CN102180710B - Method for producing rhodobacter sphaeroides microbial fertilizer from citric acid waste water - Google Patents
Method for producing rhodobacter sphaeroides microbial fertilizer from citric acid waste water Download PDFInfo
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- CN102180710B CN102180710B CN2011100301217A CN201110030121A CN102180710B CN 102180710 B CN102180710 B CN 102180710B CN 2011100301217 A CN2011100301217 A CN 2011100301217A CN 201110030121 A CN201110030121 A CN 201110030121A CN 102180710 B CN102180710 B CN 102180710B
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- citric acid
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/20—Fertilizers of biological origin, e.g. guano or fertilizers made from animal corpses
Abstract
The invention discloses a method for producing a rhodobacter sphaeroides microbial fertilizer from citric acid waste water, and belongs to the microorganism field. The method comprises the following steps: a. diluting citric acid waste water with clear water till the COD concentration is 12000-15000 mg/L, adjusting the pH to 6.5-7.5 with a sodium hydroxide aqueous solution, sterilizing the solution at 115-126 DEG C for 10-35 min; b. inoculating rhodobacter sphaeroides preserved by a slant into an erlenmeyer flask containing the citric acid waste water, performing shaking culture at 30-37 DEG C under 120-200 rpm for 24-48 hours to obtain liquid seeds; c. inoculating the liquid seeds into a fermentor containing the citric acid waste water with a volume ratio of 2%-20%, culturing the seeds in sterile air at 25-37 DEG C for 12-48 hours to obtain fermentation broth; d. determining the cultured fermentation broth by a spectrophotometer, packaging the broth when the OD660 value is 1.0-1.5 or the bacterial count reaches 109-1010 CFU/ml so as to obtain a finished product. Compared with the prior art, the invention produces a microbial fertilizer from citric acid waste water, and has the characteristics of low production cost, energy saving and environmental protection, good product application effect, etc.
Description
Technical field
The present invention relates to microorganism field, specifically a kind of method of utilizing the red bacterial micro-organism fertilizer of citric acid wastewater production class ball.
Background technology
Citric acid is widely used as a kind of important industrial chemicals and foodstuff additive, its production be take potato do or corn as raw material, successively through raw material process, fermentation, extract, the operation such as refining makes product.Mainly from neutralizer, affination water, cleaning of evaporator water and the filter wash water distribution of fermentation and abstraction process generation, COD concentration mostly is 15000～25000mg/L to citric acid wastewater, belongs to high concentrated organic wastewater.According to statistics, 1 ton of citric acid of every production produces 7.5～15 tons of high-concentration waste waters.Directly the discharging meeting will cause severe contamination to environment, process qualified discharge costly.
The red bacterium of class ball (Rhodobacter sphaeroides) belongs to the α subgroup of purple bacteria group in the bacterium territory, has metabolic way widely, can grow under multiple growth conditions.Have energy metabolism widely mechanism as it, comprising: photosynthesis, inorganic nutrients, aerobic and anaerobic respiration etc.It can also the fixed nitrogen molecule, (http://mmg.uth.tmc.edu/sphaeroides/introduction/introduction.ht ml) such as synthetic important biomolecule active substance tetrapyrrole (tetrapyrroles), chlorophyll (chlorophylls), protoheme (heme), vitamin B12.
A large amount of Research Literatures show, the red bacterium of class ball become a kind of microorganism that the industrialized developing potentiality are arranged very much (Zhao Kai, in shadow, the characteristic of the red bacterium of .1 strain class ball such as Jiang Dan and degraded SD-1750 thereof.Environmental science, 2009,30 (4): 1199-1204).Existing many pieces of articles have been reported and have been utilized the red fermentation using bacteria of class ball to produce 5-ALA (ALA); ALA can promote plant chlorophyll synthetic, strengthens stress resistance of plant, Promoting plant growth.The document of producing Coenzyme Q10 99.0 with the red bacterium of class ball is also a lot; Coenzyme Q10 99.0 is one of coenzyme that has in human body vital role, claim again ubiquinone, it plays an important role in proton displacement and electronics transmission in respiratory chain in vivo, it is the activator of cellular respiration and cellular metabolism, be also important antioxidant and nonspecific immunity strengthening agent, be widely used in clinical medicine, healthcare products and makeup.Also have bibliographical information the red fermentation using bacteria of class ball produce carotenoid, hydrogen etc.We have also found the characteristic of the red bacterium Promoting plant growth of class ball.Huge applications potentiality in view of the red bacterium of class ball, the researchist that houston, u.s.a University of Texas health science center microbiology and molecular genetics are has carried out the red bacterial genomes planning studies of class ball, examining order was completed in 2005, and functional gene research also makes significant progress.
The cultural method of the red bacterium of class ball has multiple, as: the Chinese patent (patent No.: the production method that discloses a red bacterium of kind ball 200510132627.3), its substratum is take lactic acid as main carbon source, add a small amount of inorganic salt, trace element and somatomedin, realized industrial fermentation cultivation scale.(application number: disclose a kind of method of utilizing the red bacterium of class ball to produce Coenzyme Q10 99.0 200810116206.5), its substratum adds a small amount of inorganic salt take glucose, white sugar and corn steep liquor as carbon source to Chinese patent, has realized the high yield of Coenzyme Q10 99.0.And take citric acid wastewater as the main medium raw material, the method for cultivating the red bacterium of class ball has no bibliographical information.
Summary of the invention
Technical assignment of the present invention is for above-mentioned the deficiencies in the prior art, and a kind of method that can the red bacterial micro-organism fertilizer of recycling citric acid wastewater production class ball is provided.
Bacterial strain used in the present invention is the red bacterium of class ball (Rhodobacter sphaeroides) bacterial strain by contriver's separation screening plant height effect Promoting plant growth out, this bacterial strain is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center on April 20th, 2010, deposit number is: CGMCC No.3757, depositary institution address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode: 100101.
Technical assignment of the present invention is realized in the following manner: utilize the method for the red bacterial micro-organism fertilizer of citric acid wastewater production class ball, be characterized in, the method comprises the following steps:
A, lemon acid waste water pretreatment:
First dilute citric acid wastewater to COD concentration 12000～15000mg/L with clear water, then regulate citric acid wastewater pH to 6.5-7.5 with aqueous sodium hydroxide solution, in 115～126 ℃ of sterilizations 10～35 minutes;
B, liquid seeds preparation:
The red bacterium of class ball of inoculation slant preservation in the triangular flask that the good citric acid wastewater of above-mentioned pre-treatment is housed, in 30～37 ℃, 120～200 rev/mins of shaking culture 24～48 hours obtain liquid seeds;
C, liquid state fermentation:
Volume ratio with 2%～20% was cultivated 12～48 hours in 25～37 ℃ of logical sterile airs to inoculation aforesaid liquid seed in the good citric acid wastewater fermentor tank of pre-treatment is housed, and obtained fermented liquid;
D, aftertreatment:
Above-mentioned cultured fermented liquid reaches to 10 through spectrophotometric determination OD660 value 1.0～1.5 or bacterial count
9～10
10Pack after CFU/mL, namely get the red bacterial micro-organism fertilizer of class ball finished product.
Further:
In step a, clear water used refers to tap water or well water.
In step b, the concentration of aqueous sodium hydroxide solution is 12-20N.
The method of the red bacterial micro-organism fertilizer of citric acid wastewater production class ball of utilizing of the present invention compared with prior art has following outstanding beneficial effect:
(1) utilize citric acid wastewater to produce microbial fertilizer, turn waste into wealth, both reduced the fertiliser production cost, saved again the high cost of wastewater treatment, energy-conserving and environment-protective have huge social benefit and economic benefit;
(2) microbial fertilizer of the method preparation can promote crop to absorption and the utilization of nutrient, reduces the usage quantity of chemical fertilizer, thereby improves the farm environment quality, improves the quality of agricultural-food.
Description of drawings
Accompanying drawing 1 is the design sketch of the inventive method contrast experiment that prepared microbial fertilizer is (experimental example two);
Accompanying drawing 2 is design sketchs of the inventive method contrast experiment that prepared microbial fertilizer is (experimental example three).
Embodiment
With concrete Preparation Example, experimental example, the method for the red bacterial micro-organism fertilizer of citric acid wastewater production class ball of utilizing of the present invention is described in detail below.
Preparation Example one:
To take from the Jiangsu joint 3# of biochemistry corporation,Ltd. pipeline citric acid wastewater as raw material.Waste water COD is 18000mg/L.
Production stage:
A, lemon acid waste water pretreatment:
First dilute citric acid wastewater to COD concentration 15000mg/L with tap water, then regulate wastewater pH to 6.5-7.5 with the aqueous sodium hydroxide solution that concentration is 12N, in 121 ℃ of sterilizations 20 minutes.
B, liquid seeds preparation:
The red bacterium of class ball of inoculation slant preservation in the triangular flask that the good citric acid wastewater of above-mentioned pre-treatment is housed, in 32 ℃, 150 rev/mins of shaking culture 30 hours obtain liquid seeds.
C, liquid state fermentation:
To the liquid seeds of inoculating 10% (volume ratio) in the good citric acid wastewater fermentor tank of pre-treatment is housed, cultivated 30 hours in 32 ℃ of logical sterile airs.
D, aftertreatment:
Cultured fermented liquid is through spectrophotometric determination OD660 value 1.22; Bacterial count 5 * 10
9CFU/mL, packing namely gets the red bacterial micro-organism fertilizer of class ball finished product, preserves in lucifuge, shady and cool place.
Preparation Example two:
Take the 1# pipeline lemon acid waste water of taking from the limited chemical industry of Yixing City joint biological chemistry company limited as raw material, waste water COD is 21000mg/L.
Production stage:
A, lemon acid waste water pretreatment:
First dilute citric acid wastewater to COD concentration 14000mg/L with well water, then regulate wastewater pH to 6.5-7.5 with the 18N aqueous sodium hydroxide solution, in 116 ℃ of sterilizations 30 minutes.
B, liquid seeds preparation:
The red bacterium of class ball of inoculation slant preservation in the triangular flask that the good citric acid wastewater of above-mentioned pre-treatment is housed, in 35 ℃, 180 rev/mins of shaking culture 40 hours obtain liquid seeds.
C, liquid state fermentation:
To the liquid seeds of inoculating 15% (volume ratio) in the good citric acid wastewater fermentor tank of pre-treatment is housed, cultivated 36 hours in 35 ℃ of logical sterile airs.
D, aftertreatment:
Cultured fermented liquid is through spectrophotometric determination OD660 value 1.4; Bacterial count to 8 * 10
9The CFU/mL packing namely gets the red bacterial micro-organism fertilizer of class ball finished product, preserves in lucifuge, shady and cool place.
Experimental example one;
Compare experiment take the Preparation Example one red bacterial micro-organism fertilizer of gained class ball and urea as fertilizer.
Adopt green house of vegetables residential quarter experiment, 12 experiment residential quarters that conditionally complete is identical are set, each residential quarter 60m
2, the plantation rape in the Brassica Napus Seedling phase, keeps 3 residential quarters of not applying fertilizer; The red bacterial micro-organism fertilizer of class ball is respectively being used once according to the rate of fertilizer application of 50L/ mu after the rape complete germination He after 3 weeks in 3 residential quarters; Urea is respectively being used once according to the rate of fertilizer application of 15kg/ mu after the rape complete germination He after 3 weeks in 3 residential quarters; 3 rates of fertilizer application according to 15kg/ mu are respectively being used urea once after the rape complete germination He after 3 weeks, and use microbial fertilizer once according to the rate of fertilizer application of 50L/ mu, the irrigation that each residential quarter is identical and way to manage in 1 day after using urea at every turn.The all " Invest, Then Investigate " yield and qualities of growth of rape 5.Use separately microbial fertilizer and use separately the yield of rape of urea basically identical, all than volume increase 50% left and right, residential quarter of not applying fertilizer, the ratio that two kinds of fertilizer is all used is only used volume increase 30% left and right of a kind, use separately in the rape of urea nitrate and nitrite content and exceed 50% and more than 20 times than the rape that do not apply fertilizer respectively, and use separately in the rape that microbial fertilizer and two kinds of fertilizer all uses in nitrate and nitrite content and the rape that do not apply fertilizer quite.Show the existing good effect of increasing production of this microbial fertilizer, can obviously improve again the quality of rape.
Experimental example two:
The fertile radish growth experiment that is used for of the red bacterial micro-organism of class ball of embodiment one preparation
Experiment place: Yixing City too magnificent farm
Test period: 2010.09.08--11.20
Edaphic condition: the protection of viscosity yellowish soil
The fertilising background: every mu applies farmyard manure 2000kg
Experimental technique: establish altogether 6 and test the residential quarter; 3 check plots, 20 squares of each residential quarters.
Each residential quarter of experimental group together is manured into soil with the Sa Teli Nitrogen, Phosphorus and Potassium 2kg+ class ball fertile 500ml of red bacterial micro-organism (being watered 50 times), draws evenly rear sowing of hoe.Evenly rear sowing of hoe is drawn with composite fertilizer and the water of isodose in each residential quarter of control group.
The calculating output of weighing after November 20 results, result is as follows:
The check plot mean yield is 98.4kg, the experimental group mean yield is 132.2kg, the average specific control group improves output 34.9%, and nitrate, nitrite content do not descend 1.8 times and 19.6 times than the contrast component, and obviously improve mouthfeel and the visual appearance of radish, show that this fertilizer has remarkable volume increase and improves the effect of radish quality.
Contrast experiment's effect as shown in Figure 1.
Embodiment three:
The fertile three-coloured amaranth growth experiment that is used for of the red bacterial micro-organism of class ball of embodiment two preparations
Experiment place: Wan Shi town, Jiangsu city
Experimental period: on April 4th, 2010---May 30
Soil Background: the construction site backfill soil, belong to yellowish-brown clayed soil, establish altogether 6 residential quarters, 20 squares of each residential quarters, wherein three experiments, three contrasts.
Control group and experimental group all, are sowed April 4 as base manure with Kingsoft board composite fertilizer (1kg is used in each residential quarter);
Carried out spraying test May 6; Each residential quarter of experimental group is watered 50 times with the fertile 50ml of the red bacterial micro-organism of class ball and sprays; The check plot was cut dish with pasting is weighed with the fresh water spraying of same amount in 25 days afterwards.
Result: dish 41.6kg is on average received in experimental group each residential quarter, average each the residential quarter 29.7kg of control group, volume increase 40% has significant effect of increasing production, its nitrate, nitrite content experimental group descend 1.5 times and 16 times than control group, have obviously improved the quality of three-coloured amaranth.
Contrast experiment's effect as shown in Figure 2.
Claims (2)
1. utilize the method for the red bacterial micro-organism fertilizer of citric acid wastewater production class ball, it is characterized in that the method comprises the following steps:
A, lemon acid waste water pretreatment:
First dilute citric acid wastewater to COD concentration 12000～15000mg/L with clear water, then regulate citric acid wastewater pH to 6.5-7.5 with aqueous sodium hydroxide solution, in 115～126 ℃ of sterilizations 10～35 minutes;
B, liquid seeds preparation:
The red bacterium of class ball of inoculation slant preservation in the triangular flask that the good citric acid wastewater of above-mentioned pre-treatment is housed, in 30～37 ℃, 120～200 rev/mins of shaking culture 24～48 hours obtain liquid seeds;
C, liquid state fermentation:
Volume ratio with 2%～20% was cultivated 12～48 hours in 25～37 ℃ of logical sterile airs to inoculation aforesaid liquid seed in the good citric acid wastewater fermentor tank of pre-treatment is housed, and obtained fermented liquid;
D, aftertreatment:
Above-mentioned cultured fermented liquid reaches to 10 through spectrophotometric determination OD660 value 1.0～1.5 or bacterial count
9～10
10Pack after CFU/mL, namely get the red bacterial micro-organism fertilizer of class ball finished product.
2. the method for utilizing the red bacterial micro-organism fertilizer of citric acid wastewater production class ball according to claim 1, is characterized in that: in step a, utilize the aqueous sodium hydroxide solution of 12-20N to regulate citric acid wastewater pH to 6.5-7.5.
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103931660B (en) * | 2013-01-17 | 2018-08-24 | 无锡中科活力生物技术有限公司 | Bacillus amyloliquefaciens biocontrol agent is produced using citric acid wastewater |
| CN104326792A (en) * | 2014-03-18 | 2015-02-04 | 新疆天业农业高新技术有限公司 | Special liquid organic-inorganic mixed fertilizer for rice under the mulched drip irrigation, and preparation method thereof |
| CN104531792B (en) * | 2014-12-12 | 2018-06-19 | 哈尔滨工业大学 | A kind of method for improving purple nonsulfur bacteria thalline 5-ALA yield |
| CN110105136A (en) * | 2019-05-08 | 2019-08-09 | 石河子大学 | A kind of citric acid microbial bacterial agent and its preparation method and application |
| CN110577419A (en) * | 2019-10-09 | 2019-12-17 | 徐州生物工程职业技术学院 | method for preparing burdock bio-organic fertilizer from waste of citric acid production from straws |
| CN114317357B (en) * | 2021-12-31 | 2023-12-29 | 绿奥环保科技（上海）有限公司 | Rhodobacter sphaeroides and application thereof |
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| CN101643292A (en) * | 2008-08-04 | 2010-02-10 | 唐晓明 | Method for treating waste water from citric acid-production |
| CN101787086A (en) * | 2010-02-11 | 2010-07-28 | 南京工业大学 | Device and method for hydrolyzing plant hemicellulose by fermented citric acid wastewater |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1837347A (en) * | 2005-03-25 | 2006-09-27 | 甘肃省科学院生物研究所 | Method for preparing vinegar by utilizing sewage and waste residue of starch and citrate |
| CN101333509A (en) * | 2008-07-07 | 2008-12-31 | 天辰神舟实业有限公司 | Rhodobacter sphaeroides mutant of coenzyme Q10 and culturing method |
| CN101643292A (en) * | 2008-08-04 | 2010-02-10 | 唐晓明 | Method for treating waste water from citric acid-production |
| CN101787086A (en) * | 2010-02-11 | 2010-07-28 | 南京工业大学 | Device and method for hydrolyzing plant hemicellulose by fermented citric acid wastewater |
Non-Patent Citations (2)
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| 应用光合细菌处理柠檬酸废水的试验研究;陈宁等;《中国水污染防治技术装备论文集》;19990630;第18-20,34页 * |
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