WO2016029647A1 - Active seaweed feed additive and preparation method therefor - Google Patents

Active seaweed feed additive and preparation method therefor Download PDF

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WO2016029647A1
WO2016029647A1 PCT/CN2015/071861 CN2015071861W WO2016029647A1 WO 2016029647 A1 WO2016029647 A1 WO 2016029647A1 CN 2015071861 W CN2015071861 W CN 2015071861W WO 2016029647 A1 WO2016029647 A1 WO 2016029647A1
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seaweed
feed additive
active
slurry
enzyme
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PCT/CN2015/071861
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French (fr)
Chinese (zh)
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李健
李明潭
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威海市世代海洋生物科技有限公司
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    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02PCLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87Re-use of by-products of food processing for fodder production

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  • the invention relates to the field of feed production, in particular to an active seaweed feed additive which is enzymatically hydrolyzed by a composite enzyme and a preparation method thereof.
  • seaweed plants are rich in nutrients and have functions such as medicinal and health care.
  • seaweed plants are gradually entering people's production and life as important consumables, and are widely used in the fields of industry, agriculture and medicine. This is because seaweed is a self-supporting plant that can perform photosynthesis and convert inorganic matter in the ocean into organic matter. Therefore, seaweed is a rich and low-cost feed resource for animals.
  • Seaweed has great potential for development as a feed ingredient and feed additive.
  • seaweed is rich in seaweed polysaccharides, protein, fat, vitamins, minerals and physiologically active substances with special effects. It is a raw material library for food, medicine and feed.
  • Marine plants are rich in nutrients and contain a variety of biologically active substances, which have biological activities such as enhancing immunity, disease resistance, anti-virus, and growth. Therefore, the development of marine plant feed and the promotion of animal husbandry are receiving increasing attention.
  • seaweed contains a variety of anti-nutritional ingredients, such as cellulose, sugar gum, agar, xylan and other non-starch polysaccharides and their toxins; plus aquatic animals eating seaweed are mostly lower animals, their intestinal tract is short, digestive function is poor Other factors have greatly affected the digestion and utilization of seaweed, resulting in waste of resources, and also easily cause various discomforts in aquatic animals, such as enteritis and swollen mouth.
  • Chinese patent CN 102987077 A discloses a preparation method of seaweed fermented feed, which improves the nutritive value that seaweed can absorb.
  • the preparation method still has the defects of complicated preparation process, long preparation time and high cost, which is far from meeting the market demand.
  • the present invention provides a method for preparing a seaweed biological feed additive, which is characterized in that the method is to add a microbial enzyme to an algae slurry prepared from seaweed or algae residue to obtain an enzymatic hydrolysis solution.
  • the seaweed biological feed wherein the composite microbial enzyme is composed of a plurality of cellulase, pectinase, protease, and amylase.
  • the mass percentage of the cellulase, the pectinase, the protease and the amylase in the complex microbial enzyme are: 60-100%, 0-40%, respectively. 0 to 5%, 0 to 5%, and the cellulase is derived from Trichoderma reesei, Trichoderma viride, and/or Aspergillus niger; the pectinase is derived from Rhizopus oryzae, Aspergillus niger and/or rice Aspergillus; the protease is derived from papain, bromelain and/or Bacillus subtilis; the amylase is derived from Bacillus licheniformis, Aspergillus oryzae and/or Aspergillus niger.
  • the enzyme activity of the cellulase is (18-24) ⁇ 10 4 ⁇ /g
  • the enzyme activity of the pectinase is (2-5) ⁇ 10 4 ⁇ /g
  • the protease activity of the protease is (10 to 50) ⁇ 10 4 ⁇ / g
  • the enzyme activity of the amylase is (1 to 5) ⁇ 10 4 ⁇ / g.
  • the method comprises adding a composite microbial enzyme to the seaweed slurry according to a mass ratio of the composite microbial enzyme to the seaweed slurry of (0.3 to 3):100, and the normal pressure and the temperature are 30 ⁇ .
  • the seaweed biological feed was obtained by insulating the enzyme for 6 to 30 hours under conditions of 65 ° C and pH 3 to 7.
  • the method further comprises the steps of: shearing and grinding the seaweed or seaweed slag into an algae slurry having particles having a diameter of 1 mm or less after inorganic decontamination; and driving the seaweed slurry into a high-speed dispersing kettle Mixing with water once, mixing the water with the kelp slurry, and the mass ratio of the first water addition to the kelp slurry is (30-40): 100; the mixed seaweed slurry is put into the enzymatic hydrolysis tank, the first Adding water twice and adding the composite microbial enzyme to form the mixed liquid, and the mass ratio of the total amount of the first water addition amount and the second water addition amount to the seaweed slurry is 50:100; and the mixing The liquid was stirred uniformly, and then heat-insulated and enzymatically hydrolyzed, and stirred once every hour during the enzymatic hydrolysis, and stirred for 5 minutes each time.
  • the enzymatically prepared mixture is subjected to solid-liquid separation by a backflushing belt filter, and the separated enzyme solution is concentrated in a range of 60 to 68 ° C; or, the enzymatic hydrolysis is carried out. A good mixture is dried.
  • the rotational speed of the high-speed dispersing vessel after starting is 3000 rpm, and after 3 minutes of agitation, the rotational speed of the high-speed dispersing vessel is reduced from 3000 rpm to 1000 rpm.
  • a seaweed biological feed additive is characterized in that the seaweed biological feed is prepared according to the preparation method as described above.
  • the seaweed feed further comprises one or more of fish meal, fish oil, soybean meal, tapioca starch, corn flour and powdered soybean phospholipid.
  • a seaweed biological feed in particular for feeding poultry/livestock or aquaculture, characterized in that the feed comprises a seaweed biological feed additive prepared as described above, or as The aforementioned seaweed feed additive.
  • the invention has the advantages of simple preparation method, easy operation and control, mild preparation process condition, short preparation period and low cost.
  • the invention improves the utilization rate of seaweed by adding a composite microbial enzyme to the slurry prepared from seaweed or seaweed residue, and retains more active nutrients in the seaweed, and the active nutrient component in the seaweed is not affected in the mild enzymatic hydrolysis process. Destruction is conducive to the digestion and absorption of active nutrients by animals and promotes the growth of animals.
  • 1 is a comparison diagram of hydrolysis effects of cellulase produced by different strains in the preparation method of seaweed feed additive of the present invention
  • 3 is a comparison diagram of hydrolysis effects of amylase produced by different strains in the preparation method of seaweed feed additive of the present invention
  • Figure 5 is a graph showing the effect of the amount of the compound microbial enzyme on the hydrolysis effect in the preparation method of the seaweed feed additive of the present invention
  • Fig. 6 is a graph showing the effect of the enzymatic hydrolysis time of the composite microbial enzyme on the hydrolysis effect in the preparation method of the seaweed feed additive of the present invention.
  • the method for preparing the seaweed biological feed additive or the seaweed feed of the present invention wherein the method is to add a microbial enzyme to the seaweed slurry prepared from seaweed or algae residue to carry out enzymatic hydrolysis to obtain the seaweed biological feed, wherein
  • the complex microbial enzyme consists of a plurality of cellulases, pectinase, proteases and amylases.
  • the present invention uses a cellulase produced by Trichoderma reesei, Trichoderma viride, and Aspergillus niger as a fiber.
  • the pectinase produced by Rhizopus oryzae, Aspergillus niger, Aspergillus oryzae is an alternative to pectinase; Caracapapaya protease Papain, pineapple (Ananas spp) protease (Bromelain), Bacillus subtilis protease is an alternative to protease; amylase produced by Bacillus licheniformis, Aspergillus oryzae, Aspergillus niger An alternative to amylase.
  • the mold was selected by studying the hydrolysis effect of cellulase or pectinase or protease or amylase produced by each mold on seaweed, and the hydrolysis effect of various enzymes on seaweed was evaluated by the content of alginic acid.
  • the enzyme activities and optimal enzymatic conditions of the above-mentioned molds selected for use in the present invention are shown in Table 1.
  • Figure 1 shows the hydrolysis effect of cellulase produced by Trichoderma reesei, Trichoderma viride and Aspergillus niger.
  • the hydrolysis effect is based on the amount of alginic acid released.
  • the cellulase produced by Trichoderma reesei has the best hydrolysis effect, and with the enzyme addition amount from 1 ⁇ to 5 ⁇ , the hydrolyzed alginic acid content is from 8g/L to 22g/L, which is produced by Trichoderma viride.
  • the cellulase is added from 1 ⁇ to 5 ⁇ with the enzyme, and the alginic acid content of the hydrolyzed acid is from 9g/L to 19.5g/L.
  • the cellulase produced by Aspergillus niger is hydrolyzed from 1 ⁇ to 5 ⁇ .
  • the alginic acid content is from 7 g/L to 17 g/L. Therefore, in the present invention, the cellulase produced by Trichoderma reesei is preferably used as the cellulase in the complex enzyme.
  • cellulase produced by Trichoderma viride and Aspergillus niger is also feasible.
  • Figure 2 shows the hydrolysis effect of pectinase produced by Aspergillus niger, Aspergillus oryzae and Rhizopus oryzae.
  • the hydrolysis effect is based on the amount of alginic acid released.
  • the hydrolysis of pectinase produced by Rhizopus oryzae The effect is best, with the amount of enzyme added from 1 ⁇ to 5 ⁇ , the hydrolyzed alginic acid content is 6.5g/L ⁇ 11.6g/L, produced by Aspergillus niger
  • the pectinase is added from 1 ⁇ to 5 ⁇ with the enzyme, and the hydrolyzed alginic acid content is 5.9g/L ⁇ 11g/L.
  • pectinase produced by Aspergillus oryzae is from 1 ⁇ 5 ⁇ , and the hydrolyzed seaweed The acid content is from 6.1 g/L to 13 g/L. Therefore, in the present invention, pectinase produced by Rhizopus oryzae is preferably used as a pectinase in the complex enzyme. Of course, it is also feasible to use pectinase produced by Aspergillus niger and Aspergillus oryzae.
  • Figure 3 shows the hydrolysis effect of subtilisin, papain, and bromelain.
  • the hydrolysis effect is based on the amount of alginic acid released. Among them, the hydrolysis effect of bromelain is the best, with the enzyme addition amount from 0.5 ⁇ to 2.5.
  • the hydrolyzed alginic acid content is 5.58 g/L to 6.8 g/L. When the amount is more than 1 Torr, the hydrolyzed alginic acid content is reduced to 6.7 g/L and tends to be balanced.
  • the hydrolysis effect of papain is from 0.5 ⁇ to 2.5 ⁇ , and the hydrolyzed alginic acid content is 5.89 g/L to 6.6 g/L.
  • subtilisin The hydrolysis effect of subtilisin is from 0.5 ⁇ to 2.5 ⁇ , and the hydrolyzed alginic acid content is from 5.78 g/L to 6.2 g/L. Therefore, in the present invention, bromelain is preferred as a protease in the complex enzyme. Of course, it is also possible to use subtilisin and papain.
  • Figure 4 shows the hydrolysis effect of amylase produced by Aspergillus niger, Aspergillus oryzae, Bacillus licheniformis, etc.
  • the hydrolysis effect is based on the amount of alginic acid released.
  • the hydrolysis effect of amylase produced by Bacillus licheniformis is the most Well, with the enzyme addition amount from 1 ⁇ to 5 ⁇ , the hydrolyzed alginic acid content is 5.77g/L to 6.2g/L, and the amylase produced by Aspergillus oryzae is hydrolyzed with the enzyme addition amount from 1 ⁇ to 5 ⁇ .
  • the alginic acid content is 5.6g/L to 5.88g/L
  • the amylase produced by Aspergillus niger is from 1 ⁇ to 5 ⁇
  • the hydrolyzed alginic acid content is from 5.55g/L to 5.78g/L. Therefore, in the present invention, the amylase produced by Bacillus licheniformis is preferred as the amylase in the complex enzyme, and of course, the amylase produced by Aspergillus niger and Aspergillus oryzae is also feasible.
  • a preferred embodiment of the present invention is to select a cellulase produced by Trichoderma reesei as a cellulase in a composite microbial enzyme, and select a pectinase produced by Rhizopus oryzae as a composite microbial enzyme.
  • Pectinase, bromelain was selected as the protease in the composite microbial enzyme
  • the amylase produced by Bacillus licheniformis was selected as the amylase in the composite microbial enzyme.
  • the composite microbial enzyme of the present invention is mainly cellulase, followed by pectinase, followed by protease, and amylase.
  • Table 2 The single enzyme source and properties of the preferred composite microorganisms of the present invention are shown in Table 2.
  • the present invention carried out the enzymatic hydrolysis experiments of Examples 1 to 6 in accordance with the ratios in Table 3.
  • a composite microbial enzyme of the present invention consisting of 100% cellulase, wherein the cellulase enzyme activity selected is 200,000 ⁇ g/g. 100 ml of kelp slurry was added, and the mixed microbial enzyme was added in a ratio of 0.4% by mass of the slurry prepared from seaweed or seaweed residue to form a mixed solution, and the enzymatic hydrolysis was carried out for 25 hours, and the content of alginic acid in the supernatant was determined to be 17.25 g. /L.
  • the invention relates to a composite microbial enzyme which is prepared by mixing 60% by mass of cellulase and 40% of pectinase, wherein the cellulase activity of the selected cellulase is 120,000 ⁇ g/g, pectin.
  • the enzyme activity of the enzyme was 12,000 ⁇ g/g.
  • the composite microbial enzyme was added in a ratio of 0.4% to form a mixed solution, and the enzymatic hydrolysis was carried out for 25 hours, and the content of alginic acid in the supernatant was determined to be 19.56 g/L.
  • the invention relates to a composite microbial enzyme which is prepared by mixing 60% by mass of cellulase with 35% of pectinase and 5% of protease, wherein the cellulase activity of the selected cellulase is 120,000 ⁇ . /g, the pectinase activity was 1.05 million ⁇ g/g, and the protease activity was 25,000 ⁇ g/g.
  • the invention relates to a composite microbial enzyme which is prepared by mixing 60% by mass of cellulase and 35% of pectinase and 5% of amylase, wherein the cellulase activity of the selected cellulase is 120,000. ⁇ , the pectinase activity was 1.05 million ⁇ g/g, and the amylase enzyme activity was 0.2 million ⁇ g/g.
  • the composite microbial enzyme of the invention is prepared by mixing 70% by mass of cellulase and 27% of pectinase and 3% of protease, wherein the cellulase activity of the selected cellulase is 140,000 ⁇ .
  • the enzyme activity of the pectinase was 0.81 ⁇ g/g, and the enzyme activity of the protease was 15,000 ⁇ g/g.
  • a composite microbial enzyme of the present invention which is prepared by mixing 60% by mass of cellulase with 30% of pectinase, 5% of protease and 5% of amylase, wherein cellulase is selected.
  • the enzyme activity was 120,000 ⁇ g
  • the pectinase activity was 0.81 ⁇ g/g
  • the protease activity was 25,000 ⁇ g/g
  • the amylase enzyme activity was 0.2 million ⁇ g/g.
  • amylase as an auxiliary enzyme in the complex microbial enzyme can make the fresh kelp more fully enzymatically hydrolyzed.
  • the content of crude protein in kelp is 5 to 9%, and the content is relatively low. From Table 2, it can be seen that the price of protease is relatively high. If the amount of protease used is equivalent to other enzymes, the processing cost will increase. Causes waste of resources.
  • the mass percentage of cellulase and pectinase used in the present invention is much larger than that of protease and amylase, and the basic enzymes using cellulase and pectinase as composite microbial enzymes, and protease and amylase as complex microbial enzymes are determined.
  • Auxiliary enzyme species Table 3 shows that the composite microbial enzyme composed of Example 6 has the best enzymatic hydrolysis effect on the kelp slurry. Example 6 was therefore taken as the preferred embodiment of the complex microbial enzyme ratio.
  • the composite microbial enzyme consisting of the optimal ratio in Example 6 was used to study the effect of the amount of the combined microbial enzyme on the hydrolysis effect of seaweed. Its hydrolysis effect is shown in Figure 5.
  • the alginic acid content gradually increased with the addition of the complex microbial enzyme from 0 to 0.6%; and the alginic acid content increased substantially linearly with the addition of the complex microbial enzyme from 0 to 0.3%;
  • the added amount is 0.3%, the alginic acid content is 17.5g/L; when the compounding enzyme reaches 0.4%, the alginic acid content reaches 18g/L; when the compounding enzyme is added more than 0.4%, the alginic acid content tends to Balance, no longer increase. Therefore, the amount of the complex microbial enzyme to be added of the present invention is preferably from 0.3 to 0.4% in view of overall cost and time.
  • Table 4 lists parameters such as reaction temperature, enzyme addition amount, pH value and enzymatic hydrolysis time in the enzymatic hydrolysis process of the composite microorganism of the present invention.
  • Table 5 lists the results of Examples 7-15 of the orthogonal test according to the parameters shown in Table 4. The present invention determines the preferred process parameters of the present invention from Tables 4 and 5.
  • Fig. 6 shows the effect of the enzymatic hydrolysis time on the enzymatic hydrolysis under the conditions of the preferred embodiment 11, i.e., the enzymatic hydrolysis temperature was 55 ° C, the amount of the composite biological enzyme added was 0.4%, and the pH was 5.5.
  • the enzymatic hydrolysis time increases, The alginic acid content increased gradually, and the alginic acid content increased linearly when the enzymatic hydrolysis time changed from 0 to 20 h.
  • the alginic acid content reached 20 g/L; when the enzymatic hydrolysis time was about 25 h, the algae The acid content reached a maximum, and the alginic acid content at this time was 23 g/L.
  • the enzymatic hydrolysis time of the invention is 18h.
  • the seaweed biological feed additive or seaweed biological feed of the present invention is prepared after determining the enzyme species of the complex enzyme microbial enzyme and the relevant ratio of various enzymes and the optimal enzymatic process parameters.
  • the method for preparing the seaweed biological feed additive or the seaweed biological feed of the present invention specifically comprises the following steps:
  • the seaweed slurry is cut and ground into particles having a diameter of 1 mm or less; and the seaweed or seaweed residue is ground to less than 1 mm, so that the active nutrients of the seaweed particles are fully exposed, thereby making the composite
  • the microbial enzyme is in full contact with the active material, thereby improving the hydrolysis efficiency.
  • the mixed solution after the above four steps is subjected to solid-liquid separation by a backflushing belt filter, and the separated enzymatic supernatant is concentrated in a range of 60 to 68 ° C; or The enzymatically mixed mixture is dried to obtain the seaweed biological feed additive or seaweed biological feed of the present invention.
  • the seaweed pulp and the water in order to make the seaweed slurry more uniformly mixed, the seaweed pulp and the water can be sufficiently mixed, the rotation speed of the high-speed dispersion kettle after starting is 3000 rpm, and after stirring for 3 minutes, the high-speed dispersion kettle The speed was reduced from 3000 rpm to 1000 rpm.
  • the cellulase, pectinase, protease and amylase of the present invention account for a percentage by mass of the complex microbial enzyme: 60 to 100%, 0 to 40%, 0 to 5%, 0.
  • the cellulase is derived from Trichoderma reesei, Trichoderma viride and/or Aspergillus niger; the pectinase is derived from Rhizopus oryzae, Aspergillus niger and/or Aspergillus oryzae; the protease is derived from papain, bromelain and / or Bacillus subtilis; amylase is derived from Bacillus licheniformis, Aspergillus oryzae and / or Aspergillus niger.
  • the cellulase of the present invention has an enzyme activity of (18-24) ⁇ 10 4 ⁇ g/g, and the pectinase has an enzyme activity of (2-5) ⁇ 10 4 ⁇ g/g.
  • the activity was (10 to 50) ⁇ 10 4 ⁇ /g, and the enzyme activity of amylase was (1 to 5) ⁇ 10 4 ⁇ /g.
  • the preparation method of the present invention is to add a composite microbial enzyme to the seaweed slurry according to a mass ratio of the composite microbial enzyme to the seaweed slurry of (0.3 to 3):100, and at normal pressure and temperature.
  • the algae biological feed additive or the seaweed biological feed is obtained by insulating the enzyme for 30 to 30 hours under conditions of 30 to 65 ° C and pH 3 to 7.
  • another aspect of the present invention relates to a seaweed biological feed additive or a seaweed biological feed, and the seaweed biological feed additive or the seaweed biological feed of the present invention is prepared according to the preparation method as described above. of.
  • the seaweed feed additive or seaweed biological feed of the present invention further comprises one or more of fish meal, fish oil, soybean meal, tapioca starch, corn flour and powdered soybean phospholipid.
  • the seaweed biological feed of the present invention can be prepared into a powdery or granular form or an amorphous form as needed.
  • seaweed biological feed particularly for feeding poultry/livestock or aquaculture.
  • the seaweed biological feed comprises an algae biological feed additive prepared as described above or a seaweed feed additive as described above.
  • the seaweed biological feed additive or seaweed biological feed prepared by the invention not only greatly improves the digestion and absorption rate of seaweed, reduces waste, but also enhances the immunity of the body and reduces the use of antibiotics, and is a healthy, efficient and novel breeding feed. . Moreover, the invention has simple preparation process, easy operation and short production time as compared with other preparation methods in the prior art, and is suitable for mass production in a short time.

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Abstract

A seaweed biological feed additive or a preparation method for a seaweed biological feed. The method is characterized by adding a composite microorganism enzyme into seaweed serosity that is prepared from seaweed or seaweed residues, and performing enzymolysis to obtain the seaweed biological feed additive, wherein the composite microorganism enzyme consists of several kinds of cellulase, pectinase, protease and amylase.

Description

一种活性海藻饲料添加剂及其制作方法Active seaweed feed additive and preparation method thereof 技术领域Technical field
本发明涉及饲料生产领域,尤其涉及一种利用复合酶进行酶解的活性海藻饲料添加剂及其制作方法。The invention relates to the field of feed production, in particular to an active seaweed feed additive which is enzymatically hydrolyzed by a composite enzyme and a preparation method thereof.
背景技术Background technique
我国作为农业大国和水产大国,水产品已成为人们重要的动物性蛋白消费产品,并且每年呈几何式快速增长。随着人们水产品消费水平的不断增加,其大大刺激了我国水产养殖业的快速发展,特别是以海藻为食物源的水产养殖尤为明显,比如鲍鱼、海参等。海藻类植物含有丰富的营养成份,具有药用和保健等功能,同时海藻类植物作为重要的消耗品正逐渐进入人们的生产生活中,并且广泛应用到工业、农业和医药业等领域中。这是由于海藻是自养植物,能进行光合作用,把海洋里的无机物质转化为有机物质,所以海藻是动物丰富的低成本饲料资源,海藻作为饲料原料和饲料添加剂开发潜力很大。此外,海藻含有丰富的海藻多糖、蛋白质、脂肪、维生素、矿物质以及具有特殊功效的生理活性物质,是食品、药物、饲料的原料库。海洋植物营养丰富,含有多种生物活性物质,具有增强机体免疫力、抗病、抗病毒、促进生长等生物活性。因此,开发海洋植物饲料,促进畜牧业发展正日趋受到广泛重视。As a big agricultural country and a big aquatic country, aquatic products have become an important animal protein consumption product, and it has grown rapidly every year. With the increasing consumption level of aquatic products, it has greatly stimulated the rapid development of China's aquaculture industry, especially the aquaculture with seaweed as a food source, such as abalone and sea cucumber. Seaweed plants are rich in nutrients and have functions such as medicinal and health care. At the same time, seaweed plants are gradually entering people's production and life as important consumables, and are widely used in the fields of industry, agriculture and medicine. This is because seaweed is a self-supporting plant that can perform photosynthesis and convert inorganic matter in the ocean into organic matter. Therefore, seaweed is a rich and low-cost feed resource for animals. Seaweed has great potential for development as a feed ingredient and feed additive. In addition, seaweed is rich in seaweed polysaccharides, protein, fat, vitamins, minerals and physiologically active substances with special effects. It is a raw material library for food, medicine and feed. Marine plants are rich in nutrients and contain a variety of biologically active substances, which have biological activities such as enhancing immunity, disease resistance, anti-virus, and growth. Therefore, the development of marine plant feed and the promotion of animal husbandry are receiving increasing attention.
但是,近年来,随着人口急剧增加,耕地大量减少,饲料紧缺已成为影响养殖业快速发展的严重的问题,人与动物争粮的矛盾也愈发的严重。由于海藻中含有多种抗营养成分,如纤维素、糖胶、琼脂、木聚糖等非淀粉多糖及其毒素;加之食用海藻的水产动物多为低等动物,其肠道短,消化功能差等因素,极大影响了海藻的消化利用率,造成资源浪费,也易引起水产动物的各种不适症,如肠炎、肿嘴。此外,由于我国水产养殖起步较晚,系统研究滞后,养殖技术不完善,目前处于一种粗放型养殖模式,因此投喂饲料多属凭感觉或经验来操作,造成饲料浪费严重,营养流失,最后引起水环境、池底腐败恶化等现象。因此,如何提高海藻利用率及其营养水平,并降除毒素是当前急需解决的 问题。However, in recent years, with the rapid increase in population and the large reduction of cultivated land, the shortage of feed has become a serious problem affecting the rapid development of the aquaculture industry, and the contradiction between people and animals for food has become more serious. Because seaweed contains a variety of anti-nutritional ingredients, such as cellulose, sugar gum, agar, xylan and other non-starch polysaccharides and their toxins; plus aquatic animals eating seaweed are mostly lower animals, their intestinal tract is short, digestive function is poor Other factors have greatly affected the digestion and utilization of seaweed, resulting in waste of resources, and also easily cause various discomforts in aquatic animals, such as enteritis and swollen mouth. In addition, due to the late start of aquaculture in China, the systematic research is lagging behind, the farming technology is imperfect, and it is currently in an extensive farming mode. Therefore, feeding feed is mostly operated by feeling or experience, resulting in serious feed waste and nutrient loss. Causes the water environment and the deterioration of the bottom of the pool. Therefore, how to improve the utilization rate of seaweed and its nutrient level, and reduce the toxin is urgently needed to be solved. problem.
面对上述问题,中国专利CN 102987077 A公开了一种海藻发酵饲料的制备方法,提高了海藻可吸收的营养价值。但是该制备方法仍然存在制备工艺复杂、制备时间长、成本较高等缺陷,这远不能满足市场的需求。Faced with the above problems, Chinese patent CN 102987077 A discloses a preparation method of seaweed fermented feed, which improves the nutritive value that seaweed can absorb. However, the preparation method still has the defects of complicated preparation process, long preparation time and high cost, which is far from meeting the market demand.
发明内容Summary of the invention
针对现有技术之不足,本发明提供了一种海藻生物饲料添加剂的制备方法,其特征在于,所述方法是向由海藻或海藻渣制成的海藻浆液中添加复合微生物酶进行酶解从而获得所述海藻生物饲料,其中,所述复合微生物酶由纤维素酶、果胶酶、蛋白酶和淀粉酶中的多种组成。In view of the deficiencies of the prior art, the present invention provides a method for preparing a seaweed biological feed additive, which is characterized in that the method is to add a microbial enzyme to an algae slurry prepared from seaweed or algae residue to obtain an enzymatic hydrolysis solution. The seaweed biological feed, wherein the composite microbial enzyme is composed of a plurality of cellulase, pectinase, protease, and amylase.
根据一个优选实施方式,所述纤维素酶、所述果胶酶、所述蛋白酶和所述淀粉酶在所述复合微生物酶中所占的质量百分比分别是:60~100%,0~40%,0~5%,0~5%,并且所述纤维素酶源于李氏木霉、绿色木霉和/或黑曲霉;所述果胶酶源于米根霉、黑曲霉和/或米曲霉;所述蛋白酶源于木瓜蛋白酶、菠萝蛋白酶和/或枯草杆菌;所述淀粉酶源于地衣芽孢杆菌、米曲霉和/或黑曲霉。According to a preferred embodiment, the mass percentage of the cellulase, the pectinase, the protease and the amylase in the complex microbial enzyme are: 60-100%, 0-40%, respectively. 0 to 5%, 0 to 5%, and the cellulase is derived from Trichoderma reesei, Trichoderma viride, and/or Aspergillus niger; the pectinase is derived from Rhizopus oryzae, Aspergillus niger and/or rice Aspergillus; the protease is derived from papain, bromelain and/or Bacillus subtilis; the amylase is derived from Bacillus licheniformis, Aspergillus oryzae and/or Aspergillus niger.
根据一个优选实施方式,所述纤维素酶的酶活为(18~24)×104μ/g,所述果胶酶的酶活为(2~5)×104μ/g,所述蛋白酶的酶活为(10~50)×104μ/g,所述淀粉酶的酶活为(1~5)×104μ/g。According to a preferred embodiment, the enzyme activity of the cellulase is (18-24)×10 4 μ/g, and the enzyme activity of the pectinase is (2-5)×10 4 μ/g. The protease activity of the protease is (10 to 50) × 10 4 μ / g, and the enzyme activity of the amylase is (1 to 5) × 10 4 μ / g.
根据一个优选实施方式,所述方法是向所述海藻浆液中按照复合微生物酶与海藻浆液的质量比为(0.3~3):100的比例添加复合微生物酶,并在常压、温度为30~65℃、pH3~7的条件下保温酶解6~30小时,从而获得所述海藻生物饲料。According to a preferred embodiment, the method comprises adding a composite microbial enzyme to the seaweed slurry according to a mass ratio of the composite microbial enzyme to the seaweed slurry of (0.3 to 3):100, and the normal pressure and the temperature are 30 ~. The seaweed biological feed was obtained by insulating the enzyme for 6 to 30 hours under conditions of 65 ° C and pH 3 to 7.
根据一个优选实施方式,所述方法还包括以下步骤:将海藻或海藻渣经过无机除杂后剪切并研磨成直径在1mm以下颗粒的海藻浆液;将所述海藻浆液打入高速分散釜中第一次加水搅拌,使得水与海带浆液充分混合,并且第一次加水量与所述海带浆液的质量比为(30~40):100;将混合后的海藻浆液打入酶解罐中,第二次加水并且加入所述复合微生物酶,形成所述混合液,并且第一次加水量和第二次加水量的总量与所述海藻浆液的质量比为50:100;以及将所述混合液搅拌均匀后加热保温酶解,并且在酶解过程中,每一小时搅拌一次,并且每次搅拌5分钟。 According to a preferred embodiment, the method further comprises the steps of: shearing and grinding the seaweed or seaweed slag into an algae slurry having particles having a diameter of 1 mm or less after inorganic decontamination; and driving the seaweed slurry into a high-speed dispersing kettle Mixing with water once, mixing the water with the kelp slurry, and the mass ratio of the first water addition to the kelp slurry is (30-40): 100; the mixed seaweed slurry is put into the enzymatic hydrolysis tank, the first Adding water twice and adding the composite microbial enzyme to form the mixed liquid, and the mass ratio of the total amount of the first water addition amount and the second water addition amount to the seaweed slurry is 50:100; and the mixing The liquid was stirred uniformly, and then heat-insulated and enzymatically hydrolyzed, and stirred once every hour during the enzymatic hydrolysis, and stirred for 5 minutes each time.
根据一个优选实施方式,将酶解好的混合液通过反冲带式过滤机进行固液分离,并且将分离出的酶解清液在60~68℃的范围内进行浓缩;或者,将酶解好的混合液进行干燥。According to a preferred embodiment, the enzymatically prepared mixture is subjected to solid-liquid separation by a backflushing belt filter, and the separated enzyme solution is concentrated in a range of 60 to 68 ° C; or, the enzymatic hydrolysis is carried out. A good mixture is dried.
根据一个优选实施方式,所述高速分散釜启动后的转速为3000转/分钟,搅动3分钟后,所述高速分散釜的转速由3000转/分钟降至1000转/分钟。According to a preferred embodiment, the rotational speed of the high-speed dispersing vessel after starting is 3000 rpm, and after 3 minutes of agitation, the rotational speed of the high-speed dispersing vessel is reduced from 3000 rpm to 1000 rpm.
本发明的另一方面,一种海藻生物饲料添加剂,其特征在于,所述海藻生物饲料是按照如前述的制备方法制备而成的。In another aspect of the invention, a seaweed biological feed additive is characterized in that the seaweed biological feed is prepared according to the preparation method as described above.
根据一个优选实施方式,所述海藻饲料还包括鱼粉、鱼油、豆粕、木薯淀粉、玉米粉和粉末大豆磷脂中一种或多种。According to a preferred embodiment, the seaweed feed further comprises one or more of fish meal, fish oil, soybean meal, tapioca starch, corn flour and powdered soybean phospholipid.
本发明的另一方面,一种海藻生物饲料,尤其是用于饲养家禽/家畜或水产养殖的饲料,其特征在于,所述饲料包括如前述的制备方法制备而成的海藻生物饲料添加剂或如前述的海藻饲料添加剂。In another aspect of the invention, a seaweed biological feed, in particular for feeding poultry/livestock or aquaculture, characterized in that the feed comprises a seaweed biological feed additive prepared as described above, or as The aforementioned seaweed feed additive.
本发明具有制备方法简单,易操作和控制,制备工艺条件温和,制备周期短,成本低等优点。本发明通过向海藻或海藻渣制成的浆液里添加复合微生物酶来提高海藻的利用率,保留了海藻中更多的活性营养成分,在温和的酶解过程中海藻中的活性营养成分没有遭到破坏,有利于动物对活性营养成分的消化吸收,促进动物的增长。The invention has the advantages of simple preparation method, easy operation and control, mild preparation process condition, short preparation period and low cost. The invention improves the utilization rate of seaweed by adding a composite microbial enzyme to the slurry prepared from seaweed or seaweed residue, and retains more active nutrients in the seaweed, and the active nutrient component in the seaweed is not affected in the mild enzymatic hydrolysis process. Destruction is conducive to the digestion and absorption of active nutrients by animals and promotes the growth of animals.
附图说明DRAWINGS
图1是本发明的海藻饲料添加剂制备方法中不同菌种所产纤维素酶的水解效果对照图;1 is a comparison diagram of hydrolysis effects of cellulase produced by different strains in the preparation method of seaweed feed additive of the present invention;
图2是本发明的海藻饲料添加剂制备方法中不同菌种所产果胶酶的水解效果对照图;2 is a comparison diagram of hydrolysis effects of pectinase produced by different strains in the preparation method of seaweed feed additive of the present invention;
图3是本发明的海藻饲料添加剂制备方法中不同菌种所产淀粉酶的水解效果对照图;3 is a comparison diagram of hydrolysis effects of amylase produced by different strains in the preparation method of seaweed feed additive of the present invention;
图4是本发明的海藻饲料添加剂制备方法中不同菌种所产蛋白酶的水解效果对照图;4 is a comparison diagram of hydrolysis effects of proteases produced by different strains in the preparation method of seaweed feed additive of the present invention;
图5是本发明的海藻饲料添加剂制备方法中复合微生物酶加量对水解效果的影响;和 Figure 5 is a graph showing the effect of the amount of the compound microbial enzyme on the hydrolysis effect in the preparation method of the seaweed feed additive of the present invention;
图6是本发明的海藻饲料添加剂制备方法中复合微生物酶的酶解时间对水解效果的影响。Fig. 6 is a graph showing the effect of the enzymatic hydrolysis time of the composite microbial enzyme on the hydrolysis effect in the preparation method of the seaweed feed additive of the present invention.
具体实施方式detailed description
下面结合附图和实施例进行详细说明。Detailed description will be made below with reference to the accompanying drawings and embodiments.
本发明的海藻生物饲料添加剂或海藻饲料的制备方法,该方法总的技术方案是向由海藻或海藻渣制成的海藻浆液中添加复合微生物酶进行酶解从而获得所述海藻生物饲料,其中,复合微生物酶由纤维素酶、果胶酶、蛋白酶和淀粉酶中的多种组成。The method for preparing the seaweed biological feed additive or the seaweed feed of the present invention, wherein the method is to add a microbial enzyme to the seaweed slurry prepared from seaweed or algae residue to carry out enzymatic hydrolysis to obtain the seaweed biological feed, wherein The complex microbial enzyme consists of a plurality of cellulases, pectinase, proteases and amylases.
1酶种的选取1 selection of enzymes
为了能够获得水解效果最佳的构成复合微生物酶的酶种,本发明以以李氏木霉(Trichoderma reesei)、绿色木霉(Trichoderma viride)、黑曲霉(Aspergillus niger)所产纤维素酶为纤维素酶的备选项;以米根霉(Rhizopus oryzae)、黑曲霉(Aspergillus niger)、米曲霉(Aspergillus oryzae)所产果胶酶为果胶酶的备选项;以木瓜(Caricapapaya)蛋白酶Papain、菠萝(Ananas spp)蛋白酶(Bromelain)、枯草杆菌(Bacillus subtilis)所产蛋白酶为蛋白酶的备选项;以地衣芽孢杆菌(Bacillus licheniformis)、米曲霉(Aspergillus oryzae)、黑曲霉(Aspergillus niger)所产淀粉酶为淀粉酶的备选项。通过研究每种霉菌所产纤维素酶或果胶酶或蛋白酶或淀粉酶对海藻的水解效果来选取霉菌,并且以海藻酸的含量来评判各种酶对海藻的水解效果。本发明所选用的上述霉菌的酶活以及最佳酶解条件如表1所示。In order to obtain an enzyme species constituting a composite microbial enzyme having the best hydrolysis effect, the present invention uses a cellulase produced by Trichoderma reesei, Trichoderma viride, and Aspergillus niger as a fiber. Alternative to the enzyme; the pectinase produced by Rhizopus oryzae, Aspergillus niger, Aspergillus oryzae is an alternative to pectinase; Caracapapaya protease Papain, pineapple (Ananas spp) protease (Bromelain), Bacillus subtilis protease is an alternative to protease; amylase produced by Bacillus licheniformis, Aspergillus oryzae, Aspergillus niger An alternative to amylase. The mold was selected by studying the hydrolysis effect of cellulase or pectinase or protease or amylase produced by each mold on seaweed, and the hydrolysis effect of various enzymes on seaweed was evaluated by the content of alginic acid. The enzyme activities and optimal enzymatic conditions of the above-mentioned molds selected for use in the present invention are shown in Table 1.
表1Table 1
Figure PCTCN2015071861-appb-000001
Figure PCTCN2015071861-appb-000001
Figure PCTCN2015071861-appb-000002
Figure PCTCN2015071861-appb-000002
将表1中不同来源的纤维素酶、果胶酶、蛋白酶和淀粉酶进行水解实验,测试结果附图1至图4所示。Cellulase, pectinase, protease and amylase from different sources in Table 1 were subjected to hydrolysis experiments, and the test results are shown in Figures 1 to 4.
图1示出了李氏木霉、绿色木霉及黑曲霉等菌种所产纤维素酶的水解效果对照,水解效果以释放海藻酸的量为标准。其中,李氏木霉所产纤维素酶的水解效果最好,且随着酶添加量从1‰~5‰,水解的海藻酸含量为8g/L~22g/L,绿色木霉所产的纤维素酶随酶添加量从1‰~5‰,水解的海藻酸含量为9g/L~19.5g/L,黑曲霉所产的纤维素酶随酶添加量从1‰~5‰,水解的海藻酸含量为7g/L~17g/L。因此本发明优选以李氏木霉所产纤维素酶作为复合酶中纤维素酶的选择,当然采用绿色木霉、黑曲霉所产纤维素酶也是可行的。Figure 1 shows the hydrolysis effect of cellulase produced by Trichoderma reesei, Trichoderma viride and Aspergillus niger. The hydrolysis effect is based on the amount of alginic acid released. Among them, the cellulase produced by Trichoderma reesei has the best hydrolysis effect, and with the enzyme addition amount from 1‰ to 5‰, the hydrolyzed alginic acid content is from 8g/L to 22g/L, which is produced by Trichoderma viride. The cellulase is added from 1‰ to 5‰ with the enzyme, and the alginic acid content of the hydrolyzed acid is from 9g/L to 19.5g/L. The cellulase produced by Aspergillus niger is hydrolyzed from 1‰ to 5‰. The alginic acid content is from 7 g/L to 17 g/L. Therefore, in the present invention, the cellulase produced by Trichoderma reesei is preferably used as the cellulase in the complex enzyme. Of course, cellulase produced by Trichoderma viride and Aspergillus niger is also feasible.
图2示出了黑曲霉、米曲霉及米根霉等菌种所产果胶酶的水解效果对照,水解效果以释放海藻酸的量为准,其中,米根霉所产果胶酶的水解效果最好,随酶添加量从1‰~5‰,水解的海藻酸含量为6.5g/L~11.6g/L,黑曲霉所产 果胶酶随酶添加量从1‰~5‰,水解的海藻酸含量为5.9g/L~11g/L,米曲霉所产果胶酶随酶添加量从1‰~5‰,水解的海藻酸含量为6.1g/L~13g/L。因此本发明优选以米根霉所产果胶酶作为复合酶中果胶酶的选择,当然,采用黑曲霉、米曲霉所产果胶酶也是可行的。Figure 2 shows the hydrolysis effect of pectinase produced by Aspergillus niger, Aspergillus oryzae and Rhizopus oryzae. The hydrolysis effect is based on the amount of alginic acid released. Among them, the hydrolysis of pectinase produced by Rhizopus oryzae The effect is best, with the amount of enzyme added from 1‰ to 5‰, the hydrolyzed alginic acid content is 6.5g/L~11.6g/L, produced by Aspergillus niger The pectinase is added from 1‰ to 5‰ with the enzyme, and the hydrolyzed alginic acid content is 5.9g/L~11g/L. The pectinase produced by Aspergillus oryzae is from 1‰~5‰, and the hydrolyzed seaweed The acid content is from 6.1 g/L to 13 g/L. Therefore, in the present invention, pectinase produced by Rhizopus oryzae is preferably used as a pectinase in the complex enzyme. Of course, it is also feasible to use pectinase produced by Aspergillus niger and Aspergillus oryzae.
图3示出了枯草杆菌蛋白酶、木瓜蛋白酶、菠萝蛋白酶的水解效果对照,水解效果以释放海藻酸的量为准,其中,菠萝蛋白酶的水解效果最好,随着酶添加量从0.5‰~2.5‰,水解的海藻酸含量为5.58g/L~6.8g/L,添加量超过1‰时,水解的海藻酸含量降为6.7g/L且趋于平衡。木瓜蛋白酶的水解效果随酶添加量从0.5‰~2.5‰,水解的海藻酸含量为5.89g/L~6.6g/L。枯草杆菌蛋白酶的水解效果随酶添加量从0.5‰~2.5‰,水解的海藻酸含量为5.78g/L~6.2g/L。因此本发明优选菠萝蛋白酶作为复合酶中蛋白酶的选择,当然,采用枯草杆菌蛋白酶、木瓜蛋白酶也是可行的。Figure 3 shows the hydrolysis effect of subtilisin, papain, and bromelain. The hydrolysis effect is based on the amount of alginic acid released. Among them, the hydrolysis effect of bromelain is the best, with the enzyme addition amount from 0.5‰ to 2.5. The hydrolyzed alginic acid content is 5.58 g/L to 6.8 g/L. When the amount is more than 1 Torr, the hydrolyzed alginic acid content is reduced to 6.7 g/L and tends to be balanced. The hydrolysis effect of papain is from 0.5 ‰ to 2.5 酶, and the hydrolyzed alginic acid content is 5.89 g/L to 6.6 g/L. The hydrolysis effect of subtilisin is from 0.5 ‰ to 2.5 酶, and the hydrolyzed alginic acid content is from 5.78 g/L to 6.2 g/L. Therefore, in the present invention, bromelain is preferred as a protease in the complex enzyme. Of course, it is also possible to use subtilisin and papain.
图4示出了黑曲霉、米曲霉、地衣芽孢杆菌等菌种所产淀粉酶的水解效果图,水解效果以释放海藻酸的量为准,其中,地衣芽孢杆菌所产淀粉酶的水解效果最好,随着酶添加量从1‰~5‰,水解的海藻酸含量为5.77g/L~6.2g/L,米曲霉所产淀粉酶随着酶添加量从1‰~5‰,水解的海藻酸含量为5.6g/L~5.88g/L,黑曲霉所产淀粉酶随着酶添加量从1‰~5‰,水解的海藻酸含量为5.55g/L~5.78g/L。因此本发明优选地衣芽孢杆菌所产淀粉酶作为复合酶中淀粉酶的选择,当然采用黑曲霉、米曲霉所产淀粉酶也是可行的。Figure 4 shows the hydrolysis effect of amylase produced by Aspergillus niger, Aspergillus oryzae, Bacillus licheniformis, etc. The hydrolysis effect is based on the amount of alginic acid released. Among them, the hydrolysis effect of amylase produced by Bacillus licheniformis is the most Well, with the enzyme addition amount from 1‰ to 5‰, the hydrolyzed alginic acid content is 5.77g/L to 6.2g/L, and the amylase produced by Aspergillus oryzae is hydrolyzed with the enzyme addition amount from 1‰ to 5‰. The alginic acid content is 5.6g/L to 5.88g/L, and the amylase produced by Aspergillus niger is from 1‰ to 5‰, and the hydrolyzed alginic acid content is from 5.55g/L to 5.78g/L. Therefore, in the present invention, the amylase produced by Bacillus licheniformis is preferred as the amylase in the complex enzyme, and of course, the amylase produced by Aspergillus niger and Aspergillus oryzae is also feasible.
综上所述,本发明的一个最优实施方式是,选取李氏木霉所产纤维素酶为复合微生物酶中的纤维素酶,选取米根霉所产果胶酶为复合微生物酶中的果胶酶,选取菠萝蛋白酶为复合微生物酶中的蛋白酶,选取地衣芽孢杆菌所产淀粉酶为复合微生物酶中的淀粉酶。并且,本发明的复合微生物酶以纤维素酶为主,果胶酶次之,蛋白酶又次之,淀粉酶再次之。本发明优选复合微生物的单酶来源及性能如表2中所示。In summary, a preferred embodiment of the present invention is to select a cellulase produced by Trichoderma reesei as a cellulase in a composite microbial enzyme, and select a pectinase produced by Rhizopus oryzae as a composite microbial enzyme. Pectinase, bromelain was selected as the protease in the composite microbial enzyme, and the amylase produced by Bacillus licheniformis was selected as the amylase in the composite microbial enzyme. Moreover, the composite microbial enzyme of the present invention is mainly cellulase, followed by pectinase, followed by protease, and amylase. The single enzyme source and properties of the preferred composite microorganisms of the present invention are shown in Table 2.
表2Table 2
Figure PCTCN2015071861-appb-000003
Figure PCTCN2015071861-appb-000003
Figure PCTCN2015071861-appb-000004
Figure PCTCN2015071861-appb-000004
2复合微生物酶的配比设计2 compounding design of compound microbial enzyme
本发明按照表3中的配比进行实施例1~6的酶解实验。The present invention carried out the enzymatic hydrolysis experiments of Examples 1 to 6 in accordance with the ratios in Table 3.
表3table 3
Figure PCTCN2015071861-appb-000005
Figure PCTCN2015071861-appb-000005
实施例1Example 1
本发明的一种复合微生物酶,其是由100%纤维素酶构成,其中选用的纤维素酶的酶活为20万μ/g。取100ml海带浆液,按与由海藻或海藻渣制成的浆液的质量比为0.4%的比例添加该复合微生物酶形成混合液,保温酶解25h,测定上清液中海藻酸的含量为17.25g/L。A composite microbial enzyme of the present invention consisting of 100% cellulase, wherein the cellulase enzyme activity selected is 200,000 μg/g. 100 ml of kelp slurry was added, and the mixed microbial enzyme was added in a ratio of 0.4% by mass of the slurry prepared from seaweed or seaweed residue to form a mixed solution, and the enzymatic hydrolysis was carried out for 25 hours, and the content of alginic acid in the supernatant was determined to be 17.25 g. /L.
实施例2Example 2
本发明的一种复合微生物酶,其是由质量百分比为60%的纤维素酶和40%的果胶酶混合而成,其中选用的纤维素酶的酶活为12万μ/g,果胶酶的酶活为1.2万μ/g。取100ml海带浆液,按与由海藻或海藻渣制成的浆液的质量比为 0.4%的比例添加该复合微生物酶形成混合液,保温酶解25h,测定上清液中海藻酸的含量为19.56g/L。The invention relates to a composite microbial enzyme which is prepared by mixing 60% by mass of cellulase and 40% of pectinase, wherein the cellulase activity of the selected cellulase is 120,000 μg/g, pectin. The enzyme activity of the enzyme was 12,000 μg/g. Take 100ml of kelp slurry, according to the mass ratio of the slurry made from seaweed or seaweed residue The composite microbial enzyme was added in a ratio of 0.4% to form a mixed solution, and the enzymatic hydrolysis was carried out for 25 hours, and the content of alginic acid in the supernatant was determined to be 19.56 g/L.
实施例3Example 3
本发明的一种复合微生物酶,其是由质量百分比为60%的纤维素酶和35%的果胶酶以及5%的蛋白酶混合而成,其中选用的纤维素酶的酶活为12万μ/g,果胶酶的酶活为1.05万μ/g,蛋白酶的酶活为2.5万μ/g。取100ml海带浆液,按与由海藻或海藻渣制成的浆液的质量比为0.4%的比例添加该复合微生物酶形成混合液,保温酶解25h,测定上清液中海藻酸的含量为21.32g/L。The invention relates to a composite microbial enzyme which is prepared by mixing 60% by mass of cellulase with 35% of pectinase and 5% of protease, wherein the cellulase activity of the selected cellulase is 120,000 μ. /g, the pectinase activity was 1.05 million μg/g, and the protease activity was 25,000 μg/g. 100ml of kelp slurry was added, and the mixed microbial enzyme was added in a ratio of 0.4% by mass of the slurry prepared from seaweed or seaweed residue to form a mixed solution, and the enzymatic hydrolysis was carried out for 25 hours, and the content of alginic acid in the supernatant was determined to be 21.32 g. /L.
实施例4Example 4
本发明的一种复合微生物酶,其是由质量百分比为60%的纤维素酶和35%的果胶酶以及5%的淀粉酶混合而成,其中选用的纤维素酶的酶活为12万μ,果胶酶的酶活为1.05万μ/g,淀粉酶的酶活为0.25万μ/g。取100ml海带浆液,按与由海藻或海藻渣制成的浆液的质量比为0.4%的比例添加该复合微生物酶形成混合液,保温酶解25h,测定上清液中海藻酸的含量为19.62g/L。The invention relates to a composite microbial enzyme which is prepared by mixing 60% by mass of cellulase and 35% of pectinase and 5% of amylase, wherein the cellulase activity of the selected cellulase is 120,000. μ, the pectinase activity was 1.05 million μg/g, and the amylase enzyme activity was 0.2 million μg/g. 100 ml of kelp slurry was added, and the composite microbial enzyme was added in a ratio of 0.4% by mass of the slurry prepared from seaweed or seaweed residue to form a mixed solution, and the enzymatic hydrolysis was carried out for 25 hours, and the content of alginic acid in the supernatant was determined to be 19.62 g. /L.
实施例5Example 5
本发明的一种复合微生物酶,其是由质量百分比为70%的纤维素酶和27%的果胶酶以及3%的蛋白酶混合而成,其中选用的纤维素酶的酶活为14万μ,果胶酶的酶活为0.81万μ/g,蛋白酶的酶活为1.5万μ/g。取100ml海带浆液,按与由海藻或海藻渣制成的浆液的质量比为0.4%的比例添加该复合微生物酶形成混合液,保温酶解25h,测定上清液中海藻酸的含量为22.35g/L。The composite microbial enzyme of the invention is prepared by mixing 70% by mass of cellulase and 27% of pectinase and 3% of protease, wherein the cellulase activity of the selected cellulase is 140,000 μ. The enzyme activity of the pectinase was 0.81 μg/g, and the enzyme activity of the protease was 15,000 μg/g. 100 ml of kelp slurry was added, and the mixed microbial enzyme was added in a ratio of 0.4% by mass of the slurry prepared from seaweed or seaweed residue to form a mixed solution, and the enzymatic hydrolysis was carried out for 25 hours, and the content of alginic acid in the supernatant was determined to be 22.35 g. /L.
实施例6Example 6
本发明的一种复合微生物酶,其是由质量百分比为60%的纤维素酶和30%的果胶酶、5%的蛋白酶以及5%的淀粉酶混合而成,其中选用的纤维素酶的酶活为12万μ/g,果胶酶的酶活为0.81万μ/g,蛋白酶的酶活为2.5万μ/g,淀粉酶的酶活为0.25万μ/g。取100ml海带浆液,按与由海藻或海藻渣制成的浆液的质量比为0.4%的比例添加该复合微生物酶形成混合液,保温酶解25h,测定上清液中海藻酸的含量为24.46g/L。A composite microbial enzyme of the present invention which is prepared by mixing 60% by mass of cellulase with 30% of pectinase, 5% of protease and 5% of amylase, wherein cellulase is selected. The enzyme activity was 120,000 μg, the pectinase activity was 0.81 μg/g, the protease activity was 25,000 μg/g, and the amylase enzyme activity was 0.2 million μg/g. 100 ml of kelp slurry was added, and the composite microbial enzyme was added in a ratio of 0.4% by mass of the slurry prepared from seaweed or seaweed residue to form a mixed solution, and the enzymatic hydrolysis was carried out for 25 hours, and the content of alginic acid in the supernatant was determined to be 24.46 g. /L.
结合实施例1~6的复合微生物酶对海藻的降解效果(酶解效果以释放海藻酸的量为标准)可以看出,复合微生物酶对鲜海带的酶解效果要比使用单一酶 效果好。鲜海带中除了含有纤维素、粗蛋白,还含有海带多糖,分别是褐藻胶、褐藻糖胶及褐藻淀粉。其中褐藻淀粉是一种细胞内多糖,主要由β-1,3-D-葡聚糖组成,长链上有少量β-1,6链间糖苷键存在,在海带中的含量一般为1%左右。因此复合微生物酶中选用淀粉酶作为辅助酶,可以使得鲜海带得到更充分的酶解。另,海带中粗蛋白的含量为5~9%,含量相对较低,而从表2中可以看出蛋白酶的价格比较高,若选用蛋白酶的用量跟其他酶种相当,会使得加工成本增加,造成资源浪费。因此,本发明中选用的纤维素酶、果胶酶的质量百分比远大于蛋白酶和淀粉酶,确定采用纤维素酶、果胶酶为复合微生物酶的基本酶种,蛋白酶和淀粉酶为复合微生物酶的辅助酶种。表3显示了实施例6组成的复合微生物酶对海带浆液的酶解效果最好。因此将实施例6作为复合微生物酶配比的最优实施例。Combining the compound microbial enzymes of Examples 1 to 6 to degrade algae (the enzymatic hydrolysis effect is based on the amount of released alginic acid), it can be seen that the enzymatic hydrolysis effect of the composite microbial enzyme on fresh seaweed is better than using a single enzyme. The effect is good. In addition to cellulose and crude protein, fresh seaweed also contains kelp polysaccharides, which are alginate, fucoidan and brown algae starch. Among them, brown algae starch is an intracellular polysaccharide mainly composed of β-1,3-D-glucan. There are a small amount of β-1,6 interchain glycosidic bonds on the long chain, and the content in kelp is generally 1%. about. Therefore, the use of amylase as an auxiliary enzyme in the complex microbial enzyme can make the fresh kelp more fully enzymatically hydrolyzed. In addition, the content of crude protein in kelp is 5 to 9%, and the content is relatively low. From Table 2, it can be seen that the price of protease is relatively high. If the amount of protease used is equivalent to other enzymes, the processing cost will increase. Causes waste of resources. Therefore, the mass percentage of cellulase and pectinase used in the present invention is much larger than that of protease and amylase, and the basic enzymes using cellulase and pectinase as composite microbial enzymes, and protease and amylase as complex microbial enzymes are determined. Auxiliary enzyme species. Table 3 shows that the composite microbial enzyme composed of Example 6 has the best enzymatic hydrolysis effect on the kelp slurry. Example 6 was therefore taken as the preferred embodiment of the complex microbial enzyme ratio.
3复合微生物酶加量对水解效果的影响3The effect of the amount of compound microbial enzyme on the hydrolysis
以实施例6中的最优配比组成的复合微生物酶来研究复合微生物酶的加量对海藻水解效果的影响。其水解效果如图5中所示。The composite microbial enzyme consisting of the optimal ratio in Example 6 was used to study the effect of the amount of the combined microbial enzyme on the hydrolysis effect of seaweed. Its hydrolysis effect is shown in Figure 5.
随着复合微生物酶的添加量从0~0.6%的变化,海藻酸的含量逐渐增加;并且随着复合微生物酶添加量从0~0.3%的变化,海藻酸含量呈大致线性增加;当复合酶添加量为0.3%时,海藻酸含量为17.5g/L;当复合酶添加量到达0.4%时,海藻酸含量到达18g/L;当复合酶添加量超过0.4%时,海藻酸的含量趋于平衡,不再增加。因此,综合成本和时间的考虑,本发明的复合微生物酶的添加量优选0.3~0.4%。The alginic acid content gradually increased with the addition of the complex microbial enzyme from 0 to 0.6%; and the alginic acid content increased substantially linearly with the addition of the complex microbial enzyme from 0 to 0.3%; When the added amount is 0.3%, the alginic acid content is 17.5g/L; when the compounding enzyme reaches 0.4%, the alginic acid content reaches 18g/L; when the compounding enzyme is added more than 0.4%, the alginic acid content tends to Balance, no longer increase. Therefore, the amount of the complex microbial enzyme to be added of the present invention is preferably from 0.3 to 0.4% in view of overall cost and time.
4酶解工艺参数的确定4 Determination of enzymatic process parameters
表4列出了本发明的复合微生物酶酶解过程中的反应温度、酶添加量、pH值和酶解时间等参数。表5列出了按照表4中所示参数进行正交试验的实施例7~15的结果分析。本发明通过表4和表5确定出本发明的优选工艺参数。Table 4 lists parameters such as reaction temperature, enzyme addition amount, pH value and enzymatic hydrolysis time in the enzymatic hydrolysis process of the composite microorganism of the present invention. Table 5 lists the results of Examples 7-15 of the orthogonal test according to the parameters shown in Table 4. The present invention determines the preferred process parameters of the present invention from Tables 4 and 5.
表4Table 4
Figure PCTCN2015071861-appb-000006
Figure PCTCN2015071861-appb-000006
Figure PCTCN2015071861-appb-000007
Figure PCTCN2015071861-appb-000007
表5table 5
Figure PCTCN2015071861-appb-000008
Figure PCTCN2015071861-appb-000008
取2700ml鲜海带匀浆,分装至27个250ml的三角瓶中,每瓶装量100ml,三个一组按照正交试验的实施例7~15进行实验,每组中三个样品按照一个实施例进行实验,酶解结束后测定上清液中海藻酸的含量,每个实施例中海藻酸含量取每组三个样品所测的海藻酸含量的平均值。由表3可知,实施例11为各因素的最优实施例。2700ml fresh seaweed homogenate was taken and packed into 27 250ml triangular flasks, each containing 100ml, three groups were tested according to the orthogonal test examples 7-15, three samples in each group according to one embodiment The experiment was carried out, and the content of alginic acid in the supernatant was measured after the end of the enzymatic hydrolysis. The alginic acid content in each example was taken as the average value of the alginic acid content measured in each of the three samples. As can be seen from Table 3, Embodiment 11 is a preferred embodiment of each factor.
5酶解时间对水解效果的影响5The effect of enzymatic hydrolysis time on hydrolysis
实施例16Example 16
取15L的鲜海带匀浆,将其加入酶解罐中,再加入复合微生物酶,其中复合微生物酶按照优选实施例6的配比混合,并且按照优选实施例11的工艺参数设置反应温度、复合微生物酶添加量及pH值,经测试其酶解后海藻酸含量平均值为23.2g/L。15 L of fresh seaweed homogenate was taken, added to the enzymatic hydrolysis tank, and then a composite microbial enzyme was added, wherein the composite microbial enzyme was mixed according to the ratio of the preferred embodiment 6, and the reaction temperature and the composite were set according to the process parameters of the preferred example 11. The amount of microbial enzyme added and the pH value were tested and the average alginic acid content was 23.2 g/L.
图6示出了优选实施例11的条件下,即酶解温度为55℃,复合生物酶添加量为0.4%,pH为5.5时,酶解时间对酶解效果的影响。随着酶解时间的增加, 海藻酸含量逐渐增加,并且当酶解时间从0~20h变化时,海藻酸含量呈大致线性增加,酶解时间达到18h时,海藻酸含量达到20g/L;酶解时间为25h左右时,海藻酸含量达到最大,此时的海藻酸含量为23g/L。之后随着酶解时间的增加海藻酸含量趋于平衡不再变化。综合成本考虑,本发明选用酶解时间为18h。Fig. 6 shows the effect of the enzymatic hydrolysis time on the enzymatic hydrolysis under the conditions of the preferred embodiment 11, i.e., the enzymatic hydrolysis temperature was 55 ° C, the amount of the composite biological enzyme added was 0.4%, and the pH was 5.5. As the enzymatic hydrolysis time increases, The alginic acid content increased gradually, and the alginic acid content increased linearly when the enzymatic hydrolysis time changed from 0 to 20 h. When the enzymatic hydrolysis time reached 18 h, the alginic acid content reached 20 g/L; when the enzymatic hydrolysis time was about 25 h, the algae The acid content reached a maximum, and the alginic acid content at this time was 23 g/L. Then, as the enzymatic hydrolysis time increases, the alginic acid content tends to balance and no longer changes. For the comprehensive cost consideration, the enzymatic hydrolysis time of the invention is 18h.
6海藻生物饲料的制备6 Preparation of seaweed biological feed
在确定好复合酶微生物酶的酶种及各种酶的相关配比以及最佳的酶解工艺参数之后,制备本发明的海藻生物饲料添加剂或海藻生物饲料。本发明的海藻生物饲料添加剂或海藻生物饲料的制备方法具体包括以下步骤:The seaweed biological feed additive or seaweed biological feed of the present invention is prepared after determining the enzyme species of the complex enzyme microbial enzyme and the relevant ratio of various enzymes and the optimal enzymatic process parameters. The method for preparing the seaweed biological feed additive or the seaweed biological feed of the present invention specifically comprises the following steps:
(1)将海藻或海藻渣经过无机除杂后剪切并研磨成直径在1mm以下颗粒的海藻浆液;将海藻或海藻渣研磨至1mm以下,使得海藻颗粒的活性营养物质充分暴露,从而使得复合微生物酶与活性物质充分接触,进而提高水解效率。(1) After the seaweed or algae residue is subjected to inorganic impurity removal, the seaweed slurry is cut and ground into particles having a diameter of 1 mm or less; and the seaweed or seaweed residue is ground to less than 1 mm, so that the active nutrients of the seaweed particles are fully exposed, thereby making the composite The microbial enzyme is in full contact with the active material, thereby improving the hydrolysis efficiency.
(2)将海藻浆液打入高速分散釜中第一次加水搅拌,使得水与海带浆液充分混合,并且第一次加水量与所述海带浆液的质量比为(30~40):100;进一步优选为35:100。(2) driving the seaweed slurry into the high-speed dispersion kettle for the first time, adding water and stirring, so that the water and the kelp slurry are thoroughly mixed, and the mass ratio of the first water addition to the kelp slurry is (30-40): 100; It is preferably 35:100.
(3)将混合后的海藻浆液打入酶解罐中,第二次加水并且加入预先配好的复合微生物酶,形成混合液。此时,进行第二次加水,并且第一次加水量和第二次加水量的总量与所述海藻浆液的质量比为50:100,合理控制加水量有利于控制水分含量,提高海藻饲料的营养含量。(3) The mixed seaweed slurry is put into the enzymatic hydrolysis tank, the second time water is added and the pre-formed composite microbial enzyme is added to form a mixed liquid. At this time, the second addition of water is performed, and the ratio of the total amount of the first water addition amount and the second water addition amount to the seaweed slurry is 50:100, and the reasonable control of the water addition amount is advantageous for controlling the moisture content and improving the seaweed feed. Nutritional content.
(4)将混合液搅拌均匀后加热保温酶解,并且在酶解过程中,每一小时搅拌一次,并且每次搅拌5分钟,使得海藻中的活性营养物质与复合微生物酶充分接触,提高水解效率。(4) The mixture is stirred and homogenized, then heat-insulated and enzymatically hydrolyzed, and stirred once every hour during the enzymatic hydrolysis process, and stirred for 5 minutes each time, so that the active nutrients in the seaweed are sufficiently contacted with the complex microbial enzyme to improve hydrolysis. effectiveness.
根据一个优选实施方式,在经历过上述四步骤之后的混合液通过反冲带式过滤机进行固液分离,并且将分离出的酶解清液在60~68℃的范围内进行浓缩;或者,将酶解好的混合液进行干燥,从而获得本发明的海藻生物饲料添加剂或海藻生物饲料。According to a preferred embodiment, the mixed solution after the above four steps is subjected to solid-liquid separation by a backflushing belt filter, and the separated enzymatic supernatant is concentrated in a range of 60 to 68 ° C; or The enzymatically mixed mixture is dried to obtain the seaweed biological feed additive or seaweed biological feed of the present invention.
根据一个优选实施方式,在搅拌过程中,为了使海藻浆液混合的更加均匀,使海藻浆与水能够充分混合,高速分散釜启动后的转速为3000转/分钟,搅动3分钟后,高速分散釜的转速由3000转/分钟降至1000转/分钟。 According to a preferred embodiment, in the process of stirring, in order to make the seaweed slurry more uniformly mixed, the seaweed pulp and the water can be sufficiently mixed, the rotation speed of the high-speed dispersion kettle after starting is 3000 rpm, and after stirring for 3 minutes, the high-speed dispersion kettle The speed was reduced from 3000 rpm to 1000 rpm.
根据一个优选实施方式,本发明的纤维素酶、果胶酶、蛋白酶和淀粉酶在复合微生物酶中所占的质量百分比分别是:60~100%,0~40%,0~5%,0~5%,并且纤维素酶源于李氏木霉、绿色木霉和/或黑曲霉;果胶酶源于米根霉、黑曲霉和/或米曲霉;蛋白酶源于木瓜蛋白酶、菠萝蛋白酶和/或枯草杆菌;淀粉酶源于地衣芽孢杆菌、米曲霉和/或黑曲霉。According to a preferred embodiment, the cellulase, pectinase, protease and amylase of the present invention account for a percentage by mass of the complex microbial enzyme: 60 to 100%, 0 to 40%, 0 to 5%, 0. ~5%, and the cellulase is derived from Trichoderma reesei, Trichoderma viride and/or Aspergillus niger; the pectinase is derived from Rhizopus oryzae, Aspergillus niger and/or Aspergillus oryzae; the protease is derived from papain, bromelain and / or Bacillus subtilis; amylase is derived from Bacillus licheniformis, Aspergillus oryzae and / or Aspergillus niger.
根据一个优选实施方式,本发明的纤维素酶的酶活为(18~24)×104μ/g,果胶酶的酶活为(2~5)×104μ/g,蛋白酶的酶活为(10~50)×104μ/g,淀粉酶的酶活为(1~5)×104μ/g。According to a preferred embodiment, the cellulase of the present invention has an enzyme activity of (18-24)×10 4 μg/g, and the pectinase has an enzyme activity of (2-5)×10 4 μg/g. The activity was (10 to 50) × 10 4 μ/g, and the enzyme activity of amylase was (1 to 5) × 10 4 μ/g.
根据一个优选实施方式,本发明的制备方法是向所述海藻浆液中按照复合微生物酶与海藻浆液的质量比为(0.3~3):100的比例添加复合微生物酶,并在常压、温度为30~65℃、pH3~7的条件下保温酶解6~30小时,从而获得所述海藻生物饲料添加剂或海藻生物饲料。According to a preferred embodiment, the preparation method of the present invention is to add a composite microbial enzyme to the seaweed slurry according to a mass ratio of the composite microbial enzyme to the seaweed slurry of (0.3 to 3):100, and at normal pressure and temperature. The algae biological feed additive or the seaweed biological feed is obtained by insulating the enzyme for 30 to 30 hours under conditions of 30 to 65 ° C and pH 3 to 7.
在本发明总的技术方案的指导下,本发明的另一方面涉及一种海藻生物饲料添加剂或海藻生物饲料,本发明的海藻生物饲料添加剂或海藻生物饲料是按照如前述的制备方法制备而成的。Under the guidance of the general technical scheme of the present invention, another aspect of the present invention relates to a seaweed biological feed additive or a seaweed biological feed, and the seaweed biological feed additive or the seaweed biological feed of the present invention is prepared according to the preparation method as described above. of.
并且,根据一个优选实施方式,本发明的海藻饲料添加剂或海藻生物饲料还包括鱼粉、鱼油、豆粕、木薯淀粉、玉米粉和粉末大豆磷脂中一种或多种。而且,根据需要,本发明的海藻生物饲料可以制备成粉状或颗粒状或不定形态。Moreover, according to a preferred embodiment, the seaweed feed additive or seaweed biological feed of the present invention further comprises one or more of fish meal, fish oil, soybean meal, tapioca starch, corn flour and powdered soybean phospholipid. Moreover, the seaweed biological feed of the present invention can be prepared into a powdery or granular form or an amorphous form as needed.
在本发明总的技术方案的指导下,本发明的另一方面涉及一种海藻生物饲料,尤其是用于饲养家禽/家畜或水产养殖的饲料。该海藻生物饲料包括如前述的制备方法制备而成的海藻生物饲料添加剂或如前述的海藻饲料添加剂。Under the guidance of the general technical solution of the present invention, another aspect of the present invention relates to a seaweed biological feed, particularly for feeding poultry/livestock or aquaculture. The seaweed biological feed comprises an algae biological feed additive prepared as described above or a seaweed feed additive as described above.
通过本发明制备的海藻生物饲料添加剂或海藻生物饲料既大大提高了海藻的消化吸收率,减少浪费,又可增强机体免疫力,同时减少抗生素的使用,是一种健康、高效、新型的养殖饲料。并且本发明和现有技术中的其他制备方法相比,其制备过程简单,易操作,生产时间短,适用于短时间内的大规模生产。The seaweed biological feed additive or seaweed biological feed prepared by the invention not only greatly improves the digestion and absorption rate of seaweed, reduces waste, but also enhances the immunity of the body and reduces the use of antibiotics, and is a healthy, efficient and novel breeding feed. . Moreover, the invention has simple preparation process, easy operation and short production time as compared with other preparation methods in the prior art, and is suitable for mass production in a short time.
需要注意的是,上述具体实施例是示例性的,本领域技术人员可以在本发明公开内容的启发下想出各种解决方案,而这些解决方案也都属于本发明的公开范围并落入本发明的保护范围之内。本领域技术人员应该明白,本发明说明书及其附图均为说明性而并非构成对权利要求的限制。本发明的保护范围由权 利要求及其等同物限定。 It should be noted that the above specific embodiments are exemplary, and those skilled in the art can devise various solutions inspired by the disclosure of the present invention, and these solutions are also within the scope of the present disclosure and fall into the present disclosure. Within the scope of protection of the invention. It should be understood by those skilled in the art that the claims The scope of protection of the present invention is right The requirements and their equivalents are limited.

Claims (10)

  1. 一种活性海藻饲料添加剂的制作方法,其特征在于,所述方法是向由海藻或海藻渣制成的海藻浆液中添加复合微生物酶进行酶解从而获得所述活性海藻饲料添加剂,其中,所述复合微生物酶由纤维素酶、果胶酶、蛋白酶和淀粉酶中的多种组成。A method for preparing an active seaweed feed additive, characterized in that the method comprises the steps of: adding a microbial enzyme to an algae slurry prepared from seaweed or algae residue to carry out enzymatic hydrolysis to obtain the active seaweed feed additive, wherein The complex microbial enzyme consists of a plurality of cellulases, pectinase, proteases and amylases.
  2. 如权利要求1所述的活性海藻饲料添加剂的制作方法,其特征在于,所述纤维素酶、所述果胶酶、所述蛋白酶和所述淀粉酶在所述复合微生物酶中所占的质量百分比分别是:60~100%,0~40%,0~5%,0~5%,并且The method for producing an active seaweed feed additive according to claim 1, wherein the quality of said cellulase, said pectinase, said protease and said amylase in said composite microbial enzyme The percentages are: 60 to 100%, 0 to 40%, 0 to 5%, 0 to 5%, and
    所述纤维素酶源于李氏木霉、绿色木霉和/或黑曲霉;The cellulase is derived from Trichoderma reesei, Trichoderma viride, and/or Aspergillus niger;
    所述果胶酶源于米根霉、黑曲霉和/或米曲霉;The pectinase is derived from Rhizopus oryzae, Aspergillus niger and/or Aspergillus oryzae;
    所述蛋白酶源于木瓜蛋白酶、菠萝蛋白酶和/或枯草杆菌;The protease is derived from papain, bromelain and/or Bacillus subtilis;
    所述淀粉酶源于地衣芽孢杆菌、米曲霉和/或黑曲霉。The amylase is derived from Bacillus licheniformis, Aspergillus oryzae and/or Aspergillus niger.
  3. 如权利要求2所述活性海藻饲料添加剂的制作方法,其特征在于,所述纤维素酶的酶活为(18~24)×104μ/g,所述果胶酶的酶活为(2~5)×104μ/g,所述蛋白酶的酶活为(10~50)×104μ/g,所述淀粉酶的酶活为(1~5)×104μ/g。The method for producing an active seaweed feed additive according to claim 2, wherein the cellulase has an enzyme activity of (18 to 24) × 10 4 μ/g, and the pectinase has an enzyme activity of (2). ~5)×10 4 μ/g, the protease activity of the protease is (10 to 50)×10 4 μ/g, and the enzyme activity of the amylase is (1 to 5)×10 4 μ/g.
  4. 如权利要求1所述活性海藻饲料添加剂的制作方法,其特征在于,所述方法是向所述海藻浆液中按照复合微生物酶与海藻浆液的质量比为(0.3~3):100的比例添加复合微生物酶,并在常压、温度为30~65℃、pH3~7的条件下保温酶解6~30小时,从而获得所述活性海藻饲料添加剂。The method for producing an active seaweed feed additive according to claim 1, wherein the method comprises adding a compound to the seaweed slurry according to a mass ratio of the composite microbial enzyme to the seaweed slurry of (0.3 to 3):100. The microbial enzyme is subjected to heat preservation for 6 to 30 hours under normal pressure, temperature of 30 to 65 ° C, and pH 3 to 7, thereby obtaining the active seaweed feed additive.
  5. 如权利要求4所述活性海藻饲料添加剂的制作方法,其特征在于,所述方法还包括以下步骤:The method for producing an active seaweed feed additive according to claim 4, wherein the method further comprises the steps of:
    将海藻或海藻渣经过无机除杂后剪切并研磨成直径在1mm左右颗粒的海藻浆液;The seaweed or algae residue is subjected to inorganic impurity removal, and then ground and ground into an algae slurry having a diameter of about 1 mm;
    将所述海藻浆液打入高速分散釜中第一次加水搅拌,使得水与海带浆液充 分混合,并且第一次加水量与所述海带浆液的质量比为(30~40):100;The seaweed slurry is pumped into the high-speed dispersion kettle for the first time, and the water and the kelp slurry are charged. Mixing, and the mass ratio of the first water addition to the kelp slurry is (30-40): 100;
    将混合后的海藻浆液打入酶解罐中,第二次加水并且加入所述复合微生物酶,形成所述混合液,并且第一次加水量和第二次加水量的总量与所述海藻浆液的质量比为50:100;以及The mixed seaweed slurry is driven into the enzymatic hydrolysis tank, water is added for the second time, and the composite microbial enzyme is added to form the mixed liquid, and the total amount of the first water addition amount and the second water addition amount and the seaweed The mass ratio of the slurry is 50:100;
    将所述混合液搅拌均匀后加热保温酶解,并且在酶解过程中,每一小时搅拌一次,并且每次搅拌5分钟。The mixture was stirred uniformly, and then heat-insulated and hydrolyzed, and stirred once every hour during the enzymatic hydrolysis, and stirred for 5 minutes each time.
  6. 如权利要求5所述活性海藻饲料添加剂的制作方法,其特征在于,将酶解好的混合液通过反冲带式过滤机进行固液分离,并且将分离出的酶解清液在60~68℃的范围内进行浓缩;或者,将酶解好的混合液进行干燥。The method for preparing an active seaweed feed additive according to claim 5, wherein the enzymatically mixed mixture is subjected to solid-liquid separation by a backflushing belt filter, and the separated enzymatic hydrolysate is at 60 to 68. Concentration is carried out in the range of °C; or the enzymatically prepared mixture is dried.
  7. 如权利要求5所述活性海藻饲料添加剂的制作方法,其特征在于,所述高速分散釜启动后的转速为3000转/分钟,搅动3分钟后,所述高速分散釜的转速由3000转/分钟降至1000转/分钟。The method for preparing an active seaweed feed additive according to claim 5, wherein the rotation speed of the high-speed dispersion vessel after starting is 3000 rpm, and after the agitation for 3 minutes, the rotation speed of the high-speed dispersion kettle is 3000 rpm. Dropped to 1000 rpm.
  8. 一种活性海藻饲料添加剂,其特征在于,所述活性海藻饲料添加剂是按照如权利要求1~7之一所述的制作方法制作而成的。An active seaweed feed additive, characterized in that the active seaweed feed additive is produced according to the production method according to any one of claims 1 to 7.
  9. 如权利要求8所述的活性海藻饲料添加剂,其特征在于,所述活性海藻饲料添加剂还包括鱼粉、鱼油、豆粕、木薯淀粉、玉米粉和粉末大豆磷脂中一种或多种。The active seaweed feed additive according to claim 8, wherein the active seaweed feed additive further comprises one or more of fish meal, fish oil, soybean meal, tapioca starch, corn flour, and powdered soybean phospholipid.
  10. 一种活性海藻饲料,尤其是用于饲养家禽/家畜或水产养殖的饲料,其特征在于,所述饲料包括如权利要求1~7之一所述的制作方法制作而成的活性海藻饲料添加剂或如权利要求8或9所述的活性海藻饲料添加剂。 An active seaweed feed, in particular for feeding poultry/livestock or aquaculture, characterized in that the feed comprises an active seaweed feed additive prepared by the production method according to any one of claims 1 to 7 or The active seaweed feed additive according to claim 8 or 9.
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