CN111876451B - Method for extracting amino acid from chicken manure - Google Patents

Method for extracting amino acid from chicken manure Download PDF

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CN111876451B
CN111876451B CN202010793284.XA CN202010793284A CN111876451B CN 111876451 B CN111876451 B CN 111876451B CN 202010793284 A CN202010793284 A CN 202010793284A CN 111876451 B CN111876451 B CN 111876451B
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李建芬
徐先洲
何貟
申文娟
喻玖宏
祝苑
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Wuhan Polytechnic University
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Abstract

The invention provides a method for extracting amino acid from chicken manure, which comprises the following steps: s1, preparing a chicken manure fermentation stock solution for later use; s2, adding alkaline protease into the chicken manure fermentation stock solution, and hydrolyzing to obtain a primary hydrolysate for later use; s3, centrifuging and filtering the primary hydrolysate to obtain a solution a for later use; s4, deodorizing the solution a to obtain a solution b for later use; s5, preparing a double aqueous phase system, and mixing the solution b with the double aqueous phase system to obtain a solution c with obvious double aqueous phases for later use; s6, enabling the upper organic phase of the solution c to pass through an ion exchange column, eluting with deionized water until the organic phase is colorless, and then eluting the ion exchange column with ammonia water to obtain a solution d for later use; s7, heating the solution d in a water bath, and then concentrating in vacuum to obtain the amino acid. The invention provides a method for extracting amino acid from chicken manure, which has the advantages of simple operation, rapidness, high efficiency and the like.

Description

一种从鸡粪中提取氨基酸的方法A kind of method for extracting amino acid from chicken manure

技术领域technical field

本发明涉及固废资源化领域。更具体地说,本发明涉及一种从鸡粪中提取氨基酸的方法。The invention relates to the field of solid waste recycling. More specifically, the present invention relates to a method for extracting amino acids from chicken manure.

背景技术Background technique

伴随着经济的快速发展,人民的生活水平日益提高,对肉质蛋白的需求量也日益渐升,大型养殖场的出现造成相对应的污染。由于鸡粪堆积在周围环境中,粪便中的挥发性物质会造成空气污染,引来蚊蝇以及寄生虫;鸡粪中的液体会渗透或随着降雨而进入地下水,污染周围的水源;堆积的鸡粪易结块,不利于作物的生长,影响环境。因此,环境工作者们近年来正在发展绿色环境友好的家禽粪便废弃物资源化治理手段。With the rapid economic development, people's living standards are improving day by day, and the demand for meat protein is also increasing day by day. The emergence of large farms has caused corresponding pollution. Due to the accumulation of chicken manure in the surrounding environment, the volatile substances in the manure will cause air pollution, attracting mosquitoes and parasites; the liquid in the chicken manure will penetrate or enter the groundwater with rainfall, polluting the surrounding water sources; Chicken manure is easy to agglomerate, which is not conducive to the growth of crops and affects the environment. Therefore, environmental workers are developing green and environment-friendly poultry manure waste resource management methods in recent years.

鉴于双水相系统可以提供较温和的操作条件,降低目标产物的伤害或减少小产物的不稳定/易变性。相比较传统的水-有机溶剂萃取法,双水相的界面应力远低于前者,使得被萃取的产物受到较小的伤害。由于游离的氨基酸会向聚合物的一层中聚集,有利于两相中氨基酸浓度的增加而到达分离目的。目前的文献报道中的氨基酸提取方法主要有醇提法、水提法、乙醇微波萃取法和超声波水提法等,相关的报道实例较少。施丰成等人做的从烟草工业废弃烟末中提取游离氨基酸的方法,利用PEG聚合物和硫酸铵盐形成的双水相系统并结合闪失提取器提取废弃烟末颗粒中的氨基酸,较好的纯度可以达到96%左右。In view of the fact that the two-phase system can provide milder operating conditions, reduce the damage of target products or reduce the instability/mutability of small products. Compared with the traditional water-organic solvent extraction method, the interfacial stress of the aqueous two phase is much lower than the former, so that the extracted product suffers less damage. Since the free amino acids will aggregate into one layer of the polymer, it is beneficial to increase the concentration of amino acids in the two phases to achieve the purpose of separation. The amino acid extraction methods currently reported in the literature mainly include alcohol extraction, water extraction, ethanol microwave extraction and ultrasonic water extraction, etc. There are few examples of related reports. The method for extracting free amino acids from waste tobacco powder in the tobacco industry by Shi Fengcheng et al. utilizes a two-phase system formed by PEG polymer and ammonium sulfate salt combined with a flash-loss extractor to extract amino acids in waste tobacco powder particles, with better purity. Can reach around 96%.

在鸡粪排泄物中含有16%(质量分数)左右未被及时消化和吸收的蛋白质、多肽和游离氨基酸,一座10万只养殖规模的现代化养鸡场每天可以产生8吨左右的鸡粪,及时的提取其中的氨基酸,每天的产量1吨左右。Chicken manure contains about 16% (mass fraction) of proteins, polypeptides and free amino acids that have not been digested and absorbed in time. A modern chicken farm with a breeding scale of 100,000 can produce about 8 tons of chicken manure every day. The extraction of amino acids, the daily output is about 1 ton.

发明内容SUMMARY OF THE INVENTION

本发明的目的是提供一种具有操作简单、快速高效等优点的从鸡粪中提取氨基酸的方法。The purpose of the present invention is to provide a method for extracting amino acids from chicken manure, which has the advantages of simple operation, rapidity and high efficiency.

为了实现根据本发明的这些目的和其它优点,提供了一种从鸡粪中提取氨基酸的方法,包括以下步骤:In order to achieve these objects and other advantages according to the present invention, a method for extracting amino acids from chicken manure is provided, comprising the following steps:

S1、制备鸡粪发酵原液,备用;S1, prepare chicken manure fermentation stock solution, standby;

S2、在所述鸡粪发酵原液中加入碱性蛋白酶,对其进行水解,得到初步水解液,备用;S2, in described chicken manure fermentation stock solution, add alkaline protease, it is hydrolyzed, obtain preliminary hydrolyzate, standby;

S3、将所述初步水解液离心并过滤后,去除部分不溶的杂质和蛋白酶,得到溶液a,备用,去除的杂质和蛋白酶可用于制作化肥;S3, after described preliminary hydrolyzate is centrifuged and filtered, remove partially insoluble impurities and protease to obtain solution a, for subsequent use, and the removed impurities and protease can be used to make chemical fertilizers;

S4、将所述溶液a进行除臭处理,得到溶液b,备用;S4, carry out deodorization treatment to described solution a, obtain solution b, standby;

S5、制备双水相系统,并将所述溶液b和所述双水相系统混合,得到形成明显的双水相的溶液c,备用;S5, prepare a two-phase system, and mix the solution b with the two-phase system to obtain a solution c that forms an obvious two-phase system, for subsequent use;

S6、将所述溶液c的上层有机相过离子交换柱,用去离子水洗脱至无色,再使用氨水洗脱所述离子交换柱得到溶液d,备用;S6, the upper organic phase of the solution c is passed through the ion exchange column, eluted with deionized water until colorless, and then the ion exchange column is eluted with ammonia water to obtain solution d, for subsequent use;

S7、将所述溶液d进行水浴加热,然后真空浓缩,即制得所述氨基酸。S7. The solution d is heated in a water bath, and then concentrated in vacuo to obtain the amino acid.

本发明所要解决的技术问题是未被及时分解的多肽和蛋白质、氨基酸的萃取和树脂中氨基酸的分离。蛋白酶可以水解多肽和蛋白质,双水相系统可以萃取出游离的氨基酸,强酸处理的AB-8型大孔离子交换树脂可以吸附氨基酸,浓氨水可以洗脱吸附柱上的氨基酸。The technical problem to be solved by the present invention is the extraction of polypeptides and proteins that have not been decomposed in time, the extraction of amino acids and the separation of amino acids in resins. The protease can hydrolyze polypeptides and proteins, the two-phase system can extract free amino acids, the AB-8 macroporous ion exchange resin treated with strong acid can adsorb amino acids, and concentrated ammonia water can elute the amino acids on the adsorption column.

本发明的技术原理:多肽和蛋白质在蛋白酶的充分作用下可以部分水解为更小分子的氨基酸,由于原液中含有2%的不溶悬浮物,蛋白酶会附着在上面。24-36h之内离心可以有效的去除悬浮物质和蛋白酶。由聚乙二醇与硫酸铵形成的双水相系统,在双相交界面处界面应力相比较传统的水-萃取剂较低,氨基酸会向聚合物一端扩散,发酵液中的无机盐等会向盐相扩散,达到分离目的。The technical principle of the present invention: polypeptides and proteins can be partially hydrolyzed into amino acids with smaller molecules under the sufficient action of protease. Since the stock solution contains 2% insoluble suspended matter, the protease will be attached to it. Centrifugation within 24-36h can effectively remove suspended matter and protease. In the two-phase system formed by polyethylene glycol and ammonium sulfate, the interface stress at the two-phase interface is lower than that of the traditional water-extractant, amino acids will diffuse to one end of the polymer, and inorganic salts in the fermentation broth will The salt phase diffuses to achieve the purpose of separation.

本发明聚合物与氨基酸的混合物在通过强酸处理的AB-8型大孔离子交换树脂,树脂会吸附氨基酸,用去离子水过交换柱至无色,浓氨水会洗脱氨基酸,在72℃的水浴中加速氨气的挥发可以分离出高纯度的氨基酸(浓缩温度不宜过高,防止氨基酸脱水缩合)。The mixture of the polymer and amino acid of the present invention is treated with strong acid in the AB-8 type macroporous ion exchange resin, the resin will adsorb the amino acid, deionized water is used to pass through the exchange column until colorless, and concentrated ammonia water will elute the amino acid. Accelerating the volatilization of ammonia in a water bath can separate high-purity amino acids (the concentration temperature should not be too high to prevent dehydration and condensation of amino acids).

优选的是,所述的一种从鸡粪中提取氨基酸的方法中,S2中所述碱性蛋白酶的加入量为500mL原液/5g蛋白酶,水解时间为18-30h,水解温度为28-32℃。Preferably, in the method for extracting amino acids from chicken manure, the amount of alkaline protease added in S2 is 500mL stock solution/5g protease, the hydrolysis time is 18-30h, and the hydrolysis temperature is 28-32°C .

优选的是,所述的一种从鸡粪中提取氨基酸的方法中,S2中所述碱性蛋白酶为食品级的小分子肽解酶,所述小分子肽解酶的酶活性为20万u/g。Preferably, in the method for extracting amino acids from chicken manure, the alkaline protease in S2 is a food-grade small-molecule peptidase, and the small-molecule peptidase has an enzymatic activity of 200,000 u /g.

优选的是,所述的一种从鸡粪中提取氨基酸的方法中,S3中离心条件为在7000-9000r/min的转速下离心6-8分钟。Preferably, in the method for extracting amino acids from chicken manure, the centrifugation conditions in S3 are centrifugation at a rotational speed of 7000-9000 r/min for 6-8 minutes.

优选的是,所述的一种从鸡粪中提取氨基酸的方法中,S4中使用循环真空抽滤机将所述溶液a抽滤1.5-3h后,得到所述溶液b。Preferably, in the method for extracting amino acids from chicken manure, the solution b is obtained after the solution a is suction filtered for 1.5-3 hours using a circulating vacuum filter in S4.

优选的是,所述的一种从鸡粪中提取氨基酸的方法中,所述双水相系统为PEG4000和硫酸铵溶液混合而成。Preferably, in the method for extracting amino acids from chicken manure, the two-phase system is formed by mixing PEG4000 and ammonium sulfate solution.

优选的是,所述的一种从鸡粪中提取氨基酸的方法中,所述双水相系统的制备方法如下:Preferably, in the described method for extracting amino acids from chicken manure, the preparation method of the aqueous two-phase system is as follows:

将质量分数为25-40%的PEG-4000水溶液和硫酸铵按照8-12:5.2-5.7mL/mg的体积质量比为混合而成。The PEG-4000 aqueous solution with a mass fraction of 25-40% and ammonium sulfate are mixed according to a volume-to-mass ratio of 8-12:5.2-5.7 mL/mg.

优选的是,所述的一种从鸡粪中提取氨基酸的方法中,S6中离子交换柱采用AB-8型大孔吸附树脂,所述AB-8型大孔吸附树脂使用前在浓度为1.5mol/L的盐酸中浸泡18-30h。Preferably, in the method for extracting amino acids from chicken manure, the ion exchange column in S6 adopts AB-8 type macroporous adsorption resin, and the AB-8 type macroporous adsorption resin is used at a concentration of 1.5 Soak in mol/L hydrochloric acid for 18-30h.

优选的是,所述的一种从鸡粪中提取氨基酸的方法中,S6中使用的浓氨水的浓度为1mol/L。Preferably, in the described method for extracting amino acids from chicken manure, the concentration of the concentrated ammonia water used in S6 is 1 mol/L.

优选的是,所述的一种从鸡粪中提取氨基酸的方法中,S7中将所述溶液d置于70-75℃的条件下水浴加热30-50min。Preferably, in the method for extracting amino acids from chicken manure, in S7, the solution d is placed under the condition of 70-75°C and heated in a water bath for 30-50min.

本发明的有益效果是:The beneficial effects of the present invention are:

1、本发明利用聚乙二醇和硫酸铵混合就形成双水相系统,鸡粪中的无机组分会亲近于双水相中的盐相,有机组分主要是氨基酸,亲近于聚合物相,利用游离的氨基酸和鸡粪中的其他组分在双水相系统中会向不同的组分扩散,达到分离的目的。本发明可以有效的保证游离氨基酸的活性和提取的纯度,纯度可以达到96%。1. The present invention utilizes polyethylene glycol and ammonium sulfate to mix to form a two-phase system, the inorganic components in the chicken manure will be close to the salt phase in the two-water phase, and the organic components are mainly amino acids, which are close to the polymer phase, using Free amino acids and other components in chicken manure will diffuse to different components in the two-phase system to achieve the purpose of separation. The invention can effectively ensure the activity of free amino acids and the purity of extraction, and the purity can reach 96%.

2、利用本发明的方法可以减少相对于其他化学反应而导致的氨基酸污染和变质等问题。2. The method of the present invention can reduce the problems of amino acid contamination and deterioration caused by other chemical reactions.

3、本发明提供的以双水相法萃取鸡粪中的氨基酸具备方法简便,实验药剂毒害性低和环境友好等特点,可以有效的解决家禽类动物的粪便问题,实现固废的资源化。3. The method of extracting amino acids in chicken manure by the aqueous two-phase method provided by the present invention has the characteristics of simple method, low toxicity of experimental chemicals and environmental friendliness, etc., which can effectively solve the problem of feces of poultry animals and realize the resource utilization of solid waste.

4、本发明通过加入除臭的工序,可以去除水解液中97%的挥发性、刺激性气体,同时提高氨基酸的产率,降低了酸性气体对后续实验试剂的影响。4. The present invention can remove 97% of volatile and irritating gases in the hydrolyzate by adding a deodorizing process, while improving the yield of amino acids and reducing the influence of acid gases on subsequent experimental reagents.

5、本发明使用碱性蛋白酶(食品级)可以做到水解鸡粪中未被水解的多肽和蛋白,可以提高丝氨酸和络氨酸的产率,且此蛋白酶的稳定很好,在25℃下,每三个月的酶活损失小于1%。5. The present invention uses alkaline protease (food grade) to hydrolyze unhydrolyzed polypeptides and proteins in chicken manure, which can improve the yield of serine and tyrosine, and the stability of this protease is very good. , the loss of enzyme activity every three months is less than 1%.

本发明的其它优点、目标和特征将部分通过下面的说明体现,部分还将通过对本发明的研究和实践而为本领域的技术人员所理解。Other advantages, objects, and features of the present invention will appear in part from the description that follows, and in part will be appreciated by those skilled in the art from the study and practice of the invention.

具体实施方式Detailed ways

下面结合实施例对本发明做进一步的详细说明,以令本领域技术人员参照说明书文字能够据以实施。The present invention will be further described in detail below with reference to the embodiments, so that those skilled in the art can implement according to the description.

需要说明的是,下述实施方案中所述实验方法,如无特殊说明,均为常规方法,所述试剂和材料,如无特殊说明,均可从商业途径获得。It should be noted that the experimental methods described in the following embodiments are conventional methods unless otherwise specified, and the reagents and materials can be obtained from commercial sources unless otherwise specified.

<实施例1><Example 1>

一种从鸡粪中提取氨基酸的方法,包括以下步骤:A method for extracting amino acids from chicken manure, comprising the following steps:

S1、制备鸡粪发酵原液,备用;S1, prepare chicken manure fermentation stock solution, standby;

S2、在所述鸡粪发酵原液中加入500mL原液/5g的碱性蛋白酶,于温度为28℃的条件下水解时间18h,得到初步水解液,备用,其中碱性蛋白酶为食品级的小分子肽解酶,所述小分子肽解酶的酶活性为20万u/g;S2, add the alkaline protease of 500mL stock solution/5g in the described chicken manure fermentation stock solution, under the condition that the temperature is 28 ℃, the hydrolysis time is 18h, obtain the preliminary hydrolyzate, standby, wherein the alkaline protease is the small molecular peptide of food grade lyase, the enzyme activity of the small molecule peptidase is 200,000 u/g;

S3、将所述初步水解液在7000r/min的转速下离心6分钟后过滤,得到溶液a,备用;S3, the described preliminary hydrolyzate is filtered after centrifugal 6 minutes under the rotating speed of 7000r/min, obtains solution a, for subsequent use;

S4、使用循环真空抽滤机将所述溶液a抽滤1.5h后,得到溶液b,备用;S4, after the solution a is suction filtered for 1.5h using a circulating vacuum filter, a solution b is obtained, which is for subsequent use;

S5、将质量分数为25%的PEG-4000水溶液和硫酸铵按照8:5.2mL/mg的体积质量比为混合,制备得到双水相系统,并将所述溶液b和所述双水相系统混合,得到形成明显的双水相的溶液c,备用;S5. Mix the PEG-4000 aqueous solution with a mass fraction of 25% and ammonium sulfate in a volume-to-mass ratio of 8:5.2 mL/mg to prepare a two-phase system, and combine the solution b with the two-phase system Mix, obtain the solution c that forms obvious two-aqueous phase, standby;

S6、将所述溶液c的上层有机相过离子交换柱,用去离子水洗脱至无色,再使用浓度为1mol/L的氨水洗脱所述离子交换柱得到溶液d,备用;S6, the upper organic phase of the solution c is passed through the ion exchange column, eluted to colorless with deionized water, and then the ion exchange column is eluted with the ammoniacal liquor with a concentration of 1 mol/L to obtain solution d, for subsequent use;

其中离子交换柱采用AB-8型大孔吸附树脂,所述AB-8型大孔吸附树脂使用前在浓度为1.5mol/L的盐酸中浸泡18h;The ion exchange column adopts AB-8 macroporous adsorption resin, and the AB-8 macroporous adsorption resin is soaked in hydrochloric acid with a concentration of 1.5mol/L for 18h before use;

S7、将所述溶液d置于70℃的条件下水浴加热30min,然后真空浓缩,即制得所述氨基酸。S7. The solution d is heated in a water bath at 70° C. for 30 minutes, and then concentrated in vacuo to obtain the amino acid.

<实施例2><Example 2>

一种从鸡粪中提取氨基酸的方法,包括以下步骤:A method for extracting amino acids from chicken manure, comprising the following steps:

S1、制备鸡粪发酵原液,备用;S1, prepare chicken manure fermentation stock solution, standby;

S2、在所述鸡粪发酵原液中加入500mL原液/5g的碱性蛋白酶,于温度为30℃的条件下水解时间24h,得到初步水解液,备用,其中碱性蛋白酶为食品级的小分子肽解酶,所述小分子肽解酶的酶活性为20万u/g;S2, add the alkaline protease of 500mL stock solution/5g in the described chicken manure fermentation stock solution, under the condition that the temperature is 30 ℃, the hydrolysis time is 24h, obtain the preliminary hydrolyzed solution, standby, wherein the alkaline protease is the small molecular peptide of food grade lyase, the enzyme activity of the small molecule peptidase is 200,000 u/g;

S3、将所述初步水解液在8000r/min的转速下离心7分钟后过滤,得到溶液a,备用;S3, the described preliminary hydrolyzate is filtered after centrifugal 7 minutes under the rotating speed of 8000r/min, obtains solution a, for subsequent use;

S4、使用循环真空抽滤机将所述溶液a抽滤2h后,得到溶液b,备用;S4, after the described solution a is suction filtered for 2h using a circulating vacuum filter, solution b is obtained, for subsequent use;

S5、将质量分数为30%的PEG-4000水溶液和硫酸铵按照10:5.5mL/mg的体积质量比为混合,制备得到双水相系统,并将所述溶液b和所述双水相系统混合,得到形成明显的双水相的溶液c,备用;S5. Mix the PEG-4000 aqueous solution with a mass fraction of 30% and ammonium sulfate in a volume-to-mass ratio of 10:5.5 mL/mg to prepare a two-phase system, and combine the solution b and the two-phase system Mix, obtain the solution c that forms obvious two-aqueous phase, standby;

S6、将所述溶液c的上层有机相过离子交换柱,用去离子水洗脱至无色,再使用浓度为1mol/L的氨水洗脱所述离子交换柱得到溶液d,备用;S6, the upper organic phase of the solution c is passed through the ion exchange column, eluted to colorless with deionized water, and then the ion exchange column is eluted with the ammoniacal liquor with a concentration of 1 mol/L to obtain solution d, for subsequent use;

其中离子交换柱采用AB-8型大孔吸附树脂,所述AB-8型大孔吸附树脂使用前在浓度为1.5mol/L的盐酸中浸泡24h;The ion exchange column adopts AB-8 macroporous adsorption resin, and the AB-8 macroporous adsorption resin is soaked in hydrochloric acid with a concentration of 1.5mol/L for 24h before use;

S7、将所述溶液d置于72℃的条件下水浴加热40min,然后真空浓缩,即制得所述氨基酸。S7. The solution d is placed under the condition of 72° C. and heated in a water bath for 40 min, and then concentrated in vacuo to obtain the amino acid.

<实施例3><Example 3>

一种从鸡粪中提取氨基酸的方法,包括以下步骤:A method for extracting amino acids from chicken manure, comprising the following steps:

S1、制备鸡粪发酵原液,备用;S1, prepare chicken manure fermentation stock solution, standby;

S2、在所述鸡粪发酵原液中加入500mL原液/5g的碱性蛋白酶,于温度为32℃的条件下水解时间30h,得到初步水解液,备用,其中碱性蛋白酶为食品级的小分子肽解酶,所述小分子肽解酶的酶活性为20万u/g;S2. Add 500mL stock solution/5g alkaline protease to the chicken manure fermentation stock solution, and hydrolyze for 30h under the condition that the temperature is 32°C to obtain a preliminary hydrolyzed solution for use, wherein the alkaline protease is a food-grade small molecular peptide lyase, the enzyme activity of the small molecule peptidase is 200,000 u/g;

S3、将所述初步水解液在9000r/min的转速下离心8分钟后过滤,得到溶液a,备用;S3, the preliminary hydrolyzate is filtered after centrifugal 8 minutes at the rotating speed of 9000r/min to obtain solution a, for subsequent use;

S4、使用循环真空抽滤机将所述溶液a抽滤3h后,得到溶液b,备用;S4, after the solution a is suction filtered for 3h using a circulating vacuum filter, solution b is obtained, for subsequent use;

S5、将质量分数为40%的PEG-4000水溶液和硫酸铵按照12:5.7mL/mg的体积质量比为混合,制备得到双水相系统,并将所述溶液b和所述双水相系统混合,得到形成明显的双水相的溶液c,备用;S5, the PEG-4000 aqueous solution with a mass fraction of 40% and ammonium sulfate are mixed according to the volume mass ratio of 12:5.7 mL/mg to prepare a two-phase system, and the solution b and the two-phase system are mixed Mix, obtain the solution c that forms obvious two-aqueous phase, standby;

S6、将所述溶液c的上层有机相过离子交换柱,用去离子水洗脱至无色,再使用浓度为1mol/L的氨水洗脱所述离子交换柱得到溶液d,备用;S6, the upper organic phase of the solution c is passed through the ion exchange column, eluted to colorless with deionized water, and then the ion exchange column is eluted with the ammoniacal liquor with a concentration of 1 mol/L to obtain solution d, for subsequent use;

其中离子交换柱采用AB-8型大孔吸附树脂,所述AB-8型大孔吸附树脂使用前在浓度为1.5mol/L的盐酸中浸泡30h;The ion exchange column adopts AB-8 macroporous adsorption resin, and the AB-8 macroporous adsorption resin is soaked in hydrochloric acid with a concentration of 1.5 mol/L for 30 hours before use;

S7、将所述溶液d置于75℃的条件下水浴加热50min,然后真空浓缩,即制得所述氨基酸。S7. The solution d is placed under the condition of 75°C and heated in a water bath for 50 min, and then concentrated in vacuo to obtain the amino acid.

尽管本发明的实施方案已公开如上,但其并不仅仅限于说明书和实施方式中所列运用,它完全可以被适用于各种适合本发明的领域,对于熟悉本领域的人员而言,可容易地实现另外的修改,因此在不背离权利要求及等同范围所限定的一般概念下,本发明并不限于特定的细节和这里示出与描述的实施例。Although the embodiment of the present invention has been disclosed as above, it is not limited to the application listed in the description and the embodiment, and it can be applied to various fields suitable for the present invention. For those skilled in the art, it can be easily Therefore, the invention is not limited to the specific details and embodiments shown and described herein without departing from the general concept defined by the appended claims and the scope of equivalents.

Claims (8)

1. A method for extracting amino acid from chicken manure is characterized by comprising the following steps:
s1, preparing a chicken manure fermentation stock solution for later use;
s2, adding alkaline protease into the chicken manure fermentation stock solution, and hydrolyzing the alkaline protease to obtain a primary hydrolysate for later use;
s3, centrifuging and filtering the primary hydrolysate to obtain a solution a for later use;
s4, deodorizing the solution a to obtain a solution b for later use;
s5, preparing a double aqueous phase system, and mixing the solution b with the double aqueous phase system to obtain a solution c with obvious double aqueous phases for later use;
wherein the aqueous two-phase system is formed by mixing 25-40% of PEG-4000 aqueous solution and ammonium sulfate according to the volume-mass ratio of 8-12:5.2-5.7 mL/mg;
s6, enabling the upper organic phase of the solution c to pass through an ion exchange column, eluting with deionized water until the organic phase is colorless, and then eluting the ion exchange column with ammonia water to obtain a solution d for later use;
s7, heating the solution d in a water bath, and then concentrating in vacuum to obtain the amino acid.
2. The method for extracting amino acids from chicken manure according to claim 1, wherein the alkaline protease is added in an amount of 500mL stock solution/5 g protease in S2, the hydrolysis time is 18-30h, and the hydrolysis temperature is 28-32 ℃.
3. The method for extracting amino acids from chicken manure according to claim 2, wherein the alkaline protease in S2 is a food grade small molecule peptase, and the enzymatic activity of the small molecule peptase is 20 ten thousand u/g.
4. The method as claimed in claim 1, wherein the centrifugation at S3 is performed at 7000-9000r/min for 6-8 min.
5. The method for extracting amino acid from chicken manure as claimed in claim 1, wherein the solution b is obtained after the solution a is filtered by suction for 1.5-3h in S4 using a circulating vacuum filter.
6. The method for extracting amino acid from chicken manure as claimed in any one of claims 1 to 5, wherein the ion exchange column in S6 adopts AB-8 type macroporous adsorbent resin, and the AB-8 type macroporous adsorbent resin is soaked in 1.5mol/L hydrochloric acid for 18-30h before use.
7. A process for extracting amino acids from chicken manure as claimed in any one of claims 1 to 5, wherein the concentration of concentrated ammonia used in S6 is 1 mol/L.
8. A process for extracting amino acids from chicken manure as claimed in any one of claims 1-5, wherein the solution d is heated in water bath at 70-75 ℃ for 30-50min in S7.
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