TW201922111A - Method of increasing free phosphorus in vegetable feeding stuff - Google Patents

Abstract

The present invention relates to a two-staged method of increasing free phosphorus in vegetable feeding stuff, which is characterized by treating the plant feeding stuff with Aspergillus oryzae to destroy the cell wall of the vegetable feeding stuff to release phytic acid, and then digesting the released phytic acid with phytase to release free phosphorus.

Description

Method for improving free phosphorus content in plant feed

The present invention relates to a method for increasing the free phosphorus content in a vegetable feed, and more particularly to a method for improving the free phosphorus content in a vegetable feed by a two-stage treatment using rice bran and phytase.

Phosphorus is an essential trace element for animal growth, and phosphorus deficiency leads to decreased production performance and increased mortality. Phytic acid (also known as "hexaphosphate") is a compound found primarily in plant seeds (such as soybean seeds) and is a way in which seeds are used to store phosphorus (at least 80% of the phosphorus in the grain is in the form of phytic acid). The phytic acid system consists of a muscle type inositol core covalently attached to six phosphate groups. Monogastric animals, such as poultry and pigs, cannot effectively digest the phosphoric acid in phytic acid because of the lack of enzymes required to digest phytic acid.

Phytate in plant feed not only affects the absorption of phosphorus in the digestive tract, but in the environment of stomach acid in the digestive tract of poultry, most of the phytate is soluble or can be mixed with protein, so it is digested. In the channel, a variety of different phytate mixtures can be formed. If the diet contains high levels of calcium (a general phenomenon in egg-laying diets), the protein-phytate bond will break and form a calcium-phytate precipitate. In the intestinal acid-base environment, one gram of phytic acid can bind 3 to 6 moles of calcium to form a non-soluble phytate. Once this phenomenon occurs, trace minerals may also precipitate together with the calcium-phytate mixture, so phytic acid has many harmful effects.

However, most of the phosphorus in plant feeds and seeds are in the form of phytate phosphate (phytate). Therefore, animals can only use a small amount of free phosphorus in the feed, while the phosphorus contained in phytic acid is in the animal. The essential nutrients that cannot be synthesized by themselves, therefore, the industry needs to provide phosphorus in the feed to accelerate its growth. In the past, the solution to the problem was to add an inorganic phosphorus supplement directly to the animal feed. However, the use of inorganic phosphorus supplements causes an excretion of the explosives and an increase in the amount of phosphate entering the environment, which is a major problem for environmental protection.

There are currently examples of enzymes that utilize phytase as a supplement to animal feed pellets. For example, Taiwan Patent Publication TW201512400 discloses a planting acidase as an additive for animal feed, which can increase total phosphorus contained in feed, reduce phosphorus released into the environment via excretion, and increase other minerals by adding phytase. Digestibility of substances and amino acids. In addition, studies have reported that the addition of phytase can increase the utilization of phytate phosphorus in livestock and poultry, reduce the amount of calcium hydrogen phosphate in the diet, and reduce the cost of feed formulation. At present, the conventional method of phytase is to add 500 FTU/kg of phytase to the mixed feed of livestock and poultry, which can release 40%~50% of phytate phosphorus in the diet instead of adding phytase. 0.1% of available phosphorus provided by calcium hydrogen phosphate or other sources of phosphorus, equivalent to calcium hydrogen phosphate (16% phosphorus), is about 6.25 kg/t. According to the report, in order to improve the utilization rate of feed, excessive addition of phytase and its effect (research on the excessive addition of phytase in the diet) Journal of Animal Nutrition 2017, Vol.29(2): 382-388 .

The purpose of the present invention is to focus on how to make the phosphorus content in the vegetable feed to maximize the free phosphorus content under the control of lower cost, and to generate higher economic benefits to provide animal utilization.

The present invention is based on the above object, and provides a method for treating plant feed by two-stage treatment, so that the plant feed can release a large amount of free phosphorus after treatment, thereby obtaining a feed having a higher free phosphorus content, so that the feed can be obtained. High levels of phosphorus are required for the supply of animals and meet the demands of high economic efficiency.

Accordingly, one aspect of the present invention relates to a method for improving the content of free phosphorus in a vegetable feed by a two-stage treatment comprising: in the first stage, the action of rice bran bacteria and phytic acid to decompose and destroy the surface structure of the plant cell wall To relieve the trapping of the plant cell wall outer structure, and the second stage of the plant, adding phytase, the phytic acid is efficiently distributed to release free phosphorus.

In some embodiments of the invention, the first stage is to ferment 0.1%-0.5% (w/w) of the rice blast fungus with the feed material at 30 ° C for one to three days. In a preferred embodiment of the present invention, the rice bran strain is added to the plant feed material by a fungus which is prepared by pre-synthesis with soybean at 30 ° C for one to two days.

In some embodiments of the present invention, the second stage of the phytase is cultured at 30 ° C for three to four days. In a preferred embodiment of the invention, the phytase activity is 5000 units/g.

In some embodiments of the invention, the method further comprises mixing the feedstock with water prior to the first stage.

Figure 1 shows the effect of adding phytase (Ph), bacillary dysentery (Ao) and two-stage addition (Ao->Ph) on free phosphorus in soybean hulls.

Figure 2 shows the cellulase activity after fermentation of the addition of bacillary dysentery (Ao) to the soybean hulls and two-stage addition (Ao->Ph).

The other features and advantages of the present invention are further exemplified and illustrated in the following examples, which are intended to be illustrative only and not to limit the scope of the invention.

In the first and second stages, the phytase (Ao) is combined with phytase to treat plant feed ingredients.

Preparation of bacillus: 50g (w/v) deionized water was added to 50g soybeans. After sterilization, 0.1% (w/w) Aspergillus oryzae powder was added and fermented at 30 °C for two days.芫Use.

Two-stage treatment of plant feed: The soybean shell is pulverized and used as a feed material for the test. Sterilization was carried out by adding 100% (w/v) deionized water to 50 g of crushed soybean hulls. In the first stage treatment, the bacillus prepared as described above is first added, and the fermentation is carried out at 30 ° C for one day; then the second stage treatment is carried out, and then phytase (phytase is activated, the activity is 5000 unit / g), and the culture is carried out at 30 ° C. day.

In addition, a control group was prepared: 100% (w/v) deionized water was added to 50 g of crushed soybean shell, and sterilized at 121 ° C for 15 minutes, and then left at 30 ° C for 4 days; phytase (Ph) group: 50g of crushed soybean shell was added with 100% (w/v) deionized water, sterilized, phytase was added and placed at 30 ° C for 4 days; and bacillus (Ao) group: 100% of 50 g of crushed soybean shell was added. (w/v) Deionized water was sterilized, phytase and bacillus were added, and left at 30 ° C for 4 days.

Example 2: Evaluation of free phosphorus content in plant feed by two-stage bacillary dysentery (Ao) combined with phytase

Phosphorus determination: 5g sample was taken and 45mL deionized water was added. Stirring was continued for 1 hour at room temperature to extract inorganic phosphorus. Vanadium molybdate acid was used as a reagent to combine with inorganic phosphorus to form a yellow compound, which was determined at a wavelength of 415 nm. Absorbance value; a standard curve was prepared with KH2PO4, and the concentration of the extracted inorganic phosphorus was calculated.

As shown in the results of FIG. 1, the treatment of the two-stage addition (Ao->Ph) of the present invention is compared with the treatment group in which only phytase (Ph) or bacillus (Ao) is added, and the free phosphorus in the bean shell. Released, there is a significant improvement (relative to the untreated bean hull, the feed ingredient control group).

Cellulase activity assay: 5g sample was weighed and added with 45mL 0.1M, pH=5.5 acetate buffer, 30 minutes after ice extraction, centrifuged at 4000rpm for 10 minutes, filtered with Advantec No.1 filter paper to obtain cellulase. Activity determination; 1% carboxymethyl-cellulose dissolved in pH=5.5 acetate buffer as a substrate, 0.5mL enzyme solution mixed with 0.5mL substrate solution and reacted at 37 ° C for 15 minutes, with 3mL of 3,5-dinitro water Salicylic acid (DNS) is used as a reaction terminator and color former because it forms an orange compound with reducing sugar, absorbs the absorbance at a wavelength of 540 nm, and draws a standard curve with 0.1-0.5 mg/mL glucose, and returns the equation according to the standard curve. Cellulase activity was calculated; cellulase activity was defined as 1 g of sample degraded from 1% of the cellulose matrix per minute at 37 ° C, pH = 5.5 to produce 1 μmol of glucose as 1 unit.

The results in Figure 2 show that the two treatment groups (Ao and Ao->Ph after fermentation with added bacillus) compared to the untreated soybean shell group (feed material addition group) and the phytase-only treatment group (Ph). The cellulase enzyme activity has a significant effect.

Based on the above results, the present invention first utilizes rice bran bacteria to decompose and destroy the surface structure of the plant cell wall of the plant feed, and then adds a two-stage plant feed treatment method in which phytase is separated from the phytic acid which is detached from the plant cell wall. In the process of the first stage, phytic acid can be released by first releasing the outer structure of the plant cell wall, and then phytase is efficiently decomposed, and a large amount of free phosphorus is released. By the method of the invention, the method of lower manufacturing cost can effectively improve the phosphorus content in the treated plant feed, and the utility value of the cookware industry.

Claims (5)

A two-stage treatment method for improving the content of free phosphorus in plant feed materials comprises: in the first stage, adding Aspergillus oryzae to phytic acid in a plant feed material, the surface structure of the plant cell wall is Decomposition and destruction, release the phytic acid in the outer wall of the plant cell wall, and the second stage, adding phytase (activity of 5000 units/g) to the plant feed treated in the first stage for decomposition of phytic acid Free phosphorus is released. The method of claim 1, wherein 0.1%-0.5% (w/w) of the rice blast fungus is fermented with the feed material at 30 ° C for one to three days in the first stage. The method according to claim 1 or 2, wherein the rice bran strain is added to the plant feed material by a fungus which is prepared by pre-synthesis with soybean at 30 ° C for one to two days. The method of claim 1, wherein the phytase is incubated at 30 ° C for three to four days in the second stage. The method of claim 1 further comprising mixing the feedstock with water prior to the first stage.