A flocculation clarification method for mollusc enzymatic hydrolysate
By controlling the temperature changes of shellfish enzymatic hydrolysate and employing a combination of freezing and high-temperature heating treatment, the problems of high energy consumption and flavor impact of conventional clarification technologies are solved, achieving efficient and green flocculation clarification, which is suitable for the industrial production of shellfish enzymatic hydrolysate.
Patent Information
- Application Number
- CN202311007805.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-08-11
- Publication Date
- 2025-11-04
- Estimated Expiration
- 2043-08-11
AI Technical Summary
Conventional clarification techniques are energy-intensive and can easily affect the flavor of the enzymatic hydrolysate, which is not conducive to the development and production of seasonings.
By controlling the temperature change of the shellfish enzymatic hydrolysate, flocculation and clarification can be achieved without adding flocculants by using a combination of freezing and high-temperature heating treatments. This includes freezing at -30 to -20°C for 6 to 20 hours, then heating at 90 to 100°C for 5 to 60 minutes, and finally filtering after allowing the flocs to settle completely.
It achieves efficient clarification of shellfish enzymatic hydrolysate, retains the flavor of the hydrolysate to the greatest extent, and has a wide range of applications and is easy to mass-produce industrially.
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Figure CN117016740B_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to a flocculation clarification method for shellfish enzymatic hydrolysate, belonging to the technical field of food processing. BACKGROUND
[0002] Shellfish has delicious meat, and is used to prepare umami flavoring. During the enzymatic hydrolysis of shellfish, a large amount of turbidity will be produced, which will affect the appearance and limit the further processing and utilization of the shellfish.
[0003] At present, the clarification techniques commonly used in industry include water extraction and alcohol precipitation, high-speed centrifugation, macroporous resin separation, membrane separation, etc. It is found that the water extraction and alcohol precipitation technique has a large loss of effective components in the solution, and for the enzymatic hydrolysate with incomplete impurity precipitation after one-time alcohol precipitation, multiple alcohol precipitation operations are required, which is easy to cause adverse effects on the flavor; the high-speed centrifugation technique is fast in operation, but has high energy consumption and high equipment maintenance cost; the macroporous resin separation technique has a wide application range, but the resin structure has diversity, and the resin itself has certain toxicity, and its controllability and safety need to be improved; the membrane separation technique has a series of membrane pollution problems such as easy blocking of membrane holes and easy formation of adhesion layer on the membrane surface. It can be seen that the conventional clarification techniques have high energy consumption and are easy to affect the flavor of the enzymatic hydrolysate, which is not conducive to the development and production of flavoring.
[0004] Therefore, it is necessary to develop a green, simple and convenient clarification technique to realize the clarification of the shellfish enzymatic hydrolysate. SUMMARY
[0005] [TECHNICAL PROBLEM]
[0006] The conventional clarification techniques have high energy consumption and are easy to affect the flavor of the enzymatic hydrolysate, which is not conducive to the development and production of flavoring.
[0007] [TECHNICAL SCHEME]
[0008] In order to solve the above problems, the present application only controls the temperature change of the shellfish enzymatic hydrolysate, and the flocculation clarification of the shellfish enzymatic hydrolysate can be realized without adding a flocculant. The method of the present application is simple, has a wide application range, and the prepared shellfish enzymatic hydrolysate retains the original flavor to the greatest extent.
[0009] The first object of the present application is to provide a flocculation clarification method for shellfish enzymatic hydrolysate, comprising the following steps:
[0010] The shellfish enzymatic hydrolysate is frozen at-30~ -20℃ for 6~20 h, and then heated at 90~100℃ for 5~60 min. After standing until the flocculation is completely settled, filtration is performed to obtain a clarified solution of the enzymatic hydrolysate.
[0011] In one embodiment of the present application, the freezing treatment is freezing treatment at -30℃ for 18h; and the heating treatment is heating treatment at 95℃ for 10min.
[0012] In one embodiment of the present application, the heating treatment is water bath heating.
[0013] In one embodiment of the present application, the preparation method of the shellfish enzymatic hydrolysate comprises the following steps:
[0014] (1) taking shellfish meat, washing, maturing, homogenizing to obtain shellfish meat paste;
[0015] (2) adding water to the shellfish meat paste to obtain shellfish homogenate, and then using acid protease and complex protease to enzymatically hydrolyze the shellfish homogenate, inactivating the enzyme to obtain a shellfish enzymatic hydrolysate.
[0016] In one embodiment of the present application, the shellfish in step (1) comprises one of surf clam, mussel; and the surf clam is one of surf clam, white clam or green clam.
[0017] In one embodiment of the present application, the maturing in step (1) is to add water to the shellfish meat for steaming, and the steaming time is 40-50min; after steaming, the clam juice is filtered and the water is dried.
[0018] In one embodiment of the present application, the homogenization in step (1) is to homogenize the shellfish meat after maturing with water to obtain shellfish meat paste without obvious particles and with certain fluidity; wherein the mass ratio of the shellfish meat after maturing to water is 1:1-1.5.
[0019] In one embodiment of the present application, the mass ratio of the shellfish meat paste to water in step (2) is 1:1-2.
[0020] In one embodiment of the present application, the acid protease in step (2) has an enzyme activity of 1000 U / g, which is purchased from Beijing Solabio Science and Technology Co., Ltd.; and the complex protease has an enzyme activity of 2500 U / g, which is purchased from Beijing Solabio Science and Technology Co., Ltd.
[0021] In one embodiment of the present application, the enzymatic hydrolysis in step (2) is two-step enzymatic hydrolysis, the first step is to adjust the pH of the shellfish homogenate to 2.5-3.5, add 1.2-1.5% acid protease, and enzymatically hydrolyze at 43-47℃ for 55-65min; the second step is to adjust the pH of the enzymatic hydrolysate to 6.5-7.5, add 3.9-4.1% complex protease, and enzymatically hydrolyze at 48-52℃ for 175-185min to obtain the enzymatic hydrolysate; wherein the amount of the acid protease and the complex protease is relative to the mass percentage of the cooked shellfish meat.
[0022] In one embodiment of the present application, the enzyme inactivation in step (2) is by boiling water bath for 5-15 min.
[0023] A second object of the present application is to provide a method for preparing mollusk enzymatic hydrolysate powder, which comprises the following steps: first, preparing a clear solution by the method of the present application; and then, obtaining the mollusk enzymatic hydrolysate powder by spray drying the clear solution.
[0024] In one embodiment of the present application, the spray drying is by feeding the clear solution into the spray dryer through a peristaltic pump, wherein the inlet temperature of the peristaltic pump is 130-180°C, the outlet temperature is 70-80°C, and the peristaltic pump rate is 10-30 mL / min.
[0025] A third object of the present application is to provide a method for maximizing the retention of the flavor of mollusk enzymatic hydrolysate without affecting the flavor and achieving flocculation and clarification, which comprises the following steps:
[0026] freezing the mollusk enzymatic hydrolysate at -30 to -20°C for 6-20 h, then heating at 90-100°C for 5-60 min, standing until the flocculation is completely settled, and filtering to obtain the clear solution of the mollusk enzymatic hydrolysate with the flavor unaffected.
[0027] [Advantages]
[0028] (1) The present application can achieve flocculation and clarification of mollusk enzymatic hydrolysate without adding flocculants by only controlling the temperature change of the mollusk enzymatic hydrolysate, without significantly affecting the flavor of the enzymatic hydrolysate, and has great advantages in flavor retention of flavorings.
[0029] (2) The present application can obtain mollusk enzymatic hydrolysate with high clarity by filtering or low-speed centrifugation, which is easy to realize industrial large-scale centrifugation and can be widely used in actual industrial production.
[0030] (3) The present application can achieve flocculation and clarification by low-temperature freezing and high-temperature water bath, greatly improving the clarity and transparency of the mollusk enzymatic hydrolysate. BRIEF DESCRIPTION OF DRAWINGS
[0031] Figure 1 The flow chart of the flocculation and clarification method for mollusk enzymatic hydrolysate according to the present application.
[0032] Figure 2 The flocculation effect comparison chart of the sea cucumber enzymatic hydrolysate of Example 1 (A), Comparative Example 1 (B) and Comparative Example 2 (C).
[0033] Figure 3 The flocculation effect comparison chart of the sea cucumber enzymatic hydrolysate of Example 1 (A), Comparative Example 1 (B) and Comparative Example 2 (C) at room temperature for 20 min.
[0034] Figure 4 The absorbance value (OD742) of the clam enzymolysis clarified liquid obtained by filtering the clam enzymolysis liquid of Example 1, Comparative Example 1, Comparative Example 2 and Comparative Example 3 was measured at 742 nm. 742 nm ).
[0035] Figure 5 The absorbance value (OD742) of the clam enzymolysis clarified liquid obtained by centrifuging the clam enzymolysis liquid of Example 1, Comparative Example 1, Comparative Example 2 and Comparative Example 3 at different centrifugal speeds for 5 min was measured at 742 nm. 742 nm ).
[0036] Figure 6 The radar chart of the electronic nose response value of the clam enzymolysis liquid and the clam enzymolysis clarified liquid in Example 1.
[0037] Figure 7 The absorbance value (OD742) of the clam enzymolysis clarified liquid obtained by filtering the clam enzymolysis liquid of Example 1, Example 2 with low-temperature freezing time of 6 h (A), 12 h (B) and 20 h (C) was measured at 742 nm. 742 nm ).
[0038] Figure 8 The absorbance value (OD742) of the clam enzymolysis clarified liquid obtained by filtering the clam enzymolysis liquid of Example 1, Example 3 with high-temperature water bath temperature of 80℃ (A), 90℃ (B) and 100℃ (C) was measured at 742 nm. 742 nm ).
[0039] Figure 9 The absorbance value (OD742) of the clam enzymolysis clarified liquid obtained by filtering the clam enzymolysis liquid of Example 1, Example 4 with high-temperature water bath time of 5 min (A) and 15 min (B) was measured at 742 nm. 742 nm ).
[0040] Figure 10 The absorbance value (OD742) of the mussel enzymolysis liquid and the mussel enzymolysis clarified liquid in Example 5 was measured at 742 nm. 742 nm ). DETAILED DESCRIPTION
[0041] The preferred embodiments of the present application are described below, and it should be understood that the embodiments are for better explaining the present application and are not used to limit the present application.
[0042] Test method:
[0043] 1. Determination of the clarity of the filtrate:
[0044] The clarity of the clam enzymolysis clarified liquid obtained by filtering was determined by ultraviolet spectrophotometry.
[0045] The specific test method is as follows: 10 mL of the clam enzymolysis clear liquid obtained by standing filtration was determined by UV-5100B type ultraviolet spectrophotometer to determine the absorbance (OD) at 742 nm 742 nm ).
[0046] 2. Centrifugal supernatant clarity determination:
[0047] The ultraviolet spectrophotometry was used to test the clarity of the clam enzymolysis clear liquid obtained by centrifugation at different speeds.
[0048] The specific test method is as follows: 10 mL of the clam enzymolysis clear liquid obtained by standing filtration was determined by UV-5100B type ultraviolet spectrophotometer to determine the absorbance (OD) at 742 nm 742 nm ).
[0049] 3. Flavor test:
[0050] The electronic nose test method was used to test the electronic nose of the clam enzymolysis liquid and the clam enzymolysis clear liquid.
[0051] The specific test method is as follows: 5 mL of the clam enzymolysis liquid and the clam enzymolysis clear liquid were taken into sample bottles, respectively, and were detected by the electronic nose PEN3 system after standing at room temperature for 30 min.
[0052] The detection conditions are as follows: detector cleaning time 20 s, detection time 180 s, and sample flow rate 300 mL / min. The electronic nose is equipped with W1C, W5S, W3C, W6S, W5C, W1S, W1W, W2S, W2W, and W3S, 10 sensors
[0053] Raw materials used in the examples:
[0054] The acid protease has an enzyme activity of 1000 U / g, which is purchased from Beijing Solabio Technology Co., Ltd.;
[0055] The complex protease has an enzyme activity of 2500 U / g, which is purchased from Beijing Solabio Technology Co., Ltd.;
[0056] The clam used in the examples is flower clam;
[0057] Example 1
[0058] A flocculation clarification method for shellfish enzymolysis liquid, comprising the following steps:
[0059] (1) Preparation of clam enzymolysis liquid:
[0060] Take the clam meat, wash and remove impurities, add an appropriate amount of purified water, steam for 45 min, filter the clam juice, and control the moisture content;
[0061] Take 50 g of cooked clam meat, add 50 g of purified water, homogenize until there are no obvious particles, and obtain 100 g of clam meat paste;
[0062] Take 100 g of clam meat paste, add 150 g of purified water, and stir until uniform to obtain 250 g of clam homogenate. Then adjust the pH of the clam homogenate to 3 with 6 mol / L hydrochloric acid solution, add 0.72 g of acid protease, and enzymatically hydrolyze at 45 ℃ for 1 h. Then adjust the pH of the reaction system to 7 with 6 mol / L sodium hydroxide solution, add 1.99 g of complex protease, and enzymatically hydrolyze at 50 ℃ for 3 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated by boiling water bath for 10 min to obtain 250 g of clam enzymatic hydrolysate.
[0063] (2) Clarification method:
[0064] Freeze the clam enzymatic hydrolysate at -30 ℃ for 18 h, then treat it with high-temperature water bath at 95 ℃ for 10 min, and let it stand until the flocculation is completely settled. Then filter to obtain the clarified solution of the enzymatic hydrolysate.
[0065] Comparative Example 1
[0066] Adjust the low-temperature freezing in step (2) of Example 1 to low-temperature refrigeration, specifically refrigerate at 4 ℃ for 18 h, and the others remain the same as Example 1.
[0067] Comparative Example 2
[0068] Adjust the low-temperature freezing in step (2) of Example 1 to normal temperature storage, specifically store at 25 ℃ (normal temperature) for 18 h, and the others remain the same as Example 1.
[0069] Test the performance of the obtained clarified solution of the enzymatic hydrolysate, and the test results are as follows:
[0070] Figure 2 The figure is the flocculation effect comparison of the clam enzymatic hydrolysate of Example 1 (A), Comparative Example 1 (B) and Comparative Example 2 (C). Figure 3 The figure is the flocculation effect comparison of the clam enzymatic hydrolysate of Example 1 (A), Comparative Example 1 (B) and Comparative Example 2 (C) after standing at normal temperature for 20 min. From Figure 2 and Figure 3 It can be seen that: Example 1 quickly appears flocculation, and then precipitates, realizing the clarification treatment of the enzymatic hydrolysate; while the flocculation of the clam enzymatic hydrolysate of Comparative Example 1 and Comparative Example 2 is mainly in small particles, and the flocculation clarification effect is not obvious, and the filtrate has low clarity after standing and filtering.
[0071] Comparative Example 3
[0072] Step (2) of Example 1 is omitted. The clam enzymatic hydrolysate obtained in step (1) is placed at 25°C (room temperature) and stored in the same way as in Example 1.
[0073] The results showed that the clam enzymatic hydrolysate did not flocculate, and the filtrate had low clarity after standing and filtration.
[0074] The clear solution of the obtained enzymatic hydrolysate was subjected to performance testing, and the test results are as follows:
[0075] Figure 4 The absorbance (OD) value at 742 nm of the clam enzymatic hydrolysate obtained by filtration of the clam enzymatic hydrolysate from Example 1, Comparative Example 1, Comparative Example 2, and Comparative Example 3 is shown in Figure 1. 742 nm ).from Figure 4 It can be seen that: Example 1 OD 742 nm =0.285, Comparative Example 1 OD 742 nm =1.443, Comparative Example 2 OD 742 nm =1.657, Comparative Example 3 OD 742 nm =1.715; It can be seen that the clam enzymatic hydrolysis clarification liquid obtained by static filtration in Example 1 has the highest clarity and obvious clarification effect.
[0076] Figure 5 The absorbance (OD) values at 742 nm of the clam enzymatic hydrolysates obtained by centrifuging the clam enzymatic hydrolysates of Example 1, Comparative Example 1, Comparative Example 2, and Comparative Example 3 at different centrifuge speeds for 5 min are shown. 742 nm ).from Figure 5 It can be seen that, at different centrifugation speeds, the OD of the clam enzymatic hydrolysis clarified solution obtained from centrifugation in Example 1 varies. 742 nm All were lower than those of Comparative Examples 1, 2, and 3. Taking 1000 rpm as an example, the OD of Example 1 was... 742 nm =0.843, Comparative Example 1 OD 742 nm =1.355, Comparative Example 2 OD 742 nm =1.441, Comparative Example 3 OD 742 nm =1.581. The clam enzymatic hydrolysis solution obtained by centrifugation in Example 1 had the highest clarity; the OD of the supernatant obtained by centrifugation at 3000 rpm in Example 1 was... 742 nm The OD value was 0.330. The OD values of the supernatants obtained from centrifugation at 8000 rpm for Comparative Examples 1, 2, and 3 were... 742 nm The values were 0.344, 0.382, and 0.499, respectively. The clarification effect of the supernatant obtained by centrifugation at 3000 rpm in Example 1 was superior to that of the supernatants obtained by centrifugation at 8000 rpm in Comparative Examples 1, 2, and 3, with a significant clarification effect.
[0077] Figure 6 This is a radar chart showing the electronic nose response values of the clam enzymatic hydrolysate and the clarified clam enzymatic hydrolysate from Example 1. From... Figure 6 It can be seen that the response values of the clam enzymatic hydrolysate and the clarified clam enzymatic hydrolysate are not significantly different. This indicates that the clarification method of the present invention does not have a significant impact on the flavor of the enzymatic hydrolysate.
[0078] Example 2
[0079] The low-temperature freezing time in step (2) of Example 1 was adjusted to 6 h, 12 h, and 20 h, while the other preservation methods were the same as in Example 1.
[0080] The results showed that flocculation and subsequent precipitation occurred in all cases in Example 2, thus achieving the clarification treatment of the enzymatic hydrolysate.
[0081] The clear solution of the obtained enzymatic hydrolysate was subjected to performance testing, and the test results are as follows:
[0082] Figure 7 The absorbance (OD) values at 742 nm of the clam enzymatic hydrolysate obtained by filtration of the clam enzymatic hydrolysate after low-temperature freezing times of 6 h (A), 12 h (B), and 20 h (C) in Examples 1 and 2. 742 nm ).from Figure 7 It can be seen that: Example 1 OD 742 nm =0.285, Example 2 (A) OD 742 nm =1.376, Example 2 (B) OD 742 nm =0.568, Example 2 (C) OD 742 nm =0.300, indicating that the clam enzymatic hydrolysis clarification liquid obtained by static filtration in Example 1 has the highest clarity and the most obvious clarification effect.
[0083] Example 3
[0084] Adjust the temperature of the high-temperature water bath in step (2) of Example 1 to 80℃, 90℃, and 100℃, and keep the rest the same as in Example 1.
[0085] The results showed that flocculation and subsequent precipitation occurred in all three examples, thus achieving the clarification of the enzymatic hydrolysate.
[0086] The clear solution of the obtained enzymatic hydrolysate was subjected to performance testing, and the test results are as follows:
[0087] Figure 8 The absorbance (OD) values at 742 nm of the clam enzymatic hydrolysate obtained by filtration of the clam enzymatic hydrolysate obtained from high-temperature water bath temperatures of 80℃ (A), 90℃ (B), and 100℃ (C) in Examples 1 and 3. 742 nm ).from Figure 8 It can be seen that: Example 1 OD 742 nm=0.285, Example 3 (A) OD 742 nm =0.626, Example 3 (B) OD 742 nm =0.332, Example 3 (C) OD 742 nm =0.344, indicating that the clam enzymatic hydrolysis clarification liquid obtained by static filtration in Example 1 has the highest clarity and the most obvious clarification effect.
[0088] Example 4
[0089] Adjust the high-temperature water bath time in step (2) of Example 1 to 5 min and 15 min, and keep the rest the same as in Example 1.
[0090] The results showed that flocculation and subsequent precipitation occurred in all four examples, thus achieving the clarification of the enzymatic hydrolysate.
[0091] Figure 9 The absorbance (OD) value at 742 nm of the clam enzymatic hydrolysate obtained by filtration after high-temperature water bath time of 5 min (A) and 15 min (B) in Examples 1 and 4 is shown. 742 nm ).from Figure 9 It can be seen that: Example 1 OD 742 nm =0.285, Example 4 (A) OD 742 nm =0.616, Example 4 (B) OD 742 nm =0.290, indicating that the clam enzymatic hydrolysis clarification liquid obtained by static filtration in Example 1 has the highest clarity and obvious clarification effect.
[0092] Example 5
[0093] The clams in Example 1 were replaced with mussels, while everything else remained the same as in Example 1.
[0094] The results showed that flocculation and subsequent precipitation occurred in Example 5, thus achieving the clarification of the enzymatic hydrolysate.
[0095] Figure 10 The absorbance values (OD values) at 742 nm of the mussel enzymatic hydrolysate from Example 5 and the clarified mussel enzymatic hydrolysate obtained after clarification treatment are shown. 742 nm ).from Figure 10 It can be seen that the OD of the mussel enzymatic hydrolysate... 742 nm =1.327, OD of mussel enzymatic hydrolysis clarified solution 742 nm =0.169.
[0096] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.
Claims
1. A method for flocculation and clarification of shellfish enzymatic hydrolysate, characterized in that, Includes the following steps: The shellfish enzymatic hydrolysate was frozen at -30°C for 18 h, then heated at 95°C for 10 min, allowed to stand until the flocs had completely settled, and filtered to obtain a clear solution of the enzymatic hydrolysate; the heating treatment was water bath heating. The preparation method of the shellfish enzymatic hydrolysate includes the following steps: (1) Take shellfish meat, wash, cook and homogenize to obtain shellfish meat paste; the homogenization is to add water to the cooked shellfish meat and homogenize it until there are no obvious particles and the shellfish meat paste has a certain fluidity; wherein, the mass ratio of cooked shellfish meat to water is 1:1~1.
5. (2) Water was added to the shellfish meat paste to obtain a shellfish homogenate; then acidic protease and complex protease were used to enzymatically hydrolyze the shellfish homogenate to inactivate the enzymes and obtain a shellfish hydrolysate; the mass ratio of shellfish meat paste to water was 1:1~2. The enzymatic hydrolysis is a two-step process. The first step involves adjusting the pH of the shellfish homogenate to 2.5–3.5, adding 1.2–1.5% acidic protease, and hydrolyzing at 43–47 °C for 55–65 min. The second step involves adjusting the pH of the hydrolysate to 6.5–7.5, adding 3.9–4.1% complex protease, and hydrolyzing at 48–52 °C for 175–185 min to obtain the hydrolysate.
2. The method according to claim 1, characterized in that, The enzyme inactivation in step (2) is performed by boiling water bath for 5-15 minutes.
3. A method for preparing enzymatically hydrolyzed shellfish powder, characterized in that, The method involves first preparing a clear solution using the method described in any one of claims 1 to 2; then spray-drying the clear solution to obtain shellfish enzymatic hydrolysate.
4. The method according to claim 3, characterized in that, The spray drying process involves feeding a clarified solution into a spray dryer via a peristaltic pump. The inlet temperature of the peristaltic pump is 130~180℃, the outlet temperature is 70~80℃, and the pump speed is 10~30mL / min.