JPH03112445A - Preparation of cheese-like substance using soybean protein as raw material - Google Patents

Abstract

PURPOSE:To easily obtain a cheese-like substance from soybean protein by adding lactobacillus and a soybean protein coagulase to a material containing soybean protein to effect the lactic acid fermentation and coagulation of the material, separating and removing whey from the coagulum and adding a thermoplasticity-imparting substance to the product. CONSTITUTION:Lactobacillus and a soybean protein coagulase are added in the order or at the same time to a soybean protein-containing material such as soya milk to effect the lactic acid fermentation and coagulation of soybean protein. The soybean protein coagulase is preferably an enzyme produced by 26-D7 (FERM BP-1778) which is a mutant of Bacillus licheniformis. After separating and removing whey from the obtained coagulum, a thermoplasticity- imparting substance (e.g. gelatin or gum) is added to the coagulum to obtain the objective cheese-like substance from soybean protein. The cheese-like substance has hardness, coagulation property, elasticity and chewability close to those of cheese prepared from animal milk.

Description

Detailed Description of the Invention (Field of Industrial Application) The present invention provides a cheese-like substance that has properties extremely similar to the hardness, cohesiveness, elasticity, and chewability of cheese produced from animal milk. , relates to a method of producing soybean protein using soybean protein as a raw material.

(Prior Art) Recently, the harmful effects caused by the intake of large amounts of animal foods have been viewed as a problem, and plant foods have been attracting attention.

Foods made from soybeans, which are especially highly nutritious among dietary foods, are undergoing a major review, and new soybean-based foods are being developed to replace long-known miso, soy sauce, tofu, etc. .

(Problem to be Solved by the Invention) By the way, cheese has been produced by coagulating animal proteins such as animal milk with coagulating enzymes for a long time. Production has not been carried out because a coagulating enzyme with a high coagulating ability to coagulate soybean protein cannot be obtained.

The present inventors have discovered a strain with high production ability of soybean protein coagulase, and have succeeded in producing a cheese-like substance using soybean protein as a raw material based on this strain.

Therefore, an object of the present invention is to provide a method for producing a cheese-like substance using soybean protein as a raw material.

(Means for Solving the Problems) The present invention relates to a method for producing a cheese-like substance using soybean protein as a raw material, and is characterized by each of the following production methods (1) to (3).

(1) After adding lactic acid bacteria and soybean protein coagulase in this order or simultaneously to a material containing soybean protein, lactic acid fermenting the soybean protein and coagulating it, and separating and removing whey from the resulting coagulated product. A substance that imparts thermoplasticity is added to the coagulated material.

In the production method described in items (2) and [1), the soybean protein coagulating oxygen is an enzyme produced by Bacillus licheniformis, which has low spore-forming ability and high productivity of soybean protein coagulase.

In the manufacturing method described in item +31i11, the substance imparting thermoplasticity is at least one substance selected from gelling agents, emulsion stabilizers, and thickeners.

(Actions and Effects of the Invention) According to the production method of the present invention, since lactic acid bacteria and soybean protein coagulase are used, the soybean protein in the material can be sufficiently coagulated, and the soybean protein formed during the cheese production process can be sufficiently coagulated. A coagulum is obtained which has properties similar to those of the curd (but with a different composition).

Furthermore, according to the production method of the present invention, after the whey is separated and removed from the coagulated product, a substance that imparts thermoplasticity is added to the coagulated product, so that the thermal properties vary depending on the type and amount of the substance added. shows. In other words, depending on the heating conditions, the solidified product can exhibit various states such as solidification, softening, melting, and intermediate states between these two, and can be used in the same way as cheese. The entire product can be sterilized by heating uniformly, allowing for long-term storage, and there is no risk of abnormal fermentation occurring during storage and changing the taste.

(Detailed Description of the Invention) The present invention involves using a material containing soybean protein, lactic acid fermentation of soybean protein using lactic acid bacteria, and coagulating the lactic acid-fermented soybean protein using soybean protein coagulating enzyme. This can be roughly divided into two steps: 1) separating and removing the whey from the coagulated product, and then imparting thermoplasticity to the coagulated product. Therefore, each of these items will be explained in detail below.

(1) Material containing soybean protein The material is generally a suspension of soybean protein 1, and soybean milk in which soybean protein is dissolved in water is used. The raw material for soy protein may be in any form, such as soy milk itself made during tofu production, powdered soybeans, isolated soy protein, etc., and the protein concentration of the suspension is appropriately selected depending on the desired final product. . In addition, considering the efficiency in the subsequent lactic acid fermentation and coagulation steps, the protein concentration is preferably 5 wt % or more. If necessary, fats and oils, lactose, etc. are added to this suspension and mixed uniformly.

For mixing, a high-speed stirrer or a homogenizer can be used, and hot water can also be used. Addition of fats and oils adjusts the smoothness and hardness of the final product, and addition of lactose promotes lactic acid fermentation.

Prior to adding lactic acid bacteria to a suspension of soy protein (hereinafter referred to as soy milk), the soy milk is sterilized and homogenized.

Either high-temperature sterilization or pasteurization may be used to sterilize soymilk. In high-temperature sterilization, heat treatment is performed at, for example, 80°C or higher for 10 minutes or more, and in low-temperature sterilization, heat treatment is performed at, for example, around 60°C. will be held. Note that retort sterilization, high pressure sterilization, etc. can also be used. A homogenizer is used to homogenize soy milk, which eliminates the graininess of the soy milk, especially when powdered soy protein is used, and when oil is added, it emulsifies more uniformly, making the final product even smoother. . 100-5 in homogenizer
It is preferable to homogenize soybean milk using a knife under the condition of 00 kg/Cm2.

(2) Lactic acid bacteria, lactic acid fermentation All the usual lactic acid bacteria that act as starters for lactic acid fermentation in the manufacturing process of cheese, yogurt, etc. can be used. Skim milk is generally used for culturing lactic acid bacteria, but they can be cultured in various media that do not pose a problem in contamination with foods. When lactic acid bacteria are subcultured 2 to 10 times, the acidity increases, and if the acidity is not so necessary, the bacterial cell powder can be used as is.

Lactic acid bacteria with a p114.5 to 4.7 and an acidity of around 0.75 are usually used, and are added in an amount of 1 to 10 wt% to soymilk. The amount added may be outside the above range, but even if more than the above range is added, the fermentation condition will remain the same as when the appropriate amount is added, and if less than the above range is added, the fermentation time will take longer. Too much leads to a drop in efficiency. The fermentation temperature can be set to any temperature that allows lactic acid bacteria to grow.
The temperature is preferably 10°C to 60°C in view of the increase in acidity, the growth of lactic acid bacteria, the state of protein curd formation, etc.

(3) Soybean protein coagulase Soybean protein coagulase (hereinafter referred to as coagulase) may be of any type as long as it is non-toxic and has the ability to coagulate soybean protein. The coagulating enzyme produced by the producing bacteria disclosed in the application No. 63-79208 and the patent application filed on September 27, 1999 is preferred. This producing bacterium is a mutant strain of Bacillus licheniformis, named 2607, and published in FERM No. 1778 (FERM
It has been internationally deposited with the Institute of Microbial Technology, Agency of Industrial Science and Technology as OP-1778), and the coagulating enzyme produced thereby has the following properties.

■When applied to soy milk, it coagulates soy protein and forms a smooth coagulated product without bitterness.

(2) It is purified to a single band during SDS electrophoresis, and its molecular weight is around 30,000.

■The optimum temperature for activation is approximately 80°C, and 35°C to 40°C.
It is thermally stable for 1 hour at ℃.

(2) The optimum pH for activity is on the acidic side, and the activity is almost lost on the alkaline side exceeding p17. Also, pH 4~
p) In the range of 19, even if held at 4°C for 17 hours,
At least 70% activity remains.

■Activity is inhibited by phenylmethylsulfonyl fluoride and toluenesulfonyl fluoride,
Moreover, the activity is not affected by metal ions.

The safety of the coagulation enzyme has been confirmed through administration tests on mice.

G4), addition of coagulation enzyme, coagulation The soybean protein fermented with lactic acid is coagulated by adding the coagulation enzyme. The enzyme may be added at the same time as the lactic acid bacteria, or may be added after lactic acid fermentation has progressed. Differences in the timing of addition will cause slight differences in the properties of the final product.

The amount of the enzyme added (enzyme concentration) is related to the coagulation time, but the higher the fermentation temperature during lactic acid fermentation and the higher the acid concentration, the faster the coagulation time, so the appropriate amount to be added should be selected with these factors in mind. Ru. However, even if the amount of enzyme added is increased more than necessary, the effect on the coagulation time will be small and there is a possibility that the final product will have a bitter taste. In the actual manufacturing process, lactic acid bacteria are added to soymilk and after a predetermined period of time, soymilk becomes P).
It is preferable to add the enzyme when the acidity is around 6.4 and the acidity is 0.2 to 0.25, and the amount added is 0.1 to 0.5 wt% of the 10 culture concentrate of the culture filtrate. It is preferable that there be.

A coagulated product (curd) is formed several minutes to several hours after the addition of the enzyme, and during this time it is preferable to carry out a cutting operation in which vertical and horizontal cuts are made in the card using a card knife or the like. As a result, whey leaches out from the slits in the curd, facilitating whey removal, and lactic acid fermentation further progresses in the whey, promoting curd formation.

(5) Separation and removal of whey The obtained curd or the curd undergoing the coagulation process is pressed to separate and remove whey from the curd. In addition, if it is not necessary to remove the whey sufficiently, you may wait for the whey to naturally ooze out from the curd without pressing the curd. The card is placed in a bag made of woven fabric, preferably filter cloth, and the card is pressed in this state. Further, in the press, the pressure may be increased stepwise in multiple stages, or the card may be placed in a bag and rolled out into a plate shape and then pressed. Pressure, time, temperature, etc. of the press may be appropriately selected.

In addition, in the pressing process, if the temperature is controlled at the same time as whey is removed to further advance lactic acid fermentation and promote coagulation, whey can be easily removed and hard and firm curd can be obtained.

(6) Coagulated product (curd) The obtained curd has various characteristics such as hardness, smoothness, flavor, etc. depending on the conditions of lactic acid fermentation, enzymatic coagulation, and whey separation and removal. By aging itself or this curd, it becomes a food product as a natural cheese-like substance.

The substance has no bitterness, astringency, or soybean-specific flavor, and has a cheese-like smoothness. Note that the substance can be modified as a food by adding and mixing taste, stabilizers, flavors, etc.

Depending on the purpose, the curd is aged at 5 to 50°C for a period of one day to several years; aging at low temperatures and/or short periods of time yields less mature curds; You can get something with a high degree of ripeness. Such aging makes the structure of the curd smoother and imparts a unique flavor. If curd is inoculated with mold or the like and then aged, the degree of ripening can be increased and different flavors can be imparted. Although the surface of the card is coated with wax or the like so that the surface of the card is not contaminated during aging, it is also preferable to vacuum pack the card.

(7) A substance to impart thermoplasticity is added to the curd from which the thermoplasticized whey of the coagulated product has been separated and removed, or to the curd that has undergone the aging process. The substance is at least one selected from emulsion stabilizers, gelling agents, and thickeners, and gelatin, gums, and the like are particularly preferred. The thermoplastic substance that can be used as a material may be added at the same time as, or before or after, flavoring, which is added as necessary.

This imparts thermoplasticity to the card, allowing it to soften or melt when heated. Utilizing this thermoplastic property, it is possible to uniformly heat sterilize the entire card by heat-melting or heat-softening the card, which allows the final product to be stored for a long period of time, and during storage. There is no risk that abnormal fermentation will occur and the taste of the product will change. Furthermore, the product can be used in exactly the same way as conventional cheese, and is very easy to use, for example, on pizza or toast.

In addition, by utilizing the thermoplasticity of curd, it is possible to homogenize the curd in a heated and molten state using a homogenizer, and it is also possible to easily uniformly season the curd and adjust the acidity. It is extremely advantageous industrially because it can be carried out simultaneously. It is also extremely easy to fill the final product into a container and mold it into a desired shape.

Therefore, the final product obtained is a food product as a processed cheese-like substance, and by changing the type and amount of thermoplasticizer added, hard and soft cheese-like substances of various hardnesses can be obtained. The safety of the substance has also been confirmed through administration tests on mice.

(Example) In this example, a cheese-like substance (final product) made from soybean protein is produced by the production method according to the present invention, and the quality of the obtained product is measured and the safety of the product is confirmed. We conducted a test for this purpose.

(1), Production of cheese-like substance (la), Preparation of soymilk Full-fat soybean flour is used as a raw material for soybean protein, and the same soybean flour 1
5.2 kg, vegetable oil 11.35 kg, emulsifier lecithin 0.17 kg, and lactose 0.38 kg were mixed and dissolved in 75.9 kg of water, and sterilized at 80°C for 30 minutes.
℃ and finally use a homogenizer to weigh 250 kg.
100 kg of soy milk was prepared by homogenization under the condition of /cm2.

(lb), lactic acid fermentation, coagulation 9.5wt% skim milk powder solution was inoculated with lactic acid bacteria and heated to 35°C.
Using a culture medium obtained by culturing for 16 hours in a vacuum as a starter, 3 kg of this lactic acid bacteria starter (pH 4.5 to 4.7, acidity approximately 0.75) was added to the above soymilk (pl + 6.5 to 6.8, acidity approximately O12). The inoculated lactic acid bacteria are S, thermophile, and thermophile.
us, L, bulgaricus, L, acid
It is opillus. As a result, soy milk has a pH of approximately 6.
．． 4. The acidity becomes 0.2 to 0.25, and about 0.03 wt% of soybean protein coagulase is added thereto.

The coagulation enzyme was obtained from bacterial strain 26D-7 (Bacillus lic
heniformis) in a pH 7.0 medium (16.92 g of corn starch, 16.9 g of corn starch liquor)
2g, disodium hydrogen phosphate 10.15g, water 334
0g) for 16 hours. 3,384 ml of this culture solution was filtered through Celite, and the resulting filtrate was filtered through a precision filtration membrane (pore diameter 0.2 μm).
Bacterial cells were isolated using a filtrate, and the filtrate after bacterial cell separation was concentrated 10 times using an ultrafiltration membrane. Obtained concentrate 3
33ml was added to and mixed with the soymilk undergoing lactic acid fermentation, and the mixture was kept at 40°C for 3°5 hours.

(lc) Separation and removal of whey Curds are gradually formed by the addition of a coagulating enzyme, and after the above-mentioned time has elapsed, the curds are cut vertically and horizontally using a card knife to promote whey leaching, and after 30 minutes, the curds are The whey was removed by stage pressing. Prior to pressing, the curd was housed in a filtration bag made of woven synthetic fiber cloth and rolled out into a thin plate shape so that whey could be easily removed. Hold the card in this state at 20°C10, 1kg/cm2.
Hony was removed for 2 hours at the same temperature and at 0.2 kg/cm2 overnight.

(ld), Aging of the coagulum The resulting curd weighed 35 kg, which was vacuum packed and aged at 20°C for one month, yielding a natural cheese-like substance, which was fully edible. It's something you get. In this example, soft and hard cheese-like substances were prepared using unripened curd and aged curd.

(le), a soft type cheese-like substance. Curds both before and after ripening can be used satisfactorily, and in this example, curds before ripening were used. The water content and pl+ of the curd were adjusted, and xanthan gum and locust bean gum were added to it at Oj5wt% and 0.15w, respectively.
t% was added and mixed.

Each of the above gums is excellent as an emulsion stabilizer or thickener, and when used together, they gel the curd, giving it cream-like softness and smoothness, as well as thermoplasticity, while minimizing oil separation. limited. Thereafter, the curd was sterilized by heating at 80°C for 30 minutes while stirring with a mixer, and 300 kg/kg was sterilized using a homogenizer.
Do not homogenize under conditions of cm2. It goes without saying that the obtained product can be flavored by adding various flavors, and the flavors can be added either before or after the addition of each of the above-mentioned gums.

(If) Hard type cheese-like substance In this example, aged curd, that is, a natural type cheese-like substance was used. The water content and pH of the curd were adjusted, and carrageenan, guar gum, and decaglycer monostearate were added at 0.75 wt% and 0.2 wt%, respectively.
wt% and 1 wt% were added and mixed. Carrageenan acts as a gelling agent, guar gum acts as a thickener, and decaglycer monostearate acts as an emulsifier, giving appropriate hardness and smoothness in a state of low water content, as well as thermoplasticity. To obtain a product with high hardness, the water may be evaporated using a vacuum mixer or soaked in 5% salt water. Immersing the curd in salt water makes the curd firmer and more resistant to bacterial contamination, and gives it a nice salty taste. In addition, heat sterilization is performed at 80℃ in a melted state for 30 minutes, similar to soft cheese-like substances.
In this example, the sample was treated with salt water for 0 minutes, and then stored in a refrigerator at 5° C. for 2 days to dry, thereby obtaining the final product.

(2) Product quality The product is a pure vegetable cheese-like substance that has soybean protein as its main ingredient and vegetable oil added to it. It is extremely effective as a health food for those who consume excessive cholesterol.

Among the products, the properties of the hard cheese-like substance were measured using a texturometer (RGTX-2-1M model, manufactured by Zenken Texturometer Co., Ltd.) under the following conditions. The results are shown in the table below in comparison with the characteristics of an example of domestically produced processed cheese, and the softeners and melters are also listed.

(2a), Measurement conditions by texturometer Temperature: 1
0℃ Sample thickness: 13mm Plunger: Diameter 1
8mm Clearance: 2mm Voltage=IV chart paper speed: 1500mm/min Chewing speed: 1
2 times/m1n (blank below) (2b).

Measurement Results As is clear from these results, the cheese-like substance produced according to the present invention has about 80% harderness and about 82% elasticity compared to processed cheese, and can be chewed until solid food can be swallowed. They are almost the same in chewability, which is energy, and have properties extremely similar to processed cheese. In addition, since the cheese-like substance has thermoplasticity that softens when heated to about 40°C and melts when heated to 70 to 80°C, it can be used for cooking in the same way as processed cheese.

(3) Safety of enzymes and cheese-like substances A hard cheese-like substance similar to the measurement sample in the previous section (2) is freeze-dried into powder (hereinafter referred to as powdered cheese substance), and To confirm the safety of the soybean protein coagulase used (enzyme produced by strain 26D-7), an administration test was conducted on mice under the following conditions.

(3a) Mice used for administration test of cheese-like powder substance:
DDY 4 weeks old, 0 mice (20 males, 20 mice) Feed: Mouse feed manufactured by Funabashi Farm Co., Ltd. was used as the basic feed, and the following four types of mixed feeds were prepared.

In addition, each percentage in the compounded feed is all percentage by weight.

(b) Soybean powder 5.4% + cheese-like powder substance 0.4%
Ten basic feed 94% (U), soybean powder 3. θ% + Cheese-like powder substance 3.0%
10 basic feed 94% (c), cheese-like powder substance 6. θ% 10 basic feed 94%.

(d), 100% basic feed (comparative example) Breeding method: Divide the mice into 4 groups each of males and females, with 5 males in 1 group and 5 females in 1 group. The feed was administered to the animals for 13 weeks.

Observation items Average weight of mice per two cages, feed intake,
Eye color, movement, coat gloss, and necropsy at the end of the experiment.

Test method: F-1 test using multiway analysis of variance of feed intake and body weight was performed.

(3b), Enzyme administration test mice used: DDY 4 years old, 20 males, 10 females) Feed Enzyme for mice manufactured by Nifunabashi Farm Co., Ltd.: Added to drinking water to adjust the enzyme concentration to 180 μg/ +++ I Preparation Breeding method: 5 males and 5 females are divided into 2 groups each for males and females, and one group for each male and female is provided with feed, enzyme water (PO), and a pond. Each group was administered feed and tap water for 13 weeks (comparative example).

Observation items: Average mouse weight per cage, feed intake, enzyme water or tap water intake, appearance of mice, and necropsy at the end of the experiment.

Test method: F-1 test was performed using a multi-way analysis of variance method of enzyme water intake and body weight.

(3c), Effect test on enzyme reproduction Mouse used: D
DY, 4 years old, 9 males, 3 males, 6 females> Feed: Feed water prepared by adjusting the above feed and tap water to an enzyme concentration of 30 μg/ml.

Breeding method: Obtain Fl by mating, divide it into 6 pairs, and breed F.
I got 2.

Observation items: Mouse movement, birth rate (3d), results The relationship between the administered feed and mouse body weight in the cheese-like powder substance administration test is shown in Figure 1. From this result, the body weight of mice in each group is higher in #1 (black circles) than in males (white circles), but in the cheese-like powder substance administration groups (A) to (C) and the same substance non-administration group (D). There is no significant difference in weight between the two. No abnormalities in eye color, movement, coat luster, etc. were observed in any of the mice during the breeding period, and no mice died during this period. No abnormalities were observed in the autopsy results of the mice at the end of the experiment. However, a tendency for less fat deposition was observed in the cheese-like powder substance administration groups (a) to (c).

FIG. 2 shows the relationship between the presence or absence of enzyme administration and the body weight of mice in the enzyme administration test. This result shows that there is a difference in body weight depending on whether the enzyme was administered or not. The total water intake was 10,100 O for males in the enzyme administration group (E) and 800 ml for females.
They ingested more than the non-administered group. No abnormalities were observed in the appearance of the mice during the experimental period, and no deaths occurred. No abnormalities were observed in the autopsy results at the end of rearing.

Fat deposition in the peritoneum and mesentery was observed to the same extent in both groups, regardless of whether enzymes were administered or not, but almost no subcutaneous fat was observed in the enzyme-administered group (Po).

In the enzyme effect test on reproduction, no abnormalities were observed in the appearance or behavior of the mice during the breeding period. The birth rate was slightly lower at 73% for Fl and 74% for F2, but this seems to be due to poor testing rather than the influence of the enzyme.

[Brief explanation of drawings]

FIG. 1 is a graph showing the relationship between the administration of the cheese-like substance obtained by the production method of the present invention and the body weight of mice, and FIG. It is a graph showing the relationship with body weight.

Claims (3)

[Claims] (1) After adding lactic acid bacteria and soybean protein coagulase in this order or simultaneously to a material containing soybean protein, lactic acid fermenting the soybean protein and coagulating it, and separating and removing whey from the resulting coagulated product. A method for producing a cheese-like substance using soybean protein as a raw material, which comprises adding a substance that imparts thermoplasticity to the coagulated product. (2) In the production method described in item 1, the soybean protein coagulating oxygen is an enzyme produced by Bacillus licheniformis, which has low spore-forming ability and high productivity of soybean protein coagulating enzyme. A method for producing a cheese-like substance using soybean protein as a raw material. (3) In the manufacturing method according to item 1, the substance imparting thermoplasticity is at least one substance selected from gelling agents, emulsion stabilizers, and thickeners. A method for producing a cheese-like substance using soybean protein as a raw material.