CN105861578A - Lactobacillus fermentation medium and application thereof to lactic acid production - Google Patents
Lactobacillus fermentation medium and application thereof to lactic acid production Download PDFInfo
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Abstract
The invention discloses a lactobacillus fermentation medium and application thereof to lactic acid production, and belongs to the field of microbial fermentation. In the lactobacillus fermentation medium, yeast extract is adopted as a unique organic nitrogen source, meanwhile, a proper amount of amino acid and vitamin is added, glutamic acid, phenylalanine, leucine and isoleucine are preferably selected as the amino acid, and nicotinic acid and biotin are preferably selected as the vitamin. According to the medium, nutrient substances needed for lactobacillus in the growth and metabolism process can be provided, the activity of the lactobacillus can be remarkably improved, bacterium growth can be promoted, the yield of lactic acid can be increased, and the production cost can be reduced.
Description
Technical field
The invention belongs to field of microbial fermentation, in particular to a kind of lactic acid bacteria fermentation culture medium and the application in production of lactic acid thereof.
Background technology
Lactic acid is one of three big organic acid, lactic acid and its esters and derivant be widely used in food, brewage, pharmacy, leather, Medicated cigarette, chemical industry, printing, environmental protection, the industry such as agricultural.At present, the production method of lactic acid has chemical synthesis, fermentation method, enzyme process；Wherein fermentation method has the advantage such as ease for operation, safety and becomes the main method of production of lactic acid, and fermentation method includes lactic acid bacteria fermentation and aspergillus oryzae fermentation etc..Used by numerous lactic acid enterprises owing to lactic acid bacteria fermentation production lactic acid has the advantages such as theoretical yield is high, by-product is less, energy-conservation.But, the nutritional requirement of lactobacter growth is high, in particular for the nutritional labeling that some are special.Traditional EMS culture medium (peptone 1%, Carnis Bovis seu Bubali cream 1%, yeast extract 0.5%, glucose 2%, dibasic ammonium citrate 0.2%, potassium dihydrogen phosphate 0.2%, Tween 80 0.1%, magnesium sulfate 0.058%, manganese sulfate 0.025%, sodium acetate 0.2%) although the nutritional labeling enriched containing peptone, meat extract, yeast extract etc., but wherein key growth factors composition and content are difficult to determine, therefore by-product is more, and lactic acid production is relatively low, and production efficiency is the highest.
Number of patent application is 201010581051.X, patent name discloses a kind of glutamic acid that adds for " a kind of add the method that glutamic acid improves lactic acid bacteria Biomass at low ph conditions " and improves the method for lactic acid bacteria Biomass at low ph conditions and application thereof, but the method being not directed to improve lactic acid production；Number of patent application is 200610118858.3, patent name discloses a kind of culture medium being applicable to cultivate lactic acid bacteria for " culture medium of a kind of new fermenting lactic acid ", containing peptone in described culture medium, Carnis Bovis seu Bubali cream, the multiple nitrogen such as yeast extract source, dipotassium hydrogen phosphate, dibasic ammonium citrate, sodium acetate, magnesium sulfate, manganese sulfate, ferrous sulfate, the inorganic salts such as sodium chloride, also has appropriate element-vitamine compound (such as thiamine, riboflavin, nicotinic acid, pyridoxal hydrochloride, biotin etc.), due to containing three kinds of organic nitrogen sources, medium component is complicated, production cost is higher.
Summary of the invention
In view of the deficiencies in the prior art, in order to improve the growth metabolism ability of lactic acid bacteria and reduce production of lactic acid cost, the invention discloses a kind of lactic acid bacteria fermentation special culture media.Therefore, first purpose of the present invention is to provide a kind of lactic acid bacteria fermentation culture medium.
First purpose of the present invention is achieved in that
A kind of lactic acid bacteria fermentation culture medium, it is characterised in that: in described fermentation medium, contain only yeast extract one organic nitrogen source, possibly together with appropriate aminoacid (glutamic acid, phenylalanine, leucine and isoleucine) and vitamin (nicotinic acid, biotin).
Preferably, described lactic acid bacteria fermentation culture medium contains: glucose 50-150
G/L, yeast extract 10-50 g/L, potassium dihydrogen phosphate 4-8 g/L, sodium citrate 2-5 g/L, tween 80 1-2 ml/L, magnesium sulfate 0.2-0.5 g/L, manganese sulfate 0.05-0.2 g/L, glutamic acid 50-300 mg/L, phenylalanine-3,4-quinone 0-200
Mg/L, leucine 30-200 mg/L, isoleucine 20-100 mg/L, nicotinic acid 4-20
Mg/L, biotin 0.05-0.2 mg/L, pH4.5-7.5.
In an embodiment of the present invention, the pH value adjusting described lactate fermentation culture medium is 5.5-6.5, and most preferred pH value is 6.0.
In an embodiment of the present invention, during the fermentation with sodium hydroxide, ammonia, the pH value of the calcium hydroxide described culture medium of regulation.
Second object of the present invention is the application providing any of the above-described described lactic acid bacteria fermentation culture medium in lactate fermentation.
Second object of the present invention is achieved in that
Under conditions of applicable lactobacter growth metabolism, culture medium described above is cultivated lactic acid bacteria, obtains streptococcus acidi lactici fermented solution, separate in streptococcus acidi lactici fermented solution and obtain lactic acid.
The present invention is compared with prior art, have the significant advantage that culture medium of the present invention can not only provide lactic acid bacteria nutrient substance required during growth metabolism, moreover it is possible to significantly improve lactic bacteria activity, promote thalli growth, improve lactic acid production, reduce production cost.
Detailed description of the invention
The foregoing of the present invention is described in further detail by form more by the following examples, but this should not being interpreted as, the scope of the above-mentioned theme of the present invention is only limitted to below example, and all technology realized based on foregoing of the present invention belong to the scope of the present invention.
Embodiment 1
1, preparation culture medium
(1) according to yeast extract 25 g/L, potassium dihydrogen phosphate 5 g/L, sodium citrate 4 g/L, tween 80 1 ml/L, magnesium sulfate 0.3 g/L, the ratio of manganese sulfate 0.1 g/L, weigh above nutrient media components and add the dissolving of appropriate distilled water；PH6.0,121 DEG C of sterilizing 20 min is adjusted with 1 mol/L sodium hydroxide.
(2) according to final concentration of 120 g/L of glucose, mother solution, 115 DEG C of sterilizing 20 min are prepared.
(3) according to culture medium Glutamic Acid final concentration 5 g/L, prepare mother solution, use aseptic filter membrane filtration sterilization.
(4) step (1), (2), the composition of (3) are proportionally mixed, obtain fermentation medium for lactate fermentation.
2, fermentation
Activated spawn lactobacillus rhamnosus (being purchased from Chinese industrial Culture Collection) is accessed EMS culture medium (peptone 10 g/L, Carnis Bovis seu Bubali cream 10 g/L, yeast extract 5 g/L, glucose 20 g/L, dibasic ammonium citrate 2 g/L, potassium dihydrogen phosphate 2 g/L, Tween 80 1 ml/L, magnesium sulfate 0.58 g/L, manganese sulfate 0.25 g/L, sodium acetate 2 g/L, pH6.2-6.4), at anaerobic jar 37 DEG C, cultivate 20-22h, obtain seed liquor.
By 10% inoculum concentration, seed liquor is inoculated in the 2.7 above-mentioned fermentation medium of L, it is ensured that fermentation tank anaerobic environment, stream adds 5
Mol/L ammonia spirit, maintains pH value about 6.0,40 DEG C, cultivate 40 h under 150 r/min, record lactic acid content 70 in lactic fermentation liquid
G/L, the cost of raw material being computed unit mass lactic acid is 0.0242 yuan/gram lactic acid.
Embodiment 2
1, preparation culture medium
(1) according to peptone 10 g/L, Carnis Bovis seu Bubali cream 10 g/L, yeast extract 5 g/L, dipotassium hydrogen phosphate 1.5 g/L, dibasic ammonium citrate 2.5 g/L, sodium acetate 4 g/L, magnesium sulfate 0.2 g/L, manganese sulfate 0.15 g/L, ferrous sulfate 0.01 g/L, sodium chloride 0.025 g/L, the ratio of tween 80 1.25 ml/L, weigh above nutrient media components and add the dissolving of appropriate distilled water；With 1
Mol/L sodium hydroxide adjusts pH6.0,121 DEG C of sterilizing 20 min.
(2) according to final concentration of 120 g/L of glucose, mother solution, 115 DEG C of sterilizing 20 min are prepared.
(3) final concentration of according to vitamin in culture medium: thiamine 0.045 mg/L, riboflavin 0.015
Mg/L, nicotinic acid 3.5 mg/L, pyridoxal hydrochloride 0.25
Mg/L, biotin 0.09 mg/L, prepare mother solution respectively, use aseptic filter membrane filtration sterilization.
(4) step (1), (2), the composition of (3) are proportionally mixed, obtain fermentation medium for lactate fermentation.
2, fermentation
Activated spawn lactobacillus rhamnosus (being purchased from Chinese industrial Culture Collection) is accessed EMS culture medium (peptone 10 g/L, Carnis Bovis seu Bubali cream 10 g/L, yeast extract 5 g/L, glucose 20 g/L, dibasic ammonium citrate 2 g/L, potassium dihydrogen phosphate 2 g/L, Tween 80 1 ml/L, magnesium sulfate 0.58 g/L, manganese sulfate 0.25 g/L, sodium acetate 2 g/L, pH6.2-6.4), at anaerobic jar 37 DEG C, cultivate 20-22h, obtain seed liquor.
By 10% inoculum concentration, seed liquor is inoculated in the 2.7 above-mentioned fermentation medium of L, it is ensured that fermentation tank anaerobic environment, stream adds 5
Mol/L ammonia spirit, maintains pH value about 6.0,40 DEG C, cultivate 40 h under 150 r/min；Record lactic acid content 88 in lactic fermentation liquid
G/L, the cost of raw material being computed unit mass lactic acid is 0.0197 yuan/gram lactic acid.
Embodiment 3
1, the preparation of culture medium
(1) according to yeast extract 25 g/L, potassium dihydrogen phosphate 5 g/L, sodium citrate 4 g/L, tween 80 1 ml/L, magnesium sulfate 0.3
G/L, the ratio of manganese sulfate 0.1 g/L, weigh above nutrient media components and add the dissolving of appropriate distilled water；With 1
Mol/L sodium hydroxide adjusts pH6.0,121 DEG C of sterilizing 20 min.
(2) according to final concentration of 120 g/L of glucose, mother solution, 115 DEG C of sterilizing 20 min are prepared.
(3) final concentration of according to aminoacid in culture medium and vitamin: glutamic acid 200 mg/L, phenylalanine 100
Mg/L, leucine 100 mg/L, isoleucine 50
Mg/L, prepares mother solution respectively, uses aseptic filter membrane filtration sterilization.
(4) step (1), (2), the composition of (3) are proportionally mixed, obtain fermentation medium for lactate fermentation.
2, fermentation
Activated spawn lactobacillus rhamnosus (being purchased from Chinese industrial Culture Collection) is accessed EMS culture medium (peptone 10 g/L, Carnis Bovis seu Bubali cream 10 g/L, yeast extract 5 g/L, glucose 20 g/L, dibasic ammonium citrate 2 g/L, potassium dihydrogen phosphate 2 g/L, Tween 80 1 ml/L, magnesium sulfate 0.58 g/L, manganese sulfate 0.25 g/L, sodium acetate 2 g/L, pH6.2-6.4), at anaerobic jar 37 DEG C, cultivate 20-22h, obtain seed liquor.
By 10% inoculum concentration, seed liquor is inoculated in the 2.7 above-mentioned fermentation medium of L, it is ensured that fermentation tank anaerobic environment, stream adds 5
Mol/L ammonia spirit, maintains pH value about 6.0,40 DEG C, cultivate 40 h under 150 r/min；Recording lactic acid content 90 g/L in lactic fermentation liquid, the cost of raw material being computed unit mass lactic acid is 0.0179 yuan/gram lactic acid.Ratio is under same culture conditions, and the lactic acid production only adding glutamic acid (see embodiment 1) improves 29%, and cost reduces 26%.
Embodiment 4:
1, the preparation of culture medium
(1) according to yeast extract 25 g/L, potassium dihydrogen phosphate 5 g/L, sodium citrate 4 g/L, tween 80 1 g/L, magnesium sulfate 0.3 g/L, the ratio of manganese sulfate 0.1 g/L, weigh above nutrient media components and add the dissolving of appropriate distilled water；PH6.0,121 DEG C of sterilizing 20 min is adjusted with 1 mol/L sodium hydroxide.
(2) according to final concentration of 120 g/L of glucose, mother solution, 115 DEG C of sterilizing 20 min are prepared.
(3) final concentration of according to aminoacid in culture medium and vitamin: glutamic acid 100 mg/L, phenylalanine 80
Mg/L, leucine 80 mg/L, isoleucine 40
Mg/L, nicotinic acid 8 mg/L, biotin 0.06
Mg/L, prepares mother solution respectively, uses aseptic filter membrane filtration sterilization.
(4) step (1), (2), the composition of (3) are proportionally mixed, obtain fermentation medium for lactate fermentation.
2, fermentation
Activated spawn lactobacillus rhamnosus (being purchased from Chinese industrial Culture Collection) is accessed EMS culture medium (peptone 10 g/L, Carnis Bovis seu Bubali cream 10 g/L, yeast extract 5 g/L, glucose 20 g/L, dibasic ammonium citrate 2 g/L, potassium dihydrogen phosphate 2 g/L, Tween 80 1 g/L, magnesium sulfate 0.58 g/L, manganese sulfate 0.25 g/L, sodium acetate 2 g/L, pH6.2-6.4), at anaerobic jar 37 DEG C, cultivate 20-22h, obtain seed liquor.
By 10% inoculum concentration, seed liquor being inoculated in the 2.7 above-mentioned fermentation medium of L, it is ensured that fermentation tank anaerobic environment, stream adds 5 mol/L ammonia spirits, maintains pH value about 6.0,40 DEG C, cultivate 40 under 150 r/min
H, records lactic acid content 97 g/L in lactic fermentation liquid, and the cost of raw material being computed unit mass lactic acid is 0.0167 yuan/gram lactic acid.Ratio is under same culture conditions, and the lactic acid production only adding glutamic acid (see embodiment 1) improves 38.6%, and cost reduces 31%.Ratio is under same culture conditions, and the lactic acid production adding (see embodiments 2) such as peptone, Carnis Bovis seu Bubali cream, yeast extracts improves 10%, and cost reduces 15%.
Claims (4)
1. a lactic acid bacteria fermentation culture medium and the application in production of lactic acid thereof, it is characterised in that: described fermentation training
Support in base using yeast extract as unique organic nitrogen source, add appropriate aminoacid and vitamin, the preferred paddy of aminoacid simultaneously
Propylhomoserin, phenylalanine, leucine and isoleucine, the preferred nicotinic acid of vitamin, biotin.
A kind of lactic acid bacteria fermentation culture medium the most according to claim 1 and the application in production of lactic acid thereof, it is special
Levy and be in described fermentation medium to contain: glucose 50-150g/L, yeast extract 10-50g/L, potassium dihydrogen phosphate
4-8g/L, sodium citrate 2-5g/L, tween 80 1-2g/L, magnesium sulfate 0.2-0.5g/L, manganese sulfate 0.05-0.2g/L,
Glutamic acid 50-300mg/L, phenylalanine-3,4-quinone 0-200mg/L, leucine 30-200mg/L, isoleucine 20-100
Mg/L, nicotinic acid 4-20mg/L, biotin 0.05-0.2mg/L.
A kind of lactic acid bacteria fermentation culture medium the most according to claim 1 and the application in production of lactic acid thereof, it is special
Levy and be: the pH value of described fermentation medium is 5.5-6.5.
A kind of lactic acid bacteria fermentation culture medium the most according to claim 1 and the application in production of lactic acid thereof, it is special
Levy and be: described fermentation medium can be used for lactate fermentation and produces.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109652491A (en) * | 2019-01-29 | 2019-04-19 | 润盈生物工程（上海）有限公司 | A kind of screening method for lactobacillus acidophilus amino acid requirement |
| CN113969244A (en) * | 2020-07-24 | 2022-01-25 | 中创宏远（北京）环保科技有限公司 | Preparation method and application of bacillus mojavensis |
| WO2023283821A1 (en) * | 2021-07-14 | 2023-01-19 | 万华化学集团股份有限公司 | Preparation method for and application of lactic acid |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1962851A (en) * | 2006-11-29 | 2007-05-16 | 华东理工大学 | Culture medium formulation for fermentation production of lactic acid |
| CN102643875A (en) * | 2012-04-24 | 2012-08-22 | 中国科学院微生物研究所 | Method for producing D-lactic acid by utilizing jerusalem artichoke hydrolysate |
| CN104099263A (en) * | 2013-04-11 | 2014-10-15 | 任发政 | Lactobacillus salivarius, application of Lactobacillus salivarius, extracellular polysaccharide, and preparation method and application of extracellular polysaccharide |
-
2016
- 2016-06-13 CN CN201610410472.3A patent/CN105861578A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1962851A (en) * | 2006-11-29 | 2007-05-16 | 华东理工大学 | Culture medium formulation for fermentation production of lactic acid |
| CN102643875A (en) * | 2012-04-24 | 2012-08-22 | 中国科学院微生物研究所 | Method for producing D-lactic acid by utilizing jerusalem artichoke hydrolysate |
| CN104099263A (en) * | 2013-04-11 | 2014-10-15 | 任发政 | Lactobacillus salivarius, application of Lactobacillus salivarius, extracellular polysaccharide, and preparation method and application of extracellular polysaccharide |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109652491A (en) * | 2019-01-29 | 2019-04-19 | 润盈生物工程（上海）有限公司 | A kind of screening method for lactobacillus acidophilus amino acid requirement |
| CN113969244A (en) * | 2020-07-24 | 2022-01-25 | 中创宏远（北京）环保科技有限公司 | Preparation method and application of bacillus mojavensis |
| WO2023283821A1 (en) * | 2021-07-14 | 2023-01-19 | 万华化学集团股份有限公司 | Preparation method for and application of lactic acid |
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