JPS60192545A - Production of cheese product having filiform structure - Google Patents

Abstract

PURPOSE:To obtain the titled product capable of keeping filiform structure in spite of preservation for a long period, by using a milk coagulating enzyme obtained by removing acidic protease from a crude enzymic solution produced by a specific basidiomycetes as a milk coagulating enzyme to be added to a cheese raw material. CONSTITUTION:Acidic protease present in a solution of a crude enzyme containing a milk coagulating enzyme produced by cultivating Irpex lacteus is removed therefrom to give a milk coagulating enzyme fraction, which is collected to prepare a curd. The softening of filiform structure is not caused by removing the acidic protease participating in aging of cheese during the preservation of cheese having the filiform structure.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a method for producing a cheese product having a filamentous structure;
More specifically, the present invention relates to a method for producing a cheese product capable of retaining a filamentous structure consisting of a large number of p-fibers for a long period of time.

The term "thread-like tissue" used here refers to a structure in which a piece of cheese can be torn vertically with both hands, for example, like tearing a squid, and a large number of thread-like streaks are exposed on the torn surface. .

Cheese products with such a filamentous structure have a texture similar to that of scallops or sardines, and have a mild flavor without any unpleasant taste, so they are widely accepted by consumers who do not traditionally like cheese. The demand for this natural cheese is increasing.

Traditionally, pH
Pasta consisting of curd adjusted to about 5.0 to about 5.4 in water, heated to a temperature of about 55°C to 71°C to form a soft plastic mass, and the resulting curd packaged. A method for producing filata cheese (Japanese Patent Publication No. 4B-62) is known, but this method has the disadvantage that the filamentous structure described above gradually deteriorates.

In addition, in recent years, curds with a pH of 5.5 to 5.8 have been made from cheese raw materials, kneaded in hot water, and extruded after separating the hot water to maintain the above-mentioned filamentous structure. Process for manufacturing cheese products (Special Publication No. 5B-4
No. 8145) was developed, and the product is now commercially available.

However, even in cheese products produced by the above method, the cheese ripens to some extent during storage, which may soften the filamentous structure of the cheese and gradually reduce its fibrous properties.

The inventor of the present invention, as a result of studying to eliminate the above-mentioned drawbacks during storage of cheese products having a filamentous structure, found that there was a problem with the milk-clotting enzyme used in the cheese raw material when preparing curd, and When cheese with a filamentous structure is produced from a curd prepared using a milk-clotting enzyme obtained by removing acidic protease, a proteolytic enzyme mixed in, from a crude enzyme solution containing milk-clotting enzyme produced by bacteria, this The present invention was made based on the knowledge that a product capable of retaining filamentous tissue for a long period of time can be obtained.

In other words, the present invention provides a method for producing a cheese product that can maintain its filamentous structure without damage even during long-term storage of several months by using the above-mentioned milk-clotting enzyme as a milk-clotting enzyme. The purpose is to propose the following.

The present invention will be explained in detail below.

The feature of the present invention is a method of manufacturing cheese products having a filamentous structure by kneading a mixture prepared by adding milk-clotting enzyme to cheese raw material in hot water, separating the hot water and extruding it. In the above, as the milk-clotting enzyme, Irpex
The purpose of this method is to use a milk-clotting enzyme obtained by removing acidic protease from the crude enzyme produced by A. 1acteus.

It has long been known that the basidiomycete, a type of basidiomycete, produces milk-clotting enzyme. Although this milk-clotting enzyme has excellent milk-clotting activity, it is difficult to knead curd prepared using it in hot water. For cheese with a filamentous structure obtained by pressing, separating hot water, and extruding, as mentioned in 6; become.

This phenomenon is explained by the fact that the crude enzymes produced by Asus brevifolia contain two types of enzymes: milk-clotting enzyme, which is inhibited by pepstatin, which is a specific inhibitor of acidic protease, and acidic protease, which is not inhibited by pepstatin.
% and 25%.

In other words, the acidic protease mixed in the milk-clotting enzyme produced by the milk-clotting enzyme takes part in the ripening of the cheese during storage of the cheese product having a thread-like structure produced using the milk-clotting enzyme, and causes the thread-like structure of the cheese to deteriorate. This causes the material to become soft.

From this point of view, in the present invention, a milk-clotting enzyme fraction obtained by removing acidic protease mixed therein from a solution of milk-clotting enzyme-containing crude enzyme produced by cultivating Asuba mushrooms in a conventional manner. Collect and use for preparing curd.

In the present invention, the following two methods are applied to remove acidic protease, which is a proteolytic enzyme, from the solution of the crude enzyme produced by Aspergillus chinensis.

For example, the above-mentioned crude enzyme is dissolved in an acetate buffer, the pH of the enzyme solution is adjusted to 6.3 to 7.6, and the mixture is mixed in the enzyme solution by treating it at a temperature of 30 to 40°C for within 30 minutes. It is removed by inactivating the acidic protease present.

Another method is to pass the enzyme solution through a column of phenyl ethylamine sepharose to remove the curdled milk P in the solution.
Only one gene itself is adsorbed on the column, while the acidic protease is eluted through the column, and then the milk-clotting enzyme fraction adsorbed on the column is eluted.

When the activity in the milk-clotting enzyme fraction obtained by removing acidic protease as described above was investigated, the degree of inhibition by pepstatin of the enzyme obtained by inactivating acidic protease as described above was determined. The enzyme was treated with lo-jM pepstatin at a temperature of 0°C for 15 minutes,
The results showed that the inhibition rate was 99%, and the remaining proteolytic activity was 99%. (), 6%.

That is, it was confirmed that 99% of the enzymes produced by depleting acidic protease as described above had both components exhibiting milk-clotting activity. Incidentally, with the enzyme that does not inactivate acidic protease, the inhibition rate was 71% and the remaining proteolytic activity was 29%.

Furthermore, as a result of examining the degree of inhibition by pepstatin of the enzyme obtained through the phenyl-ethylamine-Sepharose column as described above, the inhibition rate was 98%. Minutes are 98%
It has been confirmed that it exists.

In addition, the milk curd activity and proteolytic activity were measured by the following procedure.

Milk curd activity: 0.0% in skim milk at a concentration of 1O%. Add the test enzyme solution to 51111 with OlM CaC1, added at 0.5+xj! In addition, the time (seconds) until the milk curdled at 35°C was measured.

The activity unit (SU) was determined by the following formula.

In the formula, t indicates milk curdling time.

Proteolytic activity: Substrate 1.0+wj! with 0.1M glycine hydrochloride added to 1.2% hemoglobin and pll adjusted to 3.0! Test enzyme solution 0.2+nj! After reacting at 35° C. for 30 minutes, 1 ml of 0.55 M) dichloroacetic acid was added, the resulting precipitate was removed, and the absorbance of the supernatant at 28 On+* was measured.

The amount of enzyme that increases the absorbance at 280 nm by 1 under the above conditions was defined as 1 unit.

Next, each curd prepared using the milk-clotting enzyme and calf rennet preparation (hereinafter referred to as HR) obtained as described above was shaped into a cheese having a thread-like structure, and the state of the thread-like structure and the thread-like texture were measured. The results of a tissue retention test are shown. In addition,
The milk curdling activity (power 1 + li) of the sample milk curdling enzyme is 11900
S U /g and HI? That is 178.000
Since it is S U / g, the amount added to the raw milk was adjusted so that ζ would have the same titer when used.

Test method: Using fresh milk that has been partially defatted and adjusted to a fat percentage of 3.0% as raw milk, 40 kg of this raw milk was poured into two cheese vats in equal amounts and heated at 72°C for 15 seconds. After heat sterilization, the mixture was immediately cooled to 31"C. Next, 2 g of calcium chloride was added to each vat and melted, and 300 mJ of lactic acid bacteria starter was added thereto for acid ripening. Then, each raw milk was curdled. Milk enzyme (hereinafter referred to as IR) and HR were added and mixed uniformly and allowed to stand.The obtained curdled milk curd (
Cut the curd (pH 6.3) into 10 ounce cubes, warm the cut curd to 38°C while stirring gently, stir occasionally until the pll of the curd reaches 5.8, and then add the whey. Completely eliminated. While inverting the obtained card into a 20 cm square, the hydrophilicity and pH reduction of the card progressed, and when the PI (PI) reached 5.2, the 20-308
Collect the curd and raise the temperature of the whole curd while gently kneading it in warm water of about 11°C at 70°C until the temperature of the curd reaches 50-53°C.
When the temperature reached ℃, the card was slowly pulled with both hands to shape it into a thin string, then the string-shaped card was folded while maintaining the temperature, and pulled again to form a single string. This operation was repeated until the cards were sufficiently spread. (This operation is called stretching) As a result of the above test, I? It was confirmed that the curd using this method could be sufficiently spread as well as the cart using HR, so it was confirmed that it could be used satisfactorily for producing cheese products having a filamentous structure.

Next, add 60 pieces of cheese shaped into strings as described above.
As a result of examining changes over time when stored for several days, the cheese using IR showed fibrous properties and retained its filamentous structure even after 60 days, while the cheese using HR showed fibrous properties even after 45 days. There is a clear decrease in fibrous properties, and 60
After several days, it was confirmed that the retention of filamentous tissue was considerably poor.

As mentioned above, the cheese having a filamentous structure made from a curd prepared using a milk-clotting enzyme obtained by removing acidic protease from a crude enzyme produced by Aspergillus muscariae according to the present invention has excellent fibrous properties. Furthermore, since the filamentous structure can be retained even during long-term storage, the present invention can be said to have the advantage of significantly increasing the commercial value of cheese products containing the filamentous structure.

EXAMPLES The present invention will be explained in more detail with reference to Examples below.

Example 1 Irpex Iacteus ATCC After sterilizing the medium 2i adjusted to pH 5.5, Irpex Iacteus ATCC was added to the medium.
20123 strain mycelium was inoculated and heated to 40O at a temperature of 30℃.
r,p,1n,,157! Culture was carried out for 90 hours with aeration of /min.

Add ammonium sulfate to 0.8 saturation to the culture filtrate obtained as described above, and collect the precipitate by centrifugation.
This precipitate was dissolved in a small amount of distilled water and dialyzed against running water overnight, and the resulting dialyzed solution was freeze-dried to prepare rennet (crude enzyme).

Next, 50 g of the obtained rennet (crude fermentation I)
mj! of acetate buffer (0.01 M, pH 5.5), the pl of this solution was adjusted to 6.5 using O.1N sodium hydroxide solution, and kept at a temperature of 35 °C for 20 min. , the proteolytic enzyme (acidic protease) present in the above rennet was inactivated.

The results of examining the milk curdling activity and proteolytic activity of the enzyme solution obtained by the treatment as described above are shown in Table 1. As a control, the results of similarly examining the milk curdling activity and proteolytic activity of a rennet solution (crude enzyme solution) adjusted to pn 6.5 as described above and stored in water are also included. The results are shown in Table 1. ~Table 1 Cheese was prepared by the following procedure using the milk-clotting enzyme obtained as described above.

Fat percentage 2.5-3.5%, non-fat milk solids content 8.2-8.6
% raw milk was pasteurized (75°C, 15 seconds), 0.01% calcium chloride and 1.5% lactic acid bacteria starter were added, and after standing at 30°C for 0.5 to 1.5 hours, prepared as above. Add 0.0035% of milk clotting enzyme to produce curd.

This curd is cut into 5III1 cubes with a card knife and stirred with a card breaker, being careful not to crush the cut cards. After 15 minutes of stirring, when the whey separates from the curd, a portion of the whey is removed, the product temperature is raised to 34° C., and the moisture content of the curd is adjusted (generally, the moisture content will be around 45% the next morning). This process makes the card somewhat stronger. When the po of the curd reaches 5.5, a stretch test is performed to confirm that the curd spreads into a rubbery state, and then the entire amount of whey is removed.

Stack this card and squeeze it to get Card In (220 per hit)
~240 kg weight) and drain off excess whey (approximately 20 minutes). This piled card was cut into 3c+a square pieces, put into a kneading machine, and while being kneaded in hot water with a material temperature of 70°C, the hot water was gradually reduced to 1 mtt.
Extrude through the extrusion hole at around 1, and cut to a certain length. The cut pieces were immersed and heated in salt brine (20% saline) for 2 hours, dried overnight, and then packaged to obtain a cheese product. The thus obtained cheese product was stored at 5° C. for 2 months, and sensory evaluation revealed that the filamentous structure described above was completely maintained.

Example 2 A crude enzyme was prepared according to the procedure described in Example 1, and the crude enzyme was 0.1
8 with lOmA' of 0.01M vinegar Full 42# solution (ρ
115.5) Melt h1! The above solutions were dialyzed against the above buffer. The dialyzed crude enzyme solution was preliminarily diluted with phenyl ethylamine, which had been parallelized with the 1h solution described in J-1.
It was passed through a Sepharose (Wako Pure Chemical Industries-M) column (gel volume: 2 ml) and washed with the above buffer to adsorb the milk-clotting enzyme fraction onto the column, while the protein decomposition mixed in the crude enzyme solution was removed. Only the enzyme fraction was eluted.

The column was then charged with 0.01 containing 0.5 M NaCl.
The milk-clotting enzyme fraction adsorbed on the column was eluted through M acetic acid (H solution).

The milk curdling activity and proteolytic activity of the obtained milk curdling enzyme fraction are shown in Table 2.

Table 2 Next, using the milk-clotting enzyme obtained as described above, a milk powder was prepared in the same manner as described in Example 1.

The obtained cheese exhibited a highly fibrous thread-like structure, and it was observed that the above-mentioned thread-like structure was retained even after being stored for two months.

Claims (3)

[Claims] (1) A method for producing a cheese product with a filamentous structure by kneading RM-made curd by adding milk-clotting enzyme to cheese raw material in hot water, separating the hot water and extruding it.
ζ, Irpcx 1a as the above milk-clotting enzyme
1. A method for producing a cheese product that maintains a filamentous structure, characterized by using a milk-clotting enzyme produced by culturing a milk-clotting enzyme fraction (II) produced by culturing a milk-clotting enzyme. (2) A solution of crude enzyme containing milk-clotting enzyme at pH 6.3-6.
A patent claim that removes acidic protease from the crude enzyme by inactivating the acidic protease mixed in the crude enzyme-containing solution by treating it at a temperature of 30 to 40°C for within 30 minutes. The manufacturing method described in item (1). (3) A solution of the crude enzyme containing milk-clotting enzyme is passed through a phenyl-ethylamine-Sepharose column to adsorb the milk-clotting enzyme fraction in the crude enzyme solution, and the acidic protease fraction is eluted from the crude □enzyme. The manufacturing method according to claim 11, which removes acidic protease.