JP3217909B2 - Method for producing hydrolyzed seasonings - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to an amino acid seasoning (HAP) obtained by hydrolyzing an animal protein.
And a method for producing the same. More specifically, the present invention relates to a method for producing a seasoning having a high taste without causing the generation of harmful chlorinated compounds by combining hydrolysis with hydrochloric acid and protease.

[0002]

2. Description of the Related Art Seasonings obtained by hydrolyzing proteins include HVP (hydrolized vegetable protein) derived from cereal proteins and HAP (hydrolized animal protein) derived from animal proteins. There are two methods of adding hydrochloric acid and treating at a high temperature,
A method of hydrolyzing by the action of a protease is known.

When proteins are treated with hydrochloric acid at high temperatures,
Most of the peptide bonds are hydrolyzed to produce a seasoning with a high taste. On the other hand, trace amounts of oils and fats present in the protein raw material react with hydrochloric acid to produce dichloropropanol (hereinafter abbreviated as DCP) or monosaccharide. It has become clear that undesired chlorinated compounds such as chloropropanediol (hereinafter abbreviated as MCP) are produced, and this has recently become a problem from the viewpoint of food safety. As a method for producing a seasoning containing no undesirable chlorinated compounds as described above,
(A) The temperature of the hydrolysis treatment with hydrochloric acid is 60 to 97 ° C.
(Japanese Patent Application Laid-Open No. H2-135058), (b) treating the hydrolysis reaction product with hydrochloric acid at a high temperature under a weak alkaline pH (Japanese Patent Application Laid-Open No. -135056),
(C) a method of further hydrolyzing a chlorinated compound produced by treating at a slightly elevated temperature at an alkaline pH (JP-A-4-88951), and (d) chlorination produced by gel permeation chromatography A method for removing a compound (Japanese Patent Application Laid-Open No. 2-135057) has been reported. However, the one manufactured by the above method (a) has a DCP of 50 ppb or less, but the MCP has poor analytical accuracy and is described only at 10 ppm or less.
Have not been clarified as to whether the temperature is sufficiently low, and the temperature control is complicated. In the methods (b) and (c), heating is performed under alkaline conditions. However, in an alkaline aqueous solution, MCP is gradually converted to glycidol.

[0004]

Embedded image

[0005] Glycidol is also concerned about its safety similarly to MCP (H. Jackson et al., Natur.
e, Vol. 226, p. 86, (1970)) remains as a problem. Therefore, even if DCP or MCP is hydrolyzed by treating a reaction product with hydrochloric acid with an alkali, the harmfulness of the reaction product is not necessarily reduced. The method using the gel permeation chromatography of the above (d) is problematic in that when it is carried out industrially, the cost increases.

[0006] There is also a method in which protein hydrolysis is carried out by using a protease. However, in the reaction by the enzyme, the hydrolysis rate is low, and a bitter peptide is easily produced, and the taste is as good as that hydrolyzed with hydrochloric acid. It is difficult to produce a seasoning that has it. Recently, a technique has been reported in which a vegetable protein is deamidated with hydrochloric acid and then hydrolyzed with a proteolytic enzyme (JP-A-4-365451). Even if it is deamidated with hydrochloric acid and then degraded by ordinary protease, its hydrolysis rate is low and only those having poor taste can be produced.

[0007]

SUMMARY OF THE INVENTION An object of the present invention is to reduce undesired substances such as chlorinated compounds, for example, DCP by 50 p.
An object of the present invention is to develop a method for producing a hydrolyzed seasoning derived from animal protein having a high hydrolysis rate, that is, a high taste, in which the MCP is reduced to 1 ppm or less and the MCP is reduced to 1 ppm or less, preferably 100 ppb or less.

[0008]

Means for Solving the Problems As a result of intensive studies to solve the above problems, the present inventors have found that the amount of lipid contained in the starting protein is less than a certain level and that the protein is hydrolyzed by hydrochloric acid. If the rate is controlled to a certain level or less, almost no MCP is produced, and the shortage of the rate of hydrolysis by hydrochloric acid is compensated for by the hydrolysis by proteolytic enzymes to achieve the conventional rate of hydrolysis, and It was confirmed that both the taste and the flavor were inferior to those obtained by sufficiently hydrolyzing with hydrochloric acid alone, and the present invention was completed. The details are described below.

Any protein such as meat protein, animal skin protein, and bone protein can be used as a raw material protein.
However, the lipid contained in the raw protein is required to be as low as possible. At most, it is necessary to reduce the amount of lipid to 1% or less based on the weight of protein in the raw material. The lipid content can be calculated by obtaining the volume of the lipid by the Babcock method and multiplying this value by the specific gravity of the lipid (usually 0.9).

Either the hydrolysis with hydrochloric acid or the hydrolysis with proteolytic enzyme may be performed first. The amount of hydrochloric acid added to the starting protein is not particularly limited as long as it is an amount necessary for hydrolysis of the protein, and there are no particular restrictions on the temperature, pressure, reaction time, and the like of the hydrolysis. However, it is necessary to control the hydrolysis conditions so that the rate of hydrolysis by hydrochloric acid, that is, the ratio of the number of peptide bonds hydrolyzed by hydrochloric acid to the total number of peptide bonds before hydrolysis is 60% or less. FIG. 1 shows a graph of the amount of MCP generated after hydrolysis versus the hydrolysis rate. In addition, a raw material having a lipid weight of 1% or less based on the protein weight was used. As shown in FIG. 1, when a protein having a low lipid content is used, under conditions where the hydrolysis rate is 60% or less,
The MCP formation reaction did not proceed, and the MCP after the hydrolysis reaction
The content is around or below the detection limit of 50 ppb. Under the condition that MCP is not generated, DCP is not detected at all (detection limit: 50 ppb). The hydrolysis rate by the reaction is

[0011]

(Equation 1)

Thus, the total hydrolysis rate in the sample is

[0013]

(Equation 2)

[0014] DCP
And MCP can be analyzed by gas chromatography / mass fragment method. The hydrolysis rate of the conventional hydrolysis seasoning is 70 to 80%, and the shortage of the hydrolysis rate (10 to 20%) can be compensated for by the protease. As the enzyme to be used, any enzyme can be used as long as it is an enzyme that can be generally used for food use.However, when the hydrolysis-finished solution with hydrochloric acid is enzymatically treated, a large amount of salt generated when neutralizing hydrochloric acid is present. Therefore, it is desirable to use an enzyme that can work even under conditions of high salt concentration. Conversely, there is no such limitation when prior to enzymatic hydrolysis. Hereinafter, the present invention will be described in detail with reference to Examples, but the present invention is not limited thereto.

[0015]

Example 1 Pork bone 3 in a 5-liter high-pressure autoclave
600 g and 720 g of water were charged, and the temperature was raised after sealing. When the internal pressure of the autoclave reached 0.5 kg / cm ² , the air in the autoclave was evacuated, sealed again, heated until the internal pressure of the autoclave became 5 kg / cm ² , and the autoclave was boiled for 1 hour. After cooling, the liquid in the autoclave was transferred to a 5-liter separating funnel, and the upper layer oil was removed to recover 2400 g of the lower layer pork bone extract, which was concentrated by an evaporator to obtain TN% 7.7% and F- N0.39
450 g of a 1% concentrate were obtained. The lipid content of this concentrate was measured by the Babcock method and found to be 0.5% or less. The protein content of this concentrate was calculated to be about 50% by TN x 6.25, and the lipid content relative to the protein was calculated to be 1% or less. 780 g of water was added to 210 g of the concentrate to dilute it, and the pH was adjusted to 8.0 by adding a 16% sodium hydroxide solution. 0.6 L of Alcalase in this solution
4 g (manufactured by Novo) was added and reacted at 55 ° C. for 6 hours. The pH during the reaction was adjusted to always be 8.0 with a 16% sodium hydroxide solution. The reaction-terminated liquid is subjected to an ultrafiltration membrane having a molecular weight cutoff of 6000 (SIP-1013 manufactured by Asahi Kasei Corporation).
And the filtrate was concentrated to 250 g with an evaporator. The concentrated liquid has TN = 6.4%,
FN = 1.01%, and the hydrolysis rate was calculated to be 11%. The entire amount of the concentrated solution is charged into a pressure-resistant container, and concentrated hydrochloric acid 9
6 g and 140 g of water were added, and the mixture was hydrolyzed with stirring at 100 ° C. for 15 hours. After cooling, an appropriate amount of sodium carbonate (powder) was added to adjust the pH to 5.5.
o. 2 Filter with filter paper to remove insolubles, add 7.5 g of activated carbon (Carbofin, manufactured by Takeda Pharmaceutical Co., Ltd.) and add 30 g at 75 ° C.
Bleached for minutes. The decolorizing solution was treated with Toyo No. 2 Filtered with filter paper to remove activated carbon, T-N = 3.3%, F-N =
400 g of a 2.13% standard product was obtained (standard A). The hydrolysis rate with hydrochloric acid was calculated to be 58%, and the total hydrolysis rate (the ratio of TN to FN in the sample) was calculated to be 65%.

The content of the chlorinated compound contained in the sample was analyzed by the following method. 8 g of the sample was placed in a 50 ml volumetric flask, and water was added to make up to 50 ml. The whole amount was transferred to a 100-ml glass vial containing 14 g of sodium chloride, and 4 ml of diethyl ether was added, followed by vigorous stirring for 5 minutes.
The ether layer was separated, and 1 μl thereof was used to analyze DCP by gas chromatography / mass fragment method (GC / MS method). The model used was a GC made by Shimadzu Corporation.
Using MS QP-1000 in a splitless introduction system,
Column is Spelcowax10 Fused Sil
ica capillary column (0.25mmφ × 15m)
It was used. The sample injection port temperature was set at 250 ° C., and the column temperature was maintained at 70 ° C. for 1 minute after the sample injection, and then the column temperature was set at 15 minutes per minute.
The temperature was increased by 220 ° C. to 220 ° C. Helium of 0.68 kg / cm ² is used as a carrier gas, EI (Electron Impact) is used as an ionization method, and the ion source temperature is set to 250 ° C. (ionization voltage: 70 e).
V, ionization current: 60 μA). The set mass numbers were m / z = 79 and 81. The DCP content in the sample was calculated by comparing the obtained peak area with a calibration curve created from the peak area obtained for an aqueous solution containing DCP of a known concentration and treated in exactly the same manner. As a result of the analysis, the DCP content in this sample was below the detection limit of 50 ppb. MCP in the sample is also GC / M
The analysis was performed by the S method. That is, after putting 20 g of the sample into a 50 ml volumetric flask and adding water to make up to 50 ml,
It was added to a Merck Extrelut column. 200 ml of diethyl ether was passed through the column, and the eluted ether was concentrated by natural evaporation in a fume hood, and ether was added to 50 ml. 2 ml of this ether solution and 1 ml of 1 μg / ml ether solution of 1,2-hexanediol as an internal standard substance and 1 ml
10 ml of an ether solution containing 10% phenylboric acid
The volume was added to a 1-liter volumetric flask, and the volume was adjusted to 10 ml with ether. The MCP was analyzed by analyzing 2 μl by GC / MS. The apparatus used was GCMS QP-1000 manufactured by Shimadzu Corporation in a splitless introduction system, and the column was a fused Silica OV-101.
A capillary column (0.25 mmφ × 25 m) was used. The sample injection port temperature was set to 240 ° C., the column temperature was maintained at 70 ° C. for 1 minute after the sample injection, and then the temperature was raised to 240 ° C. by 15 ° C./min. The carrier gas is 1.3
kg / cm ² of helium and ionization method E
I, using an ion source temperature of 250 ° C. (ionization voltage: 70
eV, ionization current: 60 μA). The set mass number of the MCP analysis was m / z = 196, and the set mass number of the internal standard substance was m / z = 204. Set mass number m / z = 1
By comparing the ratio of the peak area of 96 to the peak area of the set mass number m / z = 204 with a calibration curve prepared from the ratio of the peak areas obtained by completely treating the aqueous solution containing the known concentration of MCP. The MCP content in the sample was calculated. As a result of the analysis, the MCP content in this sample was 0.08p
pm.

This preparation exhibited good taste and flavor.

[0018]

Example 2 780 g of water was added to 210 g (TN = 7.7%, FN = 0.38%, lipid content of protein is 1% or less) of pork bone concentrate prepared in the same manner as in Example 1 above. Was added. The pH was adjusted to 8.0 by adding an appropriate amount of a 16% sodium hydroxide solution, and 0.6 g of 4 g of alcalase was added, and the enzymatic reaction and ultrafiltration were performed in exactly the same manner as in Example 1. Concentrated hydrochloric acid was added to this solution to adjust the pH to 5.0, and then 5 g of Protease M (manufactured by Amano Pharmaceutical) was added.
To carry out a protease reaction with stirring for 6 hours. The reaction solution was subjected to ultrafiltration in the same manner as in Example 1 and concentrated by an evaporator. As a result, a concentrated solution 250 of TN = 6.3% was obtained.
g was obtained. The result of analyzing the FN of the concentrate was 1.79.
% And the hydrolysis rate was calculated to be 28%. Next, 96 g of concentrated hydrochloric acid and 140 g of water were added to the whole amount of the concentrated solution, and the mixture was charged into a pressure-resistant vessel, and hydrolyzed with stirring at 100 ° C. for 15 hours. As a result of performing the operations of neutralization and decolorization in the same manner as in Example 1, 400 g of a sample having TN = 3.2% was obtained (sample B). Since the FN of the sample was 2.24%, the hydrolysis rate with hydrochloric acid was calculated to be 58%, and the total hydrolysis rate was calculated to be 70%. DCP was not detected at all from this sample, and MCP was also 0.1 ppm.

[0019]

Example 3 750 g (TN = 7.6%, FN = 0.39%, lipid content 1% or less of protein) of 750 g of pork bone concentrate prepared in the same manner as in Example 1 and 300 g of concentrated hydrochloric acid And 450 g of water were added, and the mixture was charged into a pressure-resistant container, and stirred at 100 ° C. for 15 hours to perform hydrolysis. Example 1
Neutralization and decolorization are performed in substantially the same manner as in
= 3.3%, FN = 2.06%
0 g was obtained. The hydrolysis rate was calculated to be 60%. 500 g of water was added to 490 g of the hydrolyzed hydrochloric acid solution, and 16%
The pH was adjusted to 8.0 with sodium hydroxide. 4 g of Alcalase (0.6 L) was added and reacted at 55 ° C. for 6 hours. The pH during the reaction is always 8% with 16% sodium hydroxide.
It was controlled to be 0. The reaction-terminated liquid was filtered through a SIP-1013 ultrafiltration membrane manufactured by Asahi Kasei Corporation to remove the enzyme, and concentrated by an evaporator to obtain TN = 5.
300 g of a 1% sample was obtained (sample C). FN of this sample
Was 3.4%, so the total hydrolysis rate was calculated to be 67%. The chlorinated compounds in this sample are DCP, MCP
Both were below the detection limit (50 ppb).

[0020]

Comparative Example 1 500 g of pork bone concentrate prepared in the same manner as in Example 1 (TN = 7.7%, FN = 0.35%, lipid content of protein 1% or less) and 192 g of concentrated hydrochloric acid And 280 g of water were added, and the mixture was charged into a pressure-resistant container, and stirred at 110 ° C. for 15 hours to perform hydrolysis. Example 1
Neutralization and decolorization are performed in substantially the same manner as in
= 3.1% and FN = 1.89% to obtain 800 g of a standard (standard D). The hydrolysis rate of the sample with hydrochloric acid was 59%, and the total hydrolysis rate was calculated to be 61%. DCP and MCP in the preparation were 50 ppb or less and 5
It was 0 ppb.

[0021]

Comparative Example 2 500 g of pork bone concentrate (TN = 7.8%,
(FN = 0.36%, lipid content based on protein: 1% or less), 280 g of concentrated hydrochloric acid was added, and the mixture was stirred at 110 ° C. for 15 hours in a pressure vessel to carry out a hydrolysis reaction.
Neutralization and decolorization were carried out in the same manner as in Example 1, and TN = 4.
550 g of a standard having 7% and FN = 3.36% was obtained (standard E). The hydrolysis rate by the reaction was calculated to be 70%, and the total hydrolysis rate of the sample was calculated to be 71%. The DCP and MCP of this sample were 0.2 ppm and 31 ppm, respectively.

[0022]

[Table 1]

[0023]

Table 1 summarizes the results of various evaluations of the samples A to E. As is clear from Table 1, by using a raw material having a low lipid content and combining hydrolysis with a hydrolysis rate of 60% or less with hydrochloric acid and hydrolysis with a protease, the content of the chlorinated compound is extremely low and present. A seasoning with a high taste can be produced.

[Brief description of the drawings]

FIG. 1 is a graph showing the amount of MCP produced with respect to the hydrolysis rate of a sample obtained by hydrolyzing a pork bone concentrate (lipid content relative to protein of 1% or less) with hydrochloric acid under various conditions. The hydrolysis conditions were as follows: ○: 110 ° C., heating for 15 hours, Δ: 100
□ indicates heating at 20 ° C., and □ indicates heating at 100 ° C. for 15 hours. The hydrolysis rate was controlled by changing the amount of concentrated hydrochloric acid to be added.

──────────────────────────────────────────────────続 き Continuation of the front page (56) References JP-A-48-44455 (JP, A) JP-A-50-77568 (JP, A) JP-B-44-10145 (JP, B1) JP-B-48-48 43637 (JP, B1) JP-B-49-11078 (JP, B1) JP-B-49-25346 (JP, B1) European Patent Application Publication 495390 (EP, A1) (58) Fields investigated (Int. Cl. ⁷ , DB name) A23L 1/22-1/237

Claims (1)

(57) [Claims] 1. The lipid content is less than 1% of the protein weight.
In hydrolyzing animal protein raw materials, hydrolysis of hydrochloric acid with a hydrolysis rate of 60% or less and hydrolysis with a protease are combined , and the total hydrolysis rate is 65% or more.
Has a dichloropropanol content of 50 ppb or less, and
Monochloropropanediol content of 100 ppb or less
A method for producing a hydrolyzed seasoning, which is used as a seasoning.