CN101260389A - Enzyme producing method for white rot fungus - Google Patents
Enzyme producing method for white rot fungus Download PDFInfo
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- CN101260389A CN101260389A CNA2008100360389A CN200810036038A CN101260389A CN 101260389 A CN101260389 A CN 101260389A CN A2008100360389 A CNA2008100360389 A CN A2008100360389A CN 200810036038 A CN200810036038 A CN 200810036038A CN 101260389 A CN101260389 A CN 101260389A
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- white
- rot fungi
- producing method
- enzyme
- enzyme producing
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- Enzymes And Modification Thereof (AREA)
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Abstract
The invention relates to a white rot fungi continuous high-efficient enzyme producing method, which belongs to the environmental biotechnical field and can be applied in the condition of difficult degradation of organic waste water treatment and environmental repair. In order to realize the aims of the invention, the white rot fungi continuous high-efficient enzyme producing method uses cobs as growth carriers and supplemental nutrition substrates of white rot fungus, utilizes an air-lift fermentation apparatus and optimizes the fermentation conditions, realizes continuous high-efficient output of lignin peroxidases and manganese peroxidases, and simultaneously reduces the enzyme production cost.
Description
Technical field
Patent of the present invention relates to a kind of white-rot fungi and continues efficient enzyme producing method, belongs to the Environmental Biotechnology field, can be applied in organic wastewater with difficult degradation thereby processing and the environment remediation.
Background technology
The refractory organic industrial sewage that comprises dye chemical industry waste water has colourity height, salinity height, strong toxicity, pollutant load height and complicated component, is difficult to characteristics such as degraded, be emphasis and the difficult point that industrial wastewater pollution is administered, these trade effluents after being discharged in the environment, will not cause serious ecological damage and human health problems as not dealing carefully with.
White-rot fungi under the lignin degradation enzyme catalysis by free chain reaction to waste water from dyestuff in various organic pollutants etc. have wide spectrum, degradation function efficiently.The lignin-degrading enzymes of white rot fungus secretion mainly contains lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase (Lac), and they are quick, the efficient leitungskerns handled of refractory organic industrial sewage.How to realize the lasting efficient output of white-rot fungi lignin-degrading enzymes, be urgent problem during the through engineering approaches of white-rot fungi Environmental Biotechnology is used, the working cost of building extensive enzyme reactor treatment system, reducing Industrial Wastewater Treatment is significant.
Summary of the invention
Patent of the present invention provides a kind of white-rot fungi to continue efficient method of producing enzyme.
For realizing purpose of the present invention, white-rot fungi of the present invention continues efficient enzyme producing method and comprises: guarantee to continue to produce enzyme and reduce corn cob filler, spore liquid preparation and the fermentation parameter control method of producing the enzyme cost.
Main contents of the present invention are: utilize growing carrier and the extra-nutrition matrix of corn cob as white-rot fungi, utilize airlift fermentation equipment and optimization of fermentation conditions, realize the lasting efficient output of lignin peroxidase and manganese peroxidase, reduce simultaneously and produce the enzyme cost.Specifically may further comprise the steps:
(1) pre-treatment of corn cob: corn cob is processed into the cubes that is of a size of 0.5cm * 0.5cm * 0.5cm, sterilization is 15 minutes under 115 ℃, 0.1Mpa, put as for the sterilisable chamber internal cooling, put it in sterilisable chamber to the shaking in the bottle of limit nitrogen substratum is housed the back, makes each shake that the quantity of corn core carrier remains on 10/100ml in the bottle;
(2) white-rot fungi spore liquid preparation: inject sterilized water in the flat board, add the white-rot fungi spore powder, filter through 4 layers of lens wiping paper, the elimination mycelium gets the white-rot fungi spore liquid;
(3) corn core carrier and white-rot fungi spore liquid are fed fermentor tank, temperature remains on 37 ℃ in the fermentor tank, and air input is 1.0vvm, adds limit nitrogen substratum in fermentor tank, realizes the enzyme producing method of white-rot fungi.
The enzyme producing method of white-rot fungi provided by the invention, wherein limitting the nitrogen medium component is (L
-1): glucose 10g, ammonium tartrate 0.2g, phenylcarbinol 0.54g, KH
2PO
42.56g, MgSO
47H
2O0.71g, vitamins B
10.001g, liquid microelement 70mL, the 10mmol/L acetate buffer solution is regulated pH to 4.5; Liquid microelement (L wherein
-1) composition be: Padil 0.6g, MnSO
4H
2O 0.5g, NaCl 1g, FeSO
47H
2O 0.1g, CoCl
2H
2O 0.19g, CaCl
22H
2O 1.56g, ZnSO
47H
2O 0.1g, CuSO
45H
2O 0.1g, KAl (SO
4)
212H
2O 0.01g, HBO
30.01g, Na
2MoO
42H
2O 0.01g.
The unusual effect of patent of the present invention is: the lasting efficient output of (1) lignin peroxidase and manganese peroxidase; (2) producing the enzyme cost descends.
Embodiment
The present invention is described in detail below in conjunction with embodiment:
(1) pre-treatment of corn cob, size, charging capacity and add the time: corn cob is cleaned with clear water earlier, aftertreatment becomes to be of a size of the cubes of 0.5cm * 0.5cm * 0.5cm, with its sterilization 15 minutes under 115 ℃, 0.1Mpa, put as for allowing its cooling in the sterilisable chamber, put it in sterilisable chamber to the shaking in the bottle of limit nitrogen substratum is housed the back, makes each shake that the quantity of corn core carrier remains on 10/100ml in the bottle.
(2) white-rot fungi spore liquid preparation: inject a small amount of sterilized water (laboratory sterilized water: 100mL water is added 200mL or 250mL triangular flask, add tampon, wrapping in dull and stereotyped, the sterilization 30 minutes down of 121 ℃, 0.1Mpa), scrape spore powder with push rod, filter the elimination mycelium through 4 layers of lens wiping paper.The gained filtrate is counted with blood counting chamber.
Thalline counting step: 1) get cleaning exsiccant blood counting and pull, on nucleonics, add cover glass.2) get bacterium liquid and shake up, drip a droplet with dropper by the cover glass edge, then bacterium liquid infiltrates voluntarily, and nucleonics must not have bubble.3) sample concentration requires every little lattice to have 5-10 thalline to be advisable.4) when counting, get upper left, lower-left, upper right, bottom right and central grid (i.e. 80 little lattice) by diagonal lines.5) each sample repeat count is 2-3 time, takes the mean, by formula counting cells number/(mL)=(cell count/80 in the 80 little lattice) * 400 * 10000 * extension rate.The control of fermentation parameter:, make the interior temperature of fermentor tank remain on 37 ℃ by the attemperation control valve; By regulating air intake valve, making air input is 1.0vvm; Regularly in fermentor tank, add certain limit nitrogen substratum by detected enzyme in early stage change curve alive, make Phanerochaete chrysosporium in fermentor tank, can sustainable high efficiency produce enzyme.
Steam enters fermentor tank and heats and sterilize; Make fermentation jar temperature maintain 115 ℃, tank pressure maintains 0.1Mpa, is incubated 15 minutes; Stop steam and enter, fermentor tank naturally cools to 38 ℃; Sterile air is fed fermentor tank to provide fermentation required oxygen; Add limit nitrogen substratum 15L to fermentor tank.White-rot fungi spore and corn cob filler are put into fermentor tank; Needing to detect enzyme in the fermenting process lives and total sugar content and Changing Pattern thereof; When enzyme work reaches peak value in the fermented liquid, can discharge crude enzyme liquid, or make enzyme preparation product through purification.Nutrient solution adopts limit nitrogen substratum, and its composition is (L
-1): glucose 10g, ammonium tartrate 0.2g, phenylcarbinol 0.54g, KH
2PO
42.56g, MgSO
47H
2O 0.71g, vitamins B
10.001g, liquid microelement 70mL, the 10mmol/L acetate buffer solution is regulated pH to 4.5.Liquid microelement (L wherein
-1) composition be: Padil 0.6g, MnSO
4H
2O 0.5g, NaCl 1g, FeSO
47H
2O 0.1g, CoCl
2H
2O 0.19g, CaCl
22H
2O 1.56g, ZnSO
47H
2O0.1g, CuSO
45H
2O 0.1g, KAl (SO
4)
212H
2O 0.01g, HBO
30.01g, Na
2MoO
42H
2O 0.01g.
Claims (2)
1, a kind of enzyme producing method of white-rot fungi, is characterized in that as the growing carrier of thalline and the additional matrix of producing enzyme with corn cob:
(1) pre-treatment of corn cob: corn cob is processed into the cubes that is of a size of 0.5cm * 0.5cm * 0.5cm, sterilization is 15 minutes under 115 ℃, 0.1Mpa, put as for the sterilisable chamber internal cooling, put it in sterilisable chamber to the shaking in the bottle of limit nitrogen substratum is housed the back, makes each shake that the quantity of corn core carrier remains on 10/100ml in the bottle;
(2) white-rot fungi spore liquid preparation: inject sterilized water in the flat board, add the white-rot fungi spore powder, filter through 4 layers of lens wiping paper, the elimination mycelium gets the white-rot fungi spore liquid;
(3) corn core carrier and white-rot fungi spore liquid are fed fermentor tank, temperature remains on 37 ℃ in the fermentor tank, and air input is 1.0vvm, adds limit nitrogen substratum in fermentor tank, realizes the enzyme producing method of white-rot fungi.
2. the enzyme producing method of white-rot fungi as claimed in claim 1, it is characterized in that limitting the nitrogen medium component is (L
-1): glucose 10g, ammonium tartrate 0.2g, phenylcarbinol 0.54g, KH
2PO
42.56g, MgSO
47H
2O 0.71g, vitamins B
10.001g, liquid microelement 70mL, the 10mmol/L acetate buffer solution is regulated pH to 4.5; Liquid microelement (L wherein
-1) composition be: Padil 0.6g, MnSO
4H
2O 0.5g, NaCl 1g, FeSO
47H
2O 0.1g, CoCl
2H
2O0.19g, CaCl
22H
2O 1.56g, ZnSO
47H
2O 0.1g, CuSO
45H
2O 0.1g, KAl (SO
4)
212H
2O 0.01g, HBO
30.01g, Na
2MoO
42H
2O 0.01g.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100360389A CN101260389A (en) | 2008-04-15 | 2008-04-15 | Enzyme producing method for white rot fungus |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100360389A CN101260389A (en) | 2008-04-15 | 2008-04-15 | Enzyme producing method for white rot fungus |
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| Publication Number | Publication Date |
|---|---|
| CN101260389A true CN101260389A (en) | 2008-09-10 |
Family
ID=39961089
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|---|---|---|---|
| CNA2008100360389A Pending CN101260389A (en) | 2008-04-15 | 2008-04-15 | Enzyme producing method for white rot fungus |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101508981B (en) * | 2009-03-19 | 2011-03-30 | 华东师范大学 | Method and apparatus for duration enzyme production of biomass carrier with organic waste water treatment |
| CN104402120A (en) * | 2014-09-25 | 2015-03-11 | 湖南大学 | Treatment method of methylene blue in waste water by white rot fungi |
| CN105233458A (en) * | 2015-10-19 | 2016-01-13 | 桂林理工大学 | Method for degrading bisphenol A through white-rot fungus crude enzyme |
| CN105417724A (en) * | 2015-12-15 | 2016-03-23 | 林康艺 | Composition for degrading acid brilliant green |
| CN105417725A (en) * | 2015-12-15 | 2016-03-23 | 李国深 | Method for degrading acid brilliant green through laccase produced from white rot fungi |
| CN106638081A (en) * | 2016-10-28 | 2017-05-10 | 河南华禹环保科技有限公司 | Method for preparing filter paper fiber carrier for white-rot fungus culture |
| CN106916755A (en) * | 2017-04-07 | 2017-07-04 | 山西大学 | A kind of culture medium of suitable white-rot fungi growth |
| CN109081439A (en) * | 2018-10-26 | 2018-12-25 | 江苏哈宜环保研究院有限公司 | A method of sewage is handled using corncob as the denitrification filter pool reactor of biomass carrier and using it |
| CN114908024A (en) * | 2022-06-23 | 2022-08-16 | 合肥工业大学 | Method for promoting white rot fungi to directionally produce enzyme and degrade plastic pollutants |
-
2008
- 2008-04-15 CN CNA2008100360389A patent/CN101260389A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101508981B (en) * | 2009-03-19 | 2011-03-30 | 华东师范大学 | Method and apparatus for duration enzyme production of biomass carrier with organic waste water treatment |
| CN104402120A (en) * | 2014-09-25 | 2015-03-11 | 湖南大学 | Treatment method of methylene blue in waste water by white rot fungi |
| CN104402120B (en) * | 2014-09-25 | 2016-09-14 | 湖南大学 | The method processing methylene blue in waste with whiterot fungi |
| CN105233458A (en) * | 2015-10-19 | 2016-01-13 | 桂林理工大学 | Method for degrading bisphenol A through white-rot fungus crude enzyme |
| CN105233458B (en) * | 2015-10-19 | 2018-11-06 | 桂林理工大学 | A method of utilizing white-rot fungi crude enzyme liquid degradation bisphenol-A |
| CN105417724A (en) * | 2015-12-15 | 2016-03-23 | 林康艺 | Composition for degrading acid brilliant green |
| CN105417725A (en) * | 2015-12-15 | 2016-03-23 | 李国深 | Method for degrading acid brilliant green through laccase produced from white rot fungi |
| CN106638081A (en) * | 2016-10-28 | 2017-05-10 | 河南华禹环保科技有限公司 | Method for preparing filter paper fiber carrier for white-rot fungus culture |
| CN106916755A (en) * | 2017-04-07 | 2017-07-04 | 山西大学 | A kind of culture medium of suitable white-rot fungi growth |
| CN109081439A (en) * | 2018-10-26 | 2018-12-25 | 江苏哈宜环保研究院有限公司 | A method of sewage is handled using corncob as the denitrification filter pool reactor of biomass carrier and using it |
| CN114908024A (en) * | 2022-06-23 | 2022-08-16 | 合肥工业大学 | Method for promoting white rot fungi to directionally produce enzyme and degrade plastic pollutants |
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Open date: 20080910 |