CN104073452A - Lactic acid bacteria culture medium for producing lactase by fermentation - Google Patents
Lactic acid bacteria culture medium for producing lactase by fermentation Download PDFInfo
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- CN104073452A CN104073452A CN201410264060.4A CN201410264060A CN104073452A CN 104073452 A CN104073452 A CN 104073452A CN 201410264060 A CN201410264060 A CN 201410264060A CN 104073452 A CN104073452 A CN 104073452A
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- CN
- China
- Prior art keywords
- acid bacteria
- substratum
- culture medium
- sumylact
- milk
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title claims abstract description 42
- 235000014655 lactic acid Nutrition 0.000 title claims abstract description 21
- 241000894006 Bacteria Species 0.000 title claims abstract description 20
- 239000001963 growth medium Substances 0.000 title abstract description 9
- 108010005774 beta-Galactosidase Proteins 0.000 title abstract description 8
- 238000000855 fermentation Methods 0.000 title abstract description 7
- 230000004151 fermentation Effects 0.000 title abstract description 7
- 102100026189 Beta-galactosidase Human genes 0.000 title abstract description 4
- 108010059881 Lactase Proteins 0.000 title abstract description 4
- 229940116108 lactase Drugs 0.000 title abstract description 4
- 239000004310 lactic acid Substances 0.000 title abstract description 4
- 239000000843 powder Substances 0.000 claims abstract description 28
- 239000005862 Whey Substances 0.000 claims abstract description 24
- 102000007544 Whey Proteins Human genes 0.000 claims abstract description 24
- 108010046377 Whey Proteins Proteins 0.000 claims abstract description 24
- 239000000306 component Substances 0.000 claims abstract description 10
- 229910017053 inorganic salt Inorganic materials 0.000 claims abstract description 3
- 235000013343 vitamin Nutrition 0.000 claims abstract description 3
- 239000011782 vitamin Substances 0.000 claims abstract description 3
- 229930003231 vitamin Natural products 0.000 claims abstract description 3
- 229940088594 vitamin Drugs 0.000 claims abstract description 3
- 150000003722 vitamin derivatives Chemical class 0.000 claims abstract description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 10
- 238000012262 fermentative production Methods 0.000 claims description 10
- 241000186660 Lactobacillus Species 0.000 claims description 8
- 235000019156 vitamin B Nutrition 0.000 claims description 8
- 239000011720 vitamin B Substances 0.000 claims description 8
- 229940039696 lactobacillus Drugs 0.000 claims description 4
- 239000012533 medium component Substances 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 4
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- LFVGISIMTYGQHF-UHFFFAOYSA-N ammonium dihydrogen phosphate Chemical compound [NH4+].OP(O)([O-])=O LFVGISIMTYGQHF-UHFFFAOYSA-N 0.000 abstract 1
- 229940041514 candida albicans extract Drugs 0.000 abstract 1
- 238000005457 optimization Methods 0.000 abstract 1
- 239000012138 yeast extract Substances 0.000 abstract 1
- 238000002474 experimental method Methods 0.000 description 11
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 8
- 239000008101 lactose Substances 0.000 description 8
- 230000001580 bacterial effect Effects 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 230000035764 nutrition Effects 0.000 description 3
- 230000000050 nutritive effect Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical group N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 241000186604 Lactobacillus reuteri Species 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 108010089934 carbohydrase Proteins 0.000 description 2
- 235000013351 cheese Nutrition 0.000 description 2
- 235000013365 dairy product Nutrition 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 229940001882 lactobacillus reuteri Drugs 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 2
- 239000001393 triammonium citrate Substances 0.000 description 2
- 235000011046 triammonium citrate Nutrition 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229930003270 Vitamin B Chemical group 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- -1 extractum carnis Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a lactic acid bacteria culture medium for producing lactase by fermentation. The culture medium components comprise whey powder, yeast extract powder, NH4H2PO4; and the culture medium further comprises an inorganic salt and vitamin. The culture medium disclosed by the invention can provide good growth for lactic acid bacteria and can produce high-activity lactase; and meanwhile, compared with the culture medium before optimization, the culture medium disclosed by the invention has production cost which is greatly lowered by 50%.
Description
Technical field
The invention belongs to biological technical field, relate to milk-acid bacteria substratum, especially a kind of milk-acid bacteria substratum of fermentative production Sumylact L.
Background technology
Sumylact L, can catalysis lactose hydrolysis be semi-lactosi and glucose, also has the effect of transgalactosidation simultaneously.Sumylact L is widely used, and in dairy industry, Sumylact L is usually used in producing low-lactose dairy product and oligomeric galactose.Adopt the problems such as Sumylact L hydrolyzes lactose can also effectively reduce that lactose crystn, whey in freeze concentration milk-product separated out.In food processing process, add sugariness, solubleness and local flavor that a certain amount of Sumylact L can also improve food.Aspect environment protection, Sumylact L can decompose the lactose in whey, slows down the pollution that higher biological oxygen demand causes water body after wheys draining.
The natural origin of Sumylact L is very extensive, comprises many animals, plant and microorganism.Many milk-acid bacterias can produce Sumylact L, but common milk-acid bacteria galactopoiesis carbohydrase substratum is lactose MRS substratum at present, and its formula is lactose 20g/L, peptone 10g/L, and extractum carnis 8g/L, yeast soaks powder 4g/L, CH
3cOONa5g/L, Triammonium citrate 2g/L, MgSO
40.2g/L, K
2hPO
42g/L, MnSO
40.05g/L, Tween-801mL.This substratum nutritive substance is abundant, but the Sumylact L vigor that bacterial strain produces is not high, and this substratum fermentation costs is too high.
Whey is to produce the by product of the milk-product such as cheese, and the large appointment of every production 1kg cheese produces 9L whey, and annual generation the in the whole world exceedes 100,000,000 6 thousand ten thousand tons of wheys.Whey contains in milk 55% nutritive ingredient, comprise lactose (4.5～6.0%w/v), protein (0.6～1.1%w/v), fat (0.06～0.5%w/v) and mineral salt (0.8～1.0%w/v), and the multiple nutrients material such as lactic acid, citric acid, inorganic nitrogen, vitamin B group.But at present a large amount of wheys are directly discharged as waste water, severe contamination ecotope, and a large amount of nutritive ingredients that contain in whey are wasted.
Summary of the invention
The object of the present invention is to provide a kind of can be cost-saving, be suitable for cultivating milk-acid bacteria, can significantly improve again the milk-acid bacteria substratum of the fermentative production Sumylact L of lactase fermentation level.Substratum provided by the invention is suitable for utilizing lactobacillus (Lactobacilli) fermentative production Sumylact L, not only can meet the basic nutrition demand of lactobacter growth, can also significantly improve Sumylact L vigor.
The present invention is achieved by the following technical solutions:
A milk-acid bacteria substratum for fermentative production Sumylact L, described medium component is included as whey powder, yeast soaks powder, NH
4h
2pO
4.
And, in described substratum, also comprise inorganic salt and VITAMIN.
And described substratum is made up of following component:
(1) whey powder 12.5～17.5g/L;
(2) yeast soaks powder 4.5～6g/L;
(3)NH
4H
2PO
42.5～4.5g/L；
(4) vitamins B
14.5～7.5mg/L
(5)CH
3COONa5g/L；
(6)MgSO
40.2g/L；
(7)MnSO
40.05g/L；
(8)Tween-801ml/L。
And the pH value of described substratum is 6.0～6.5.
And described milk-acid bacteria is selected from lactobacillus (Lactobacilli).
The invention has the beneficial effects as follows:
1, the present invention utilizes whey powder to produce the Major Nutrient material of Sumylact L substratum as lactobacillus-fermented, not only can meet the basic nutrition demand of lactobacter growth, reduces production costs, and also can effectively reduce the pollution of whey to environment.
2, the present invention utilizes whey powder successfully to replace the multiple nutrients material in MRS substratum, and produce the Major Nutrient material of Sumylact L using whey powder as lactobacillus-fermented, basal culture medium not only can meet the basic nutrition demand of lactobacter growth, has significantly improved Sumylact L vigor.
3, the application has removed the expensive raw material such as extractum carnis, peptone, Triammonium citrate in MRS substratum, use instead cheap inorganic nitrogen-sourced, and the substratum that you mention is on MRS basis, add the raw materials such as whey powder, cost is far away higher than our new formula, add VITMAIN B1 simultaneously, improved Sumylact L output.
Brief description of the drawings:
Fig. 1 is whey powder concentration experiment of single factor result of the present invention; Wherein, when whey powder is 15g/L, Sumylact L vigor is the highest;
Fig. 2 is that yeast of the present invention soaks powder concentration experiment of single factor result; Wherein, when yeast soaks powder and is 5g/L, Sumylact L vigor is the highest;
Fig. 3 is NH of the present invention
4h
2pO
4concentration experiment of single factor result; Wherein, NH
4h
2pO
4during for 4g/L, Sumylact L vigor is the highest;
Fig. 4 is vitamins B of the present invention
1concentration experiment of single factor result; Wherein, vitamins B
1during for 5mg/L, Sumylact L vigor is the highest.
Embodiment:
Mode below by embodiment further illustrates the present invention, but does not therefore limit the present invention in described scope of embodiments.
Milk-acid bacteria of the present invention is selected lactobacillus (Lactobacilli).
The present invention is inoculated into (capacity 250mL triangular flask) in 100mL different fermentations substratum by cultured seed liquor with the inoculum size of 1% (v/v), 35 DEG C leave standstill cultivation 24h, measure beta-galactosidase enzymes vigor, and determine the optimum concn of the each component of substratum according to the size of enzyme activity.
1, in substratum, add respectively different concns whey powder (final concentration is respectively 10g/L, 15g/L, 20g/L, 25g/L, 30g/L), all the other compositions and content are constant.35 DEG C leave standstill cultivation 24h, measure Sumylact L vigor.
2, on the basis of above-mentioned experiment, other components unchanged of whey powder substratum, research different concns yeast soaks the impact of powder (final concentration is respectively 2g/L, 3g/L, 4g/L, 5g/L, 6g/L) on bacterial strain product beta-galactosidase enzymes vigor.
3, on the basis of above-mentioned experiment, other components unchanged of whey powder substratum, research different concns NH
4h
2pO
4(final concentration is respectively 2g/L, 3g/L, 4g/L, 5g/L, 6g/L) produces the impact of beta-galactosidase enzymes vigor on bacterial strain.
4, on the basis of above-mentioned experiment, other components unchanged of whey powder substratum, research different concns vitamins B
1(ultimate density is respectively 0mg/L, 5mg/L, 10mg/L, 15mg/L, 20mg/L) produces the impact of beta-galactosidase enzymes vigor on bacterial strain.
5, orthogonal test: on the basis of above-mentioned experiment, in conjunction with above-mentioned experiment of single factor, choose whey powder, NH
4h
2pO
4, yeast soaks powder, vitamins B
1for research object, design 4 factor 4 horizontal quadrature tests (table 1), determine best medium formula.Orthogonal experiments is in table 2.
Table 14 factor 4 horizontal quadrature test design
Table 2 orthogonal experiments
Each component that when concrete fermentation, the milk-acid bacteria galactopoiesis carbohydrase substratum of the present embodiment comprises is: whey powder 17.5g/L, yeast soak powder 6g/L, NH
4h
2pO
44.5g/L, vitamins B
1(VitB1) 7.5mg/L, CH
3cOONa5g/L, MgSO
40.2g/L, MnSO
40.05g/L, Tween-801ml/L.
Its preparation method is as follows: the amount with 1L is configured, and accurately weighs above each component, adds 1L distilled water, fully stirs, and dissolves completely to each component, and it is stand-by after 6.5,115 DEG C of sterilizing 20min regulating Medium's PH Value.
After fermentation, detect testing data:
Adopt above substratum, leave standstill and cultivate lactobacillus reuteri (Lactobacillus reuteri) 24h at 35 DEG C, the enzyme activity that the Sumylact L vigor of its generation produces than initial lactose-MRS substratum is high by 56.6%, and culture medium cost in this example is only 51.3% of lactose-MRS substratum.
Claims (5)
1. a milk-acid bacteria substratum for fermentative production Sumylact L, is characterized in that, described medium component is included as whey powder, yeast soaks powder, NH
4h
2pO
4.
2. the milk-acid bacteria substratum of fermentative production Sumylact L according to claim 1, is characterized in that: in described substratum, also comprise inorganic salt and VITAMIN.
3. the milk-acid bacteria substratum of fermentative production Sumylact L according to claim 2, is characterized in that: described substratum is made up of following component:
(1) whey powder 12.5～17.5g/L;
(2) yeast soaks powder 4.5～6g/L;
(3)NH
4H
2PO
42.5～4.5g/L；
(4) vitamins B
14.5～7.5mg/L
(5)CH
3COONa5g/L；
(6)MgSO
40.2g/L；
(7)MnSO
40.05g/L；
(8)Tween-801ml/L。
4. the milk-acid bacteria substratum of fermentative production Sumylact L according to claim 1, is characterized in that: the pH value of described substratum is 6.0～6.5.
5. the milk-acid bacteria substratum of fermentative production Sumylact L according to claim 1, is characterized in that: described milk-acid bacteria is selected from lactobacillus (Lactobacilli).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410264060.4A CN104073452B (en) | 2014-06-13 | 2014-06-13 | The lactic acid bacteria culturing medium of fermenting and producing Lactose enzyme |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410264060.4A CN104073452B (en) | 2014-06-13 | 2014-06-13 | The lactic acid bacteria culturing medium of fermenting and producing Lactose enzyme |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104073452A true CN104073452A (en) | 2014-10-01 |
| CN104073452B CN104073452B (en) | 2016-09-28 |
Family
ID=51595059
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410264060.4A Expired - Fee Related CN104073452B (en) | 2014-06-13 | 2014-06-13 | The lactic acid bacteria culturing medium of fermenting and producing Lactose enzyme |
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| Country | Link |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112704116A (en) * | 2020-12-10 | 2021-04-27 | 红原牦牛乳业有限责任公司 | Low-lactose A2 yak milk separation and purification system and separation and purification method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1916170A (en) * | 2006-09-08 | 2007-02-21 | 肖雯娟 | Method for producing lactase of neutral liquid |
| CN101358215A (en) * | 2008-09-24 | 2009-02-04 | 东北农业大学 | Method for producing L-lactic acid by fermentation of whey |
-
2014
- 2014-06-13 CN CN201410264060.4A patent/CN104073452B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1916170A (en) * | 2006-09-08 | 2007-02-21 | 肖雯娟 | Method for producing lactase of neutral liquid |
| CN101358215A (en) * | 2008-09-24 | 2009-02-04 | 东北农业大学 | Method for producing L-lactic acid by fermentation of whey |
Non-Patent Citations (2)
| Title |
|---|
| SONIA A DE BALES ET AL: "Production of lactase by Candida pseudotropicalis grown in whey", 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》, vol. 37, no. 6, 30 June 1979 (1979-06-30) * |
| 李艾黎 等: "利用乳清培养基生产乳品发酵剂的研究", 《食品科学》, vol. 27, no. 4, 30 April 2006 (2006-04-30) * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112704116A (en) * | 2020-12-10 | 2021-04-27 | 红原牦牛乳业有限责任公司 | Low-lactose A2 yak milk separation and purification system and separation and purification method |
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|---|---|
| CN104073452B (en) | 2016-09-28 |
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