JP2003012389A - Liquid fertilizer containing peptides and amino acid and method of manufacturing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a liquid fertilizer and a method of manufacturing the liquid fertilizer having preferable fertilizer response, and little growth inhibition. SOLUTION: The liquid fertilizer is characterized in that in the liquid fertilizer containing peptides and amino acid, nitrogen in the peptides is 30-70 mass% of total amount of nitrogen in the peptides and the amino acid.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a liquid fertilizer and a method for producing the same, and more particularly to a liquid fertilizer containing peptides and amino acids and a method for producing the same.

[0002]

2. Description of the Related Art Conventionally, 16 elements of hydrogen, carbon, oxygen, nitrogen, phosphoric acid, potassium, calcium, magnesium, sulfur, iron, manganese, boron, copper, zinc, molybdenum and chlorine are generally required for plant growth. It is said that Of these, hydrogen, carbon, and oxygen are required in large amounts, but it is not necessary to artificially give them as fertilizer by replenishing them from the air or water. On the other hand, nitrogen is said to be a fertilizer together with phosphate and potassium for producing chlorophyll, enzymes, plant hormones, etc. in addition to proteins, which are the main components of cells forming plants.

In recent years, with the advance of petrochemicals, fertilization mainly on chemical fertilizers has been carried out, and the use amount of organic fertilizers and fertilizers containing organic substances has decreased. However, since fertilization biased toward such chemical fertilizers causes a decline in soil fertility,
Recently, organic fertilizers or organic fertilizers have been receiving attention. The active ingredient of organic fertilizers is mainly nitrogen content, and as a typical one of them, animal organic fertilizers such as fish dregs, meat dregs, bone meal, horn horn dust, and leather dust, rapeseed oil dregs, soybean oil dregs, Known plant-based organic fertilizers such as cottonseed oil dregs, waste-based organic fertilizers such as dried fungi and sludge are known.

Extraction residue of gelatin, which is one of typical animal organic substances, or glue or by-product produced in the production process thereof (hereinafter, this extraction residue is extracted with hot water at higher temperature, hydrolyzed, or protein Organic matter containing (including extraction products including those obtained by degrading enzymes) contains amino acids having a remarkable fertilizing effect such as proline, glycine, glutamic acid, and aspartic acid, but it is used as a fertilizer. Also, the fertilizing effect was insufficient. When the present inventor conducted a fertilization test on gelatin and the extraction residue of gelatin with a commercial chemical fertilizer, the expected fertilization effect was not obtained. Further, these animal organic substances have low water-solubility, and when they are used in combination with other fertilizing components such as phosphoric acid and potash components, they tend to precipitate and are difficult to use as raw materials for liquid fertilizers.

On the other hand, fertilizers using amino acids obtained by hydrolyzing animal and plant proteins and amino acids obtained by chemical synthesis or fermentation are described in, for example, JP-A-11-228276,
7-10670, 6-40786, 6-24
No. 884, No. 5-330965, No. 5-330968
No. 2001-89278 and 2001-8927.
9, ibid 2000-327471, ibid 2000-1780
91, ibid 2000-26181, ibid 2001-8927.
9 etc. These amino acids have small molecular weights and have improved water solubility.

However, although fertilizers using such low molecular weight amino acids can be expected to have a rapid effect, the fertilizer effect does not continue. In addition, when the present inventor conducted a fertilization test using these commercially available chemical fertilizers, some of the adapted plant species showed a growth disorder as seen during excessive fertilization.

Therefore, there is a demand for the development of a liquid fertilizer having a positive fertilizing effect without any growth damage to crops.

[0008]

SUMMARY OF THE INVENTION It is an object of the present invention to provide a liquid fertilizer having a suitable fertilization rate and less growth damage, and a method for producing the same.

[0009]

The above object of the present invention can be achieved by the following means.

1. A liquid fertilizer containing peptides and amino acids, wherein the nitrogen content of the peptides is 30 to 70% by mass of the total nitrogen content of the peptides and amino acids.

2. The liquid fertilizer according to the above 1, wherein the total amino nitrogen content of the peptides and amino acids is 1% by mass or more.

3. Phosphoric acid (P ₂ O ₅ ) component, potassium (K ₂ O)
The liquid fertilizer according to 1 or 2 above, wherein the components are 2 to 7% by mass and 2 to 7% by mass, respectively.

4. Manufacture of liquid fertilizer characterized by manufacturing the liquid fertilizer according to any one of 1 to 3 above using peptides and amino acids obtained by hydrolyzing gelatin, glue or an extraction residue by-produced in the manufacturing process thereof. Method.

5. The fats and oils contained in peptides and amino acids obtained by hydrolyzing gelatin, glue, or an extraction residue produced as a by-product in the production process are
The method for producing a liquid fertilizer according to the above item 4, characterized in that the content is at most% by mass.

The present invention will be described in detail below.

As a result of earnest research in view of such circumstances, the present inventor has obtained the following knowledge and completed the present invention.

That is, the peptides are decomposed mainly in the soil by the action of microorganisms, and a part of them is mineralized to first produce ammonia, and then the ammonia is changed to nitric acid. This mineralization rate is slower as the molecular weight of the peptides is higher, and this is mainly because the expected fertilization effect was not obtained in the fertilization test of gelatin and the extraction residue of gelatin described above. Most of amino acids (including peptides with extremely small molecular weight) are not mineralized and are directly absorbed from the leaf surface or roots, which contributes to the rapid effect.
In addition, if the rate of mineralization is very fast, such as urea,
Before the nitrogen component is absorbed by agricultural products, a large part is lost due to leaching and denitrification, which may reduce the utilization rate of fertilizer. Therefore, in order to have a suitable fertilization rate, it is necessary to select the mixing ratio of peptides and amino acids in order to control the mineralization of nitrogen components. In addition, a part of the liquid fertilizer is directly applied to the leaf surface, concentrated due to the drying of water, and the growth of osmotic pressure may cause growth failure. It is considered that the increase in osmotic pressure is also related to the molecular weight of peptides.

As a result of further examination based on the above findings,
The object of the present invention was found to be achieved in a liquid fertilizer containing peptides and amino acids by adjusting the nitrogen content of the peptides to 30 to 70% by mass of the total nitrogen content of the peptides and amino acids. . In order to further develop the effects of the present invention, the total amino nitrogen content of the peptides and amino acids should be 1% by mass or more, the phosphoric acid (P ₂ O ₅ ) component, and the potassium (K ₂ It is preferable that the O) components are 2 to 7% by mass and 2 to 7% by mass, respectively.

In the production method, peptides, amino acids obtained by hydrolyzing gelatin, glue, or an extraction residue produced as a by-product in the production process are used, and the oil and fat content thereof is 5% by mass or less of the peptides and amino acids. It is preferable.

Next, the peptides and amino acids used in the present invention will be described in detail. Among the polypeptides produced by dehydration polycondensation of the carboxyl group and amino group of L-α-amino acids, those having a molecular weight of more than 5000 are called proteins, and those having a molecular weight of 5,000 or less are called peptides in the present invention. Peptides include simple peptides consisting only of polypeptide chains and complex peptides containing sugar, fat, and phosphorus, and both peptides are included in the present invention.
In the present invention, the molecular weight of the peptides is preferably 2 to 50, more preferably 2 to 30, and even more preferably 2 to 10 in terms of the degree of polymerization of amino acids. When the molecular weight is larger than this, the water solubility and the fertilizing effect are slow, and, for example, when it is used in combination with other fertilizing components, precipitation tends to occur.

The types of amino acids used in the present invention include glycine, alanine, valine, leucine, isoleucine, proline, hydroxyproline, arginine, lysine, hydroxylysine, aspartic acid, glutamic acid, serine, threonine, tyrosine, phenylalanine, histidine. , Methionine and cystine. Further, a mixture of these can also be preferably used.

The starting materials for peptides and amino acids are:
Animal protein such as fish debris, meat debris, bone meal, hoof horn meal, leather meal, and vegetable organic matter such as rapeseed oil residue, soybean oil residue, cottonseed oil residue can be used. A raw extraction residue is preferred.

To obtain peptides and amino acids, these raw materials may be hydrolyzed with an inorganic acid such as hydrochloric acid, sulfuric acid or nitric acid, neutralized and desalted by a conventional method. In addition, as the amino acid, an amino acid obtained by chemical synthesis or fermentation can also be used.

When composing a liquid fertilizer using these peptides and amino acids, it is necessary that the nitrogen content of the peptides is 30 to 70 mass% of the total nitrogen content of the peptides and amino acids. If it is less than this range, growth failure tends to occur, and if it exceeds this range, the rate of fertilization is insufficient. In addition, it is preferable that the total amino nitrogen content of peptides and amino acids in the total nitrogen content is 1% by mass or more. 1
If it is less than mass%, the effect of amino nitrogen is weak. The upper limit is not particularly limited, and can be selected in consideration of solubility and precipitability.

In order to determine the total nitrogen content of peptides and amino acids, ammonia obtained by hydrolyzing this solution may be titrated with an acid by a standard method. To determine the nitrogen content of only amino acids in a mixed solution of peptides and amino acids, according to the formol titration method, after neutralizing this solution, the amino group of the amino acids is lost as an oxymethyl derivative by addition of formalin, Titrate with caustic soda standard solution.

The fat content of the liquid fertilizer of the present invention is preferably about 2.0 mass% or less. If the oil and fat content exceeds this range, the fertilizing effect is reduced and the components may change during storage, which is not preferable. Therefore, when using peptides or amino acids obtained by hydrolyzing gelatin, glue, or an extraction residue produced as a by-product in the production process thereof, the oil and fat content is preferably 5% by mass or less. When the amount of fats and oils is high, gelatin, glue or an extraction residue by-produced in the production process thereof can be defatted by a chitosan coagulation sedimentation method, an ion exchange resin method or a fats and oils-adsorptive filter medium filtration method.

Known gelatin may be used as a preferable starting material for the peptides and amino acids used in the present invention, or glue or an extraction residue by-produced in the production process thereof. Gelatin is a color matching the skin and bones of animals.
It is obtained by extracting Gen with hot water and drying. The gelatin extraction step is usually carried out 3 to 5 times by gradually increasing the temperature of the hot extraction water to about 45 to 80 ° C. As the gelatin used as the raw material of the fertilizer of the present invention, gelatin extracted with high temperature water is used in terms of price. The oil and fat content of such gelatin is as high as several mass% although it depends on the oil and fat content of the raw material collagen itself. Also,
The residue after such extraction further contains collagen, and a gelatin extraction residue can be obtained by extraction with hotter hot water, hydrolysis, or decomposition with a proteolytic enzyme. This residue may contain high fats and oils, up to 20% by weight.

[0028] Gelatin, glue or an extraction residue produced as a by-product in the production process thereof contains collagen protein as a main component and contains fats and oils, water, Ca and the like. This collagen protein is
It is composed of amino acids such as glycine, proline, hydroxyproline, glutamic acid, alanine, arginine, aspartic acid, lysine, leucine, serine and valine in descending order of content, and the content of proline and hydroxyproline is higher than that of other proteins.

As described above, gelatin, glue or an extraction residue by-produced in the production process thereof is derived from animals, but these contain amino acids important for plants. For example, in the body metabolism of paddy rice, proline is accumulated during flower bud formation and particle settling time, and in plants such as eggplant where vegetative growth and reproductive growth are performed at the same time, proline increases in the amino acid balance at the flower bud site. In vivo metabolic system uptake experiments have shown that proline is taken up in large amounts during the reproductive growth period.

As described above, gelatin, glue, or the extraction residue by-produced in the production process thereof originally has a very excellent fertilizing component. When peptides and amino acids obtained by hydrolyzing gelatin, glue, or an extraction residue produced as a by-product in the manufacturing process are fertilized, the active ingredients peptides and amino acids are gradually decomposed by bacteria in the ground, and a considerable part is converted to ammonia. Is absorbed by plants in the form of nitrate or nitrate,
Some amino acids or peptides are absorbed as they are. Mineralization test of peptides extracted from gelatin extraction residue by proteolytic enzyme (Organic nitrogen compounds such as proteins contained in organic matter are decomposed by the action of soil microorganisms to determine the amount of inorganic nitrogen compound ammonia or nitric acid produced. (Measured), it was slightly larger in 1 week and almost the same in 2 weeks as compared with steamed hoof horn powder, which is a typical animal-based surficial fertilizer.

In comparison with rapeseed meal, which is a typical plant organic fertilizer, it was slightly smaller in 1 week and almost the same in 2 weeks. In this way, the peptides extracted from the gelatin extraction residue by proteolytic enzymes have the same mineralization rate as steamed hoof horn powder or rapeseed meal, but there are differences in fertilization between amino acids and peptides. There is a part that is absorbed by, which is considered to be the cause.

The fats and oils contained in gelatin, glue or the extraction residue produced as a by-product in the production process thereof are mainly composed of esters of higher fatty acids and their hydrolysates. The fatty acid composition is
Oleic acid, palmitic acid, stearic acid, palmitoleic acid, linoleic acid, myristic acid, and the like are characterized in that the unsaturated fatty acids that are easily oxidized are as high as 70%.

The fats and oils contained in gelatin, glue or an extraction residue produced as a by-product in the production process are quantified as the fats and oils after extraction with hexane (gelatin test method for photography, 7th edition, 22nd edition). (Refer to section, Issued by the Joint Council on Photographic Gelatin Testing Law). Gelatin, glue or the fat and oil contained in peptides and amino acids obtained by hydrolyzing the extraction residue by-produced in the production process can also be used to quantify the components extracted with hexane as fat and oil. it can.

To degrease gelatin, glue or an extraction residue by-produced in the production process thereof, a coagulation sedimentation method using a polymer coagulant such as chitosan, an ion exchange resin method, a high-performance oil adsorbent method, pulp filtration Method, heating floatation method, activated carbon method, pH
There is a coagulation sedimentation method by adjustment and the like, and these methods can be combined in consideration of desired product properties and cost.
Of these, the coagulation sedimentation method using chitosan, the ion exchange resin removal method and the high performance oil adsorbent method are particularly preferable. In addition,
Chitosan is a polysaccharide containing an amino group obtained by hydrolyzing chitin, which is an organic skeletal substance of arthropods, annelids, mollusks, etc., with concentrated alkali.

In order to degrease chitosan, an aqueous solution of chitosan is added to and stirred with an aqueous solution of gelatin, glue or an extraction residue by-produced in the production process. The amount of chitosan added is 0.03 to 1.0 part by mass, preferably 0.1 to 0.5 part by mass, relative to 100 parts by mass of gelatin, glue or an extraction residue produced as a by-product in the production process.

The standard operating conditions are 40 to 60 ° C.
Then, an aqueous solution of 5 to 10% by mass of gelatin, glue or an extraction residue by-produced in the manufacturing process is prepared, and 0.1 to
1.0 mass% chitosan aqueous solution or water dispersion is added and stirred for 0.2 to 1.0 hour. When left to stand for 0.1 to 0.5 hours, flocs having a high oil content are formed, which are removed by centrifugation. Since the degreasing effect can be improved by increasing the amount of chitosan used, the amount of chitosan used is adjusted according to the oil and fat content of the raw material.

When degreasing with an ion exchange resin,
It is preferable to use a strong acid cation exchange resin and a strong salt cation exchange resin in combination by a known method. For example, gelatin, glue, or an extraction residue by-produced in the manufacturing process 1
20 to 150 L of ion exchange resin is used for 00 kg.

The standard operating conditions are 40 to 60 ° C.
Then, an aqueous solution of 5 to 10% by mass of gelatin, glue or an extraction residue by-produced in the manufacturing process is prepared, and the space velocity (SV) is 2-1 in an ion exchange column filled with a strongly acidic ion exchange resin.
Pass through with 5. The effluent is similarly treated with a strongly basic ion exchange resin. Strongly acidic ion exchange resins include Diaion SK-1B, Amberlite 1R-120, and 1R-
122, XE-100, Dowex 50 and the like can be used as appropriate. Strongly basic ion exchange resins include Diaion SA-10A, SA-11A, SA-20
A, Amberlite 1RA-400, IRA-40
1, IRA-410, IRA-411, Dowex 50 and the like can be used.

The high-performance oil adsorbent is a product for oil pollution and environmental pollution, and is a material that specifically adsorbs oil and fat. There are sheets, rolls, and powders, which are commercially available from Organo Corporation, Mitsui Toatsu Chemicals, Inc., etc.

The defatted gelatin thus obtained, glue or the fertilizer containing peptides and amino acids obtained by hydrolyzing the extraction residue by-produced in the production process thereof may be used as a simple fertilizer. Alternatively, other fertilizing components may be added and mixed for use.

A simple fertilizer containing peptides and amino acids
In case of fertilizer, phosphoric acid and potassium content are insufficient.
Therefore, fertilizers containing it are generally preferred, and phosphoric acid (P
_TwoO ₅) Ingredient, potash (K_TwoO) component is 2 to 7 mass each
%, 2 to 7 mass% is preferable.

The fertilizer of the present invention can be used for various plants. Depending on the target plant, chitosans, water-soluble vitamins, nucleic acids, organic bases and other trace elements which are plant cell active ingredients may be used. It is preferable to add magnesium, sulfur, iron, manganese, boron or the like.

The amount of fertilizer applied varies depending on the amount of fertilizer components in the land, the type of source fertilizer and the type of applied fertilizer, the type of plant applied, the season, etc., but it can usually be used in the range of 0.1 to 10 g / m ² . The standard amount used is 0.5-2 g / m ² .

[0044]

EXAMPLES Next, the present invention will be described more specifically based on examples, but the present invention is not limited to these.

Example 1 Phosphoric acid (P ₂ O ₅ ) component and potassium (K ₂ O) component were 4% by mass, the total amino nitrogen content of peptides and amino acids was 2% by mass, and gelatin was hydrolyzed. The ratio of the nitrogen content of peptides to the total content of nitrogen content of peptides and amino acids is adjusted to 20, 40, 60, 80 by adjusting the degree.
Liquid fertilizers 1, 2, 3, and 4 were prepared using the peptides and amino acids whose mass% was changed.

This liquid fertilizer was diluted 150, 200, 250 and 300 times into strawberry, eggplant, cherry tomato and cucumber and sprayed on the leaf surface. As a result, liquid fertilizer 1 having a ratio of the nitrogen content of peptides to the total content of nitrogen content of peptides and amino acids was 20 mass% and was grown 150-, 200-, and 250-fold on eggplant, cherry tomato, and cucumber leaves. I got an obstacle. All the other liquid fertilizers were diluted with all vegetables and showed no leaf damage. For strawberries, eggplants, cherry tomatoes, cucumbers, etc., where fruits are repeatedly harvested, it is necessary to give a liquid fertilizer from the leaf surface, which has a long cultivation period and is effective for maintaining fruit and tree growth at the same time as flower bud formation. There are many cases. However, the results of this experiment showed that fertilizers with a low ratio of nitrogen content of peptides and nitrogen content of peptides and amino acids (high nitrogen content of amino acids as nitrogen content) were applied to the foliage from the viewpoint of growth failure. It indicates that spraying should be avoided.

Regarding the fertilizing rate, the ratio of the nitrogen content of peptides to the total nitrogen content of peptides and amino acids was 7
Although 0% by mass of liquid fertilizer 4 was diluted 200, 250 and 300 times and was insufficient in all vegetables, it was confirmed that other liquid fertilizers contributed to the growth immediately after application, and the expected fertilizer It was effective. That is, in the case of strawberries, shoots emerge, in the case of eggplants, fruit enlargement is observed, in the case of cherry tomatoes, the fruit shape becomes larger, in the case of cucumber, the shoots grow stronger, and flower bud formation is better. became. In addition, in the case of cherry tomatoes, it was evaluated that the taste was improved.

[Example 2] In the liquid fertilizer 2 of Example 1, peptides and amino acids having a total amino nitrogen content of 0.5, 2.0 and 4% by mass were used, and the total nitrogen content was 7% by mass. Urea is added to make the liquid fertilizer 5, 6, 7
Was produced.

As a result of evaluation of growth disorders in the same manner as in Example 1, 150 and 2 were obtained for liquid fertilizer 5 in which the total amino nitrogen content of peptides and amino acids was 0.5% by mass.
At a dilution of 00 or 250 times, the growth of eggplant, cherry tomato and cucumber leaves was impaired. The liquid fertilizers 6 and 7 having high amino nitrogen content were all vegetables and diluted, and no leaf growth failure was observed. This is probably because the mineralization rate of urea is much faster than that of amino nitrogen.

Example 3 The gelatin extraction residue was decomposed with an alkaline protease, filtered, concentrated and dried. This oil and fat content was 8.3 mass%. This 6 mass% aqueous solution 1
Add 45 L of 0.5 mass% chitosan aqueous solution to KL, and add 45
Stir at 1 ° C for 1 hour. After standing for 1 hour, the solid content was removed by centrifugation, concentrated and dried. The oil and fat content of the gelatin extraction residue degreased in this manner was 1.5% by mass. This was hydrolyzed, neutralized and desalted with hydrochloric acid by a conventional method to obtain a mixed solution of peptides and amino acids. In this solution, the nitrogen content of the peptides was 50 mass% of the total nitrogen content of the peptides and amino acids, and the oil and fat content was 1.2 mass% of the peptides and amino acids.

Next, ammonium nitrate, nitric acid, phosphoric acid, dipotassium phosphate, diammonium phosphate and potassium chloride were added to the obtained mixture of peptides and amino acids to give an amino nitrogen content of 2.0% by mass, A liquid fertilizer 8 was prepared by adjusting the total nitrogen content to 5 mass%, the phosphoric acid content (P ₂ O ₅ ) to 5 mass%, and the potassium content (K ₂ O) to 5 mass%. Further, the gelatin extraction residue before being defatted with chitosan was similarly hydrolyzed to obtain a mixed solution of peptides and amino acids. In this solution, the nitrogen content of the peptides was 50 mass% of the total nitrogen content of the peptides and amino acids, and the oil and fat content was 6.5 mass% of the peptides and amino acids.
A liquid fertilizer 9 for comparison was prepared in the same manner using this mixed liquid. The obtained liquid fertilizers 8 and 9 and a commercially available lawn fertilizer "organic advanced advanced chemicals No. 100" (Mitsui East pressure fertilizer department
Co., Ltd., total nitrogen content 10% by mass, phosphoric acid content (P ₂ O ₅ ) 10
A fertilization test was conducted on Western turf using 100% by mass of potassium and 10% by mass of potassium (K ₂ O). The evaluation results are shown in Table 1.

(Test condition) Fertilization rate 5g / m^Two (However, "10
No. 0 "is 2.5 g / m^Two) Temperature 15 ~ 20 ℃ Watering every 2 days Growth period 60 days.

(Evaluation Criteria) The growth elongation, hardness, color and luster of leaves / stems are comprehensively judged by visual inspection, and the growth roots of the evaluation roots are evaluated according to the five grades of ◎, ○, △, × and XX from the better one. Elongation and spread are comprehensively judged visually, and evaluated in 5 grades of ◎, ○, △, ×, and XX from the better.

[0054]       [Table 1]   (Growth test result)   Fertilizer name Degreasing treatment Day 10 Day 60                                 Growth of leaves and stems Growth of leaves and stems Growth of roots   Liquid fertilizer 8 Yes 〇 ◎ ◎   Liquid fertilizer 9 None △ ○ ○   Advanced Kasei No. 100 × 〇 〇

The degreased liquid fertilizer 8 showed a fertilizing effect in a short period after fertilization as compared with the liquid fertilizer 9 without degreasing, and the fertilizing effect continued even after 2 months. No growth damage was observed with any of the fertilizers.

[0056]

Industrial Applicability According to the present invention, it is possible to provide a liquid fertilizer having a suitable rate of fertilization and less growth damage, and a method for producing the same.

Claims (5)

[Claims] 1. A liquid fertilizer containing peptides and amino acids, wherein the nitrogen content of the peptides is 30 to 70 mass% of the total nitrogen content of the peptides and amino acids. 2. The liquid fertilizer according to claim 1, wherein the total amino nitrogen content of the peptides and amino acids is 1% by mass or more. 3. The phosphoric acid (P ₂ O ₅ ) component and the potassium (K ₂ O) component are 2 to 7% by mass and 2 to 7% by mass, respectively, according to claim 1 or 2. Liquid fertilizer. 4. The liquid fertilizer according to any one of claims 1 to 3, which is produced by using gelatin, glue, or peptides and amino acids obtained by hydrolyzing an extraction residue by-produced in the production process thereof. And a method for producing liquid fertilizer. 5. The oil and fat content of peptides and amino acids obtained by hydrolyzing gelatin, glue or an extraction residue by-produced in the production process thereof is 5% by mass or less of the peptides and amino acids. Item 4. A method for producing a liquid fertilizer according to Item 4.