CN113151042B - Rice acid fermentation process for producing L-lactic acid and ethyl acetate and special bacteria thereof - Google Patents
Rice acid fermentation process for producing L-lactic acid and ethyl acetate and special bacteria thereof Download PDFInfo
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- CN113151042B CN113151042B CN202110149574.5A CN202110149574A CN113151042B CN 113151042 B CN113151042 B CN 113151042B CN 202110149574 A CN202110149574 A CN 202110149574A CN 113151042 B CN113151042 B CN 113151042B
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- 235000007164 Oryza sativa Nutrition 0.000 title claims abstract description 187
- 235000009566 rice Nutrition 0.000 title claims abstract description 186
- 239000002253 acid Substances 0.000 title claims abstract description 84
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 title claims abstract description 78
- 238000000855 fermentation Methods 0.000 title claims abstract description 61
- 230000004151 fermentation Effects 0.000 title claims abstract description 61
- JVTAAEKCZFNVCJ-REOHCLBHSA-N L-lactic acid Chemical compound C[C@H](O)C(O)=O JVTAAEKCZFNVCJ-REOHCLBHSA-N 0.000 title claims abstract description 58
- 241000894006 Bacteria Species 0.000 title claims abstract description 36
- 240000007594 Oryza sativa Species 0.000 title description 147
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- DLRVVLDZNNYCBX-ZZFZYMBESA-N beta-melibiose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)O1 DLRVVLDZNNYCBX-ZZFZYMBESA-N 0.000 description 1
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- 235000011194 food seasoning agent Nutrition 0.000 description 1
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- 235000002906 tartaric acid Nutrition 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L23/00—Soups; Sauces; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
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- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/165—Paracasei
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Abstract
The invention belongs to the technical field of food, and particularly relates to a rice acid fermentation process for producing L-lactic acid and ethyl acetate and special bacteria thereof. The special bacteria are Lactobacillus paracasei CCTCC NO: M2021074 and Kluyveromyces marxianus CCTCC NO: M2021073. The invention adopts lactobacillus paracasei CCTCC NO: M2021074 and Kluyveromyces marxianus CCTCC NO: M2021073 to ferment and produce the rice acid, and the process comprises the following steps: pretreating raw materials; boiling; pasting; liquefying; saccharifying; blending and sterilizing; inoculating bacteria; and (5) fermenting. The obtained rice acid has L-lactic acid content of 3.45g/100g, and lactic acid bacteria content of 1.3-1.4 × 108About cfu/ml; the taste and flavor are good, and the sensory evaluation is high; the antioxidant property is achieved, and the DPPH clearance rate can reach about 46%; the selenium element content is 0.05-0.07 mg/kg.
Description
Technical Field
The invention belongs to the technical field of food, and particularly relates to a rice acid fermentation process for producing L-lactic acid and ethyl acetate and special bacteria thereof.
Background
The acidic food mainly comprises soybean products, fermented fish products, fermented vegetables, fermented bread, porridge and fermented beverage. Most of the acidic foods are mainly fermented by lactic acid bacteria, have rich sour and aroma components, the lactic acid bacteria in the initial stage of fermentation provide favorable environments for the later stage fermentation including alcohol and ester production, and the existence of yeast provides nutrient substances for the lactic acid bacteria; thereby contributing to the unique taste and aroma of acidic foods.
Sour soup is divided into red sour soup and white sour soup, wherein the red sour soup mainly takes tomatoes and red peppers as raw materials, the white sour soup is also called rice acid, and sticky rice or rice, flour and the like are taken as fermentation raw materials. The rice acid is a flavoring mainly containing lactic acid. However, the rice acid is not only eaten as a seasoning, but also can be used as a probiotic product, namely rice acid ferment. Enzyme products are rich in probiotic components and are being accepted and liked by more and more people. The research and development of the key control points of the texture forming mechanism and the processing technology are very deficient.
At present, the preparation of the rice acid still stays in the traditional process stage of a small workshop, the scale is small, the product standard is not uniform, the fermentation time is long, and the flavor of the food is difficult to reproduce. Due to lack of systematic research on souring strains, microorganism indexes and the like, the sour soup has unstable sourness, so that the sour soup has large flavor difference and even is rotten, the product quality is extremely difficult to stabilize, and the sour soup becomes a key technology bottleneck seriously restricting the leap-type development of industries or enterprises.
Selenium is a micronutrient essential to the human body, and is recommended to be consumed in a daily diet of 55 mug for adults, which is an important component of selenoprotein. The deficiency of selenium can lead to several diseases, such as keshan disease. At present, selenium element is lacked in many places in the world, and selenoprotein cannot be well expressed in human bodies. The selenium rice contains rich selenium element, so the rice acid ferment with the probiotic effect is researched based on the selenium rice as the raw material.
On the other hand, the pure fermentation strain shows different metabolic activities, growth rates, adaptability to substrates, fermentation flavor and the like after being separated from the fermented food; even different strains of the same strain have different properties in all aspects, and the metabolite of the fermentation strain directly influences the final quality of the fermentation product. Therefore, there is a need to study the fermentation process of rice acid ferment under the action of probiotic functional strains screened from rice acid system.
Disclosure of Invention
The invention provides a rice acid fermentation process for producing L-lactic acid and ethyl acetate and special bacteria thereof by researching microorganisms and processes for fermenting rice acid soup, solves the problems of long production period and unstable product quality of the rice acid in the prior art, improves the flavor of the product, and increases the content of trace element selenium in the rice acid; the method is realized by the following technical scheme:
the special bacteria for producing the L-lactic acid and the ethyl acetate through the rice acid fermentation are Lactobacillus paracasei (Lactobacillus paracasei) H4-11 and Kluyveromyces marxianus (Kluyveromyces marxianus) L1-1; lactobacillus paracasei (Lactob-a cillus paracasei) H4-11, which has been preserved in China center for type culture Collection with the preservation number of CCTCC NO: M2021074 and the preservation date of 2021 year, 1 month and 15 days; kluyveromyces marxianus L1-1 has been preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of M2021073 and the preservation date of 2021 year, 1 month and 15 days.
The lactobacillus paracasei H4-11 is cultured on an MRS plate for 48-72 hours to form a bacterial colony with the diameter of 0.5-2 mm, milky white, round, smooth and moist surface, and neat and convex edge. Acid and bile salt resistance, obvious acid production performance and short rod shape under a microscope. Gram-positive, non-spore, slender and curved bacilli, sometimes in the shape of sphere rods or rods, arranged in a grid or chain, without power, some with bipolar staining; can ferment fructose, galactose and glucose, cannot utilize melibiose, raffinose and xylose, cannot decompose arginine to produce ammonia, and the produced lactic acid has L-shaped optical rotation.
Kluyveromyces marxianus L1-1 is cultured on YPD and PDA plates for 3-5 d to form a colony with the diameter of 2.0-5.0 mm, red color, oval shape, smooth and moist surface, regular edge and protrusion. Inoculating into white glutinous rice soup to reduce pH to 3.69. Has fruity and mellow flavor.
The invention inoculates lactobacillus paracasei H4-11 and Kluyveromyces marxianus L1-1 in rice soup raw materials for culture and fermentation, wherein the inoculation amount of lactobacillus paracasei H4-11 is 7.80 multiplied by 108cfu/ml, the inoculation amount of Kluyveromyces marxianus L1-1 is 1.10 × 107cfu/ml; the two fermented rice soup materials have abundant viable bacteria amount of 1.58-1.65 × 10 Lactobacillus paracasei H4-11 for 24 hr8cfu/ml, viable count of Kluyveromyces marxianus L1-1 is 2.05-3.05 × 107cfu/ml; the two strains are proved to have good acid resistance to the rice acid fermentation system.
The invention also provides a rice acid fermentation process for producing L-lactic acid and ethyl acetate, which adopts the raw materials of rice soup fermented by lactobacillus paracasei H4-11 and Kluyveromyces marxianus L1-1.
Preferably, the rice acid fermentation process for producing L-lactic acid and ethyl acetate comprises the following processes:
(1) pretreatment of raw materials: pulverizing glutinous rice or selenium rice at high speed, mixing, sieving with 80-100 mesh sieve twice, and making into rice flour;
(2) boiling: mixing rice flour with water, boiling while stirring;
(3) pasting: putting the boiled rice water into a water bath kettle for gelatinization while stirring; so as to prevent local deterioration of gelatinized liquid caused by nonuniform gelatinization; gelatinizing for 30-40min at 50-60 deg.C;
(4) liquefaction: adding high temperature alpha-amylase 0.5-2.0% of the gelatinized rice soup, and liquefying at 85-95 deg.C for 30-40 min;
(5) blending and sterilizing: adding adjuvants into the liquefied rice soup, and pasteurizing at 95 deg.C for 10-20 min;
(6) inoculating bacteria: cooling rice soup to 25-40 deg.C, inoculating Lactobacillus paracasei H4-11 and Kluyveromyces marxianus L1-1 into rice soup;
(7) fermentation: and (3) fermenting the rice acid inoculated with the strain in a constant-temperature incubator to obtain rice acid soup.
Preferably, in the step (2), the mixture of rice flour and water has a rice flour content of 0.5-8.0%.
Preferably, in the step (5), the auxiliary material is one of honey and lithocarpus polystachyus rehd extract, the addition amount of the honey is 2.0-10.0% of the rice water, and the addition amount of the lithocarpus polystachyus rehd extract is 1.0-5.0% of the rice water.
Preferably, in the step (6), the inoculation amount of the lactobacillus paracasei H4-11 is 6-12%, and the inoculation amount of the kluyveromyces marxianus L1-1 is 0.1-2.0%.
Preferably, in the step (7), the temperature of the constant-temperature incubator is 25-40 ℃, and the fermentation time is 48-96 h.
Preferably, the method further comprises saccharifying the rice soup before sterilizing, wherein saccharifying enzyme with a mass of 0.02-0.04% is added into the liquefied rice soup, the saccharifying time is 1-2h, and the saccharifying temperature is 50-60 ℃.
Preferably, in the step (4), the high-temperature alpha-amylase can be replaced by low-temperature alpha-amylase or medium-temperature alpha-amylase, and the corresponding liquefaction temperature is adjusted to a temperature suitable for enzymolysis of the low-temperature alpha-amylase or the medium-temperature alpha-amylase.
In the step (1), the raw material for preparing the rice acid can also be quinoa, coix seed, brown rice, black glutinous rice, millet and red rice.
The water bath kettle used for gelatinization in the step (3) can be replaced by a heating appliance such as an induction cooker or an electric ceramic cooker.
In the step (6), the inoculation concentrations of Lactobacillus paracasei H4-11 and Kluyveromyces marxianus L1-1 can be adjusted according to the required acidity.
The invention has the beneficial effects that:
the invention adopts lactobacillus paracasei H4-11 and Kluyveromyces marxianus L1-1 as fermentation bacteria of the rice acid; the L-lactic acid produced by the lactobacillus paracasei H4-11 fermented rice acid has strong performance, and simultaneously, a small amount of tartaric acid, acetic acid, succinic acid and the like can be produced, so that the probiotic function of the rice acid is improved; kluyveromyces marxianus L1-1 is used for fermenting rice acid to produce ester fragrance components such as ethyl acetate, ethyl propionate, ethyl acetate, ethanol and the like, and plays an important role in enhancing fragrance of rice acid ferment; and has better oxidation resistance. Selenium rice is used as a raw material of the rice acid, and after fermentation treatment is carried out on lactobacillus paracasei H4-11 and Kluyveromyces marxianus L1-1, the trace element selenium in the selenium rice can be fully maintained in the rice acid. The obtained rice acid has L-lactic acid content of about 3.45g/100g, and lactic acid bacteria content of 1.3-1.4 × 108About cfu/ml; the taste and flavor are good, and the sensory evaluation is high; the antioxidant property is achieved, and the DPPH clearance rate can reach about 46%; the detection on the content of selenium element is 0.05-0.07 mg/kg, which meets the requirement that the content of selenium in the rice processed from the selenium-rich rice in the selenium-rich rice paddy (GB/T22499-2008) is between 0.04 mg/kg and 0.30 mg/kg.
Detailed Description
The technical solution of the present invention is further defined below with reference to the specific embodiments, but the scope of the claims is not limited to the description.
Example 1
A rice acid fermentation process for producing L-lactic acid and ethyl acetate comprises the following steps:
(1) pretreatment of raw materials: pulverizing selenium rice at high speed, mixing, sieving with 80 mesh sieve twice, and making into rice flour; (2) boiling: mixing rice flour with water, wherein the content of the mixed rice flour is 8%, heating and boiling while stirring;
(3) pasting: putting the boiled rice water into a water bath kettle for gelatinization while stirring; so as to prevent local deterioration of gelatinized liquid caused by nonuniform gelatinization; gelatinizing time is 30min, and gelatinizing temperature is 60 ℃;
(4) liquefaction: adding 1.0 wt% of high temperature alpha-amylase into gelatinized rice soup, and liquefying for 30min at 95 deg.C;
(5) blending and sterilizing: adding honey with the mass of 5.0% into the liquefied rice soup, and performing pasteurization at 95 ℃ for 20 minutes;
(6) inoculating bacteria: cooling rice soup to 30 deg.C, mixing 8.0% (0.78 × 10) of Lactobacillus paracasei H4-11 and Kluyveromyces marxianus L1-18CFU/ml) and 1.0% (0.11X 10)7CFU/ml) into the rice soup;
(7) fermentation: and (3) fermenting the rice acid inoculated with the strain in a constant-temperature incubator at 35 ℃ for 96 hours to obtain rice acid soup.
Example 2
A rice acid fermentation process for producing L-lactic acid and ethyl acetate comprises the following steps:
(1) pretreatment of raw materials: pulverizing selenium rice at high speed, mixing, sieving with 80 mesh sieve twice, and making into rice flour;
(2) boiling: mixing rice flour with water, wherein the content of the mixed rice flour is 8%, heating and boiling while stirring;
(3) pasting: putting the boiled rice water into a water bath kettle for gelatinization while stirring; so as to prevent local deterioration of gelatinized liquid caused by nonuniform gelatinization; gelatinizing for 30min at 60 deg.C;
(4) liquefaction: adding 1.0 wt% of high temperature alpha-amylase into gelatinized rice soup, and liquefying for 30min at 95 deg.C;
(5) saccharification: adding 0.02 wt% of saccharifying enzyme into the liquefied rice soup, wherein the saccharifying time is 2h, and the saccharifying temperature is 60 ℃;
(6) blending and sterilizing: adding honey with the mass of 5.0% into the liquefied rice soup, and performing pasteurization at 95 ℃ for 20 minutes;
(7) inoculating bacteria: cooling rice soup to 30 deg.C, mixing 8.0% (strain concentration of 0.78 × 10) of Lactobacillus paracasei H4-11 and Kluyveromyces marxianus L1-18cfu/ml) and 1.0% (strain concentration 0.11X 10)7cfu/ml) into the rice soup;
(8) fermentation: and (3) fermenting the rice acid inoculated with the strain in a constant-temperature incubator at 35 ℃ for 96 hours to obtain rice acid soup.
Example 3
Example 3 differs from example 2 in that not honey but lithocarpus polystachyus rehd extract is added in step (6).
Example 4
A rice acid fermentation process for producing L-lactic acid and ethyl acetate comprises the following steps:
(1) pretreatment of raw materials: pulverizing selenium rice at high speed, mixing, sieving with 100 mesh sieve twice, and making into rice flour;
(2) boiling: mixing rice flour with water, wherein the content of the mixed rice flour is 4%, heating and boiling while stirring;
(3) pasting: putting the boiled rice water into a water bath kettle for gelatinization while stirring; so as to prevent local deterioration of gelatinized liquid caused by nonuniform gelatinization; gelatinizing for 40min at 50 deg.C;
(4) liquefaction: adding high-temperature alpha-amylase accounting for 2.0% of the mass of the gelatinized rice soup into the gelatinized rice soup for liquefaction, wherein the liquefaction time is 40min and the liquefaction temperature is 85 ℃;
(5) saccharification: adding 0.04% of saccharifying enzyme into the liquefied rice soup, wherein the saccharifying time is 1h, and the saccharifying temperature is 60 ℃;
(6) blending and sterilizing: adding 10.0% of honey into the liquefied rice soup, and pasteurizing at 95 deg.C for 20 min;
(7) inoculating bacteria: cooling rice soup to 30 deg.C, adding 6% (strain concentration of 0.48 × 10) of Lactobacillus paracasei H4-11 and Kluyveromyces marxianus L1-18cfu/ml) and 0.1% (strain concentration 0.19X 10)6cfu/ml) into the rice water;
(8) fermentation: and (3) fermenting the rice acid inoculated with the strain in a constant-temperature incubator at 25 ℃ for 96h to obtain rice acid soup.
Example 5
A rice acid fermentation process for producing L-lactic acid and ethyl acetate comprises the following steps:
(1) pretreatment of raw materials: pulverizing selenium rice at high speed, mixing, sieving with 100 mesh sieve twice, and making into rice flour;
(2) boiling: mixing rice flour with water, wherein the content of the mixed rice flour is 4%, heating and boiling while stirring;
(3) pasting: putting the boiled rice water into a water bath kettle for gelatinization while stirring; so as to prevent local deterioration of gelatinized liquid caused by nonuniform gelatinization; gelatinizing for 40min at 50 deg.C;
(4) liquefaction: adding medium temperature alpha-amylase 2.0% of the gelatinized rice soup by mass into the gelatinized rice soup for liquefaction, wherein the liquefaction time is 40min, and the liquefaction temperature is 45 ℃;
(5) saccharification: adding 0.01% of saccharifying enzyme into the liquefied rice soup, wherein the saccharifying time is 1h, and the saccharifying temperature is 60 ℃;
(6) blending and sterilizing: adding honey with the mass of 5.0% into the liquefied rice soup, and performing pasteurization at 95 ℃ for 20 minutes;
(7) inoculating bacteria: cooling rice soup to 30 deg.C, adding 12% of Lactobacillus paracasei H4-11 and Kluyveromyces marxianus L1-1 (strain concentration is 0.93 × 10)8cfu/ml) and 2.0% (strain concentration 0.25X 10)7cfu/ml) into the rice soup;
(8) and (3) fermentation: and (3) fermenting the rice acid inoculated with the strain in a constant-temperature incubator at 40 ℃ for 96 hours to obtain rice acid soup.
Example 6
Example 6 differs from example 2 in that quinoa was used as a raw material, unlike the raw material used.
Control group 1
A rice acid fermentation process comprises the following steps:
(1) pretreatment of raw materials: pulverizing selenium rice at high speed, mixing, sieving with 80 mesh sieve twice, and making into rice flour;
(2) boiling: mixing rice flour with water, wherein the content of the mixed rice flour is 4%, heating and boiling while stirring;
(3) pasting: putting the boiled rice water into a water bath kettle for gelatinization while stirring; so as to prevent local deterioration of gelatinized liquid caused by nonuniform gelatinization; gelatinizing for 30min at 60 deg.C;
(4) liquefaction: adding high-temperature alpha-amylase accounting for 0.5 percent of the mass of the gelatinized rice soup into the gelatinized rice soup for liquefaction, wherein the liquefaction time is 30min and the liquefaction temperature is 95 ℃;
(5) saccharification: adding saccharifying enzyme with the mass of 0.01% into the liquefied rice soup, wherein the saccharifying time is 2h, and the saccharifying temperature is 60 ℃;
(6) and (3) sterilization: pasteurizing the saccharified rice soup at 95 ℃ for 20 minutes;
(7) inoculating bacteria: cooling rice soup to 30 deg.C, adding 4.0% (strain concentration of 0.39 × 10) of Lactobacillus paracasei H4-11 and Kluyveromyces marxianus L1-18cfu/ml) and 0.5% (strain concentration 0.05X 10)7cfu/ml) into the rice soup;
(8) fermentation: and (3) fermenting the rice acid inoculated with the strain in a constant-temperature incubator at 35 ℃ for 48 hours to obtain rice acid soup.
Control group 2
A rice acid fermentation process comprises the following steps:
(1) pretreatment of raw materials: pulverizing selenium rice at high speed, mixing, sieving with 80 mesh sieve twice, and making into rice flour;
(2) boiling: mixing rice flour with water, wherein the content of the mixed rice flour is 4%, heating and boiling while stirring;
(3) pasting: putting the boiled rice water into a water bath kettle for gelatinization while stirring; so as to prevent local deterioration of gelatinized liquid caused by nonuniform gelatinization; gelatinizing for 30min at 60 deg.C;
(4) liquefaction: adding high-temperature alpha-amylase accounting for 0.5 percent of the mass of the gelatinized rice soup into the gelatinized rice soup for liquefaction, wherein the liquefaction time is 30min and the liquefaction temperature is 95 ℃;
(5) saccharification: adding 0.01% of saccharifying enzyme into the liquefied rice soup, wherein the saccharifying time is 2h, and the saccharifying temperature is 60 ℃;
(6) fermentation: and (3) fermenting the saccharified rice soup in a constant-temperature incubator at 35 ℃ for 96 hours to obtain the rice sour soup.
Control group 3
The control group 3 is different from example 2 in that the only inoculated species in step (7) is Lactobacillus paracasei H4-11.
Control group 4
The control group 4 is different from example 2 in that the only inoculated strain in step (7) is Kluyveromyces marxianus L1-1.
Test examples
The rice acid of the examples and the control group is taken for layering comparison: the index of each rice acid sample is measured for 3 times; sensory scoring 10 points were used, and 5 volunteers were invited to taste scoring. The comparative results are shown in the following table.
TABLE 1 analysis of the evaluation index of different Miric acids
As can be seen from the data in Table 1, in example 2, the fermented rice acid ferment prepared from honey has high L-lactic acid yield, contains abundant lactic acid bacteria and yeast, has the highest sensory score, and has good taste substances and ester fragrance components; for example 3, the rice acid prepared by the polystachy has good oxidation resistance, and the DPPH clearance rate reaches 54.51%; the comparative group 1 has slightly inferior performance indexes compared with the above ones due to short fermentation time, and the fermentation time is an important condition affecting the comprehensive evaluation index of the fermented rice acid ferment; the control group 2 is naturally fermented rice acid, and the acid production capacity, the oxidation resistance, the viable bacteria amount and the sensory index of the naturally fermented rice acid are relatively poor, so that the feasibility of the evaluation of the probiotic effect and the flavor index of the rice acid ferment in the invention is more prominent. The control group 3 and the control group 4 adopt one of two strains for fermentation, and compared with the example 2, the content of L-lactic acid and the content of probiotics are lower, which shows that the two strains adopted by the invention have good synergistic effect.
It should be noted that the above examples and test examples are only for further illustration and understanding of the technical solutions of the present invention, and are not to be construed as further limitations of the technical solutions of the present invention, and the invention which does not highlight essential features and significant advances made by those skilled in the art still belongs to the protection scope of the present invention.
Sequence listing
Sequence listing
<110> Guizhou university
<120> rice acid fermentation process for producing L-lactic acid and ethyl acetate and special bacteria thereof
<130>
<160> 2
<170> PatentIn version 3.3
<210> 1
<211> 951
<212>DNA
<213> Artificial sequence
<400> 1 (Lactobacillus paracasei H4-11)
cggcgggggg ggtctataat gcagtcgaac gagttctcgt tgatgatcgg 50
tgcttgcacc gagattcaac atggaacgag tggcggacgg gtgagtaaca 100
cgtgggtaac ctgcccttaa gtgggggata acatttggaa acagatgcta 150
ataccgcata gatccaagaa ccgcatggtt cttggctgaa agatggcgta 200
agctatcgct tttggatgga cccgcggcgt attagctagt tggtgaggta 250
atggctcacc aaggcgatga tacgtagccg aactgagagg ttgatcggcc 300
acattgggac tgagacacgg cccaaactcc tacgggaggc agcagtaggg 350
aatcttccac aatggacgca agtctgatgg agcaacgccg cgtgagtgaa 400
gaaggctttc gggtcgtaaa actctgttgt tggagaagaa tggtcggcag 450
agtaactgtt gtcggcgtga cggtatccaa ccagaaagcc acggctaact 500
acgtgccagc agccgcggta atacgtaggt ggcaagcgtt atccggattt 550
attgggcgta aagcgagcgc aggcggtttt ttaagtctga tgtgaaagcc 600
ctcggcttaa ccgaggaagc gcatcggaaa ctgggaaact tgagtgcaga 650
agaggacagt ggaactccat gtgtagcggt gaaatgcgta gatatatgga 700
agaacaccag tggcgaaggc ggctgtctgg tctgtaactg acgctgaggc 750
tcgaaagcat gggtagcgaa caggattaga taccctggta gtccatgccg 800
taaacgatga atgctaggtg ttggagggtt tccgcccttc agtgccgcag 850
ctaacgcatt aagcattccg cctggggagt acgaccgcaa ggttgaaact 900
caaaggaatt gacgggggcc cgcacaagcg gtggagcatg tggtttaatt c 951
<210> 2
<211> 551
<212>DNA
<213> Artificial sequence
<400> 2 (Kluyveromyces marxianus L1-1)
acacgggtta tgcttagtac ggcgagtgaa gcggcaaaag ctcaaatttg 50
aaatctggcg tcttcgacgt ccgagttgta atttgaagaa ggcgactttg 100
tagctggtcc ttgtctatgt tccttggaac aggacgtcat agagggtgag 150
aatcccgtgt ggcgaggatc ccagttattt gtaaagtgct ttcgacgagt 200
cgagttgttt gggaatgcag ctctaagtgg gtggtaaatt ccatctaaag 250
ctaaatattg gcgagagacc gatagcgaac aagtacagtg atggaaagat 300
gaaaagaact ttgaaaagag agtgaaaaag tacgtgaaat tgttgaaagg 350
gaagggcatt tgatcagaca tggcgtttgc ttcggctttc gctgggccag 400
catcagtttt agcggttgga taaatcctcg ggaatgtggc tctgcttcgg 450
tagagtgtta tagcccgtgg gaatacagcc agctgggact gaggattgcg 500
acttttgtca aggatgctgg cgtaatggtt aaatgccgcc cgtcttgaac c 551
Claims (10)
1. The special bacterium for producing the L-lactic acid and the ethyl acetate through the rice acid fermentation is characterized in that the special bacterium is lactobacillus paracasei (L) (L-lactic acid and ethyl acetate)Lactobacillus paracasei) H4-11 and Kluyveromyces marxianus (K)Kluyveromyces marxianus) L1-1; lactobacillus paracasei: (Lactobacillus paracasei) H4-11, which is preserved in China center for type culture Collection with the preservation number of CCTCC NO: M2021074, and the preservation date of 2021 year, 1 month and 15 days; kluyveromyces marxianus (K.), (Kluyveromyces marxianus) L1-1, which has been preserved in China center for type culture Collection with the preservation number of CCTCC NO: M2021073, and the preservation date of 2021 year, 1 month and 15 days.
2. The use of the special bacteria for rice acid fermentation for producing L-lactic acid and ethyl acetate in the rice acid fermentation process as claimed in claim 1, wherein the rice soup raw material treated by fermentation with Lactobacillus paracasei H4-11 and Kluyveromyces marxianus L1-1 is used.
3. The application of the special rice acid fermentation bacteria for producing L-lactic acid and ethyl acetate in the rice acid fermentation process as claimed in claim 2, characterized by comprising the following steps:
(1) pretreatment of raw materials: pulverizing glutinous rice or selenium rice at high speed, mixing, sieving with 80-100 mesh sieve twice, and making into rice flour;
(2) boiling: mixing rice flour with water, boiling while stirring;
(3) pasting: putting the boiled rice water into a water bath kettle for gelatinization while stirring; so as to prevent local deterioration of gelatinized liquid caused by nonuniform gelatinization; gelatinizing for 30-40min at 50-60 deg.C;
(4) liquefaction: adding high temperature alpha-amylase 0.5-2.0% of the gelatinized rice soup, and liquefying at 85-95 deg.C for 30-40 min;
(5) blending and sterilizing: adding adjuvants into the liquefied rice soup, and pasteurizing at 95 deg.C for 10-20 min;
(6) inoculating bacteria: inoculating Lactobacillus paracasei H4-11 and Kluyveromyces marxianus L1-1 into rice soup when the rice soup is cooled to 25-40 deg.C;
(7) fermentation: and (3) fermenting the rice acid inoculated with the strain in a constant-temperature incubator to obtain rice acid soup.
4. The application of the special rice acid fermentation bacterium for producing L-lactic acid and ethyl acetate in the rice acid fermentation process as claimed in claim 3, wherein in the step (2), the content of rice flour in the mixture of rice flour and water is 0.5-8.0%.
5. The application of the special rice acid fermentation bacteria for producing L-lactic acid and ethyl acetate in the rice acid fermentation process as claimed in claim 3, wherein in the step (3), the gelatinization time is 30-40min, and the gelatinization temperature is 50-60 ℃.
6. The application of the special rice acid fermentation bacteria for producing L-lactic acid and ethyl acetate in the rice acid fermentation process, as claimed in claim 3, wherein in step (5), the auxiliary material is one of honey and lithocarpus polystachyus rehd extract, the addition amount of honey is 2.0-10.0% of the mass of rice water, and the addition amount of lithocarpus polystachyus rehd extract is 1.0-5.0% of the mass of rice water.
7. The application of the special rice acid fermentation bacteria for producing L-lactic acid and ethyl acetate in the rice acid fermentation process in the step (6) is characterized in that the inoculation amount of the lactobacillus paracasei H4-11 is 6-12%, and the inoculation amount of the Kluyveromyces marxianus L1-1 is 0.1-2.0%.
8. The application of the special rice acid fermentation bacteria for producing L-lactic acid and ethyl acetate in the rice acid fermentation process as claimed in claim 3, wherein in the step (7), the temperature of the constant-temperature incubator is 25-40 ℃, and the fermentation time is 48-96 h.
9. The application of the special rice acid fermentation bacteria for producing L-lactic acid and ethyl acetate in the rice acid fermentation process as claimed in claim 3, further comprising saccharifying the liquefied rice soup before sterilization, wherein saccharifying enzyme with the mass of 0.02-0.04% is added into the liquefied rice soup, the saccharifying time is 1-2h, and the saccharifying temperature is 50-60 ℃.
10. The application of the special rice acid fermentation bacteria for producing L-lactic acid and ethyl acetate in the rice acid fermentation process according to claim 3, wherein in the step (4), the high-temperature alpha-amylase can be replaced by low-temperature alpha-amylase or medium-temperature alpha-amylase, and the corresponding liquefaction temperature is adjusted to be suitable for enzymolysis of the low-temperature alpha-amylase or the medium-temperature alpha-amylase.
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