CN1590534A - Seawater vibrio culture medium - Google Patents
Seawater vibrio culture medium Download PDFInfo
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- CN1590534A CN1590534A CN 200310111862 CN200310111862A CN1590534A CN 1590534 A CN1590534 A CN 1590534A CN 200310111862 CN200310111862 CN 200310111862 CN 200310111862 A CN200310111862 A CN 200310111862A CN 1590534 A CN1590534 A CN 1590534A
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- Prior art keywords
- substratum
- peptone
- seawater
- tryptones
- glucose
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- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- 241000607598 Vibrio Species 0.000 title claims abstract description 18
- 239000013535 sea water Substances 0.000 title claims description 17
- 239000001963 growth medium Substances 0.000 title abstract description 10
- 239000001888 Peptone Substances 0.000 claims abstract description 16
- 108010080698 Peptones Proteins 0.000 claims abstract description 16
- 235000019319 peptone Nutrition 0.000 claims abstract description 16
- 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 claims abstract description 15
- 239000008103 glucose Substances 0.000 claims abstract description 15
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims abstract description 14
- 229940041514 candida albicans extract Drugs 0.000 claims abstract description 11
- 239000012138 yeast extract Substances 0.000 claims abstract description 11
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract description 3
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical compound CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 claims description 25
- 239000007788 liquid Substances 0.000 claims description 24
- 108010046845 tryptones Proteins 0.000 claims description 22
- 206010047400 Vibrio infections Diseases 0.000 claims description 16
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 claims description 11
- 235000015097 nutrients Nutrition 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 3
- 238000012258 culturing Methods 0.000 abstract description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 abstract 2
- 244000068988 Glycine max Species 0.000 abstract 1
- 235000010469 Glycine max Nutrition 0.000 abstract 1
- 239000011780 sodium chloride Substances 0.000 abstract 1
- 239000012137 tryptone Substances 0.000 abstract 1
- 241000894006 Bacteria Species 0.000 description 26
- 230000001580 bacterial effect Effects 0.000 description 11
- 210000000496 pancreas Anatomy 0.000 description 11
- 238000000034 method Methods 0.000 description 10
- 239000002609 medium Substances 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 6
- 241000607594 Vibrio alginolyticus Species 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000012807 shake-flask culturing Methods 0.000 description 4
- 238000011177 media preparation Methods 0.000 description 3
- 239000012533 medium component Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 230000029058 respiratory gaseous exchange Effects 0.000 description 3
- -1 Glucose Dipotassium hydrogen phosphate Sodium-chlor Chemical compound 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 239000008399 tap water Substances 0.000 description 2
- 235000020679 tap water Nutrition 0.000 description 2
- 241000607618 Vibrio harveyi Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000000306 component Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a culture medium for culturing sea vibrio, which is a solution prepared from tryptone, peptone, soybean peptone, yeast extract, sodium chloride, glucose, dipotassium hydrogen phosphate and water according to a certain proportion.
Description
Technical field
The present invention relates to a kind of substratum that the seawater vibrios is cultivated that is used for, be applicable in the biotechnology fields such as seawater vibrios fermentation.
Background technology
Because the nutritional requirement of seawater vibrios is higher, the substratum that is used for the cultivation of seawater vibrios mainly contains pancreas soy peptone substratum (TSB), 2216E etc., culture effect with the former pancreas soy peptone substratum is good, is made up of Tryptones 17g/L, soy peptone 3g/L, glucose 2.5g/L, dipotassium hydrogen phosphate 2.5g/L, sodium-chlor 15g/L.Because the Tryptones consumption as nitrogenous source in this substratum is big, the medium preparation cost is higher relatively, the cost height that mass-producing is cultivated.Therefore, it is very necessary cultivating for the mass-producing that provides a kind of lower-cost substratum to be used for the seawater vibrios.
Peptone is widely used in microbiological culture media, and the source is abundant, and cost is more much lower than Tryptones, replaces Tryptones can reduce the preparation cost of substratum significantly with peptone.
Summary of the invention
The objective of the invention is to improve pancreas soy peptone substratum, reduce the medium preparation cost of seawater vibrios, improve the biological yield of seawater vibrios liquid culture.
The technical scheme of the object of the invention is, partly replaces Tryptones with peptone, adds a small amount of yeast extract paste, improves the component concentration of glucose and dipotassium hydrogen phosphate.Culture medium prescription is: Tryptones 5g/L, peptone 15g/L, soy peptone 3g/L, yeast extract paste 1g/L, glucose 4g/L, dipotassium hydrogen phosphate 5g/L, sodium-chlor 15g/L.
Adopt substratum of the present invention to have following tangible characteristics and beneficial effect:
1, reduce the usage quantity of Tryptones greatly, reduced the medium preparation cost, this culture medium cost is 70% of pancreas soy peptone (TSB) substratum;
2, improved the biological yield that the seawater vibrios is cultivated, the biological yield of this culture medium culturing seawater vibrios is higher more than 30% than pancreas soy peptone substratum (TSB);
Embodiment
By the following examples the present invention is described in further detail.
Embodiment one: the optimization that pancreas soy peptone substratum is formed
1, gets Tryptones 20,17,14 g/L, soy peptone 9,6,3g/L, glucose 7.5,5,2.5g/L, dipotassium hydrogen phosphate 7.5,5,2.5g/L, 4 factors, high, medium and low 3 levels design orthogonal experiment;
2, listed by table 1 (orthogonal experiment requirement), with the liquid nutrient medium of 9 kinds of different concns of ordinary method preparation, all medium pHs 7.5, sodium-chlor is 15g/L, and water is tap water, is sub-packed in the triangular flask of 250mL, every bottle of 50mL liquid nutrient medium, 121 ℃, sterilization in 20 minutes;
3, picking seawater vibrios breathes out the fresh inclined-plane of Vickers bacterium (Vibrio harveyi) bacterial strain (EpGY020401) kind, inoculate in the sterilized pancreas soy peptone liquid nutrient medium (100mL/500mL triangular flask), and 28 ℃, 180r/m, shake-flask culture 16 hours;
4, cultured bacterium liquid is inoculated in 9 kinds of substratum that step 2 prepared with the 5mL/ bottle, 28 ℃, 180r/m, shake-flask culture 24 hours;
5, with the method for step 4 a cultured bacterium liquid with plate count, measure the viable count of bacterium liquid, the result is as shown in table 1, by the extreme difference calculating of various levels, determines that Tryptones 20g/L, soy peptone 3g/L, glucose 5g/L, dipotassium hydrogen phosphate 5g/L, sodium-chlor 15g/L are for more excellent;
Table 1 pancreas soy peptone medium component is optimized orthogonal experiment
| Substratum | Composition (g/L) | Bacterium liquid viable count * 10 8cfu/mL | ||||
| Tryptones | Soy peptone | Glucose | Dipotassium hydrogen phosphate | Sodium-chlor | ||
| ????1 | ??20 | ??9 | ????7.5 | ??7.5 | ??15 | ???37 |
| ????2 | ??20 | ??6 | ????5.0 | ??5.0 | ??15 | ???243 |
| ????3 | ??20 | ??3 | ????2.5 | ??2.5 | ??15 | ???223 |
| ????4 | ??17 | ??6 | ????7.5 | ??2.5 | ??15 | ???0 |
| ????5 | ??17 | ??3 | ????5.0 | ??7.5 | ??15 | ???231 |
| ????6 | ??17 | ??9 | ????2.5 | ??5.0 | ??15 | ???207 |
| ????7 | ??14 | ??3 | ????7.5 | ??5.0 | ??15 | ???0 |
| ????8 | ??14 | ??9 | ????5.0 | ??2.5 | ??15 | ???18 |
| ????9 | ??14 | ??6 | ????2.5 | ??7.5 | ??15 | ???32 |
| High | ??503 | ??262 | ????37 | ??300 | ||
| In | ??408 | ??275 | ????492 | ??450 | ||
| Low | ??50 | ??454 | ????462 | ??241 | ||
| The suitableeest | ??20 | ??3 | ????5.0 | ??5.0 | ??15 | |
0 when being plate count, in dilution 10
7Do not detect bacterium colony on the flat board doubly;
Embodiment two: pancreas soy peptone substratum compares with the culture effect of TSB substratum after improving
1, prepare embodiment one determined substratum and TSB substratum respectively, method is with embodiment one;
2, with two kinds of culture medium inoculated seawater vibrios (breathing out Vickers bacterium EpGY020401 bacterial strain), carry out the comparison of culture effect, cultured bacterium liquid is measured the viable count of bacterium liquid with the method for plate count, and method is with embodiment one;
3, as shown in table 2, the bacterium liquid viable count of the TSB substratum after the improvement can reach 2.88 * 10
10Cfu/mL, and former TSB substratum is 1.61 * 10
10Cfu/mL, biological yield improves 79%;
Table 2TSB substratum with improvement TSB substratum relatively
| Substratum (g/L) | Tryptones | Soy peptone | Glucose | Dipotassium hydrogen phosphate | Sodium-chlor | Bacterium liquid viable count (cfu/mL) |
| TSB | ??17 | 3 | 2.5 | 2.5 | 15 | 1.61×10 10 |
| Improvement TSB | ??20 | 3 | 5.0 | 5.0 | 15 | 2.88×10 10 |
Embodiment three: reduce the raw materials cost experiment of improved culture medium
1, gets Tryptones+peptone 15+5,10+10,5+15g/L (promptly using peptone instead of part Tryptones), soy peptone 3,2,1g/L, glucose 4,3,2g/L, yeast extract paste 3,2,1g/L (adding a small amount of yeast extract paste), 4 factors, high, medium and low 3 levels design orthogonal experiment;
2, listed by table 3-1,3-2 (orthogonal experiment requirement), liquid nutrient medium with 9 kinds of different concns of ordinary method preparation, all substratum dipotassium hydrogen phosphate 5g/L, pH7.5, sodium-chlor are 15g/L, and water is tap water, be sub-packed in the triangular flask of 250mL, every bottle of 50mL liquid nutrient medium, 121 ℃, sterilization in 20 minutes;
3, picking seawater vibrios breathes out Vickers bacteria strain (EpGY020401) and the fresh inclined-plane of vibrio alginolyticus (Vibrioalginoluticus) bacterial strain (SpGY020601 bacterial strain) kind respectively, inoculate in the sterilized pancreas soy peptone liquid nutrient medium (100mL/500mL triangular flask), 28 ℃, 180r/m, shake-flask culture 16 hours;
4, cultured bacterium liquid is inoculated in 9 kinds of substratum that step 2 prepared with the 5mL/ bottle, 28 ℃, 180r/m, shake-flask culture 24 hours;
5, with the method for step 4 a cultured bacterium liquid with plate count, measure the viable count of bacterium liquid, the result is shown in table 3-1,3-2, extreme difference by various levels calculates, comprehensive twice experimental result, Tryptones 5g/L+ peptone 15g/L, soy peptone 3g/L, glucose 4g/L, yeast extract paste 1g/L are for more excellent; Determine that the Optimum of culture medium proportioning is: Tryptones 5g/L, peptone 15g/L, soy peptone 3g/L, yeast extract paste 1g/L, glucose 4g/L, dipotassium hydrogen phosphate 5g/L, sodium-chlor 15g/L, pH7.5.
Table 3-1 medium component is optimized (vibrio alginolyticus SpGY020601 bacterial strain)
| Substratum | Composition (g/L) | Bacterium liquid viable count * 10 8cfu/mL | |||
| Tryptones+peptone | Soy peptone | Yeast extract paste | Glucose | ||
| ??1 | 15+5 | ??3 | ??5 | ??4 | ???216 |
| ??2 | 15+5 | ??2 | ??3 | ??3 | ???170 |
| ??3 | 15+5 | ??1 | ??1 | ??2 | ???201 |
| ??4 | 10+10 | ??2 | ??5 | ??2 | ???168 |
| ??5 | 10+10 | ??1 | ??3 | ??4 | ???198 |
| ??6 | 10+10 | ??3 | ??1 | ??3 | ???174 |
| ??7 | 5+15 | ??1 | ??5 | ??3 | ???173 |
| ??8 | 5+15 | ??3 | ??3 | ??2 | ???237 |
| ??9 | 5+15 | ??2 | ??1 | ??4 | ???225 |
| High | 587 | ??627 | ??557 | ??639 | |
| In | 540 | ??563 | ??605 | ??517 | |
| Low | 635 | ??572 | ??600 | ??606 | |
| The suitableeest | 5+15 | ??3 | ??2 | ??4 | |
Dipotassium hydrogen phosphate 5g/L, sodium-chlor 15g/L, pH7.5.
Table 3-2 medium component is optimized (breathing out Vickers bacterium EpGY020401 bacterial strain)
| Substratum | Composition (g/L) | Bacterium liquid viable count * 10 8cfu/mL | |||
| Tryptones+peptone | Soy peptone | Yeast extract paste | Glucose | ||
| 1 | 15+5 | ??3 | ??5 | ??4 | ?205 |
| 2 | 15+5 | ??2 | ??3 | ??3 | ?207 |
| 3 | 15+5 | ??1 | ??1 | ??2 | ?206 |
| 4 | 10+10 | ??2 | ??5 | ??2 | ?206 |
| 5 | 10+10 | ??1 | ??3 | ??4 | ?211 |
| 6 | 10+10 | ??3 | ??1 | ??3 | ?222 |
| 7 | 5+15 | ??1 | ??5 | ??3 | ?211 |
| 8 | 5+15 | ??3 | ??3 | ??2 | ?286 |
| 9 | 5+15 | ??2 | ??1 | ??4 | ?288 |
| High | 618 | ??713 | ??622 | ??704 | |
| By | 639 | ??701 | ??704 | ??640 | |
| Low | 785 | ??648 | ??716 | ??698 | |
| The suitableeest | 5+15 | ??3 | ??1 | ??4 | |
Dipotassium hydrogen phosphate 5g/L, sodium-chlor 15g/L 15g/L, pH7.5.
Embodiment four: pancreas soy peptone substratum compares with the culture effect of TSB substratum after improving
1, prepare embodiment three determined substratum and TSB substratum respectively, method is with embodiment one;
Optimize substratum: Tryptones 5g/L, peptone 15g/L, soy peptone 3g/L, yeast extract paste 1g/L, glucose 4g/L, dipotassium hydrogen phosphate 5g/L, sodium-chlor 15g/L, pH7.5;
TSB substratum: Tryptones 17g/L, soy peptone 3g/L, glucose 2.5g/L, dipotassium hydrogen phosphate 2.5g/L, sodium-chlor 15g/L, pH7.5;
2, two kinds of culture medium inoculated seawater vibrios (breathing out Vickers bacterium EpGY020401 bacterial strain, vibrio alginolyticus SpGY020601 bacterial strain) carry out the comparison of culture effect, and cultured bacterium liquid is measured the viable count of bacterium liquid with the method for plate count, and method is with embodiment one;
3, as shown in table 4, the bacterium liquid viable bacteria number average of the substratum after the improvement is breathed out Vickers bacterium EpGY020401 bacterial strain and can be reached 2.95 * 10 than TSB substratum height
10Cfu/mL is than TSB substratum (2.28 * 10
10Cfu/mL), biological yield improves 29.4%; Vibrio alginolyticus SpGY020601 bacterial strain can reach 2.71 * 10
10Cfu/mL is than TSB substratum (2.02 * 10
10Cfu/mL), biological yield improves 34.2%;
Table 4 is optimized substratum with the TSB substratum relatively
| Bacterial strain | Substratum | Bacterium liquid viable count (cfu/mL) |
| Breathe out Vickers bacterium EpGY020401 | Optimize substratum | 2.95×10 10 |
| The TSB substratum | 2.28×10 10 | |
| Vibrio alginolyticus SpGY020601 | Optimize substratum | 2.71×10 10 |
| The TSB substratum | 2.02×10 10 |
Claims (4)
1, the liquid nutrient medium of a kind of seawater vibrios, main component consists of: Tryptones, peptone, soy peptone, glucose, dipotassium hydrogen phosphate, sodium-chlor is characterized in that: partly replace Tryptones with peptone.
2, substratum according to claim 1 is characterized in that: Tryptones 5g/L, peptone 15g/L.
3, substratum according to claim 1 is characterized in that: add the quick growth that a small amount of yeast extract paste promotes the seawater vibrios.
4, according to the described substratum of claim 1 to 3, it is characterized in that: glucose content is that 4g/L, dipotassium hydrogen phosphate content are 5g/L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200310111862 CN1590534A (en) | 2003-10-22 | 2003-10-22 | Seawater vibrio culture medium |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200310111862 CN1590534A (en) | 2003-10-22 | 2003-10-22 | Seawater vibrio culture medium |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1590534A true CN1590534A (en) | 2005-03-09 |
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|---|---|---|---|
| CN 200310111862 Pending CN1590534A (en) | 2003-10-22 | 2003-10-22 | Seawater vibrio culture medium |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111676179A (en) * | 2020-07-27 | 2020-09-18 | 北京市食品安全监控和风险评估中心（北京市食品检验所） | Parahemolytic vibrio culture medium, culture method and application |
-
2003
- 2003-10-22 CN CN 200310111862 patent/CN1590534A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111676179A (en) * | 2020-07-27 | 2020-09-18 | 北京市食品安全监控和风险评估中心（北京市食品检验所） | Parahemolytic vibrio culture medium, culture method and application |
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