CN104388367A - Fermentation medium for phthalic acid dibutyl degrading bacteria - Google Patents

Fermentation medium for phthalic acid dibutyl degrading bacteria Download PDF

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CN104388367A
CN104388367A CN201410798420.9A CN201410798420A CN104388367A CN 104388367 A CN104388367 A CN 104388367A CN 201410798420 A CN201410798420 A CN 201410798420A CN 104388367 A CN104388367 A CN 104388367A
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dibutyl phthalate
fermention medium
degradation bacterium
fermentation medium
phthalate degradation
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CN104388367B (en
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张颖
孙鑫
***
王蕾
陶月
冯程程
段淑伟
常琴
成毅
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Northeast Agricultural University
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Northeast Agricultural University
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    • 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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Abstract

The invention relates to a fermentation medium, in particular to a fermentation medium for phthalic acid dibutyl degrading bacteria, and aims to solve the problems of high production cost, long fermentation period and low biomass of the conventional fermentation medium for phthalic acid dibutyl degrading bacteria. The fermentation medium is prepared from corn flour, soybean flour, K2HPO4.3H2O, MgSO4.7H2O, NaCl, FeCl3.6H2O, CaCl2.2H2O, and distilled water. The biomass of the fermentation medium after being fermented is 2*1010-4*1010CFU/mL, and the production cost is low. The fermentation medium is used for fermenting the phthalic acid dibutyl degrading bacteria.

Description

A kind of fermention medium of dibutyl phthalate degradation bacterium
Technical field
The present invention relates to a kind of fermention medium.
Background technology
Main based on simple plastic greenhouse and membrane covering in industrialized agriculture.Dibutyl phthalate compounds is using advantages such as its plasticity-are strong as softening agent, be widely used in plastic sheeting for farm use, only be connected with hydrogen bond or Van der Waals force between its with polyolefins plastic molecules, very easily escape in environment, and at water body, air, in soil, bed mud, enrichment accumulation, wherein higher with dibutyl phthalate (DBP) content.The carinogenicity of dibutyl phthalate compounds, teratogenecity and mutagenicity make it once after entering soil and water, bring severe contamination to each ecotope.Microorganism remediation technology is regarded as the most effectively repairing means.
Minimal medium and LB substratum is mainly contained at present for the substratum cultivating dibutyl phthalate degradation bacterium.Be mainly that carbon source is utilized with dibutyl phthalate through verification experimental verification dibutyl phthalate degradation bacterium.
Normally used minimal medium is by the K of 1g 2hPO 43H 2the MgSO of O, 0.8g 47H 2the NH of NaCl, 0.5g of O, 1g 4nO 3, 0.075g FeCl 36H 2the CaCl of O, 0.1g 22H 2the distilled water of O and 1000mL and the dibutyl phthalate composition of 100mg/L.LB substratum soaks powder, the peptone of 10g by the yeast of 5g, the distilled water composition of NaCl and 1000mL of 10g.
Minimal medium according to market value preparation 1L needs cost 0.309 yuan, and from the viable bacteria biomass cost that it produces, often producing 1,000 hundred million bacterium colonies needs cost 0.0498 yuan; The LB substratum of preparation 1L needs cost 2.3 yuan, and often producing 1,000 hundred million bacterium colonies needs cost 0.153 yuan.
As can be seen here, although minimal medium advantage of lower cost incubation time is long, general need 2 to 3 days, although and the short relative cost of LB substratum culture cycle is higher, be difficult to use in actual production bacterial strain.
Summary of the invention
The present invention will solve existing dibutyl phthalate degradation bacterium substratum to there is the problem that production cost is high, fermentation period is long and biomass is low, provides a kind of fermention medium of dibutyl phthalate degradation bacterium.
The fermention medium of dibutyl phthalate degradation bacterium of the present invention is by 23.5 ~ 24.5g/L Semen Maydis powder, 37 ~ 38g/L analysis for soybean powder, 0.5 ~ 1.2g/L K 2hPO 43H 2o, 0.2 ~ 0.6g/L MgSO 47H 2o, 0.2 ~ 0.6g/L NaCl, 0.4 ~ 0.5g/LFeCl 36H 2o, 0.05 ~ 0.15g/L CaCl 22H 2o and distilled water composition.
The fermention medium of dibutyl phthalate degradation bacterium of the present invention uses after preparing after 121 DEG C of sterilizing 30min.
After the fermention medium of dibutyl phthalate degradation bacterium connects bacterium 30 ~ 35 DEG C, rotating speed cultivates 24h under being 125r/min.
Semen Maydis powder is corn deep processing product, and analysis for soybean powder is soya bean deep processing product, and the Semen Maydis powder adopted in the present invention and analysis for soybean powder Dou Shi the Northeast are than the agricultural byproducts being easier to the cheapness obtained, and greatly reduce production cost, fermentation period is short is only 24h.
In the present invention, dibutyl phthalate degradation bacterium fermention medium is 2 × 10 through fermentation artifact amount 10~ 4 × 10 10cFU/mL, improves an order of magnitude than minimal medium, is doubled than LB substratum; From production cost, substratum of the present invention often produces 1,000 hundred million bacterium colonies needs cost 0.0131 yuan, reduces 73.69%, reduce 91.45% than LB substratum than minimal medium.
Embodiment
Technical solution of the present invention is not limited to following cited embodiment, also comprises the arbitrary combination between each embodiment.
Embodiment one: the fermention medium of present embodiment dibutyl phthalate degradation bacterium is by 23.5 ~ 24.5g/L Semen Maydis powder, 37 ~ 38g/L analysis for soybean powder, 0.5 ~ 1.2g/L K 2hPO 43H 2o, 0.2 ~ 0.6g/L MgSO 47H 2o, 0.2 ~ 0.6g/LNaCl, 0.4 ~ 0.5g/L FeCl 36H 2o, 0.05 ~ 0.15g/L CaCl 22H 2o and distilled water composition.
Embodiment two: present embodiment and embodiment one unlike: the fermention medium of dibutyl phthalate degradation bacterium is by 23.9g/L Semen Maydis powder, 37.5g/L analysis for soybean powder, 0.8g/L K 2hPO 43H 2o, 0.4g/LMgSO 47H 2o, 0.4g/LNaCl, 0.428g/L FeCl 36H 2o, 0.1g/L CaCl 22H 2o and distilled water composition.
Embodiment three: present embodiment and embodiment one unlike: the fermention medium of dibutyl phthalate degradation bacterium is by 23.6g/L Semen Maydis powder, 37.2g/L analysis for soybean powder, 1g/L K 2hPO 43H 2o, 0.3g/LMgSO 47H 2o, 0.3g/LNaCl, 0.45g/L FeCl 36H 2o, 0.08g/L CaCl 22H 2o and distilled water composition.
Embodiment four: present embodiment and embodiment one unlike: the fermention medium of dibutyl phthalate degradation bacterium is by 24.2g/L Semen Maydis powder, 37.5g/L analysis for soybean powder, 0.7g/L K 2hPO 43H 2o, 0.5g/LMgSO 47H 2o, 0.5g/LNaCl, 0.43g/L FeCl 36H 2o, 0.12g/L CaCl 22H 2o and distilled water composition.
Embodiment five: one of present embodiment and embodiment one to four unlike: the fermention medium of described dibutyl phthalate degradation bacterium uses after preparing after 121 DEG C of sterilizing 30min.Other is identical with one of embodiment one to four.
Embodiment six: after one of present embodiment and embodiment one to five connect bacterium unlike the fermention medium of: described dibutyl phthalate degradation bacterium 30 ~ 35 DEG C, rotating speed cultivates 24h under being the condition of 125r/min.Other is identical with one of embodiment one to five.
For verifying beneficial effect of the present invention, carry out following experiment:
Experiment 1:
Be incorporated with in the 50mL triangular flask of minimal medium by single bacterium colony of dibutyl phthalate degradation bacterium, liquid amount is 30mL, and culture condition is: pH value is 7, and temperature is 35 DEG C, and rotating speed is 125rpm, shaking culture 3 days; By the concentration dilution of the bacteria suspension of acquisition to OD 600value is after 0.909, the by volume inoculum size access of percentage composition 0.2% is equipped with in the 150mL triangular flask of dibutyl phthalate degradation bacterium fermention medium, LB substratum and minimal medium, liquid amount is for being 50mL, fermentation condition is pH value is 7, temperature 35 DEG C, rotating speed 125rpm, cultivates 24 ~ 48h, above substratum all needs 121 DEG C, uses after sterilizing 30min.
Described dibutyl phthalate degradation bacterium is pseudomonas (Pseudomonassp.) DNB-S1, be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, preservation date is on November 6th, 2014, and deposit number is CGMCC No.9526.
Wherein the fermention medium of dibutyl phthalate degradation bacterium is by 23.9g/L Semen Maydis powder, 37.5g/L analysis for soybean powder, 0.8g/LK 2hPO 43H 2o, 0.4g/LMgSO 47H 2o, 0.4g/LNaCl, 0.428g/LFeCl 36H 2o, 0.1g/L CaCl 22H 2o and distilled water composition.
Minimal medium is by the K of 1g 2hPO 43H 2the MgSO of O, 0.8g 47H 2the NH of NaCl, 0.5g of O, 1g 4nO 3, 0.075g FeCl 36H 2the CaCl of O, 0.1g 22H 2the dibutyl phthalate of O, 100mg/L and the distilled water composition of 1000mL.
After fermentation culture 24h, the biomass of fermention medium is 3.3 × 10 10the biomass of CFU/mL, LB substratum is 1.5 × 10 10cFU/mL, after cultivation 48h, the biomass of minimal medium is 6.2 × 10 9cFU/mL.
The biomass of fermention medium improves an order of magnitude than minimal medium, is doubled than LB substratum; From production cost, fermention medium of the present invention often produces 1,000 hundred million bacterium colonies needs cost 0.0131 yuan, reduces 73.69%, reduce 91.45% than LB substratum than minimal medium.

Claims (6)

1. a fermention medium for dibutyl phthalate degradation bacterium, is characterized in that this fermention medium is by 23.5 ~ 24.5g/L Semen Maydis powder, 37 ~ 38g/L analysis for soybean powder, 0.5 ~ 1.2g/L K 2hPO 43H 2o, 0.2 ~ 0.6g/L MgSO 47H 2o, 0.2 ~ 0.6g/L NaCl, 0.4 ~ 0.5g/L FeCl 36H 2o, 0.05 ~ 0.15g/L CaCl 22H 2o and distilled water composition.
2. the fermention medium of a kind of dibutyl phthalate degradation bacterium according to claim 1, is characterized in that the fermention medium of dibutyl phthalate degradation bacterium is by 23.9g/L Semen Maydis powder, 37.5g/L analysis for soybean powder, 0.8g/LK 2hPO 43H 2o, 0.4g/L MgSO 47H 2o, 0.4g/LNaCl, 0.428g/L FeCl 36H 2o, 0.1g/L CaCl 22H 2o and distilled water composition.
3. the fermention medium of a kind of dibutyl phthalate degradation bacterium according to claim 1, is characterized in that the fermention medium of dibutyl phthalate degradation bacterium is by 23.6g/L Semen Maydis powder, 37.2g/L analysis for soybean powder, 1g/L K 2hPO 43H 2o, 0.3g/L MgSO 47H 2o, 0.3g/LNaCl, 0.45g/L FeCl 36H 2o, 0.08g/L CaCl 22H 2o and distilled water composition.
4. the fermention medium of a kind of dibutyl phthalate degradation bacterium according to claim 1, is characterized in that the fermention medium of dibutyl phthalate degradation bacterium is by 24.2g/L Semen Maydis powder, 37.5g/L analysis for soybean powder, 0.7g/LK 2hPO 43H 2o, 0.5g/L MgSO 47H 2o, 0.5g/LNaCl, 0.43g/L FeCl 36H 2o, 0.12g/L CaCl 22H 2o and distilled water composition.
5. the fermention medium of a kind of dibutyl phthalate degradation bacterium according to claim 1, is characterized in that the fermention medium of described dibutyl phthalate degradation bacterium uses after preparing after 121 DEG C of sterilizing 30min.
6. the fermention medium of a kind of dibutyl phthalate degradation bacterium according to claim 1, it is characterized in that the fermention medium of described dibutyl phthalate degradation bacterium connect bacterium after 30 ~ 35 DEG C, rotating speed cultivates 24h under being the condition of 125r/min.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108048371A (en) * 2018-01-29 2018-05-18 东北农业大学 A kind of multifunctional agriculture soil remediation microbial inoculum and its preparation method and application
CN108486009A (en) * 2018-03-26 2018-09-04 东北农业大学 The own ester degradation bacteria of one plant of phthalic acid two (2- ethyls) and its cultural method and application
CN109207400A (en) * 2018-09-26 2019-01-15 东北农业大学 The composite bacteria agent and biodegrading process of phthalic acid ester in a kind of efficient degradation black earth

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CN102250791A (en) * 2011-05-28 2011-11-23 青岛科技大学 Method for producing bacillus subtilis microbial inoculum by taking soybean cake meal as major raw material
CN102391971A (en) * 2011-11-21 2012-03-28 东北农业大学 Fermentation medium of atrazine degrading bacterium DNS32

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Publication number Priority date Publication date Assignee Title
CN102250791A (en) * 2011-05-28 2011-11-23 青岛科技大学 Method for producing bacillus subtilis microbial inoculum by taking soybean cake meal as major raw material
CN102391971A (en) * 2011-11-21 2012-03-28 东北农业大学 Fermentation medium of atrazine degrading bacterium DNS32

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108048371A (en) * 2018-01-29 2018-05-18 东北农业大学 A kind of multifunctional agriculture soil remediation microbial inoculum and its preparation method and application
CN108486009A (en) * 2018-03-26 2018-09-04 东北农业大学 The own ester degradation bacteria of one plant of phthalic acid two (2- ethyls) and its cultural method and application
CN109207400A (en) * 2018-09-26 2019-01-15 东北农业大学 The composite bacteria agent and biodegrading process of phthalic acid ester in a kind of efficient degradation black earth

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