Method for producing lactic acid bacteria formulation and lactic acid bacteria bacteriocin using ethanol draff liquid

A technology of lactic acid bacteria bacteriocin and waste liquid, applied in microorganism-based methods, biochemical equipment and methods, bacteria and other directions, can solve problems such as waste of resources, and achieve the effects of increasing utilization value, extending industrial chain and reducing production costs.

Inactive Publication Date: 2006-07-26

大连春天生物技术有限公司

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AI-Extracted Technical Summary

Problems solved by technology

[0018] Aiming at the above problems, the present invention provides a method for producing lactic acid bacteria preparations and lactic acid bacteria bacteriocins by using alc...

Abstract

This invention discloses a method for preparing lactobacillus preparation and lactobacillus bacteriocino with waste alcohol distiller liquid, and it belongs to bio-preparation prosessing sphere, comprising steps of: get corn alcohol distiller liquid after alcoholic fermentation; solid-liquid separate the alcohol distiller liquid to get clear licquid; obtain expand-culture medium from the clear licquid; expand culture bacterial gradually; obtain fermention medium from the clear licquid; ferment the expand-culture bacterial in fermention medium; lastly separate fermention liquid to get lactobacillus preparation and lactobacillus bacteriocino. which has high added value and is produced by using alcohol distiller liquid, especially corn alcohol distiller liquid. It can prolongs industrial chain of corn processing; can improves corn's value in use; can recycle resource and can depress the manufacturing cost of lactobacillus preparation and lactobacillus bacteriocino.

Application Domain

BacteriaMicroorganism based processes

Technology Topic

BacteriocinFermentation broth +4

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Examples

Experimental program(2)

Example Embodiment

[0050] Example 1 Production of Lactic Acid Bacteria Preparation by Submerged Fermentation of Corn Alcohol Waste Grains Liquid

[0051] 1. Preparation of the filtrate of corn alcohol waste grains

[0052] Collect fresh alcohol waste grains, centrifuge it in a horizontal centrifuge (WL450 produced by Sichuan Jiangbei Machinery Factory), separate solid and liquid, take the separated liquid and filter it through a plate and frame filter, and collect the filtrate, which is the alcohol waste grains filtrate.

[0053] 2. Process conditions of lactic acid bacteria fermentation

[0054] 1. Expansion process of lactic acid bacteria:

[0055] (1) Production strain:

[0056] Lactobacillus gasseri 1-7;

[0057] Lactobacillus crispatus 23-5;

[0058] Enterococcus faecalis Entercococcus faecalis 23-7;

[0059] Freeze-dried, preserved strains, provided by the Institute of Industrial Microbiology, Liaoning Normal University.

[0060] Proportion of liquid culture medium: add 0.5% yeast extract to the filtrate of corn alcohol waste grains; add 2% sucrose; pH 6.0-6.5; 15 pounds and 30 minutes for sterilization.

[0061] (2) Activation of the mother seed in the test tube: The mother seed was respectively inoculated into a test tube containing 10ml of liquid culture medium, incubated at 30°C for 24 hours, and verified by microscopy and bevel coating to confirm the purity.

[0062] (3) Expansion of Erlenmeyer flask: According to the inoculation amount of 5%, the test tube is inserted into a 150ml Erlenmeyer flask containing 100ml of liquid culture medium, and cultivated at 30°C for 24 hours. Then according to the inoculation amount of 5%, the 90ml strains in the 150ml Erlenmeyer flask were inserted into the 5000ml Erlenmeyer flask containing 3000ml liquid culture medium. Incubate at 30°C for 24 hours.

[0063] (4) Seed pot expansion: according to 5% of the inoculation amount, inoculate the Erlenmeyer flask strains into the seed pot; fill with nitrogen to ensure that the pot pressure is 0.3kg/cm 2; Intermittent stirring at 320 rpm; stirring every 2 hours for 5 minutes.

[0064] 2. Fermentation medium and fermentation conditions

[0065] (1) Fermentation medium: add yeast extract 0.5% to the filtrate of corn alcohol spent grains; 2% sucrose; 1% glucose; 0.2% potassium dihydrogen phosphate; pH 5.5-6.0.

[0066] (2) Fermentation conditions: After conventional sterilization, with 5% of the inoculum amount, the bacteria of bacteriococcus for 6 hours and bacillus for 24 hours are connected to the fermentation tank, cultivated at 30°C; intermittently stirred at 350 rpm /Min, stir for 5 minutes every 2 hours, fill with sterile nitrogen, keep the tank pressure 0.3kg/cm 2 , Coccus cultured for 20 hours, Bacillus cultured for 36 hours. At the end of fermentation, the pH was adjusted to 2 with HCl, and it was allowed to stand overnight.

[0067] 3. Preparation of lactic acid bacteria powder

[0068] The fermentation broth left overnight is filtered or centrifuged to collect the bacteria; skimmed milk or casein hydrolysate is added as a protective agent for lactic acid bacteria, and then vacuum freeze-drying or low-temperature vacuum drying is used to make bacterial powder, which is lactic acid bacteria jelly Dry powder.

[0069] The pH of the filtrate is adjusted to 3.5 by adding ammonia water. This filtrate can be used as a diluent of the bacterial powder. Before use, the bacterial powder and the diluent are mixed and filled into a special spray bottle air bag to make a spray formulation. Or separately fill the bacterial powder into a capsule to make a suppository.

Example Embodiment

[0070] Example 2 Production of lactic acid bacteria by submerged fermentation of corn alcohol waste grains

[0071] 1. Preparation of the filtrate of corn alcohol waste grains

[0072] Collect fresh alcohol waste grains, centrifuge it in a horizontal centrifuge (WL450 produced by Sichuan Jiangbei Machinery Factory), separate solid and liquid, take the separated liquid and filter it through a plate and frame filter, and collect the filtrate, which is the alcohol waste grains filtrate.

[0073] 2. Process conditions of lactic acid bacteria fermentation

[0074] 1. Expansion process of lactic acid bacteria:

[0075] (1) Production strain:

[0076] Lactococcus lactis. LN992;

[0077] Lactobacillus brevis;

[0078] Freeze-drying and preservation of bacteria were provided by the Institute of Industrial Microbiology, Liaoning Normal University.

[0079] (2) Liquid culture medium:

[0080] Add 0.5% yeast extract to the filtrate of corn alcohol spent grains; add 0.5% sucrose; PH5.5-6.0 15 pounds 30 minutes sterilization.

[0081] (3) Activation of the mother seed in the test tube: The mother seed was respectively inoculated into a test tube containing 10ml of liquid medium, cultured at 30°C for 24 hours, and the pure one was confirmed by microscopic examination and bevel coating.

[0082] (4) Expansion of Erlenmeyer flask: According to the inoculation amount of 5%, the test tube is inserted into a 150ml Erlenmeyer flask containing 100ml of liquid culture medium, and cultivated at 30°C for 24 hours. Then according to the inoculation amount of 5%, the 90ml strains in the 150ml Erlenmeyer flask were inserted into the 5000ml Erlenmeyer flask containing 3000ml liquid culture medium. Incubate at 30°C for 24 hours.

[0083] (5) Seed pot expansion: according to 5% of the inoculation amount, the Lactobacillus brevis strains in the Erlenmeyer flask are connected to the seed pot; according to 3% of the inoculation amount, the lactobacillus strains in the Erlenmeyer flask are connected to the seed tank. Into the seed pot, expand separately. Nitrogen filling to keep tank pressure 0.3kg/cm2 , Cultivation at 30°C, intermittent stirring at 320 rpm, stirring every 2 hours for 5 minutes, culture of Lactobacillus brevis for 24 hours, and culture of Streptococcus lactis for 6 hours.

[0084] 2. Fermentation medium and fermentation process

[0085] (1) Preparation of fermentation medium: add 0.5% yeast extract to the filtrate of corn alcohol spent grains; 0.5% sucrose; 0.2% potassium dihydrogen phosphate; PH5.5-6.0

[0086] (2) Fermentation process: After conventional sterilization, with 5% of the inoculum amount, the 24 hours of bacteriococcus Lactobacillus brevis was connected to the fermentor, and the bacterium was 6 hours at 3% of the inoculum. Inoculate Streptococcus lactis into another fermenter, culture at 30°C; stir intermittently at 350 rpm, stirring every 2 hours for 5 minutes, fill with nitrogen, and maintain a tank pressure of 0.3 kg/cm 2 , Coccus culture for 20 hours, Bacillus culture for 36 hours, when the fermentation is over, adjust the pH to 2 with HCl, and let it stand overnight.

[0087] 3. Preparation of lactic acid bacteria bacteriocins

[0088] (1) Plate and frame filtration: the PH value of 2, the fermentation broth of Lactobacillus brevis and the fermentation broth of Streptococcus lactis, which are left to stand overnight, are filtered through the plate and frame, and the filtrate is collected separately.

[0089] (2) Ultrafiltration membrane filtration. For the first time, select an ultrafiltration membrane that can retain substances above 20,000 molecular weight, perform ultrafiltration, and collect the filtrate of Streptococcus lactis and the fermentation broth of Lactobacillus brevis respectively. They are respectively subjected to a second membrane filtration with a molecular weight of more than 2000 intercepted, and the concentrated liquid is collected separately, and this concentrated liquid is the material with a molecular weight between 2000 and 20,000.

[0090] (3) Spray drying, take the concentrate of (2) above and add skimmed milk powder or denatured dextrin and NaCl to make the moisture content about 50%. Use centrifugal spray dryer to adjust the inlet air temperature to 230°C, the outlet air temperature to 90°C, the spray volume is 1000kg/h, and the product volume is 500kg/h.

[0091] (4) The two products of crushed packaging can be regarded as fermentation metabolites of lactic acid bacteria, and they also have the same function of broad-spectrum bacteriocins.

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Improve use value
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Method for producing lactic acid bacteria formulation and lactic acid bacteria bacteriocin using ethanol draff liquid

AI-Extracted Technical Summary

Problems solved by technology

Abstract

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Examples

Example Embodiment

Example Embodiment

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Description & Claims & Application Information

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