroma score is reduced, and the reason for the over-hydrolysis of pork is considered.

In conclusion, the addition amount of the protease which is the most suitable for the invention is 2.0 percent of the pork mass.

EXAMPLE 3 Effect of enzymatic hydrolysis temperature on fragrance production

Respectively adding 10g of minced and heat-treated pork serving as a substrate into 6 triangular flasks with the volume ratio of 1:1, and adjusting the pH to 7.0. Adding 2% of papain and flavourzyme into 6 triangular bottles respectively, placing in a shaking incubator (120r/min), reacting at 40 deg.C, 45 deg.C, 50 deg.C, 55 deg.C, 60 deg.C, 65 deg.C for 2h, and placing the triangular bottles in a constant temperature water bath (90 deg.C, 15min) for inactivating enzyme. Adding Maillard reaction substrates, namely 3% of xylose, 1.5% of glucose, 0.5% of VB1, 0.1% of L-cysteine, 0.5% of glycine, 0.5% of plant protein hydrolysate, 0.5% of yeast extract, 4% of salt, 2.5% of ginger powder, 1% of onion powder, 0.1% of anise, 0.2% of 5' -flavor nucleotide disodium and 3.5% of soy sauce into the enzymatic hydrolysate, reacting for 1h at 100 ℃, and cooling to 35-45 ℃ to be tested.

The catalytic action of the enzyme is greatly influenced by temperature, and the activity of the protease is different at different temperatures, so that the proper enzymolysis temperature needs to be selected.

TABLE 6 influence of the enzymolysis temperature on the aroma-producing effect

As can be seen from Table 6, when the enzymolysis temperature is 40-65 ℃, the fragrance score shows a trend that the fragrance score increases firstly and then decreases along with the increase of the temperature on the whole. When the enzymolysis temperature is about 55 ℃, the activity of the compound protease is the highest, and the content of free amino acid nitrogen is the highest and is 0.46 g/mL. The final product obtained by the Maillard reaction of the enzymolysis liquid has the highest fragrance score of 8.8, and the spare rib flavor and the fermented soybean flavor are both strong and have good harmony. Therefore, 55 ℃ is the optimal enzymolysis temperature of the compound protease. When the enzymolysis temperature is 45-55 ℃, the content of free amino acid nitrogen increases along with the temperature rise. When the temperature exceeds 55 ℃ and even higher, the content of free amino acid nitrogen decreases. It is considered that the closer the enzymolysis temperature is to the optimum temperature, the higher the activity of the protease, the better the hydrolysis effect, and the higher the content of free amino acid nitrogen.

In summary, 55 ℃ is the optimal enzymolysis temperature of the invention.

Example 4 Effect of enzymatic hydrolysis time on fragrance production

Adding 10g of minced and heat-treated pork serving as a substrate into 7 triangular bottles with the volume ratio of 1:1, and adjusting the pH to 7.0. Adding 2% of papain and flavourzyme into 7 triangular flasks respectively, placing the triangular flasks in a shaking incubator (55 ℃, 120r/min), reacting for 1h, 1.5h, 2h, 2.5h, 3h, 3.5h and 4h respectively, and then placing the triangular flasks in a constant-temperature water bath kettle (90 ℃, 15min) for enzyme deactivation. Adding Maillard reaction substrates, namely 3% of xylose, 1.5% of glucose, 0.5% of VB1, 0.1% of L-cysteine, 0.5% of glycine, 0.5% of plant protein hydrolysate, 0.5% of yeast extract, 4% of salt, 2.5% of ginger powder, 1% of onion powder, 0.1% of anise, 0.2% of 5' -flavor nucleotide disodium and 3.5% of soy sauce into the enzymatic hydrolysate, reacting for 1h at 100 ℃, and cooling to 35-45 ℃ to be tested.

Under the optimal condition, the enzyme can fully react with the substrate to generate the flavor-enhancing substance component, so that the single flavor is overcome, and the enzymolysis time is a very critical factor for the enzymolysis effect. If the hydrolysis time of the protease is too short, the reaction is insufficient, and the effect of enhancing the aroma of the odor cannot be achieved; on the contrary, if the hydrolysis time is too long, the hydrolysis is excessive and an offensive odor is generated. Therefore, controlling the time of the enzymatic hydrolysis is critical to obtain the optimal product.

TABLE 7 Effect of enzymatic hydrolysis time on fragrance production

As can be seen from Table 7, the enzymolysis time is 2-6 h, and the content of free amino acid nitrogen increases and the fragrance score gradually increases along with the increase of the enzymolysis time in the first 2.5h of enzymolysis. When the enzymolysis time is 2.5h, the content of free amino acid nitrogen is 0.48g/mL, and the fragrance score reaches the highest value. When the enzymolysis time exceeds 2.5h, the content of free amino acid nitrogen still decreases after a small increase, and the aroma score begins to decrease.

Presumably, the reason for the above phenomena is that when the enzymolysis reaches 2.5h, the hydrolysis reaction reaches the optimal state, the hydrolyzed substrate has good aroma-increasing effect, and the black bean-flavor sparerib base material has strong sparerib flavor and black bean flavor, good harmony and no foreign odor; when the enzymolysis time reaches 3.5h, the phenomenon of over hydrolysis occurs, so that the product has bad flavor; when the enzymolysis time reaches 4 hours, the protease can be hydrolyzed, so that the enzyme activity is reduced, and the small amino acid peptides in the enzymolysis liquid are subjected to reverse reaction, so that the enzymolysis effect is poor.

In conclusion, the optimal enzymolysis time of the invention is 2.5 h.

Example 5 Effect of enzymatic pH on fragrance production

Respectively adding 10g of minced and heat-treated pork serving as a substrate into 5 triangular bottles with the volume ratio of 1:1 of water. Adjusting pH to 6.0, 6.5, 7.0, 7.5 and 8.0 in 5 triangular flasks respectively, adding 2% of papain and flavourzyme, placing in a shaking incubator (55 ℃, 120r/min), reacting for 2.5h, and placing the triangular flasks in a constant temperature water bath (90 ℃, 15min) for enzyme deactivation. Adding Maillard reaction substrates, namely 3% of xylose, 1.5% of glucose, 0.5% of VB1, 0.1% of L-cysteine, 0.5% of glycine, 0.5% of plant protein hydrolysate, 0.5% of yeast extract, 4% of salt, 2.5% of ginger powder, 1% of onion powder, 0.1% of anise, 0.2% of 5' -flavor nucleotide disodium and 3.5% of soy sauce into the enzymatic hydrolysate, reacting for 1h at 100 ℃, and cooling to 35-45 ℃ to be tested.

The protease hydrolyzes protein, the product after enzymolysis is the main ingredient for increasing the flavor, and different pH values can influence the hydrolysis of the protease, thereby influencing the modification effect on the flavor of the black bean-flavor spare rib base material. If the reaction system is slightly alkaline, hydroxyl can react with unsaturated fatty acid to generate salt, and substances for enhancing the fragrance can be reduced; if the system is too acid-too-alkali, the enzyme will be inactivated. At the same time, pH also affects the dissociation of the substrate and the groups involved in the enzymatic activity center, and thus the binding and catalysis of the enzyme to the substrate. Therefore, hydrolysis under acidic conditions or neutral and alkaline conditions has different influences on the preparation of the black bean-flavor rib flavor essence.

TABLE 8 Effect of pH of the enzymatic hydrolysis on fragrance production

As can be seen from Table 8, when the pH of the reaction system increased in a gradient manner, the content of free amino acid nitrogen and the fragrance score increased continuously, and the fragrance of the product became stronger and smoother. When the pH is 7.0, the content of free amino acid nitrogen is 0.48g/100mL, the score is 9.0 min at most, and the product obtained after Maillard reaction of the enzymolysis substrate has rich sparerib flavor and fermented soybean flavor, the harmony of the sparerib flavor and the fermented soybean flavor is optimal, and the flavor is rich. When the pH exceeds 7.0, the overall score begins to decrease, and the spare rib flavor and fermented soybean flavor aroma decrease.

The reason for the above phenomenon is presumed to be that when the pH is less than 7.0, the whole reaction system is acidic, the enzyme may be inactivated and not work, and some substances with better flavor are not hydrolyzed, so that the spare rib flavor and the fermented soybean flavor are not prominent; when the pH value is 7.0, the enzyme is in neutral, so that the enzyme reaction is facilitated, and a flavor substance with better harmony is generated; when the pH is more than 7.0, the reaction system is biased to be alkaline and the flavor is decreased.

In conclusion, the optimum enzymolysis pH of the invention is 7.0.

EXAMPLE 6 Effect of feed liquid ratio on fragrance production

Adding 10g minced and heat treated pork as substrate into 5 50ml triangular bottles, respectively, adding water at a material-liquid ratio of 2:1, 1.5:1, 1:1.5, and 1:2, and adjusting pH to 7.0. Adding 2% of papain and flavourzyme into 5 triangular flasks respectively, placing in a shaking incubator (55 ℃, 120r/min), reacting for 2.5h, and placing the triangular flasks in a constant-temperature water bath kettle (90 ℃, 15min) for enzyme deactivation. Adding Maillard reaction substrates, namely 3% of xylose, 1.5% of glucose, 0.5% of VB1, 0.1% of L-cysteine, 0.5% of glycine, 0.5% of plant protein hydrolysate, 0.5% of yeast extract, 4% of salt, 2.5% of ginger powder, 1% of onion powder, 0.1% of anise, 0.2% of 5' -flavor nucleotide disodium and 3.5% of soy sauce into the enzymatic hydrolysate, reacting for 1h at 100 ℃, and cooling to 35-45 ℃ to be tested.

The feed-to-liquid ratio is responsive to substrate concentration and the extent of substrate and enzyme contact. The higher the feed-liquid ratio is, the higher the substrate content is, the thicker the whole reaction system is, the substrate is not contacted with enzyme uniformly, the incomplete reaction of the substrate is caused, and the hydrolysis degree is low; the smaller the feed-to-liquid ratio, the lower the substrate content, and the corresponding decrease in enzyme concentration, resulting in a decrease in hydrolysis efficiency.

TABLE 9 influence of enzymolyzed substrate feed liquid ratio on fragrance-producing effect

As can be seen from table 9, when the feed-liquid ratio is greater than 1:1, the content of free amino acid nitrogen increases with the decrease of the feed-liquid ratio, the fragrance score increases, the foreign odor decreases, and the fragrance harmony increases; when the ratio of the material to the liquid is less than 1:1, the content of free amino acid nitrogen is reduced along with the reduction of the ratio of the material to the liquid, the fragrance score is reduced, and fishy smell can appear; when the feed-liquid ratio is equal to 1:1, the content of free amino acid nitrogen is high and is 0.48g/100mL, the fragrance score reaches the highest score of 8.9, and the product obtained by carrying out Maillard reaction on the enzymolysis substrate has rich sparerib taste and fermented soybean fragrance, good harmony and rich and abundant flavor.

In conclusion, the optimal enzymolysis feed liquid ratio is 1:1 (v/v).

Example 7 Effect of xylose addition on fragrance-producing Effect

Respectively adding 10g of minced and heat-treated pork serving as a substrate into 5 triangular bottles with the volume ratio of 1:1, adjusting the pH value to 7.0, adding 2% of papain and flavourzyme, placing the mixture into a shaking incubator (55 ℃, 120r/min), reacting for 2.5 hours, and placing the triangular flask into a constant-temperature water bath kettle (90 ℃, 15min) for enzyme deactivation. Adding different amounts of xylose, 1.5 percent of glucose, 0.5 percent of VB1, 0.1 percent of L-cysteine, 0.5 percent of glycine, 0.5 percent of plant protein hydrolysate, 0.5 percent of yeast extract, 4 percent of salt, 2.5 percent of ginger powder, 1 percent of onion powder, 0.1 percent of anise, 0.2 percent of 5' -flavor nucleotide disodium and 3.5 percent of soy sauce into the enzymatic hydrolysate respectively, wherein the xylose in 5 triangular bottles respectively accounts for 2 percent, 2.5 percent, 3 percent, 3.5 percent and 4 percent of the pork mass, reacting for 1 hour at 100 ℃, cooling to 35-45 ℃ and detecting.

Xylose has high reaction activity in the Maillard reaction and also has partial physiological functions of dietary fiber. A small amount of xylose is added into the food, so that a good health-care effect can be embodied. The amount of xylose added has a great influence on the flavor of the product after the Maillard reaction, and therefore the following study was made on the amount of xylose added.

TABLE 10 influence of xylose addition on fragrance-producing Effect

As can be seen from table 10, when the addition amount of xylose was increased in a gradient manner, the pork rib flavor and fermented soybean flavor were increased, and the flavor score was increased. When the adding amount reaches 2.5% of the pork quality, the fragrance score reaches the highest score of 8.9, the product sparerib has the most prominent taste, the fermented soybean has strong fragrance and good harmony, and no fishy smell or pungent smell appears. When the addition amount of xylose is more than or equal to 2.5 percent of the pork quality, the spare rib flavor and the fermented soybean flavor begin to weaken, and more intense pungent flavor appears.

The reason for the above phenomenon is presumed to be that, with the increase of xylose, a rich substrate is provided for Maillard reaction, and with the disappearance of amino acid and some free sugar, some flavor substances are generated, so that the flavor and aroma are more durable and the aroma is pure. However, when the addition amount of xylose exceeds a certain limit, the browning reaction is excessive, and the excessive browning does not cause the Maillard reaction product to generate an attractive color, but causes the Maillard reaction product to lose the original organoleptic properties and generate a scorched and smelly pungent smell.

Comprehensively, the optimum addition amount of the xylose is 2.5 percent of the pork quality.

EXAMPLE 8 Effect of xylose and glucose ratios on flavor Effect

Respectively adding 10g of minced and heat-treated pork serving as a substrate into 5 triangular bottles with the volume ratio of 1:1, adjusting the pH value to 7.0, adding 2% of papain and flavourzyme, placing the mixture into a shaking incubator (55 ℃, 120r/min), reacting for 2.5 hours, and placing the triangular flask into a constant-temperature water bath kettle (90 ℃, 15min) for enzyme deactivation. Adding 2.5% of xylose, different amounts of glucose, 0.5% of VB1, 0.1% of L-cysteine, 0.5% of glycine, 0.5% of plant protein hydrolysate, 0.5% of yeast extract, 4% of salt, 2.5% of ginger powder, 1% of onion powder, 0.1% of anise, 0.2% of disodium 5' -ribonucleotide and 3.5% of soy sauce into the enzymolysis liquid respectively, wherein the adding amounts of the glucose in 5 triangular bottles are respectively xylose: the glucose is 3: 1. 2: 1. 1:1. 1: 2. 1: reacting at 3,100 deg.C for 1h, cooling to 35-45 deg.C, and testing.

The compounding ratio of xylose and glucose has great influence on the whole system of the Maillard reaction, thereby having important influence on the formation of flavor substances. Xylose contributes greatly to the meat flavor of the aroma compared to glucose, whereas glucose contributes greatly to the scorched aroma of the aroma. And simultaneously, glucose and xylose are used, so that the fragrance is richer. The experiment shows that the optimal addition amount of xylose is 2.5% of the pork quality, the xylose dosage can be fixed, the glucose dosage can be changed, the influence of the compounding ratio of reducing sugar on the aroma production effect is researched, and the optimal compounding ratio is selected.

TABLE 11 influence of xylose to glucose ratio on fragrance-producing Effect

As can be seen from table 11, when xylose: glucose is less than 2:1, the pork rib flavor becomes light, the fermented soybean flavor also becomes light, the fermented soybean flavor is not prominent, and pungent smell exists. When the ratio of xylose: glucose is greater than 2:1, with the reduction of the compound ratio, the spare rib flavor can gradually become thick and mellow, the black bean flavor is strong, the obtained flavor effect is rich, and the harmony is better. When the ratio of xylose: the mass ratio of glucose is equal to 2:1, the obtained black bean-flavored sparerib base has the most abundant flavor, the most intense sparerib flavor, the prominent black bean flavor and good harmony.

The reason for the above phenomenon is presumed to be that, as the glucose content decreases, the complementary tendency of xylose and glucose in the system becomes more and more effective, and the undesirable flavor substances gradually decrease, thereby producing a flavor-producing substance having a better flavor. When the ratio of xylose: glucose is greater than 2: in case 1, it is considered that the short peptides, amino acids, etc. in the pig bone protein hydrolysate are consumed up due to the excessive content of reducing sugar in the system, or the degradation of reducing sugar, etc. in the maillard reaction system and the generation of caramel pigment, etc. are involved, so that the prepared fragrant-based sparerib flavor and fermented bean flavor are weakened, and pungent smell appears.

Taken together, the optimum xylose of the invention: the mass ratio of glucose is 2: 1.

example 9 Effect of amino acid type on fragrance-producing Effect

Respectively adding 10g of minced and heat-treated pork serving as a substrate into 6 triangular flasks with the volume ratio of 1:1, adjusting the pH value to 7.0, adding 2% of papain and flavourzyme, placing the mixture into a shaking incubator (55 ℃, 120r/min), reacting for 2.5 hours, and placing the triangular flask into a constant-temperature water bath kettle (90 ℃, 15min) for enzyme deactivation. Adding xylose accounting for 2.5 percent of the mass of pork, glucose accounting for 1.5 percent of the mass of the pork, VB1 accounting for 0.5 percent of the mass of the pork, different amino acids, plant protein hydrolysate accounting for 0.5 percent of the mass of the pork, yeast extract accounting for 0.5 percent of the mass of the pork, salt accounting for 4 percent of the mass of the pork, ginger powder accounting for 2.5 percent of the mass of the pork, onion powder accounting for 1 percent of the mass of the pork, star anise accounting for 0.1 percent of the mass of the pork, disodium ribonucleotide accounting for 0.2 percent of the mass of the pork and soy sauce accounting for 3.5 percent of the mass of the pork, wherein the amino acids in 6 triangular bottles are respectively matched with 6L-proline, DL-methionine, glycine, proline and proline, DL-methionine, react for 1 hour at the temperature of 100 ℃, and are cooled to 35 to 45 ℃ to.

The Maillard reaction of L-cysteine, L-proline, DL-methionine and glycine with pork can produce thick and mellow flavor substances. After xylose, glucose and L-cysteine react together, the variety of aroma compounds can be increased, and good meat aroma can be generated. Because L-cysteine has a large influence on the flavor of the meat flavor, the L-cysteine is selected, and L-proline, DL-methionine and glycine are selected to be matched with the L-cysteine to enrich the flavor of the meat flavor. In the preliminary experiment, the fragrance of the fragrant fermented soybeans obtained by matching 0.2% of L-cysteine with 0.5% of other amino acids is more prominent, so that the experiment selects 0.2% of L-cysteine to match with 0.5% of other amino acids.

TABLE 12 influence of the amino acid type on the fragrance-producing Effect

As can be seen from Table 12, when L-cysteine was separately combined with L-proline or glycine, the Maillard reaction yielded a rich and pure flavor of the base pork chop, and a good flavor of fermented soybean. Wherein, when the L-cysteine is only matched with the glycine, the effect is the best, and the fragrance score is 8.9. However, when L-cysteine, L-proline and glycine are used in combination, the product after Maillard reaction has a reduced odor score and an offensive odor. It is presumed that these three amino acids produce an undesirable flavor substance in the Maillard reaction and affect the flavor-producing effect.

When L-cysteine or L-cysteine, glycine and DL-methionine are matched, the fragrance base after Maillard reaction has bitter taste, and the obtained fragrance score is low, presumably because the pork enzymolysis liquid used in the research already contains more methionine, and the addition of DL-methionine causes relative excess to influence the flavor formation.

Comprehensively, the most suitable amino acid of the invention is selected to be 0.2 percent of L-cysteine and 0.5 percent of glycine for compounding.

Example 10 Influence of the addition of L-cysteine on the fragrance-producing effect

Respectively adding 10g of minced and heat-treated pork serving as a substrate into 5 triangular bottles with the volume ratio of 1:1, adjusting the pH value to 7.0, adding 2% of papain and flavourzyme, placing the mixture into a shaking incubator (55 ℃, 120r/min), reacting for 2.5 hours, and placing the triangular flask into a constant-temperature water bath kettle (90 ℃, 15min) for enzyme deactivation. Adding xylose accounting for 2.5 percent of the pork mass, glucose accounting for 1.25 percent of the pork mass, VB1 accounting for 0.5 percent of the pork mass, L-cysteine accounting for different quantities, glycine accounting for 0.5 percent of the pork mass, vegetable protein hydrolysate accounting for 0.5 percent of the vegetable protein hydrolysate, yeast extract accounting for 0.5 percent of the vegetable protein hydrolysate, salt accounting for 4 percent of the vegetable protein hydrolysate, ginger powder accounting for 2.5 percent of the vegetable protein hydrolysate, onion powder accounting for 1 percent of the vegetable protein hydrolysate, anise accounting for 0.1 percent of the vegetable protein hydrolysate, disodium 5' -ribonucleotide accounting for 0.2 percent of the vegetable protein hydrolysate and soy sauce accounting for 3.5 percent of the vegetable protein hydrolysate, wherein the L-cysteine accounting for 5 triangular bottles respectively accounts for 0, 0.1 percent, 0.2 percent, 0.3 percent and 0.4 percent.

L-cysteine plays a leading role in the synthesis of some important meat flavor substances in the Maillard reaction, and a proper amount of cysteine can generate soft meat flavor.

TABLE 13 influence of the amount of L-cysteine added on the fragrance-producing effect

As can be seen from Table 13, when the amount of cysteine added is less than 0.2% of the pork mass, the pork rib flavor is enhanced and becomes rich with the increase of the amount of cysteine, the fermented soybean flavor becomes more and more prominent, and the whole flavor becomes harmonious and full. When the addition amount of cysteine is equal to 0.2 percent of the quality of pork, the fragrance score reaches the highest score of 9.2, and the black bean-flavor sparerib base sparerib flavor and black bean flavor are strong and prominent, have good harmony and have no foreign flavor and pungent flavor. When the amount of cysteine added is more than 0.2% by mass of pork, offensive odor and sulfur-flavored pungent odor are increased, and the pork rib flavor and fat flavor are masked, so that the overall flavor is reduced and harmony is deteriorated, and it is presumed that the cysteine content is too high or excessively decomposed to generate excessive ammonia, hydrogen sulfide, acetaldehyde and the like, thereby giving off a bad flavor.

In conclusion, the optimal dosage of the L-cysteine is 0.2 percent of the pork quality.

Example 11 influence of Maillard reaction time on fragrance-producing Effect

Respectively adding 10g of minced and heat-treated pork serving as a substrate into 5 triangular bottles with the volume ratio of 1:1, adjusting the pH value to 7.0, adding 2% of papain and flavourzyme, placing the mixture into a shaking incubator (55 ℃, 120r/min), reacting for 2.5 hours, and placing the triangular flask into a constant-temperature water bath kettle (90 ℃, 15min) for enzyme deactivation. Adding xylose accounting for 2.5 percent of the weight of pork, glucose accounting for 1.25 percent of the weight of the pork, VB1 accounting for 0.5 percent of the pork, L-cysteine accounting for 0.2 percent of the pork, glycine accounting for 0.5 percent of the pork, vegetable protein hydrolysate accounting for 0.5 percent of the pork, yeast extract accounting for 0.5 percent of the pork, salt accounting for 4 percent of the pork, ginger powder accounting for 2.5 percent of the pork, onion powder accounting for 1 percent of the pork, star anise accounting for 0.1 percent of the pork, disodium 5' -ribonucleotide accounting for 0.2 percent of the pork and soy sauce accounting for 3.5 percent of the pork, reacting substrates in 5 triangular bottles at 100 ℃ for 30min, 45min, 60min, 75min and 90min, and cooling to 35-45 ℃.

The reaction time is an important factor influencing the Maillard reaction, and the reaction time is too short and insufficient, so that the generated flavor substances are few and the fragrance is light; too long reaction time can result in excessive reaction, easy occurrence of excessive caramelization reaction, occurrence of uncomfortable scorched bitter taste and poor flavor. Therefore, the selection of an appropriate maillard reaction time is of great significance for the formation of aroma.

TABLE 14 influence of Maillard reaction time on fragrance production

As can be seen from table 14, when the reaction time is less than 60min, the score of aroma shows a rising trend with the increase of the maillard reaction time, the aroma is continuously enhanced, but the spare rib aroma and the fermented soybean aroma are not prominent enough, and the fishy smell or the sour strange taste gradually disappears. When the Maillard reaction time is 60min, the fragrance score reaches the highest score of 9.1, and the prepared black bean-flavored sparerib base material has the advantages of prominent and saturated fragrance and good harmony. When the Maillard reaction time is more than 60min, the fragrance score is reduced along with the rise of the Maillard reaction temperature, and the bad flavors such as scorch bitter and miscellaneous flavors appear.

In conclusion, the optimal Maillard reaction time of the invention is 60 min.

EXAMPLE 12 Effect of Maillard reaction temperature on fragrance Effect

Respectively adding 10g of minced and heat-treated pork serving as a substrate into 5 triangular bottles with the volume ratio of 1:1, adjusting the pH value to 7.0, adding 2% of papain and flavourzyme, placing the mixture into a shaking incubator (55 ℃, 120r/min), reacting for 2.5 hours, and placing the triangular flask into a constant-temperature water bath kettle (90 ℃, 15min) for enzyme deactivation. Adding xylose 2.5 wt%, glucose 1.25 wt%, VB1 0.5 wt%, L-cysteine 0.1 wt%, glycine 0.5 wt%, plant protein hydrolysate 0.5 wt%, yeast extract 0.5 wt%, salt 4 wt%, ginger powder 2.5 wt%, onion powder 1 wt%, anise 0.1 wt%, disodium 5' -ribonucleotide 0.2 wt%, soy sauce 3.5 wt% and 5 triangular bottle substrate into the enzymolysis liquid, reacting at 90 deg.c, 95 deg.c, 100 deg.c, 105 deg.c and 110 deg.c for 1 hr, and cooling to 35-45 deg.c for measurement.

The maillard reaction temperature is highly likely to affect the reaction path, the concentration of the reactants, the optical activity and the activity. In general, the reaction rate increases as the reaction temperature increases, but excessively high temperatures destroy the saccharide and the amino acid. Different temperatures have a great effect on the maillard reaction products and unsuitable temperatures may produce some unpleasant odours. Therefore, the selection of an appropriate maillard reaction time is of great importance for the formation of the aroma.

TABLE 15 influence of Maillard reaction temperature on fragrance production

As can be seen from Table 15, the Maillard reaction temperature has a very important influence on the fragrance production effect, and the fragrance score tends to increase and decrease along with the increase of the Maillard reaction temperature, and the fragrance score fluctuates greatly between 95 ℃ and 100 ℃.

When the Maillard reaction temperature is lower than 100 ℃, the color of the base is orange red, the pork rib flavor and the black bean flavor are not full enough, and the optimal reaction state is not reached due to the slow Maillard reaction rate. When the Maillard reaction temperature is equal to 100 ℃, the color of the fragrant base is reddish brown, the fragrance score reaches the highest score of 9.0 minutes, and the prepared black bean-flavor sparerib base material has full and prominent characteristic fragrance, good harmony and no peculiar smell or pungent smell. When the Maillard reaction temperature is higher than 100 ℃, the color of the aroma base is brownish black, the base material has obvious burnt flavor and pungent flavor, the spare rib flavor and the fermented soybean flavor of the base material are covered, the harmony is poor, the reason is supposed to be that the temperature is too high, some saccharides and amino acids are damaged or the Maillard reaction is too fast and the degree is too high, and therefore, the bad flavor substances are generated.

In conclusion, the optimal Maillard reaction temperature of the invention is 100 ℃.

Example 13

The embodiment provides a preparation method of a black bean-flavor spare rib flavor, which comprises the following steps:

1. enzymolysis: cleaning fresh pork hind leg meat, mincing, treating with boiling water for 5min, weighing the treated pork, adding water according to the feed-liquid ratio of 1:1 to prepare a mixed solution, adjusting the pH to 7.0, adding a papain and flavourzyme mixed enzyme accounting for 2% of the pork by mass, carrying out enzymolysis at 55 ℃ for 2.5h, carrying out enzyme deactivation at 90 ℃, and carrying out 15min to obtain an enzymolysis solution of the pork.

2. Maillard reaction: adding xylose and glucose which are 2.5 percent of the pork mass into the enzymolysis liquid, wherein the ratio of xylose: the mass ratio of glucose is 2: adding 0.2 percent of L-cysteine and 0.5 percent of glycine by mass of pork for compounding, adding 0.5 percent of VB1, 0.5 percent of plant protein hydrolysate and 0.5 percent of yeast extract by mass percent, carrying out Maillard reaction for 60min at the temperature of 100 ℃, and cooling to 35-45 ℃ to obtain the black bean-flavor spare rib flavor.

3. Blending: adding 4% of salt, 2.5% of ginger powder, 1% of onion powder, 0.1% of anise, 0.2% of 5' -flavour nucleotide disodium and 3.5% of soy sauce in volume percentage into the material obtained by the Maillard reaction for blending to obtain the black bean-flavor sparerib spice.

Evaluation of the pork rib flavor with fermented soybean flavor prepared in example 13 revealed that the content of free amino acid nitrogen was 0.62(g/100mL), the flavor score was 9.5, the pork rib flavor was strong, the fermented soybean flavor was strong, the harmony was good, and there were no fishy smell and pungent smell. Therefore, the best fermented soybean-flavor spare rib flavor can be obtained by adopting the preparation method of example 13, and example 13 is the best scheme for preparing the fermented soybean-flavor spare rib flavor of the invention.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. A preparation method of a black bean-flavor rib-flavor spice is characterized by comprising the following steps:

s2: maillard reaction: adding a Maillard reaction substrate into the enzymatic hydrolysate in the step (1), and reacting for 30-90 min;

s3: blending: adding salt, ginger powder, onion powder, anise, 5' -flavour nucleotide disodium and soy sauce into the material prepared in the step (2) for blending to obtain the black bean-flavor rib flavor spice.

2. The method according to claim 1, wherein the protease is selected from one or more of the following enzyme classes: papain, compound protease, hydrolytic protease, flavourzyme and compound protease.

3. The method according to claim 2, wherein the protease is a mixed enzyme of papain and flavourzyme.

4. The method according to claim 1, wherein the protease is added in an amount of 1 to 3% by mass based on the pork.

5. The preparation method according to claim 1, wherein the enzymolysis temperature is 40-65 ℃; the enzymolysis time is 1-4 h.

6. The method of claim 1, wherein the Maillard reaction substrate comprises xylose, glucose, L-cysteine, L-proline, DL-methionine, glycine.

7. The method of claim 1, wherein the Maillard reaction substrate comprises xylose, glucose, L-cysteine, glycine.

8. The preparation method according to claim 7, wherein the mass ratio of xylose to glucose is 1-3: 1-3; the addition amount of the L-cysteine is 0.1-0.4% of the mass of the pork.

9. The method according to claim 1, wherein the Maillard reaction time is 30 to 90 min; the Maillard reaction temperature is 90-110 ℃.

10. The method of claim 1, wherein the Maillard reaction substrate further comprises VB1, a plant protein hydrolysate, and a yeast extract.