Oil-soluble lipase preparation as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of enzyme preparations, and particularly provides an oil-soluble lipase preparation which comprises lipase powder and an emulsifier in a mass ratio of 1000: (6.5-46), the lipase powder comprises lipase and a stabilizer. The invention also provides a preparation method of the oil-soluble lipase preparation, and the oil-soluble lipase preparation prepared by the method provided by the invention has a good effect of emulsifying an oil-water system, and can promote lipolysis and greatly improve the utilization rate of lipase.

Description

Technical Field

[0001] The invention belongs to the technical field of enzyme preparations, and in particular relates to an oil-soluble lipase preparation and a preparation method and application thereof. Background Art

[0002] Lipase is widely present in animals, plants and microorganisms. The pancreas and adipose tissue of higher animals contain more lipase in animals. Intestinal fluid contains a small amount of lipase to supplement the insufficient fat digestion of pancreatic lipase. The gastric juice of carnivorous animals contains a small amount of butyric acid glycerol esterase. In animals, various lipases control the processes of digestion, absorption, fat reconstruction and lipoprotein metabolism; the lipase content in bacteria, fungi and yeast is even richer.

[0003] Emulsifier is an auxiliary agent that can stabilize emulsions and improve mixing, molding, drying and other processes. Adding emulsifiers to feed can improve the characteristics of oil-water mixing, prevent emulsion precipitation and oil-water separation, thereby improving the stability of the feed. Emulsifiers can also promote the dispersion of various additives in the feed, thereby improving the miscibility of the feed. In addition, emulsifiers can increase the sticky substances in the feed during feed processing, thereby improving the molding of the feed. Emulsifiers can make the water in the feed more evenly distributed, thereby improving the dryness of the feed.

[0004] Common emulsifiers are usually divided into the following categories: phospholipid emulsifiers: including soybean lecithin, lecithin and lysolecithin, etc.; food emulsifiers: including monoglycerides, sucrose esters, etc., which were first used in food, and now some are used in feed; chemical emulsifiers: including span, tween, etc., this type of emulsifier has a good emulsification effect, but has not yet been used in feed; bile acid emulsifiers: bile acid emulsifiers are a series of steroid substances produced during the metabolism of cholesterol in animals. They have surface activity. One end of its molecular structure is an alkyl group, which is lipophilic and can combine with oils and fats. The other end is a hydroxyl group and a carboxyl group, which is hydrophilic. The most important function of bile acid in animals is to promote the digestion and absorption of fats and fat-soluble substances.

[0005] Emulsifiers acting on feed can play a bridging role between fat and pancreatic lipase, increase the contact area between fat and pancreatic lipase, and thus create conditions for fat to be decomposed by pancreatic lipase. Existing feed emulsifiers such as phospholipids are also fats. When the amount added is small, the emulsification effect is poor. When the amount added is large, it is not easy for animals to absorb. Monoglycerides, sucrose esters, bile acids, etc. are insoluble in water, and the pure emulsification effect of oil and water is very poor. The oil-water separation time is very short, resulting in uneven mixing with lipase and oil, and the oil cannot be decomposed into fatty acids with maximum efficiency. Summary of the invention

[0006] The purpose of the present invention is to overcome the problem that when the existing feed-type emulsifier acts on lipase, the emulsification effect is poor, resulting in uneven mixing of lipase and oil, and the oil cannot be decomposed into fatty acids with maximum efficiency.

[0007] To this end, the present invention provides an oil-soluble lipase preparation, comprising a lipase powder and an emulsifier in a mass ratio of 1000:(6.5-46); the lipase powder comprises a lipase, a stabilizer and a soluble carrier.

[0008] Specifically, the emulsifier includes soybean lecithin, bile acid, and sucrose fatty acid ester in a mass ratio of (5-25): (0.5-1): (1-20).

[0009] Specifically, the hlb value of the above emulsifier is 6-8.

[0010] Specifically, the stabilizer includes trehalose or pullulan; and the soluble carrier includes maltodextrin.

[0011] The present invention also provides a method for preparing the above oil-soluble lipase preparation, comprising the following steps:

[0012] (1) taking liquid lipase, adding a stabilizer and a soluble carrier, and drying to obtain lipase powder;

[0013] (2) Mixing lipase powder with an emulsifier to obtain an oil-soluble lipase preparation.

[0014] Specifically, the above step (1) is as follows: taking liquid lipase with an enzyme activity of 5000-100000u / ml, adding 10-25% of a soluble carrier and 2-5% of a stabilizer based on the mass of the liquid lipase, and drying to obtain lipase powder.

[0015] Specifically, the above step (2) is specifically as follows: mixing the powder with an emulsifier to obtain an emulsified powder, mixing the emulsified powder with a carrier and sodium sulfate in a mass ratio of (2-4): (1-5): (1-2), adding a binder, wet granulating, and drying to obtain an oil-soluble lipase preparation.

[0016] Specifically, the carrier includes corn starch.

[0017] Specifically, based on the sum of the mass of the emulsified powder, the carrier and the sodium sulfate, the added amount of the above-mentioned binder is 15%-20%.

[0018] Specifically, in the above step (2), the moisture content of the oil-soluble lipase preparation after drying is controlled to be below 5%.

[0019] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0020] The oil-soluble lipase preparation provided by the invention has good effect of emulsifying oil-water system, can promote fat decomposition, and greatly improve the utilization rate of lipase. DETAILED DESCRIPTION

[0021] The technical scheme in the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Although the representative embodiments of the present invention have been described in detail, it will be understood by those skilled in the art that various modifications and changes can be made to the present invention without departing from the scope of the present invention. Therefore, the scope of the present invention should not be limited to the embodiments, but should be limited by the attached claims and their equivalents.

[0022] The invention provides an oil-soluble lipase preparation, comprising a lipase powder and an emulsifier in a mass ratio of 1000:(6.5-46); the lipase powder comprises a lipase, a stabilizer and a soluble carrier.

[0023] The emulsifier preferably comprises soybean lecithin, bile acid, and sucrose fatty acid ester in a mass ratio of 1000:(5-25):(0.5-1):(1-20), and the HLB value of the soybean lecithin, bile acid, and sucrose fatty acid ester after mixing is 6-8.

[0024] The stabilizer preferably includes trehalose or pullulan; the soluble carrier preferably includes maltodextrin.

[0025] The present invention also provides a method for preparing the above oil-soluble lipase preparation, comprising the following steps:

[0026] (1) taking a liquid lipase with an enzyme activity of 5000-100000 u / ml, adding 2-5% of a stabilizer and 10-25% of a soluble carrier based on the mass of the liquid lipase, and then drying to obtain a lipase powder;

[0027] (2) mixing lipase powder, soybean lecithin, bile acid, and sucrose fatty acid ester in a mass ratio of 1000:(5-25):(0.5-1):(1-20) to obtain an emulsified powder; wherein the ratio of soybean lecithin, bile acid, and sucrose fatty acid ester is such that the HLB value of the mixed emulsifier is within the range of 6-8;

[0028] Take the emulsified powder, mix it with the carrier and sodium sulfate in the mass ratio of (2-4): (1-5): (1-2), add 15%-20% of the binder, use a wet mixer to mix to obtain a granulation soft material, put the soft material into a swing granulator or a screw extruder for granulation, and obtain wet granules. The binder is preferably a maltodextrin aqueous solution with a mass concentration of 10% or a sodium carboxymethyl cellulose aqueous solution with a mass concentration of 1%.

[0029] The wet granules are dried, the drying temperature is controlled at 45-55° C., and the moisture content of the granules is controlled below 5%, to obtain an oil-soluble lipase preparation.

[0030] The oil-soluble lipase preparation of the present invention, its preparation method and application effects are studied below through specific examples.

[0031] Embodiment 1:

[0032] This embodiment provides an oil-soluble lipase preparation, which is prepared by the following steps:

[0033] (1) taking liquid lipase, adding 25% maltodextrin and 2% trehalose based on the mass of the lipase therein, and drying the liquid lipase using a spray drying tower to obtain a lipase powder;

[0034] (2) mixing lipase powder, soybean lecithin, bile acid, and sucrose fatty acid ester (hlb value 1) in a mass ratio of 1000:25:1:5 to obtain an emulsified powder; wherein the hlb value of the emulsifier after the soybean lecithin, bile acid, and sucrose fatty acid ester are mixed is 7.0;

[0035] Take the emulsified powder, mix it with corn starch and sodium sulfate in a mass ratio of 4:3:3, add 15% of the binder, use a wet mixer to mix to obtain a granulation soft material, put the soft material into a swing granulator for granulation, and obtain wet granules. The binder is a 10% maltodextrin aqueous solution.

[0036] The wet granules were dried using a fluidized bed dryer, the drying temperature was controlled at 45°C, and the granule moisture content was controlled at 4.5%, to obtain an oil-soluble lipase preparation.

[0037] Embodiment 2:

[0038] This embodiment provides an oil-soluble lipase preparation, which is prepared by the following steps:

[0039] (1) taking liquid lipase, adding 10% maltodextrin and 5% trehalose based on the mass of the lipase therein, and drying the liquid lipase using a spray drying tower to obtain a lipase powder;

[0040] (2) mixing lipase powder, soybean lecithin, bile acid, and sucrose fatty acid ester (hlb value 1) in a mass ratio of 1000:20:0.5:5 to obtain an emulsified powder; wherein the hlb value of the emulsifier after the soybean lecithin, bile acid, and sucrose fatty acid ester are mixed is 6.4;

[0041] Take the emulsified powder, mix it with corn starch and sodium sulfate in a mass ratio of 4:2:4, add 20% of the binder, use a wet mixer to mix to obtain a granulation soft material, put the soft material into a swing granulator for granulation, and obtain wet granules. The binder is a 1% sodium carboxymethyl cellulose aqueous solution.

[0042] The wet granules were dried using a fluidized bed dryer, the drying temperature was controlled at 45°C, and the granule moisture content was controlled at 4.7%, to obtain an oil-soluble lipase preparation.

[0043] Embodiment 3:

[0044] This embodiment provides an oil-soluble lipase preparation, which is prepared by the following steps:

[0045] (1) taking liquid lipase, adding 15% maltodextrin and 5% pullulan based on the mass of the lipase therein, and drying the liquid lipase using a spray drying tower to obtain a lipase powder;

[0046] (2) mixing lipase powder, soybean lecithin, bile acid, and sucrose fatty acid ester (hlb value 1) in a mass ratio of 1000:10:0.5:1 to obtain an emulsified powder; wherein the hlb value of the emulsifier after the soybean lecithin, bile acid, and sucrose fatty acid ester are mixed is 8.0;

[0047] Take the emulsified powder, mix it with corn starch and sodium sulfate in a mass ratio of 4:4:2, add 15% of the binder, use a wet mixer to mix to obtain a granulation soft material, put the soft material into a swing granulator for granulation, and obtain wet granules. The binder is a 1% sodium carboxymethyl cellulose aqueous solution.

[0048] The wet granules were dried using a fluidized bed dryer, the drying temperature was controlled at 55°C, and the granule moisture content was controlled at 3.4%, to obtain an oil-soluble lipase preparation.

[0049] Comparative Example 1:

[0050] This comparative example provides a lipase preparation, which is prepared by the following steps:

[0051] (1) Take liquid lipase, add 25% maltodextrin and 2% trehalose, and dry it using a spray dryer to obtain lipase powder.

[0052] (2) Take lipase powder, mix it with corn starch and sodium sulfate in a mass ratio of 4:3:3, add 15% of a binder (10% maltodextrin aqueous solution), and mix it using a wet mixer to obtain a granulation soft material. Put the soft material into a swing granulator to obtain wet granules.

[0053] The wet granules were dried using a fluidized bed dryer, the drying temperature was controlled at 45°C, and the granule moisture was controlled at an actual measured 4.8%, to obtain a lipase preparation.

[0054] Comparative Example 2:

[0055] This comparative example provides a lipase preparation, which is prepared by the following steps:

[0056] (1) Take liquid lipase, add 25% maltodextrin and 2% trehalose, and dry it using a spray dryer to obtain lipase powder.

[0057] (2) Take lipase powder, mix it with corn starch and sodium sulfate in a mass ratio of 4:2:4, add 15% of a binder (1% sodium carboxymethyl cellulose aqueous solution), and mix it using a wet mixer to obtain a granulation soft material. The soft material is put into a swing granulator to obtain wet granules.

[0058] The wet granules were dried using a fluidized bed dryer, the drying temperature was controlled at 55° C., and the granule moisture was controlled at an actual measured 3.4% to obtain a lipase preparation.

[0059] Comparative Example 3:

[0060] This comparative example provides a lipase preparation, which is prepared by the following steps:

[0061] (1) taking liquid lipase, adding 25% maltodextrin and 2% trehalose based on the mass of the lipase therein, and drying the liquid lipase using a spray drying tower to obtain a lipase powder;

[0062] (2) mixing lipase powder, soybean lecithin, bile acid, and sucrose fatty acid ester (hlb value 1) in a mass ratio of 1000:20:1:10 to obtain an emulsified powder; wherein the hlb value of the emulsifier after the soybean lecithin, bile acid, and sucrose fatty acid ester are mixed is 5;

[0063] Take the emulsified powder, mix it with corn starch and sodium sulfate in a mass ratio of 4:3:3, add 15% of the binder, use a wet mixer to mix to obtain a granulation soft material, put the soft material into a swing granulator for granulation, and obtain wet granules. The binder is a 10% maltodextrin aqueous solution.

[0064] The wet granules were dried using a fluidized bed dryer, the drying temperature was controlled at 45°C, and the granule moisture content was controlled at 4.5%, to obtain an oil-soluble lipase preparation.

[0065] Comparative Example 4:

[0066] This comparative example provides a lipase preparation, which is prepared by the following steps:

[0067] (1) taking liquid lipase, adding 25% maltodextrin and 2% trehalose based on the mass of the lipase therein, and drying the liquid lipase using a spray drying tower to obtain a lipase powder;

[0068] (2) mixing lipase powder, soybean lecithin, bile acid, and sucrose fatty acid ester (hlb value 1) in a mass ratio of 1000:20:2:5 to obtain an emulsified powder; wherein the hlb value of the emulsifier after the soybean lecithin, bile acid, and sucrose fatty acid ester are mixed is 9;

[0069] Take the emulsified powder, mix it with corn starch and sodium sulfate in a mass ratio of 4:3:3, add 15% of the binder, use a wet mixer to mix to obtain a granulation soft material, put the soft material into a swing granulator for granulation, and obtain wet granules. The binder is a 10% maltodextrin aqueous solution.

[0070] The wet granules were dried using a fluidized bed dryer, the drying temperature was controlled at 45°C, and the granule moisture content was controlled at 4.5%, to obtain an oil-soluble lipase preparation.

[0071] Embodiment 4:

[0072] In this example, the lipase preparations obtained in Examples 1-3 and Comparative Examples 1-4 were evaluated for application, and the evaluation method was as follows.

[0073] 1. Evaluation of emulsification performance

[0074] In the experimental group, 10 g of each lipase preparation provided in Examples 1-3 was added to 100 ml of a 1:1 oil-water mixed solution, stirred using a high-speed homogenizer until a uniform phase was formed, and allowed to stand at room temperature to observe the time for oil-water separation to occur.

[0075] The control group took 10g of each lipase preparation provided in Comparative Examples 1-4, added them to 100ml of a 1:1 oil-water mixed solution, stirred with a high-speed homogenizer until a uniform phase was formed, and left to stand at room temperature to observe the time for oil-water stratification. At the same time, 10g of the lipase preparation provided in Comparative Examples 1-2 was compounded with soybean lecithin, sucrose fatty acid ester (selected with an HLB value of 7), and bile acid, respectively, added to 100ml of a 1:1 oil-water mixed solution, stirred with a high-speed homogenizer until a uniform phase was formed, and left to stand at room temperature to observe the time for oil-water stratification.

[0076] The experimental results are shown in Table 1.

[0077] Table 1 Emulsification performance evaluation results

[0078]

[0079]

[0080] It can be seen from the results in Table 1 that the emulsification performance of the experimental group is much stronger than that of the control group. The emulsification effect of the lipase preparation provided by the present invention is greatly improved compared with the conventional technical route.

[0081] 2. Evaluation of the effect of lipase application

[0082] Take the lipase preparations provided in Examples 1-3 and Comparative Examples 1-4 and a commercially available lipase powder enzyme preparation (Source Leaf Bio S10035, Comparative Example 5), dilute the enzyme activity to 10u / g, take 1g of the diluted sample and add it to 10g of an oil-water mixed solution (oil-water ratio of 1:1), use a high-speed homogenizer to stir until a uniform phase is formed, place it in a 40°C constant temperature water bath to react, and use 100% ethanol solution to terminate the reaction at 10min, 60min, 120min, and 240min, respectively. After adding 2 drops of phenolphthalein, use 0.1mol / L KOH solution to titrate, and record the amount of KOH solution required for titration. The experimental results are shown in Table 2.

[0083] Table 2 Evaluation results of lipase application effect

[0084] 10min 60min 120min 240min Example 1 2.3ml 5.5ml 8.4ml 10.5ml Example 2 2.1ml 5.6ml 9ml 11.7ml Example 3 2.5ml 6.1ml 9.3ml 12.1ml Comparative Example 1 1.4ml 2.8ml 3.8ml 4.7ml Comparative Example 2 1.8ml 3.9ml 5.2ml 6.0ml Comparative Example 3 1.9ml 4.6ml 7.7ml 8.3ml Comparative Example 4 1.8ml 4.2ml 7.2ml 7.9ml Comparative Example 5 1.5ml 3.2ml 4.4ml 5.1ml

[0085] As can be seen from Table 2, the efficiency of the lipase preparation provided by the embodiment of the present invention in decomposing fat into fatty acids is much higher than that of the control group. The application effect of the lipase preparation prepared by the method of the present invention is greatly improved compared with the conventional technical route, and the utilization rate of lipase can be greatly improved.

[0086] The above examples are merely illustrative of the present invention and do not constitute a limitation on the protection scope of the present invention. All designs that are the same or similar to the present invention fall within the protection scope of the present invention.

Claims

1. An oil-soluble lipase preparation, characterized in that: The invention comprises lipase powder and emulsifier in a mass ratio of 1000:(6.5-46); the lipase powder comprises lipase, a stabilizer and a soluble carrier.

2. The oil-soluble lipase preparation according to claim 1, characterized in that: The emulsifier comprises soybean lecithin, bile acid and sucrose fatty acid ester in a mass ratio of (5-25):(0.5-1):(1-20).

3. The oil-soluble lipase preparation according to claim 2, characterized in that: The hlb value of the emulsifier is 6-8.

4. The method for preparing the oil-soluble lipase preparation according to any one of claims 1 to 3, characterized in that: The following steps are involved: (1) taking liquid lipase, adding a stabilizer and a soluble carrier, and drying the mixture to obtain lipase powder; (2) Mixing lipase powder with an emulsifier to obtain an oil-soluble lipase preparation.

5. The method for preparing the oil-soluble lipase preparation according to claim 4, characterized in that: The step (1) specifically comprises: taking liquid lipase with an enzyme activity of 5000-100000u / ml, adding 10-25% of a soluble carrier and 2-5% of a stabilizer based on the mass of the liquid lipase, and drying to obtain lipase powder.

6. The method for preparing the oil-soluble lipase preparation according to claim 4, characterized in that: The step (2) specifically comprises: mixing the powder with an emulsifier to obtain an emulsified powder, mixing the emulsified powder with a carrier and sodium sulfate in a mass ratio of (2-4): (1-5): (1-2), adding a binder, wet granulating, and drying to obtain an oil-soluble lipase preparation.

7. The method for preparing the oil-soluble lipase preparation according to claim 6, characterized in that: The carrier includes corn starch.

8. The method for preparing the oil-soluble lipase preparation according to claim 6, characterized in that: The amount of the binder added is 15%-20% based on the sum of the mass of the emulsified powder, the carrier and the sodium sulfate.

9. The method for preparing the oil-soluble lipase preparation according to claim 6, characterized in that: In the step (2), the moisture content of the oil-soluble lipase preparation after drying is controlled below 5%.

10. Use of the oil-soluble lipase preparation according to any one of claims 1 to 3 in feed additives.

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