CN108300673A - A kind of recoverying and utilizing method of Paenibacillus polymyxa fermented waste fluid - Google Patents
A kind of recoverying and utilizing method of Paenibacillus polymyxa fermented waste fluid Download PDFInfo
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- CN108300673A CN108300673A CN201711454635.9A CN201711454635A CN108300673A CN 108300673 A CN108300673 A CN 108300673A CN 201711454635 A CN201711454635 A CN 201711454635A CN 108300673 A CN108300673 A CN 108300673A
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Abstract
The invention discloses a kind of recoverying and utilizing methods of Paenibacillus polymyxa fermented waste fluid, and this approach includes the following steps：(1) it is added 0.05~0.1% chitosan by weight in the Paenibacillus polymyxa fermented waste fluid obtained to centrifugation, centrifuging and taking supernatant again after stirring flocculation；(2) 0.001~0.005% amylase is added by weight into supernatant, digests 15~60 minutes；(3) supernatant after enzymolysis is crossed into cation exchange column, obtained product is added in fermentation of bacillus culture medium and is used as fermentation synergist, which can reduce fermentation of bacillus culture medium dosage, improve spore forming rate.
Description
Technical field
The invention belongs to field of microbial fermentation.More particularly to the recycling side of Paenibacillus polymyxa fermented waste fluid
Method, the invention further relates to a kind of fermentation of bacillus synergist and fermentation synergisting methods.
Background technology
Paenibacillus polymyxa can generate various active metabolite during carrying out liquid fermentation, such as polypeptide, fat
Peptide, polysaccharide, albumen, nucleic acid, pyrazine, aldehydes matter, and most of active material properties are stablized.It ferments in Paenibacillus polymyxa
Last handling process in, heavy bacterium mud is usually collected by centrifugation, carries out spray drying and prepares Paenibacillus polymyxa bacterium powder, and
The fermented liquid supernatant of centrifugation puts into sewage disposal system usually as waste liquid, and this processing method not only wastes big in supernatant
The ingredient of amount, and serious pollution is caused to environment, it is unfavorable for energy-saving and emission-reduction.
Invention content
The purpose of the present invention is the disadvantages described above for the prior art, a kind of Paenibacillus polymyxa fermented waste fluid is provided
Recoverying and utilizing method, it is a further object to provide a kind of fermentation of bacillus synergist, which is added to
In the fermentation medium of bacillus, culture medium dosage can be reduced, improve spore forming rate.
Above-mentioned purpose is achieved through the following technical solutions：
A kind of recoverying and utilizing method of Paenibacillus polymyxa fermented waste fluid, includes the following steps：
(1) 0.05~0.1% chitosan is added by weight in the Paenibacillus polymyxa fermented waste fluid obtained to centrifugation, stirs
Mix centrifuging and taking supernatant again after flocculation；
(2) 0.001~0.005% amylase is added by weight into supernatant, digests 15~60 minutes；
(3) supernatant after enzymolysis is crossed into cation exchange column, obtained product is added to fermentation of bacillus culture
It is used as fermentation synergist in base.
A kind of fermentation of bacillus synergist, the fermentation of bacillus synergist are Paenibacillus polymyxa fermented waste fluids
Recycling product, the recoverying and utilizing method includes the following steps：
(1) 0.05~0.1% chitosan is added by weight in the Paenibacillus polymyxa fermented waste fluid obtained to centrifugation, stirs
Mix centrifuging and taking supernatant again after flocculation；
(2) 0.001~0.005% amylase is added by weight into supernatant, digests 15~60 minutes；
(3) supernatant after enzymolysis is crossed into cation exchange column, obtained product is added to fermentation of bacillus culture
It is used as fermentation synergist in base.
Preferably, the bacillus is bacillus subtilis or bacillus amyloliquefaciens or bacillus pumilus.
A kind of fermentation of bacillus synergisting method：The fermentation synergist is added in fermentation of bacillus culture medium.
Preferably, the bacillus is bacillus subtilis or bacillus amyloliquefaciens or bacillus pumilus.
It is further preferred that the fermentation synergist accounts for the 40-60% of fermentation of bacillus culture medium total weight.
The beneficial effects of the invention are as follows：
The present invention removes the macromolecule impurity in fermented waste fluid by flocculation first, then digests macromolecules starch at small
Molecule exchanges finally by cation and removes the substance for influencing fermentation of bacillus, finally realizes Paenibacillus polymyxa hair
The recycling of ferment waste liquid,
Be conducive to fermentation of bacillus shape rich in total nitrogen, ammonia nitrogen, total reducing sugar, reduced sugar, Soluble phosphorus etc. are a variety of in recovery product
At the nutriment of gemma, which can be used for bacillus subtilis, bacillus amyloliquefaciens, bacillus pumilus
In the fermented and cultured of equal microorganisms, to reduce the dosage of fermentation medium, increases spore forming rate, be a kind of excellent micro- life
Object fermentation synergist.
The present invention realizes the secondary recovery utilization to Paenibacillus polymyxa fermented waste fluid, reduces the row of pollutant
It puts, realizes the recycling of resource, there is larger economic and social benefit.
Specific implementation mode
It is that technical scheme of the present invention is expanded on further in conjunction with the embodiments below.It should be noted that of the invention
Embodiment for the present invention only have illustration, without restriction effect.Involved various experimental implementations, example in the present invention
Such as Paenibacillus polymyxa fermentation culture method is the ordinary skill in the art, the part being not particularly illustrated in text, this
The those of ordinary skill in field be referred to the present patent application day before various common tool books, scientific and technical literature or relevant theory
Bright book, handbook etc. are practiced.
Embodiment 1
After Paenibacillus polymyxa fermented and cultured, heavy bacterium mud is collected by centrifugation, it is more to carry out spray drying preparation
Viscous series bacillus bacterium powder, the fermented liquid supernatant of centrifugation are recycled processing according to the following steps：
(1) 0.05% chitosan, stirring 10 is added in the Paenibacillus polymyxa fermented waste fluid obtained to centrifugation by weight
Centrifuging and taking supernatant again after minute；
(2) 0.001% amylase is added by weight into supernatant, digests 60 minutes；
(3) supernatant after enzymolysis is crossed after highly acid polystyrene cation exchange column and is added to fermentation of bacillus training
It supports in base.
Embodiment 2
(1) 0.1% chitosan is added in the Paenibacillus polymyxa fermented waste fluid obtained to centrifugation, after stirring 1 minute again
Secondary centrifuging and taking supernatant；
(2) 0.005% amylase is added into supernatant, digests 15 minutes；
(3) supernatant after enzymolysis is crossed after highly acid polystyrene cation exchange column and is added to fermentation of bacillus training
It supports in base.
Embodiment 3
(1) 0.08% chitosan is added in the Paenibacillus polymyxa fermented waste fluid obtained to centrifugation, stirs after five minutes again
Secondary centrifuging and taking supernatant；
(2) 0.003% amylase is added into supernatant, digests 30 minutes；
(3) supernatant after enzymolysis is crossed after highly acid polystyrene cation exchange column and is added to fermentation of bacillus training
It supports in base.
Physical and chemical index analysis is carried out to the treatment fluid in embodiment 1-3, wherein total nitrogen is detected using triumphant formula nitriding, amino
Nitrogen is detected using formol titration, and total reducing sugar uses anthrone measuring method, reduced sugar that Iron mud, Soluble phosphorus is used to use P-Mo blue
Colorimetric method, protein degree refer to the ratio of ammonia nitrogen and total nitrogen.Testing result see the table below.
Table 1 recycles the physical and chemical index analysis of product
| Embodiment | Total nitrogen | Ammonia nitrogen | Total reducing sugar | Reduced sugar | Soluble phosphorus | Degree of hydrolysis |
| 1 | 4.72% | 1.12% | 4.33% | 1.08% | 0.05% | 23.72% |
| 2 | 5.01% | 2.09% | 4.26% | 1.75% | 0.06% | 41.72% |
| 3 | 4.25% | 1.62% | 3.98% | 0.94% | 0.06% | 38.12% |
Embodiment 4 recycles application of the product in bacillus amyloliquefaciens fermentation
Comparative example：
Fermentation tank culture medium (weight percent)：Starch 2.5%, dregs of beans 2%, yeast extract 0.5%, corn steep liquor 1%,
MgSO40.005%, MnSO40.005%, KH2PO40.1%, polyethers 0.01%, water are surplus.
Fermentation parameter：PH is adjusted between 7.0-7.2,0.11-0.12Mpa sterilizing 30min, bacillus amyloliquefaciens seed
Culture solution inoculum concentration 8%, the ventilatory capacity of 1VVM.
Test example：
Fermentation tank culture medium：Starch, dregs of beans, yeast extract, corn steep liquor, MgSO4、MnSO4、KH2PO4, polyethers be pair
The 80% of ratio dosage, Paenibacillus polymyxa fermented waste fluid recycle product 50%, and water is surplus.
Fermentation parameter is identical as comparative example.
Fermentation respectively takes two tank zymotic fluids to do cell count and gemma counting to 12h, 16h, 20h, calculates spore forming rate, knot
Fruit see the table below.
Table 2 recycles influence of the product to bacillus amyloliquefaciens fermentation spore forming rate
As can be seen from the above results, the bud that Paenibacillus polymyxa fermented waste fluid recycles product is added in culture medium
Spore formation rate is significantly higher than control, illustrates that the recovery product is a kind of bacillus amyloliquefaciens fermentation synergist.
Embodiment 5 recycles application of the product in fermentation of bacillus subtilis
Comparative example：
Fermentation tank culture medium (weight percent)：Starch 1%, corn flour 0.5%, glucose sugar 2%, dregs of beans 2.7%,
CaCl20.005%, MgSO40.005%, MnSO40.005%, KH2PO40.1%, polyethers 0.01%, water are surplus.
Fermentation parameter：Between adjusting pH7.0-7.2,0.07-0.08MPa sterilizing 30min, bacillus subtilis seed culture fluid
Inoculum concentration 10%, 30 DEG C, the ventilatory capacity of 1VVM.
Test example：
Fermentation tank culture medium：Starch, corn flour, glucose sugar, dregs of beans, CaCl2、MgSO4、MnSO4、KH2PO4, polyethers be
The 60% of comparative example dosage, Paenibacillus polymyxa fermented waste fluid recycle product 40%, and water is surplus.
Fermentation parameter is identical as comparative example.
By the method in embodiment 4, zymotic fluid 12h, 16h, 20h cell number and gemma number are detected, calculates spore forming rate,
As a result it see the table below.
Table 3 recycles influence of the product to fermentation of bacillus subtilis spore forming rate
| Spore forming rate | 12h | 16h | 20h |
| Test example | 2.16% | 12.68% | 89.62% |
| Reference examples | 1.53% | 9.22% | 71.70% |
As can be seen from the above results, the bud that Paenibacillus polymyxa fermented waste fluid recycles product is added in culture medium
Spore formation rate is significantly higher than control, illustrates that the recovery product is a kind of fermentation of bacillus subtilis synergist.
Embodiment 6 recycles application of the product in bacillus pumilus is fermented
Comparative example：
Fermentation tank culture medium (weight percent)：Starch 1%, peanut meal 2.5%, corn flour 2%, corn steep liquor 2%,
CaCl20.005%, MgSO40.005%, MnSO40.005%, KH2PO40.1%, polyethers 0.01%, water are surplus.
Fermentation parameter：Between adjusting pH7.0-7.2,0.11-0.12MPa sterilizing 30min, bacillus pumilus seed culture fluid
Inoculum concentration 5%, 30 DEG C, the ventilatory capacity of 1VVM.
Test example：
Fermentation tank culture medium：Starch, peanut meal, corn flour, corn steep liquor, CaCl2、MgSO4、MnSO4、KH2PO4, polyethers it is equal
It is the 70% of comparative example dosage, Paenibacillus polymyxa fermented waste fluid recycles product 60%, and water is surplus.
Fermentation parameter is identical as comparative example.
By the method in embodiment 4, detection zymotic fluid 20h, for 24 hours, 28h cell numbers and gemma number, calculate spore forming rate,
As a result it see the table below.
Table 4 recycles influence of the product to bacillus pumilus fermentation spore forming rate
| Spore forming rate | 20h | 24h | 28h |
| Test example | 12.1% | 36.41% | 86.56% |
| Reference examples | 11.5% | 27.37% | 68.63% |
As can be seen from the above results, the bud that Paenibacillus polymyxa fermented waste fluid recycles product is added in culture medium
Spore formation rate is significantly higher than control, illustrates that the recovery product is a kind of bacillus pumilus fermentation synergist.
Claims (6)
1. a kind of recoverying and utilizing method of Paenibacillus polymyxa fermented waste fluid, it is characterised in that include the following steps：
(1) 0.05~0.1% chitosan, stirring wadding is added in the Paenibacillus polymyxa fermented waste fluid obtained to centrifugation by weight
Centrifuging and taking supernatant again after solidifying；
(2) 0.001~0.005% amylase is added by weight into supernatant, digests 15~60 minutes；
(3) supernatant after enzymolysis is crossed into cation exchange column, obtained product is added in fermentation of bacillus culture medium
As fermentation synergist.
2. a kind of fermentation of bacillus synergist, it is characterised in that：The fermentation of bacillus synergist is more viscous class gemma bars
The recycling product of bacterium fermented waste fluid, the recoverying and utilizing method include the following steps：
(1) 0.05~0.1% chitosan, stirring wadding is added in the Paenibacillus polymyxa fermented waste fluid obtained to centrifugation by weight
Centrifuging and taking supernatant again after solidifying；
(2) 0.001~0.005% amylase is added by weight into supernatant, digests 15~60 minutes；
(3) supernatant after enzymolysis is crossed into cation exchange column, obtained product is added in fermentation of bacillus culture medium
As fermentation synergist.
3. fermentation of bacillus synergist as claimed in claim 2, it is characterised in that：The bacillus is bacillus subtilis
Or bacillus amyloliquefaciens or bacillus pumilus.
4. a kind of fermentation of bacillus synergisting method, it is characterised in that：Claim 2 is added in fermentation of bacillus culture medium
The fermentation synergist.
5. fermentation of bacillus synergisting method as claimed in claim 4, it is characterised in that：The bacillus is withered grass gemma
Bacillus or bacillus amyloliquefaciens or bacillus pumilus.
6. fermentation of bacillus synergisting method as described in claim 4 or 5, it is characterised in that：The fermentation synergist accounts for bud
The 40-60% of spore bacillus fermentation culture medium total weight.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109796394A (en) * | 2018-12-27 | 2019-05-24 | 武汉科诺生物科技股份有限公司 | A method of extracting auxin from Paenibacillus polymyxa fermentation liquid |
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| WO2015089183A2 (en) * | 2013-12-11 | 2015-06-18 | Ohio State Innovation Foundation | System for delivery of microbial inoculants and related materials and methods |
| CN106011005A (en) * | 2016-05-20 | 2016-10-12 | 东莞市保得生物工程有限公司 | Bacillus amyloliquefaciens T600 and preparation method and application of microbial agent |
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| CN101008017A (en) * | 2006-01-23 | 2007-08-01 | 徐文超 | Bacillus subtilis (Ehrenberg)Cohn fermented product and its production method and uses |
| WO2015089183A2 (en) * | 2013-12-11 | 2015-06-18 | Ohio State Innovation Foundation | System for delivery of microbial inoculants and related materials and methods |
| CN106011005A (en) * | 2016-05-20 | 2016-10-12 | 东莞市保得生物工程有限公司 | Bacillus amyloliquefaciens T600 and preparation method and application of microbial agent |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109796394A (en) * | 2018-12-27 | 2019-05-24 | 武汉科诺生物科技股份有限公司 | A method of extracting auxin from Paenibacillus polymyxa fermentation liquid |
| CN109796394B (en) * | 2018-12-27 | 2021-05-18 | 武汉科诺生物科技股份有限公司 | Method for extracting plant growth hormone from paenibacillus polymyxa fermentation liquor |
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