Method for extracting umami substances from morchella esculenta and application of umami substances

Abstract

The invention provides a method for extracting a ladder delicious substance from morchella armata, which comprises the following steps of: preparing a morchella armata powder raw material, cooking at normal pressure, performing ultrasonic-assisted extraction, performing enzymolysis treatment, performing centrifugal separation, and concentrating an extracting solution under reduced pressure. And drying the obtained concentrated solution, and preparing into powder for later use. The invention also provides the morchella esculenta umami substance obtained by the method. The invention also provides an application of the morchella esculenta umami substance in preparation of a flavoring agent or a flavoring peptide ingredient product. The method not only optimizes the extraction efficiency of umami substances, but also can improve the sensory attributes of the product, has wide application potential, and is especially suitable for the development of food seasonings and flavor-enhancing peptide products.

Description

Technical Field

[0001] The invention belongs to the field of food processing and relates to Morchella importuna, in particular to a method for extracting umami substances from Morchella importuna and application thereof. Background Art

[0002] Morchella tiliacea is a precious edible fungus, which is widely used in food due to its unique flavor, umami taste and rich nutrients. The umami taste of Morchella tiliacea mainly comes from compounds such as amino acids, nucleotides and small molecule peptides. As the main ingredients of natural condiments, these substances have extremely high value for deep processing and utilization. Different extraction methods have a significant impact on the retention and conversion of umami substances. Traditional extraction methods including hot water extraction, acid hydrolysis and autolysis have been widely used, while high pressure, enzyme-assisted, ultrasound-assisted and microwave-assisted technologies have attracted much attention because they can improve solubility and maintain sensory and nutritional properties.

[0003] Studies have found that cooking leads to a decrease in the free amino acids in Oyster mushrooms, while enzymatic hydrolysis promotes the release of free amino acids in Lentinus edodes. Progress has been made in the application of ultrasound-assisted extraction technology in dried straw mushrooms and the like. Morchella esculenta is rich in a variety of umami substances such as amino acids, nucleotides, and small molecule peptides, which are the main source of its umami taste. However, existing umami substance extraction technologies, such as hot water extraction and acid hydrolysis, have problems such as insufficient release of umami substances and low extraction efficiency, resulting in the natural umami taste of Morchella esculenta not being fully utilized. Therefore, there is an urgent need for an improved extraction method to improve the extraction efficiency and product quality of umami substances in Morchella esculenta. Summary of the invention

[0004] In view of the above-mentioned technical problems in the prior art, the present invention provides a method for extracting umami substances from Morchella tangata and its application. The method for extracting umami substances from Morchella tangata and its application are intended to solve the technical problems of insufficient release of umami substances and low extraction efficiency in the prior art method for extracting umami substances from Morchella tangata.

[0005] The present invention provides a method for extracting umami substances from Morchella fasciata, comprising the following steps:

[0006] 1) preparing a raw material of Morchella esculenta powder, drying fresh Morchella esculenta to a moisture content of 10-13%, then crushing into fine powder, filtering through a 70-90 mesh sieve, and setting aside;

[0007] 2) a step of steaming at 100°C under normal pressure, wherein the dried Morchella truncatula powder obtained in step 1) is mixed with water in a mass ratio of

[0008] Mix in a ratio of 1g:(8-12)ml; steam at 100℃ under normal pressure for 0.5-2 hours;

[0009] 3) an ultrasonic-assisted extraction step, wherein the solution after being boiled at 100° C. under normal pressure is subjected to ultrasonic extraction, with an ultrasonic power of 350 W, an ultrasonic time of 10 to 30 min, and a pulse ultrasonic working / interval time of (3 to 7) s / 1 s to obtain an ultrasonic extract;

[0010] 4) an enzymatic hydrolysis step, adding 3000-6000 U / g of enzyme to the ultrasonic extract of step 3), wherein the enzyme is flavor protease, neutral protease, papain, edible fungus hydrolase, and the mass ratio of the flavor protease, neutral protease, papain, and edible fungus hydrolase is 1:1:1:1, and enzymatic hydrolysis is performed at 45-55°C

[0011] 1 to 3 hours; after the reaction, the enzyme is inactivated by heating;

[0012] 5) a centrifugation step, centrifuging the enzymatic hydrolyzate from step 4) and collecting the supernatant;

[0013] 6) a step of concentrating the extract under reduced pressure, wherein the supernatant collected in step 5) is concentrated under reduced pressure to obtain an extract rich in umami components;

[0014] 7) The extract rich in umami components obtained in step 6) is dried to prepare a powder for later use.

[0015] The present invention also provides the umami substance of Morchella tiliacea obtained by the method.

[0016] The present invention also provides the use of the Morchella tiliacea umami substance in preparing a seasoning or a flavor-enhancing peptide ingredient product.

[0017] By adopting the above method of the present invention, enzymatic hydrolysis can significantly increase the content of umami amino acids (such as glutamic acid and aspartic acid) and umami nucleotides (such as 5'-GMP and 5'-AMP) in Morchella tiliacea, enhance the umami intensity of the product, and is suitable for developing high-quality food condiments.

[0018] The present invention significantly increases the content of umami amino acids and nucleotides through processing steps such as normal pressure cooking, ultrasound-assisted extraction, enzymatic hydrolysis and reduced pressure concentration. The method not only optimizes the extraction efficiency of umami substances, but also improves the sensory properties of the product, has a wide range of application potential, and is particularly suitable for the development of food seasonings and flavor-enhancing peptide products.

[0019] Compared with the prior art, the technical effect of the present invention is positive and obvious.

[0020] 1. The release rate of umami substances in Morchella tiliacea was improved through enzymatic hydrolysis, especially the content of umami amino acids and nucleotides was significantly increased.

[0021] 2. The combined use of flavor protease, papain, neutral protease and edible fungus hydrolase can effectively improve the release of amino acids and optimize the extraction efficiency.

[0022] 3. The extract can enhance the umami taste and flavor of food in the production of seasonings and flavor-enhancing peptides, and has broad application prospects.

[0023] 4. The ultrasonic-assisted extraction method is used to significantly increase the content of umami substances, improve the flavor and quality of the product, and is suitable for large-scale industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 : 5'-nucleotide content of Morchella tadalafil extracts under different treatments; Note: Different letters above the column indicate significant differences (P<0.05).

[0025] Figure 2 : EUC values ​​and sensory scores of Morchella tataricola extracts treated with different methods; Note: Different letters on the columns indicate significant differences (P<0.05). DETAILED DESCRIPTION

[0026] Example 1: Preparation and drying of Morchella edulis

[0027] Fresh Morchella tiliacea fruiting bodies purchased from Qianxi County, Guizhou Province were dried using a hot air dryer at 50°C until the moisture content of the wet basis was 13%. The dried Morchella tiliacea was crushed, filtered through an 80-mesh sieve, and set aside.

[0028] Example 2: Extraction of umami substances from Morchella oleracea by 100°C normal pressure cooking, ultrasound-assisted and single enzymatic hydrolysis

[0029] Take 50g of Morchella edulis powder, add 500mL of deionized water (solid-liquid ratio is 1:10), cook at 100℃ for 1 hour, and then perform ultrasonic-assisted extraction, wherein the ultrasonic power is 350W, the ultrasonic time is 15min, and the working / interval time of pulsed ultrasound is 5s / 1s. Then, flavor protease, neutral protease, papain, and edible fungus hydrolase are added according to the enzyme activity of 4000U / g, and stirred for enzymolysis at 50℃ for 2 hours. After the enzymolysis is completed, the solution is heated to 100℃ and kept for 15 minutes to inactivate the enzyme. Subsequently, the solution is centrifuged at 8000r / min for 15 minutes, and four enzymolysis supernatants are collected for use.

[0030] Example 3: Extraction of umami substances from Morchella edulis by ultrasound-assisted and combined enzymatic hydrolysis

[0031] Take 50g of Morchella edulis powder, add 500mL of deionized water (solid-liquid ratio is 1:10), cook at 100℃ for 1 hour, and then perform ultrasonic-assisted extraction, where the ultrasonic power is 350W, the ultrasonic time is 15min, and the working / interval time of pulsed ultrasound is 5s / 1s. Then, according to the enzyme activity of 1000U / g, flavor protease, neutral protease, papain, and edible fungus hydrolase are added step by step, and four enzymes are used for complex enzymolysis. Stir the enzymolysis at 50℃ for 2 hours. After the enzymolysis is completed, heat the solution to 100℃ and keep it for 15 minutes to inactivate the enzyme. Subsequently, centrifuge the solution at 8000r / min for 15 minutes, and collect the supernatant for use.

[0032] Example 4: Concentration and drying of extract

[0033] The supernatant is concentrated by a reduced pressure evaporation device until the volume is reduced by 50%, thereby obtaining an extract rich in umami substances. The extract can be directly used in seasonings, or further dried by a spray drying device to produce a powdered seasoning.

[0034] Example 5: Umami substance analysis and sensory evaluation

[0035] Detection of free amino acids

[0036] The free amino acid content of the Morchella oleracea extract was determined according to the method in accordance with GB 5009.124-2016 National Food Safety Standard - Determination of Amino Acids in Foods.

[0037] Detection of 5'-nucleotides

[0038] The 5'-nucleotides in the extract were determined by reverse phase high performance liquid chromatography.

[0039] Equal freshness EUC calculation

[0040] EUC value refers to the concentration of monosodium glutamate (MSG) equivalent to the umami intensity presented by umami amino acids and umami nucleotides in the food system. The larger the EUC value, the stronger the umami taste. The calculation formula is as follows:

[0041] Y=∑a i b i +1218(∑a i b i )(∑a j b j )(1)

[0042] Where: Y is the EUC value of the mixture; a i is the mass fraction of umami amino acid (Asp or Glu) 100g·g-1 ; a j The mass of umami 5'-nucleotide (5'-AMP, 5'-GMP, 5'-IMP or 5'-XMP) is 100 g·g -1 ; b i is the relative umami concentration of each umami amino acid compared with MSG (Asp, 0.077; Glu, 1.000); b j is the relative umami concentration of each umami 5'-nucleotide compared with 5'-IMP (5'-IMP, 1.0; 5'-GMP, 2.3; 5'-XMP, 0.61; 5'-AMP, 0.18); 1218 is the synergistic constant.

[0043] Analysis of flavor contribution of flavoring substances:

[0044] Taste contribution analysis (TAV) of flavor substances is usually used to evaluate the impact of individual flavor compounds (free amino acids, nucleotides, organic bases and related compounds, sugars, organic acids and inorganic ions) on taste. When TAV is greater than 1, it is considered that the compound contributes to the taste of the sample, and the larger the value, the greater the contribution. The formula is as follows:

[0045]

[0046] Where: C is the content of the substance in the sample / (mg·g -1 ); T is the taste threshold of the substance / (mg·g -1 )

[0047] Sensory evaluation

[0048] The samples were evaluated by scoring method using 0.35% monosodium glutamate solution as the standard. The temperature of the sensory evaluation room was controlled at (25±2)℃, and the evaluation team consisted of 15 members. The sample scores ranged from 0 to 10, with 0 indicating that the sample tested had no umami taste, and 10 indicating that the sample had the strongest umami taste. The umami intensity of 0.35% monosodium glutamate standard solution was 5 points. The samples were compared with the standard and then scored and evaluated.

[0049] Example 6: Analysis and sensory evaluation of umami substances in Morchella tiliacea

[0050] Free amino acids are an important component of the umami flavor of edible fungi, which not only give edible fungi a unique and delicious flavor, but also enhance their pleasant taste. According to their flavor characteristics, they can be divided into four categories: umami amino acids include Asp and Glu; sweet ones include threonine (Thr), serine (Ser), glycine (Gly), alanine (Ala) and proline (Pro); bitter ones include valine (Val), methionine (Met), isoleucine (Ile), leucine (Leu), phenylalanine (Phe), histidine (His) and arginine (Arg); tasteless amino acids include cysteine ​​(Cys), tyrosine (Tyr) and lysine (Lys)

[0051] Table 1 shows the content of free amino acids in the extract of Morchella esculenta under different treatments. The results showed that the effects of different treatments on the free amino acid content of Morchella esculenta were significantly different (P<0.05). Among them, the total amount of free amino acids in the FP treatment group was the highest, followed by the EFH and NP groups, while the difference between the APC and HPC groups was not significant, and the total amount of free amino acids in the UAH group was the lowest. The content of umami and sweet amino acids in the FP extract was significantly higher than that in the other treatment groups, followed by the EFH group, and the UAH group had the lowest content. Tyrosine was not detected in all samples.

[0052] Table 1 Free amino acid content

[0053]

[0054]

[0055]

[0056] Note: Different lowercase letters in the same column indicate significant differences, P<0.05

[0057] Table 2 shows the content of soluble peptides in the extract of Morchella tiliacea under different treatments. The soluble peptide content of the FP treatment group was the highest, followed by the EFH and NP groups. The soluble peptide content of the APC treatment group was the lowest.

[0058] Table 2 Soluble peptide content

[0059]

[0060] In addition to umami amino acids, 5'-nucleotides in edible mushrooms have also been found to be important contributors to umami taste. 5'-nucleotides mainly include 5'-GMP, 5'-IMP, 5'-AMP, 5'-CMP and 5'-UMP. Among them, 5'-IMP and 5'-GMP have a synergistic effect in enhancing the umami taste of glutamate, while 5'-UMP and 5'-CMP themselves have weak umami taste and usually need to be used in combination with glutamate (MSG).

[0061] like Figure 1 As shown in the results, there were significant differences in the effects of different extraction methods on the release of 5'-nucleotides (P < 0.05). The total amount of 5'-nucleotides and the content of 5'-GMP in the UAH group reached the highest, while the total amount of flavor nucleotides in the four groups after enzymatic hydrolysis was relatively low and not significantly different from each other. APC and HPC were relatively beneficial to the release of nucleotides. The content of 5'-AMP in the cooking liquor was significantly higher than that in the enzyme-treated liquor. No 5'-IMP was detected in any of the treatment groups.

[0062] Embodiment 7:

[0063] TAV was used to objectively evaluate the contribution of each flavor substance to the umami taste of the extract. The results showed that in Table 2, there were 14 flavor substances with TAV values ​​greater than 1 in the FP and EFH extracts, 13 in the NP extract, 12 in the PAP extract, and only 2 in the APC extract. It is speculated that the reason why the APC extract has a TAV value greater than 1 and fewer flavor substances is that the temperature and pressure are low during conventional hot water extraction, resulting in a decrease in the extraction efficiency, which in turn leads to a decrease in the content of flavor substances. The TAV value of glutamate in the FP sample is the highest among all extracts, indicating that glutamate is the substance that contributes most significantly to umami taste. In addition, the TAV values ​​of 5'-GMP in all extracts are greater than 1, indicating that it contributes significantly to the umami taste of each extract and is one of the key substances affecting the overall umami characteristics. In summary, glutamate and 5'-GMP, as the main umami substances, play an important role in the overall umami characteristics of Morchella tiliacea extracts.

[0064] Table 2 Effects of treatment methods on free amino acids and nucleotides TAV in Morchella tiliacea extract

[0065]

[0066] EUC is often used to evaluate the umami taste of edible mushrooms such as shiitake mushrooms. The EUC values ​​of Morchella esculenta under different extraction treatments are as follows: Figure 2 As shown: EUC values ​​of different extracts vary significantly. EUC values ​​range from 47.25 to 190.12 g MSG·100 g -1 Among them, the EUC value of FP sample was the highest, which was (190.12±7.47) g MSG·100 g -1 , while the EUC value of Morchella edulis after boiling was the lowest (47.25±1.78) g MSG·100g-1. According to the different EUC values, there are four levels: (1)>1000 g MSG·100 g -1 , (2) 100~1000g MSG·100g -1 , (3) 10~100g MSG·100g -1, (4) <10g MSG·100g -1

[25] The extract of Morchella edulis treated by enzymatic hydrolysis was the second level (<10g MSG·100g -1 The extracts of Morchella tiliacea treated by other methods were all at the third level (10-100 g MSG·100 g -1 ). This indicates that the umami characteristics of the enzymatically treated samples are more significant, and the EUC values ​​of the FP and EFH treatments are similar.

[0067] Sensory scores of Morchella esculenta under different treatments. The results showed that the sensory score of the sample treated with FP was the highest, followed by EFH, and the score of the sample treated with UAH was the lowest. Combining the EUC value with the sensory evaluation results, it can be concluded that the umami taste of Morchella esculenta treated with FP was the strongest. This shows that FP treatment can not only significantly improve the umami intensity of Morchella esculenta, but also improve its sensory attributes, and has a high application potential.

Claims

1. A method for extracting umami substances from Morchella oleracea, characterized in that The following steps are involved: 1) preparing a raw material of Morchella esculenta powder, drying fresh Morchella esculenta to a moisture content of 10-13%, then crushing into fine powder, filtering through a 70-90 mesh sieve, and setting aside; 2) a step of steaming at 100° C. under normal pressure, mixing the dried Morchella truncatula powder obtained in step 1) with water in a mass ratio of 1 g: (8-12) ml; steaming at 100° C. under normal pressure for 0.5-2 hours; 3) an ultrasonic-assisted extraction step, wherein the solution after being boiled at 100° C. under normal pressure is subjected to ultrasonic extraction, with an ultrasonic power of 350 W, an ultrasonic time of 10 to 30 min, and a pulse ultrasonic working / interval time of (3 to 7) s / 1 s to obtain an ultrasonic extract; 4) an enzymatic hydrolysis step, adding 3000-6000 U / g of enzyme to the ultrasonic extract of step 3), wherein the enzyme is flavor protease, neutral protease, papain, edible fungus hydrolase, and the mass ratio of the flavor protease, neutral protease, papain, and edible fungus hydrolase is 1:1:1:1, and enzymolysis is performed at 45-55° C. for 1-3 hours; after the reaction, the enzyme is inactivated by heating; 5) a centrifugation step, centrifuging the enzymatic hydrolyzate from step 4) and collecting the supernatant; 6) a step of concentrating the extract under reduced pressure, wherein the supernatant collected in step 5) is concentrated under reduced pressure to obtain an extract rich in umami components; 7) The extract rich in umami components obtained in step 6) is dried to prepare a powder for later use.

2. The umami substance of Morchella tiliacea obtained by the method according to claim 1.

3. Use of the umami substance of Morchella tiliacea according to claim 2 in the preparation of seasoning or flavor-enhancing peptide ingredient products.

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