CN102618527A - Method for recycling production of paenibacillus polymyxa microbial agent from citric acid wastewater - Google Patents
Method for recycling production of paenibacillus polymyxa microbial agent from citric acid wastewater Download PDFInfo
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- CN102618527A CN102618527A CN2011100292788A CN201110029278A CN102618527A CN 102618527 A CN102618527 A CN 102618527A CN 2011100292788 A CN2011100292788 A CN 2011100292788A CN 201110029278 A CN201110029278 A CN 201110029278A CN 102618527 A CN102618527 A CN 102618527A
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Abstract
The invention relates to a method for recycling production of a paenibacillus polymyxa microbial agent from citric acid wastewater, and belongs to the technical field of microbial agents and wastewater recycling. The method adopts main medium raw materials of high-concentration organic wastewater produced in citric acid production and utilizes a liquid-state fermentation method to produce the paenibacillus polymyxa microbial agent. The method comprises the following steps of 1, carrying out pH value adjustment and sterilization of citric acid wastewater, 2, inoculating a triangular flask with paenibacillus polymyxa subjected to slant preservation, and carrying out shaking culture to obtain a liquid strain, 3, inoculating a fermentation tank containing a citric acid wastewater medium with the liquid strain, and carrying out aseptic air culture to obtain liquid fermentation products, and 4, mixing the liquid fermentation products and bio-activated carbon to obtain the paenibacillus polymyxa microbial agent which is solid. The paenibacillus polymyxa microbial agent obtained by the method has the characteristics of low production cost and good environmental benefits, can obviously improves soil environment quality, a crop yield and crop quality, and can control a plurality of diseases of plants. The method provides an approach for effective recycling utilization of a mass of high-concentration wastewater produced in citric acid production, and reduces pollutant discharge.
Description
Technical field:
The invention belongs to microbiobacterial agent and waste water reclaiming technical field.
Background technology:
Hydrocerol A is widely used in industries such as food, medicine, chemical industry as a kind of important chemical material and foodstuff additive.At present, in the nearly various schools of thinkers of state-owned citric acid production factory, about 800,000 tons of total annual capacity is maximum citric acid production state and an export State of the whole world, the proportion of export volume in global Hydrocerol A total trade volume surpasses 60%.The production of Hydrocerol A is to do or corn is a raw material with potato, successively through raw material handle, fermentation, extract, operation such as refining makes product.Citric acid wastewater is mainly from the neutralizer, affination water, cleaning of evaporator water and the filter wash water distribution that ferment and abstraction process produces.Mainly contain starch, protein, glucide, various organic acid, ammonia nitrogen and fat etc. in the waste water, COD concentration is 20000～30000mg/L, belongs to high concentrated organic wastewater.According to statistics, 1 ton of Hydrocerol A of every production produces 7.5～15 tons of high-concentration waste waters.Directly the discharging meeting will cause severe contamination to environment, and it is costly to handle qualified discharge.
Paenibacillus polymyxa (Paenibacillus polymyxa) distributes extensively at occurring in nature, is a kind of important biocontrol microorganisms, and various plants fungi, Micobial Disease are all had the better prevention effect.Wang Zhiwen etc. have reported that a strain Paenibacillus polymyxa Cp-S316 bacterial strain has resistance of wide spectrum to plant-animal fungi, the bacteriosis original, utilize absorption, the desorption performance of macroporous adsorbent resin to the antifungus active substance of many sticking type of spore bacillus Cp-S316 generation, filter out its optimum solution vapor; This active substance has certain tolerance to heat, UV stable to trypsinase, Proteinase K, and is responsive to the chloroform part; Stable to acid, to alkali responsive (Wang Zhiwen, Yuan Shitao; He Liang etc., the extraction of Paenibacillus polymyxa Cp-S316 antifungus active substance and part property research thereof, agricultural environment science journal; 2007,26 (4): 1464-1468).(Granted publication number: report is cultivated CN 1169942C) Paenibacillus polymyxa HY96-2 and fermented supernatant fluid thereof can be used for preventing vegetative bacteria property bacterial wilt to Chinese patent; But also can be used to prevent and treat the plant seedling blight, samping off, and cucumber fusarium axysporum; Tomato wilt; The eggplant blight, root rot, and plant had fairly obvious promotion growth effect.Chinese patent (Granted publication number: disclose Paenibacillus polymyxa CN1186441C) and can be used as a kind of composition decomposing straw in the microbe fertilizer microbial inoculum; Effectively agricultural waste is too much on the one hand in the solution agriculture prodn; Polluted-water and environment, difficult treatment, chemical fertilizer on the other hand; Agricultural chemicals uses too much, the contradiction of soil deterioration.(Granted publication number: disclosing Paenibacilluspolymyxa Jaas cd CN 1325635C) can get in the plant materials under lower concentration Chinese patent, and can field planting in plant materials, breeding and conduction; Plant is had the growth of promotion simultaneously, improve output, the effect of disease resistance particularly has good control effect to soil-borne vascular bundle disease.
The working method of Paenibacillus polymyxa has multiple, as: (publication number: disclose a kind of working method of Paenibacillus polymyxa CN 1844360A), its substratum is bean dregs to Chinese patent.Chinese patent (Granted publication number: use cooked meat medium CN 1186441C) as the Paenibacillus polymyxa fermention medium.The method that with the citric acid wastewater is main medium raw material production Paenibacillus polymyxa microbiobacterial agent is not seen bibliographical information.
Summary of the invention:
The purpose of this invention is to provide a kind of method of can the recycling citric acid wastewater producing the Paenibacillus polymyxa microbiobacterial agent.
Bacterial strain used in the present invention is to imitate Paenibacillus polymyxa (Paenibacillus polymyxa) the EBL-06 bacterial strain that promotes plant-growth, antagonism phytopathogen by the plant height that contriver's separation screening comes out; This bacterial strain is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center on February 26th, 2008; Deposit number is: CGMCC No.2377; Depositary institution address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City; Institute of Microorganism, Academia Sinica, postcode: 100101.
The working method of Paenibacillus polymyxa microbiobacterial agent of the present invention is following:
1, lemon acid waste water pretreatment: with pH to 6.5～7.5 of aqueous sodium hydroxide solution adjusting waste water, in 115～126 ℃ of sterilizations 10～35 minutes.
2, liquid seeds preparation: the Paenibacillus polymyxa of inoculation slant preservation in the triangular flask that the good citric acid wastewater of pre-treatment is housed, in 30～37 ℃, 120～200 rev/mins of shaking culture 24～48 hours obtain liquid seeds.
3, liquid state fermentation: the inoculation volume ratio is 2%～15% liquid seeds in the good citric acid wastewater fermentor tank of pre-treatment is housed, and in 25～35 ℃, logical sterile air was cultivated 12-48 hour; Cultured fermented liquid is through count plate, and viable count reaches 5 * 10
8More than the CFU/ milliliter.
4, fungicide preparation: the biological carbon to cross 30 mesh sieves is a carrier, and according to fermented liquid: biological carbon is 1: 1.2～5 weight ratio, and fermented liquid is added in the biological carbon; Thorough mixing is even, obtains solid-state microbial inoculum, and air seasoning to microbial inoculum water cut is below 30%; Packing leaves shady and cool dry place in.
Embodiment:
The Paenibacillus polymyxa microbiobacterial agent of being produced by technical scheme of the present invention can promote crop to nutrient absorbing and utilization; Reduce the usage quantity of chemical fertilizer; Can effectively prevent and treat multiple fungi plant disease, reduce the usage quantity of agricultural chemicals, thereby improve the farm environment quality; And production cost is low, has effectively utilized waste water, reduces pollutant emission.Embodiment is following:
Embodiment 1: citric acid wastewater is produced the Paenibacillus polymyxa microbiobacterial agent
Lemon acid waste water pretreatment: regulate wastewater pH to 7.0 with aqueous sodium hydroxide solution, in 121 ℃ of sterilizations 20 minutes.
Liquid seeds preparation: the Paenibacillus polymyxa of inoculation slant preservation in the triangular flask that the good citric acid wastewater of pre-treatment is housed, in 32 ℃, 150 rev/mins of shaking culture 30 hours obtain liquid seeds.
Liquid state fermentation: in the good citric acid wastewater fermentor tank of pre-treatment is housed, inoculate the liquid seeds of 5% (volume ratio), in 32 ℃, logical sterile air was cultivated 16 hours; Cultured fermented liquid is through count plate, and viable count reaches 3 * 10
9The CFU/ milliliter.
Fungicide preparation: the biological carbon to cross 30 mesh sieves is a carrier, and according to fermented liquid: biological carbon is 1: 2 a weight ratio, and fermented liquid is added in the biological carbon; Thorough mixing is even, obtains solid-state microbial inoculum, and air seasoning to microbial inoculum water cut is below 30%; Packing leaves shady and cool dry place in.
Embodiment 2: the Paenibacillus polymyxa microbiobacterial agent is applied to rape production
Adopt the experiment of green house of vegetables sub-district, 8 experiment sub-districts that conditionally complete is identical are set, each sub-district 60m
2, 2 sub-districts are 1 group of parallel test, are divided into into 4 groups, the plantation rape, and in the rape seedling phase, do not apply fertilizer and compare in the 1st group of 2 sub-districts; The 2nd group of 2 sub-district are respectively imposing the Paenibacillus polymyxa microbiobacterial agent once according to 50 kilograms/mu consumption behind the rape complete germination He after 3 weeks; Urea is respectively being used once according to 15 kilograms/mu dose behind the rape complete germination He after 3 weeks in the 3rd group of 2 sub-districts; The 4th group 2 doses according to 15 kilograms/mu are respectively being used urea once behind the rape complete germination He after 3 weeks, and after using urea, impose microbiobacterial agent once, irrigation that each sub-district is identical and way to manage according to 50 kilograms/mu dose in 1 day at every turn.Growth of rape 5 all " Invest, Then Investigate " yield and qualities.Use microbiobacterial agent separately and use the yield of rape basically identical of urea separately, all than the test plot raising the output about 60% of not applying fertilizer; Two kinds of comparison photograph sub-district raising the output about 90% that fertilizer is all used; Nitrate salt and nitrite content be not to apply fertilizer and to use the minimum of microbiobacterial agent separately in the rape; Be less than 1/3 of national standard value of limiting the quantity of; Use use again behind the urea microbiobacterial agent also less than national standard limit the quantity of 1/2, nitrate salt and nitrite content all exceed standard seriously in the rape of urea and use separately.Show the existing good effect of increasing production of this microbiobacterial agent, can obviously improve the quality of rape again.
Embodiment 3: the Paenibacillus polymyxa microbiobacterial agent prevents the cucumber samping off
The cucumber samping off is the disease that is caused by fungal infection.Gather the soil in the warmhouse booth of planting cucumber for many years, mix, in the flowerpot of packing into, 2 groups of flowerpots are set, every group 10 basin in 15 ℃～16 ℃ greenhouse, keeps higher levels of humidity and lower intensity of illumination.The 1st group of direct sowing cucumber seed of flowerpot, 30 in every basin is as contrast.Admix weight ratio in the 2nd multicast kind cucumber seeds forward direction soil and be 0.2% Paenibacillus polymyxa microbiobacterial agent, 30 seeds of every then basin sowing.The back statistics that comes up is total, after seedling grows 1 true leaf, and statistics lodging seedling sum before 2 true leaves.The result shows that contrast seedling lodging rate surpasses 50%, and adds the experimental group of microbial inoculum, and the seedling lodging rate is lower than 10%.
Claims (2)
1. method of utilizing citric acid wastewater to produce Paenibacillus polymyxa (Paenibacillus polymyxa) microbiobacterial agent; It is characterized in that; This method is to be that the main medium raw material carries out liquid state fermentation and produces the Paenibacillus polymyxa microbiobacterial agent with the high concentrated organic wastewater that produces in the citric acid production process, comprises the steps:
(1) lemon acid waste water pretreatment: with pH to 6.5～7.5 of aqueous sodium hydroxide solution adjusting waste water, in 115～126 ℃ of sterilizations 10～35 minutes;
(2) liquid seeds preparation: the Paenibacillus polymyxa of inoculation slant preservation in the triangular flask that the good citric acid wastewater of pre-treatment is housed, in 30～37 ℃, 120～200 rev/mins of shaking culture 24～48 hours obtain liquid seeds;
(3) liquid state fermentation: the inoculation volume ratio is 2%～15% liquid seeds in the good citric acid wastewater fermentor tank of pre-treatment is housed, and in 25～35 ℃, logical sterile air was cultivated 12-48 hour; Cultured fermented liquid is through count plate, and viable count reaches 5 * 10
8More than the CFU/ milliliter;
(4) fungicide preparation: the biological carbon to cross 30 mesh sieves is a carrier, and according to fermented liquid: biological carbon is 1: 1.2～5 weight ratio, and fermented liquid is added in the biological carbon; Thorough mixing is even, obtains solid-state microbial inoculum, and air seasoning to microbial inoculum water cut is below 30%; Packing leaves shady and cool dry place in.
2. the Paenibacillus polymyxa described in the claim 1 (Paenibacillus polymyxa); It is characterized in that; This bacterial strain is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center on February 26th, 2008, and deposit number is: CGMCC No.2377.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103931660A (en) * | 2013-01-17 | 2014-07-23 | 无锡中科活力生物技术有限公司 | Method utilizing critic acid wastewater to produce bacillus amyloliquefaciens bio-control inoculant |
| CN104016733A (en) * | 2013-03-03 | 2014-09-03 | 湖北宜都科泰生物工程有限公司 | Multifunctional bio-organic fertilizer produced by utilizing resource recovery of yeast wastewater |
| 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 |
| CN110358701A (en) * | 2019-05-25 | 2019-10-22 | 甘肃省科学院生物研究所 | A kind of Paenibacillus polymyxa microbial inoculum and its production method |
| CN111470889A (en) * | 2020-05-28 | 2020-07-31 | 上海绿乐生物科技有限公司 | Special microbial agent for soil improvement and preparation method thereof |
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| CN101519639A (en) * | 2008-02-27 | 2009-09-02 | 中国科学院生态环境研究中心 | Paenibacillus polymyxa for preventing and treating plant fungal diseases and production method thereof |
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2011
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Patent Citations (3)
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| WO2008091416A2 (en) * | 2006-09-28 | 2008-07-31 | The Ohio State University Research Foundation | Antibiotic antimicrobial agents and methods of their use |
| CN101519639A (en) * | 2008-02-27 | 2009-09-02 | 中国科学院生态环境研究中心 | Paenibacillus polymyxa for preventing and treating plant fungal diseases and production method thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103931660A (en) * | 2013-01-17 | 2014-07-23 | 无锡中科活力生物技术有限公司 | Method utilizing critic acid wastewater to produce bacillus amyloliquefaciens bio-control inoculant |
| 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 |
| CN104016733A (en) * | 2013-03-03 | 2014-09-03 | 湖北宜都科泰生物工程有限公司 | Multifunctional bio-organic fertilizer produced by utilizing resource recovery of yeast wastewater |
| 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 |
| CN110358701A (en) * | 2019-05-25 | 2019-10-22 | 甘肃省科学院生物研究所 | A kind of Paenibacillus polymyxa microbial inoculum and its production method |
| CN110358701B (en) * | 2019-05-25 | 2022-12-16 | 甘肃省科学院生物研究所 | Paenibacillus polymyxa microbial agent and production method thereof |
| CN111470889A (en) * | 2020-05-28 | 2020-07-31 | 上海绿乐生物科技有限公司 | Special microbial agent for soil improvement and preparation method thereof |
| CN111470889B (en) * | 2020-05-28 | 2022-03-08 | 上海绿乐生物科技有限公司 | Special microbial agent for soil improvement and preparation method thereof |
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Application publication date: 20120801 |