JPS60156380A - New strain belonging to genus lactobacillus and its use - Google Patents

Abstract

PURPOSE:A novel strain Lactobacillus psychrophilus belonging to the genus Lactobacillus, capable of producing L-lactic acid from glucose by hetero type fermentation at the fittest growth temperature of <= a specific temperature, providing a high-quality meat food by the use of it. CONSTITUTION:New strains Lactobacillus psychrophilus (N-10 strain) and Lactobacillus deliciosofaciens (D-3 strain) belonging to the genus Lactobacillus are obtained by a well-known screening means. These strains produce L-lactic acid from glucose by hetero type fermentation at the fittest growth temperature of <=20 deg.C. These microorganisms have improved action as a starter in preparation of processed edible meat product, a seed mold is added to a raw material meat before aging, which is aged, to give a high-quality meat food (e.g., ham). These new strains are obtained by isolation from various raw ham and by purification.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to new bacterial species belonging to the Lactobacillus sp. and a method for producing meat foods using them.

More specifically, the present invention relates to new bacterial species Lactobacillus cyclophilus and Lactobacillus derrinosofaciens, and a method for producing meat foods of good quality using these microorganisms.

Meat foods are abundant in variety and consumption is increasing, but if they are to be handled as commercial products, a constant supply of good quality is required.

However, at present, this point is not necessarily sufficient.

An example of a problem where variation in product quality is a problem is meat products (eg, raw ham) that are not heat-treated during the manufacturing process. In other words, the type of raw pork.

Due to differences in production lots, the odor, flavor, and color of the finished raw ham may vary, resulting in variations in product quality, making it impossible to obtain a product of consistently high quality. However, solving these problems has become a challenge in this industry as well.

In order to obtain microorganisms useful as starters in the production of raw ham, which has many problems, the present inventors investigated the microbial distribution of raw ham in detail, isolated and purified 52 strains of microorganisms, and When raw ham was produced by adding this to raw meat during salting, it was found that two of the strains added to the isolated bacteria produced significantly better quality raw ham, and each was ranked N-10.
strain and D-3 strain.

When various mycological properties of these two isolated strains were examined, it was found that although both belong to the genus Lactobacillus, they are different from any known bacterial species and are new bacterial species.

The present invention was completed based on the above findings.

That is, the present invention provides (1) Lactobacillus cyclophilus which produces L-lactic acid from siglucose by hetero-fermentation at an optimal growth temperature of 20°C or lower, (2) DT from glucose by homo-fermentation, - Lactobacillus deliciosophaciens, which produces lactic acid and uses ribose but not gluconic acid as a carbon source; (3) Lactobacillus cyclophyllus or (
and) A method for producing meat foods characterized by adding and aging Lactobacillus delinosofaciens inoculum.

The N-10 strain and D-3 strain of the present invention were found in a certain raw ham after investigating the biological distribution of various raw hams, and were found in a known screening for Lactobacillus bacteria. It is obtained by means.

The mycological properties of strain N-10 and strain D-3 are as follows.

1. Mycological properties of strain N-10 (1) Morphological properties The cells are rods with a size of 0.6-0.7 x 1.2-2.0 μm and are non-motile. No spores are formed and Durham staining is positive. No cellular pleomorphism and acid fasting properties.

(2) Growth status on various media 1) Meat juice agar medium: Poor growth, forming white colonies.

11) Peptone-yeast extract-glucose agar medium; moderate growth, forming white colonies.

111) Peptone-yeast extract-glucose liquid medium medium growth, whole medium becomes cloudy.

iv) Broth gelatin puncture culture; grows to the bottom.

Gelatin is liquefied.

V) Lidmus milk; slightly acidified. No coagulation or peptonization.

(3) Physiological properties i) Catalase reaction; negative ii) Oxidase reaction; negative 1ii) MR test; positive iv) VP testro positive V) Reduction of nitrate; negative vi) Formation of indole; negative vii) Hydrogen sulfide Negative - Starch hydrolysis; weakly positive ix) Utilization of citric acid; Negative X) Production of pigment; None Xi) Urease; Negative The optimum temperature range is 10°C to 12°C.

xiii) Growth pH 1 Grows in the range of pH 4,5 to 8.0 0xLv) Attitude towards oxygen 1 Facultative anaerobic xv) O-F
Test N Fermentatively produce acid. No gas is produced.

Calm) Production of acid from sugar; glucose, mannose.

Produces acids from fructose, galactose, maltose, sucrose, trehalose, mannitol, glycerin, riconic acid, ribose and starch. Produces a small amount of acid from lactose and sorbitol.

Xylose, arabinose, and inosit do not produce acids.

xvn) Vitamin requirements; requires nicotinamide, riboflavin and calcium pantothenate. Thiamine, folic acid and pyridoxal are not required.

Once) Tolerance to salt; Tolerant to 9% salt
g) Generate lactic acid.

Mycological properties of strain Il, D-3 (1) Morphological properties The cells are rods with a size of 0.5-0.6 x 1-1-1.5 μm and are non-motile. be. It is pleomorphic, and initially appears as a planar bacillus, but later the cells become curved and loop-shaped. Durham staining is positive. No spore formation and anti-acid properties.

(2) Growth status in various media 1) Broth agar; very poor growth.

ii) Peptone-yeast extract glucose agar; poor growth, forming white colonies.

1ii) Peptone-yeast extract-glucose liquid medium;
Moderate growth, entire medium becomes cloudy.

iv) Broth gelatin puncture culture; grows to the bottom.

Gelatin does not liquefy.

v)! J) Mass milk; slightly acidified. No coagulation or peptonization.

(3) Physiological properties 1) Catalase reaction; negative 11) Oxidase reaction; negative 1ii) MR test; negative iv) VP test; negative; ) Production of hydrogen sulfide; Negative viii) Hydrolysis of starch; Negative ix) Utilization of citric acid Negative X) Formation of pigment i None xi) Urease reaction Negative It does not grow at 41°C. The optimum temperature range is 20'C to 25C.

xtii) Growth pJGrows in the range of 4.5 to 8-0 (7).

X1lv) Attitude towards oxygen; facultative anaerobic xv) O-
F test: Acid is produced by fermentation. No gas is produced.

Producing acids from sugars; producing acids from glucose, fructose, mannose, galactose, xylose and ribose. Arabinose, malt, sucrose, lactose, trehalose, sorbitol.

mannit, inosit, glycerin, starch.

Gluconic acid and esculin do not produce acids.

ml) Auxotrophy: It has a complex auxotrophy and requires the addition of yeast extract or meat extract to the growth medium.

Tolerance to 6% salt; Tolerance to 6% salt.

th) Tolerance to ethanol; Tolerance to 5% ethanol.

XX) Production of gas from gluconic acid i Negative xd) Production of lactic acid from glucose; In homo-fermentation: +J-) Lactic acid and D (→ Lactic acid are produced in approximately equal amounts.

Based on the above mycological properties, the Purge Aids Manual of Determinative Batatteriology (
Bergey's Manual of Determinology
-1 Native Bacteriology) 8th edition (1974), N-10 strain and D
Both of the -8 strains are Durham-positive, non-motile bacilli, do not form spores, are negative for the catalase reaction, grow facultatively anaerobically, and produce a significant amount of lactic acid from glucose, so Lactobacillus ( It was revealed that it was a bacterial species of the genus Lactobacillus. However, N-10
The optimum temperature for growth of the strain is below 20℃, especially between 10℃ and 12℃.
It is a psychrophilic bacterium that does not grow at 37℃, and it produces lactic acid, ethanol, and acetic acid from siglucose through heterozygous fermentation, but the lactic acid produced is L
(1) It can be clearly distinguished from any known species belonging to the genus Lactobacillus due to its unique property of containing only lactic acid.

In addition, the D-8 strain converts glucose to DL-
Lactobacillus calpertus has characteristics such as producing lactic acid, growing at 15°C, using ribose as a carbon source and not mannitol, and observing curved cells.
Curbatus). However, D
The -8 strain differs from Lactobacillus calpartas in the following mycological properties, and the two can be clearly distinguished from each other in these properties.

1) Use xylose as a carbon source and do not use gluconic acid, maltose, salicin, or estarin.

2) The optimum temperature for growth is 20 to 25°C, which is lower than the optimum temperature for Lactobacillus calvertus, which is 82 to 37°C.

That is, the bacteriological properties of the D-a strain are characterized by the fact that it produces DL-lactic acid from siglucose through homozygous fermentation, and uses ribose as a carbon source, but does not use gluconic acid.

According to the above results, both strains N-10 and D-a can be distinguished from any known bacterial species of the genus Lactobacillus, so both strains are recognized as new bacterial species.
The strain is called Lactobacillus cyclophilus (Lactobacillus-us cyclophilus), named after its psychrophilic nature.
philus), and the D-a strain is Lactobaeillus deliciosophaciens, named after its properties that improve the quality of processed meat products.
In addition to the above-mentioned mycological properties, the microorganism according to the present invention, which has been named ``S.

Higashi 1-chome 1-3) with accession number yERnP-7naθyu, and D-3 strain with accession number FEBM to the same institute.
It has been deposited as P-7yθ.

In addition, the N-10 Research Institute (Eng.FO, 2-17-85 Jusaki-cho, Yodogawa-ku, Osaka-shi, Osaka Prefecture)
The strain has been deposited with accession number IF0 14815, and the D-a strain has been deposited with accession number UFO14a14.

Next, the method for producing meat foods of the present invention includes Lactobacillus spp.
Cyclophilus or (and) Lactobacillus deliciosophaciens is used, and the above-mentioned N-10 strain and D-8 strain are preferably used. Furthermore, it is well known that the properties of microorganisms are generally not always constant and can easily mutate either naturally or artificially, and the cases of strains H-10 and D-a are exceptions to this. Rather, they are easily mutated by artificial mutagenic means such as irradiation with ultraviolet rays, Anything that works well as a starter can be used.

The microorganism belonging to the new bacterial species of the present invention was obtained by separating it from %Sa raw ham, but it can be easily obtained by restoring the dried sample of the deposited strain.

The microorganism according to the present invention is cultured using a medium containing a nutrient source used by the microorganism. The medium may be semi-synthetic or natural, solid or liquid, but liquid medium containing natural nutrients is usually preferred. The culture temperature is preferably 10 to 25°C. As the carbon source added to the culture medium, any carbon source that can be used by this bacterial species can be used. Other nutrients added include peptone, meat extract, yeast extract, and the like. For culture, static culture in a liquid medium is suitable. The culture period is usually about 2 to 5 days. The bacterial cells are usually collected by centrifuging or filtering the culture.

The meat foods targeted in the production method of the present invention refer to raw meat such as animal meat and poultry, and salted meat and heated products produced using these raw meats as raw materials.

Examples of the meat include pork, beef, and horse meat.

Examples of such meat include sheep, goat, rabbit, and whale meat, and examples of poultry include chicken and turkey.

Processed products manufactured using the raw meat include, for example, ham, sausage, and bacon.

In the present invention, the timing of adding microorganisms to meat foods is as follows:
It may be added at the raw material stage of the target meat food, at an appropriate stage in each manufacturing process such as salting or kneading, or at any of various process stages. For example, to add it to raw meat, there is a method in which the inoculum of the present invention is rubbed into raw meat. To add it to processed products, for example, for ham, sausage, bacon, etc., in the process of adding a salting agent during salting,
It may be added to raw meat by an injection method, a pickling method, a soaking method, etc. in accordance with conventionally known addition procedures for salting agents. Furthermore, if the meat is not salted, it is preferable to add it when mixing auxiliary raw materials.

The amount of microorganisms added in the present invention is appropriately selected depending on the type of meat food, manufacturing conditions, etc., but is usually 1f per 1f of meat.
03 or more, preferably 105 or more.

Meat with biological additives is usually heated to around ’20°C without heating it.
In the following, this is done by storing it in an unfrozen state. The number of days for aging is appropriately selected depending on the aging temperature and the desired quality improvement effect, but it is sufficient that the period during which the added microorganisms can act on the meat is 3 days or more, preferably 5 days or more.

Each of the microorganisms Lactobacillus cyclophilus and Lactobacillus deliciosophaciens may be used alone or in combination.

According to the production method of the present invention, meat foods of good quality can be obtained. It has a better odor and mature flavor than those obtained by conventional manufacturing methods, and its color is bright pink or strong red), improving its commercial value.

Furthermore, even if there is some variation in the quality of the raw meat, the quality of the product can be improved to a certain level, so the variation in quality between production lots is reduced, which is advantageous for industrial production.

Strains N-10 and D-3 do not universally exist in natural raw ham, and even when they do exist, they coexist with a wide variety of microorganisms, and the number of them is less than 102 per 11 parts of meat. only found. According to the present invention, cured ham containing the above-mentioned bacterial strains in its natural state is also of good quality! It!
! ÷ is obtained.

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

Experimental example 1 Raw ham raw material pork 1 piece, salt 451, caster sugar 10g
, 0.2 f of sodium nitrite and 0.5 g of sodium L-ascorbate were sprinkled on the surface of the meat, and then 10 g of the suspension of the microorganisms shown in each sample group in Table 1 was sprinkled on the surface of the meat. Mixed evenly.

Next, each sample section was salted at 5°C for 8 days, and then salted using a conventional method.
After smoking at 18-20°C for 7 days, the hams were dried in the sun at 15-18°C for 7 days and packaged to produce 5 types of raw ham.

Table-1 * Indicates the number of microorganisms per 1f of pork raw material.

After storing the five raw ham samples obtained at 5°C for 7 days, a panel of 20 people conducted a sensory test using a ranking method to determine the "desirability of smell/flavor" and "good color tone." . The results are shown in Table-2.

Table 2 Note: The numbers in Table 1 represent the total rank, and the smaller the number, the more favorable or better it is.

According to the results in Table 2, isolates N-10 strain, D-3 strain, and samples A8, 4, and 5 in which these two strains were used together are clearly different from the control group (sample A1) and the conventional starter addition amount (
Compared to sample A2), the odor, flavor, and color tone were both preferable and good.

Example 1 To 10 kQ of raw meat for raw ham, 500 F of common salt, 10 crystal glucose (1, 2 g of sodium nitrite, 5 g of sodium ascorbate, and 100 tt of water suspension of microorganisms shown in Table 8) were added. The surface of the ham was covered with salt and salted, and raw ham was produced using the method shown in Experimental Example 1 below.

When this raw ham was subjected to a sensory test by a panel of 20 people for ``desirability of smell and flavor'' and ``good color tone,'' it was found that the smell and flavor of the invention plot were clearly better than the control plot. , and the color tone was also good.

(hereinafter referred to as “Kinpaku”) Table-8 *Same as Table-1.

Example 2 4 pieces of lean pork, 10kg of lean beef and 252F salt.

Potassium nitrate I4I/, 7 g of sodium L-ascorbate was added, and the mixture was salted at 2°C for 4 days.

Next, 6 kg of this salted meat and pork fat were cut into pieces the size of rice grains using a cutter.

14 pieces of shredded salted pork, 5 to 9 pieces of pork fat, 160 f of spice mix, 20 f of caster sugar, 60 g of sodium glutamate,
Condensed phosphate preparation 60g, rum 20g, starch: 36
0'! , and 320 f of ice water were added and mixed with a mixer. This mixed sausage raw material was divided into four equal parts, and 40 vtl of a water suspension of the microorganisms shown in Table 4 was added thereto and further mixed gently. Next, fill the fibrous casing and
After drying at 0℃ for 8 days and smoking for 2 days, 10-15℃
After leaving it for 30 days, it was wrapped in vinylidene chloride film to make dry sausage. When this dry sausage was subjected to a sensory test on the desirability of odor and flavor and good color tone by Spring L'L/20 people, it was found that the inventive plot clearly had a better odor and better color than the control plot.
The flavor was favorable and the color was still good.

(Margin below) Table-4 *Same as Table-1.

Claims (3)

[Claims] (1) Lactobacillus cyclophilus that produces L-lactic acid from siglucose by Peter type fermentation at an optimal growth temperature of 20°C or lower (2) Lactobacillus deliciosofaciens produces DL-lactic acid from glucone by homofermentation and uses ribose as a carbon source but does not use gluconic acid. (3) A method for producing a meat food, which comprises adding Lactobacillus cyclophyllus or (and) Lactobacillus deliciosophaciens inoculum to unripe meat raw material and aging the meat.