JP7489178B2 - Natural cheese and its manufacturing method - Google Patents

Description

The present invention relates to natural cheese and a method for producing the same. In particular, the present invention relates to natural cheese having a nutty aroma and a method for producing the same.

The nutty aroma in cheese (hereinafter sometimes referred to as nutty aroma) is thought to be largely due to branched-chain amino acid-derived components such as 3-methylbutanal and isovaleric acid, and these components are produced by microbial enzymes using branched-chain amino acids as raw materials during cheese production and ripening.
It is known that in cheese, the branched-chain amino acid leucine (isoleucine, valine) is converted to a branched-chain keto acid intermediate by branched-chain amino acid aminotransferase, and then branched-chain aldehydes and branched-chain alcohols (3-methylbutanal and 3-methyl-1-butanol) or branched-chain organic acids (isovaleric acid) are produced (Non-Patent Documents 1 and 2).

Patent document 1 discloses a mixed medium of two or more microbial strains, at least some of which are individually selected based on their ability to perform part of an enzymatic pathway, and the individually selected microbial strains together form a complete pathway for a desired flavor compound. It also discloses an enhanced or special flavor medium, the starter medium comprising a combination of two or more microbial strains that have little or no ability to convert amino acids into volatile compounds to the extent that they form the desired flavor, but which, when used together, convert amino acids into volatile compounds under appropriate conditions to give the desired flavor. Specifically, the present invention discloses a starter medium for cheese production comprising a combination of two Lactococcus strains, in particular Lactococcus cremoris strain B1157 and Lactococcus cremoris strain SK110, and a starter medium for cheese production comprising a combination of a Brevibacterium strain and a Staphylococcus strain, in particular Brevibacterium casei strain B1392 and Staphylococcus saprophyticus strain B1144.

JP 2004-501633 A

GARCIA-CAYUELA, Tomas, et al. Expression in Lactococcus lactis of functional genes related to amino acid catabolism and cheese aroma formation is influenced by branched chain amino acids. International journal of food microbiology, 2012, 159.3: 207-213. THIERRY, Anne; RICHOUX, Romain; KERJEAN, Jean-Rene. Isovaleric acid is mainly produced by Propionibacterium freudenreichii in Swiss cheese. International dairy journal, 2004, 14.9: 801-807.

The objective of the present invention is to provide a new natural cheese with a nutty aroma and a method for producing the cheese.

In order to solve the above problems, the inventors have conducted extensive research and found that a novel natural cheese having a nutty aroma can be produced by using a bacterium belonging to the genus Propionibacterium and a bacterium belonging to the genus Lactococcus lactis as starters and maturing the cheese under specific conditions, which led to the completion of the present invention.
(1) A food product containing bacteria of the genus Propionibacterium and bacteria of the genus Lactococcus species lactis, having a moisture content of 55% by weight or less, a salt content of 0.3% by weight or more and 6.0% by weight or less, and containing 3-methylbutanal of 0.5 ppm or more, 3-methyl-1-butanol of 10 ppm or more, and isovaleric acid of 5.0 ppm or more;
A natural cheese characterized by:
(2) adding bacteria of the genus Propionibacterium and bacteria of the genus Lactococcus lactis to the raw milk;
A pre-aging heating step of heating at 55°C or less;
A method for producing natural cheese, comprising the steps of:
(3) The method for producing natural cheese according to claim 1, further comprising a post-ripening heating step at 15°C or higher and 25°C or lower.

The present invention provides a novel natural cheese having a nutty aroma and a method for producing the cheese.

The concentrations (relative values) of 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid in the culture medium in which propionic acid bacteria and/or lactic acid bacteria were cultured are shown. The concentration of α-KIC in the culture medium in which propionic acid bacteria and/or lactic acid bacteria were cultured is shown. The concentrations of 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid in the sample cheese are shown. This shows the changes over time in the concentrations of 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid in prototype cheese during the warming period after ripening.

The natural cheese of the present invention will be described in detail below.
(Natural cheese)
The natural cheese of the present invention contains bacteria of the genus Propionibacterium and bacteria of the genus Lactococcus species lactis, and contains 0.5 ppm or more of 3-methylbutanal, 10 ppm or more of 3-methyl-1-butanol, and 5.0 ppm or more of isovaleric acid.
The natural cheese of the present invention is a so-called semi-hard type or hard type, and has a pH of about 5.2 to 6.0, a moisture content of about 30% to 55%, a fat content of about 25 to 30%, a protein content of about 25 to 35%, and a salt content of 0.3% by weight or more and 6.0% by weight or less.
As described above, the concentration of 3-methyl-1-butanol is sufficient as long as it is 10 ppm or more, but is preferably 20 ppm or more, more preferably 30 ppm or more, and even more preferably 40 ppm or more.
As described above, the concentration of 3-methylbutanal is sufficient as long as it is 0.5 ppm or more, but it is preferably 1.0 ppm or more, more preferably 2.5 ppm or more, and even more preferably 4.0 ppm or more.
As described above, the isovaleric acid content may be 5.0 ppm or more, but is preferably 6.0 ppm or more, more preferably 7.0 ppm or more, and even more preferably 8.0 ppm or more.

(Method of producing natural cheese)
The process for producing natural cheese generally involves the following steps:
The process includes the steps of heat sterilizing the raw milk, cooling, adding a starter, adding rennet to coagulate the raw milk, cutting the coagulated product, heating and stirring until a predetermined pH is reached (hereinafter also referred to as the pre-aging heating step), discharging the whey and packing it into molds, salting, and maturing at a predetermined temperature. The production method of the present invention is characterized by combining a specific bacterium as a starter and carrying out the pre-aging heating step at a specific temperature.
A specific embodiment of the method for producing natural cheese of the present invention will be described below.
Raw milk with a milk fat content of about 2.8% is heat sterilized (75°C for 15 seconds, 65°C for 30 minutes, etc.), cooled to about 30°C, and calcium chloride and a starter are added.
After the starter is added, rennet is added to coagulate the raw milk. The coagulated product is cut and then heated and stirred at about 35°C to 55°C until the pH is 6.4 or less (hereinafter referred to as the pre-aging heating process), after which the whey is drained and molded. Salt is added to the desired salt content, and the product is aged at about 10°C for about six months.
As mentioned above, the pre-ripening heating step may be carried out at any temperature between about 35°C and 55°C. However, since Lactococcus lactis is a mesophilic lactic acid bacterium, the temperature is preferably 35°C to 50°C, and even more preferably 35°C to 45°C.
Cheese that has been aged for about six months can be subjected to a further aging step at about 15 to 25° C. for several weeks (hereinafter also referred to as a post-aging heating step), which can adjust the flavor to a desired level by further increasing the nutty aroma, etc. As described above, the post-aging heating step may be performed at about 15 to 25° C. for several weeks, more preferably at 18 to 23° C. for several weeks, and even more preferably at 20° C. for two weeks.

(Bacteria used)
In the production of the natural cheese of the present invention, a bacterium belonging to the genus Propionibacterium and a bacterium belonging to the genus Lactococcus lactis are used as starters. Examples of the bacterium belonging to the genus Propionibacterium (propionic acid bacterium, hereinafter also referred to simply as PAB) include Propionibacterium freudenreichii subsp. shermanii, Propionibacterium freudenreichii subsp. freudenreichii, Propionibacterium freudenreichii subsp. Examples of the bacteria that can be used include Propionibacterium globosum, Propionibacterium jensenii, Propionibacterium acidipropionici, and Propionibacterium thoenii, with Propionibacterium freudenreichii subsp. shermanii being preferred, and Propionibacterium freudenreichii subsp. shermanii SBT3197 (NITE P-03001) being more preferred.
It is preferable to use Lc. lactis subsp. latis as the bacterium of the Lactococcus genus lactis species. Although there is no particular limitation on Lc. lactis subsp. latis, preferred examples include Lc. lactis subsp. latis SBT1223 (NITE-P-03003) and Lc. lactis subsp. latis SBT1220 (NITE-P-03002).
In order to produce natural cheese containing the propionic acid bacteria of the present invention and the lactic acid bacteria of the Lactococcus lactis species and containing not less than predetermined amounts of 3-methyl-1-butanol, 3-methylbutanal, and isovaleric acid, it is preferable to select an appropriate combination of the propionic acid bacteria and the Lactococcus lactis species.
As the propionic acid bacteria, bacteria with high α-ketoisocaproic acid (α-KIC) production reaction activity are preferable, and among Lactococcus lactis species, bacteria with high branched-chain α-keto acid decarboxylase activity, i.e., bacteria with high activity to produce 3-methyl-1-butanol and 3-methylbutanal from α-KIC, are preferable.
By allowing these bacteria to coexist and selecting a combination that has higher 3-methyl-1-butanol, 3-methylbutanal, and isovaleric acid-producing activity than each bacteria alone in liquid culture, the same effects can be obtained in cheese production.

(Method of analyzing aroma components)
3-Methyl-1-butanol, 3-methylbutanal, and isovaleric acid can be quantified by GC-MS as described in the Examples.

The following describes in detail the examples of the present invention, but the present invention is not limited to these.

[Test Example 1] Screening of Propionic Acid Bacteria and Lactic Acid Bacteria (1) Cultivation Method of Propionic Acid Bacteria and Lactic Acid Bacteria Seven strains of propionic acid bacteria and four strains of lactic acid bacteria shown in Tables 1 and 2 were cultivated. Frozen stocks of each propionic acid bacteria and freeze-dried ampoules of each lactic acid bacteria were subjected to SCI (Single Colony Isolation), and inoculated into 10 mL of M17 liquid medium sterilized by autoclave, and cultured at 30°C for about 18 hours in a container under anaerobic conditions using Anaeropack Kenki (Mitsubishi Gas Chemical). Next, 1% (v/v) of this culture solution was inoculated into 900 mL of M17 liquid medium, and cultured under the same anaerobic conditions. The bacterial cells in the logarithmic growth phase were cooled on ice and centrifuged at 6,000 x g, 4°C, for 15 minutes to collect the bacteria. The collected cells were washed twice with chilled milli-Q water, centrifuged, and resuspended in milli-Q water to an OD ₆₀₀ of about 80.

(2) Measurement of products produced by Leu metabolism in propionic acid bacteria or lactic acid bacteria cells The products produced by Leu metabolism were measured at the levels of #1-11 shown in Table 3. Each cell suspension was suspended in an amount equivalent to OD ₆₀₀ :10 in terms of final turbidity in a reaction buffer (a mixture of 60 mM phosphate buffer (pH 6.8), 10 mM α-ketoglutaric acid (α-KG), 50 uM pyridoxal 5-phosphate, 50 uM thiamine pyrophosphate chloride, and 5 mM Leu). The amount of each component in the reaction buffer was adjusted to the final concentration after addition of the cell suspension. Pyridoxal 5-phosphate was adjusted to 1 mg/mL conc., and thiamine pyrophosphate chloride was adjusted to 100 mg/mL conc., and mixed. These mixtures were incubated at 30°C and sampled 48 hours after the start of the reaction. The amount of α-ketoisocaproic acid in the reaction mixture was measured using LC-MS, and the amounts of 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid were measured using GC-MS. The methods for each measurement are described below.

(3) Analytical Method of Each Component (3-1) Analytical Method of α-Ketoisocaproic Acid The frozen resting cell reaction solution was thawed, mixed with 300 mM oxalic acid aqueous solution at a ratio of 9:1, and centrifuged at 8,000×g for 15 minutes at 4° C. The supernatant was ultrafiltered with a centrifugal filter unit (Nanosep 3K) at 12,000 rpm for 60 minutes, and the permeate was subjected to LC/MS analysis (negative mode/SIM mode), and the concentration of α-ketoisocaproic acid was quantified by the absolute calibration curve method from the peak area of the ion at m/z 129.

(3-2) Analysis method of 3-methylbutanal, 3-methyl-1-butanol, isovaleric acid 2 mL of the analysis sample was placed in a 20 mL vial, the headspace was replaced with high-purity nitrogen, and then the vial was sealed with a screw cap with a septum. The target components were collected in a collector (Carbopack B&X/ShincarbonX) by the dynamic headspace method using nitrogen gas. The target components of the collector were then heated and desorbed, and GC-MS (scan mode) measurement was performed. The oven temperature rise conditions of the GC-MS were 40°C-3 minutes hold → 20°C/min → 250°C-10 minutes hold, and the measurement was performed. The amount of each component produced was compared from the relative peak areas of the obtained ions of m/z 58 (3-methylbutanal), m/z 70 (3-methyl-1-butanol), and m/z 60 (isovaleric acid).

(4) Measurement results of Leu metabolic activity of propionic acid bacteria or lactic acid bacteria (4-1) Measurement results of 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid concentrations The relative values of 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid concentrations of propionic acid bacteria and lactic acid bacteria alone (#1 to #11) 48 hours after the start of the reaction are shown in Figure 1A.
The horizontal axis shows the relative value with the largest peak area among the 11 levels for each component taken as 100%. From this result, for the propionic acid bacteria alone, ATCC6207 ^T , the type strain of P. freudenreichii, had the highest detected amount of isovaleric acid among the seven strains.
In addition, P. shermanii SBT3197 detected high amounts of 3-methylbutanal and 3-methyl-1-butanol, approximately 23 times (3-methylbutanal) and approximately 3 times (3-methyl-1-butanol) higher than the average levels detected using PABs other than SBT3197.
In the results for the lactococci alone, the amount of isovaleric acid detected was the highest in ATCC19435 ^T , the standard strain of Lc. lactis, and the amount of 3-methylbutanal and 3-methyl-1-butanol detected was the highest in Lc. lactis SBT1220. On the other hand, Lc. cremoris ATCC19257 ^T did not produce the same level of Leu-derived components as Lc. lactis.

(4-2) Measurement results of α-KIC concentration The amount of α-KIC detected in the reaction solution is shown in Figure 2A. α-KIC is a precursor of Leu-derived aroma components, and is a compound generated by deamination of Leu. For this reason, it is considered that a high α-KIC concentration indicates that the reaction subsequent to α-KIC is the rate-limiting step in the reaction for producing Leu-derived aroma components, and conversely, a low α-KIC concentration indicates that the deamination reaction, which is the reaction for producing α-KIC, is the rate-limiting step.
From Fig. 2A, it was considered that propionic acid bacteria generally had relatively high α-KIC concentrations and high α-KIC synthesis activity when used alone, while the subsequent reactions were the rate-limiting step in the synthesis reaction of aroma components derived from Leu. Among them, P. shermanii SBT3197 in particular had a very high α-KIC concentration and was considered to be a strain with high α-KIC synthesis activity. Furthermore, Lactococcus lactic acid bacteria had relatively low α-KIC concentrations, and the α-KIC synthesis reaction was considered to be the rate-limiting step in the synthesis reaction of aroma components derived from Leu. In particular, Lactococcus lactis SBT1220 and SBT1223 had low α-KIC concentrations and high reaction activity after α-KIC.

(5) Discussion From the above, it was speculated that the production of Leu-derived 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid was promoted by producing cheese using a combination of propionic acid bacteria, particularly P. shermanii SBT3197, and Lactococcus lactic acid bacteria, particularly Lc. lactis SBT1223.

[Test Example 2] Leu metabolic activity test in the coexistence of propionic acid bacteria and lactic acid bacteria Production of each aroma component was measured in the same manner as in Test Example 1, except that propionic acid bacteria with high α-KIC concentration in Test Example 1 were combined with various lactic acid bacteria.
(1) Measurement results of 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid concentrations Figure 1B shows the measurement results of 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid in the coexistence of P. shermanii SBT3197 and Lactococcus lactic acid bacteria. As a control, the test results of P. shermanii ATCC9614 ^T and Lactococcus lactic acid bacteria are also shown. The horizontal axis is expressed as a relative value, with the level with the largest peak area among 14 levels of each component taken as 100%. In the case of lactic acid bacteria alone, the detection amount of the three components of Lc. cremoris SBT19257 ^T was low, and the detection amount of 3-methylbutanal and 3-methyl-1-butanol was high for the three strains of Lc. lactis.
Regarding the effect of coexistence of propionic acid bacteria and lactic acid bacteria, the combination of P. shermanii SBT3197 and Lc. lactis showed high detection amounts of 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid, and the combination of P. shermanii ATCC9614 ^T and Lc. lactis showed that the detection amounts of 3-methyl-1-butanol and isovaleric acid were higher than the sum of the detection amounts when each was reacted alone.
From the above, it was found that the coexistence of propionic acid bacteria and Lactococcus genus lactic acid bacteria promotes the production of 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid. In particular, the combination of P. shermanii SBT3197 and Lc. lactis SBT1220 or SBT1223 showed significantly higher detection levels of the three components than the test results for the control P. shermanii ATCC9614 ^T.

(2) Measurement results of α-KIC concentration Figure 2B shows the amount of α-KIC detected in the reaction solution in the presence of propionic acid bacteria and Lactococcus lactic acid bacteria. As a result, the concentration of α-KIC accumulated in the reaction solution in the presence of P. shermanii alone was reduced by the presence of P. shermanii and Lc. lactis.
This indicates that the combination of propionic acid bacteria with relatively high α-KIC production activity and Lc. lactis with high activity in reactions after α-KIC has the effect of enhancing aroma component production. In particular, the combination of P. shermanii SBT3197 and Lc. lactis was considered to have a significantly high effect of enhancing aroma component production.

[Test Example 3] Cheese Production Example 1
(1) Test Method Natural cheese was produced using L. lactis SBT1223 and P. shermanii SBT3197 according to the following method.
The raw milk was adjusted to a milk fat content of 2.8% and pasteurized at 75°C for 15 seconds. It was then cooled to 30°C and calcium chloride (0.01 w/w%) was added. P. shermanii, L. lactis, and a commercially available starter for cheese production were then added. The combination of starters is shown in Table 4.
After the addition of these starters, calf rennet was added to a concentration of 0.024 IMCU/mL to coagulate the raw milk. The coagulated product was cut into a rectangular shape and heated with stirring at the temperature shown in Table 4 (heating temperature before aging) until the pH became 6.4 or less, after which the whey was drained, molded, and squeezed. After salting, the cheese was aged at 10°C for 6 months, and the aroma components were analyzed by the method described below. The moisture content of the resulting cheese was 39%, and the salt content was 1.5%.

(2) Analysis method of 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid in the prototype 0.2 g of the sample after 6 months of aging was weighed into a 10 mL vial, the headspace was replaced with high-purity nitrogen, and the vial was sealed with a screw cap with a septum. Each vial was set on a sample tray with a cooling function (10 ° C.), and the target components were collected in a collector (Tenax TA and Carbopack B & X / Shincarbon X) by the dynamic headspace method using nitrogen gas. Then, the target components of the collector were heated and desorbed, and GC-MS (scan mode) measurement was performed. The oven temperature rise conditions of the GC-MS were 40 ° C.-3 minutes hold → 20 ° C. / min → 250 ° C.-10 minutes hold, and the measurement was performed. The concentrations of each component were quantified by the standard addition method from the peak areas of the obtained ions of m/z 58 (3-methylbutanal), m/z 70 (3-methyl-1-butanol), and m/z 60 (isovaleric acid).

(3) Test results The concentrations of the three components, 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid, in the cheese are shown in Figure 3. The 3197L level had the highest concentration of 3-methylbutanal, at 0.8 ppm, which was more than twice as high as that of 9614L, which used P. shermanii ATCC9614 ^T as the propionic acid bacterium. The 3197LH level, which uses the same strain combination as 3197L but is heated at a higher temperature, also had a high concentration of 0.6 ppm.
In addition, 3-methyl-1-butanol was detected at a concentration of over 40 ppm at the 3197L level, the highest concentration.
The concentration of isovaleric acid was also higher than other combinations, at 8.1 ppm at the 3197L level and 7.1 ppm at the 3197LH level.

(Discussion)
These results confirmed that the test-tube results were reproduced in actual cheese.
On the other hand, the results of this study revealed that the concentration of aroma components also differs depending on the temperature during the heating and stirring process during the manufacturing process (heating temperature before aging). This is thought to be because the mesophilic lactic acid bacteria Lc. lactis decreases during the heating process at high temperatures.

[Test Example 4] Cheese Production Example 2
(1) Test Method The influence of the heating step after ripening on the amount of aroma components was studied for the 3197L and 3197LH cheeses produced in Production Example 1.
That is, 3197L and 3197LH were prepared in the same manner as in Cheese Production Example 1, and aged at 10°C for 6 months. An additional aging process was then added to the 3197L and 3197LH cheeses, which were aged at 20°C for 2 weeks (post-aging heating process), and the aroma components after aging were measured (Figure 4).

(2) Analysis results of 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid in the prototype cheeses In 3197L, the addition of a post-aging heating process of aging at 20°C for 2 weeks resulted in an increase in all three components, 3-methylbutanal, 3-methyl-1-butanol, and isovaleric acid. In particular, 3-methylbutanal increased by about 5 times. On the other hand, in 3197LH, although 3-methylbutanal and 3-methyl-1-butanol increased, the degree of increase was smaller than that in 3197L. Also, no increase in isovaleric acid was observed.

(3) Flavor evaluation of prototype cheeses Nutty aroma was evaluated for 3197L and 3197LH aged for 6 months at 10° C., and 3197L and 3197LH produced by aging for 2 weeks at 20° C. followed by an additional heating step. The evaluation was performed by 10 members of the cheese development team.
As a result, all cheeses were evaluated as having a nutty aroma. The nutty aroma was weakest in 3197LH, followed by 3197LH (heating process after aging), 3197L, and 3197L (heating process after aging). The flavor of the cheese was also preferred in 3197L, with 3197L (heating process after aging) being the most preferred.

(4) Discussion From the above, it was found that cheese with a strong nutty aroma and a pleasant flavor can be obtained by using a starter containing a combination of P. shermanii SBT3197 and Lc. lactis SBT1223 and performing the pre-ripening heating step at about 35 to 45° C. In addition, it was found that cheese with a strong nutty aroma and a pleasant flavor can be obtained by performing the post-ripening heating step at about 15 to 25° C.

The present invention provides a novel natural cheese with a nutty aroma and a method for producing the cheese.

[Reference to deposited biological material]
(1) Propionibacterium freudenreichii subsp. shermanii SBT3197
Name and address of the depository institution to which the biological material was deposited: National Institute of Technology and Evaluation, Patent Biological Deposit Center, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan (postal code: 292-0818)
Date of deposit of biological material at the depository in Loi: July 8, 2019 Accession number given by the depository in Hai: NITE P-03001

(2) Lactococcus lactis subsp. Latis SBT1220
Name and address of the depository institution to which the biological material was deposited: National Institute of Technology and Evaluation, Patent Biological Deposit Center, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan (postal code: 292-0818)
Date of deposit of biological material at the depository in Loi: July 8, 2019 Accession number given by the depository in Hai: NITE P-03002

(3) Lactococcus lactis subsp. Latis SBT1223
Name and address of the depository institution to which the biological material was deposited: National Institute of Technology and Evaluation, Patent Biological Deposit Center, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan (postal code: 292-0818)
Date of deposit of biological material at the depository in Loi: July 8, 2019 Accession number given by the depository in Hai: NITE P-03003

Claims (3)

A natural cheese (excluding pasta filata cheese and said natural cheese which has been aged for one month or less) which contains bacteria of the genus Propionibacterium and bacteria of the genus Lactococcus, species Lactis, has a moisture content of 55% by weight or less, a salt content of 0.3% by weight or more and 6.0% by weight or less, and also has 3-methylbutanal of 0.5 ppm or more, 3-methyl-1-butanol of 10 ppm or more, and isovaleric acid of 5.0 ppm or more. A method for producing natural cheese having a moisture content of 55% by weight or less, a salt content of 0.3% by weight or more and 6.0% by weight or less, 3-methylbutanal of 0.5 ppm or more, 3-methyl-1-butanol of 10 ppm or more, and isovaleric acid of 5.0 ppm or more, the method comprising the steps of adding bacteria of the genus Propionibacterium and bacteria of the genus Lactococcus lactis to raw material milk, and a pre-ripening heating step of heating at 55°C or less (however, this does not include a production method including a pasta filata step and a production method for said natural cheese in which the ripening period is one month or less ). The method for producing natural cheese according to claim 2 further comprises a post-ripening heating step at 15°C to 25°C.