CN102174583A - Method for producing biological humic acid by mixed fermentation - Google Patents
Method for producing biological humic acid by mixed fermentation Download PDFInfo
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- CN102174583A CN102174583A CN2011100420080A CN201110042008A CN102174583A CN 102174583 A CN102174583 A CN 102174583A CN 2011100420080 A CN2011100420080 A CN 2011100420080A CN 201110042008 A CN201110042008 A CN 201110042008A CN 102174583 A CN102174583 A CN 102174583A
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Abstract
The invention provides a method for producing biological humic acid by mixed fermentation. In the method, the biological humic acid is produced by the solid fermentation of bacillus subtilis and paenibacillus polymyxa. In the invention, by means of a production process including the steps of breeding and developing excellent productive strains and carrying out mixed solid fermentation, the production capacity and the quality level of biological humic acid products are increased, the produced biological humic acid products not only are rich in active humic acid (fulvic acid) but also contain probiotics, and a foundation is laid for realizing industrial production of the biological humic acid.
Description
Technical field
The invention belongs to the using microbe field, more specifically relate to a kind of method of utilizing mixed fungus fermentation to produce biological humic.
Technical background
Natural humic acid mainly is present among soil, river silt, oceanic sediment, brown coal and the weathered coal, is generated after long-term biological chemistry or physical chemistry of the earth effect by the plant and animal residues of death.The organic materials of different humify degree is adopted in the production of present domestic humic acids mostly, and for example peat, brown coal, weathered coal etc. extract by the soda acid extraction process, extraction yield is low, what obtain is the mineral humic acidss, and its activated humic acid---xanthohumic acid content is low, and does not contain probiotics.So far the manufacturing enterprise and the product that also do not have biology (or mineral) humic acids abroad.In eighties of last century the early 1990s, China North China once had the people to produce humic acid substance by straw granulated slag stack retting legal system, once caused scientific circles' sensation, b referred to as " knowledge nature; surmount nature ", had opened up the new industry of humic acids---the biological xanthohumic acid industry of distinct Chinese characteristics thus; But owing to continue to use local method stack retting technology always, yield poorly, unstable product quality, and product extracts xanthohumic acid by chemical technique again again, thereby lost the important component in the product---beneficial microorganism.
Up to the present, China's biological humic industry is scarcely out of swaddling-clothes, and fails to form the great effect to humic acids industry, more fail to bring into play its aspect the national economic development and ecological, environmental protective should be powerful.China North China (mainly being Hebei province) is the source region of biological xanthohumic acid, existing pilot scale (producing dry-matter per year below 500 tons) 5-6 family of enterprise.But the still artificial heap of these factories is built fermentation method, yields poorly, unstable product quality, and some production link (as fermentation, drying) still depends on natural condition.
Luzhou Bio-Chemical Co., Ltd., Zhao'an County, Fujian Prov. adopts indigenous flora complex ferment bagasse to produce biological humic, not only contains xanthohumic acid in the product, and has kept probiotics, makes product function more become perfect.But owing to adopt the indigenous flora fermentation, bacterial classification lacks the cultivation and the management of science, morphs easily, fails and death, has influenced quality product and output; And processing condition such as fermentative production await further to improve.
China humic acids TIA and experts and scholars attach great importance to the development of biological humic industry, pay close attention to the sector new results new technology always.In recent years, repeatedly advocate the sector and answer mass-producing, mechanize, should promote product function, realize the industrialization production of biological humic with directed bacterial classification.
Summary of the invention
The invention provides a kind of method of utilizing mixed fungus fermentation to produce biological humic, this method promotes the output and the quality of biological humic product by seed selection and the good production bacterial classification of exploitation.
To achieve these goals, the technical scheme taked of the present invention is: utilize subtilis and Paenibacillus polymyxa hybrid bacterial strain solid fermentation to produce biological humic.
The main component of the fermention medium of wherein said solid fermentation is bagasse and wheat bran.
The ratio of bagasse and wheat bran is 1:1 ~ 1.5:1(w/w) in the fermention medium.
The ratio of subtilis and Paenibacillus polymyxa is 2:1(v/v).
The cell concn of subtilis bacteria suspension and Paenibacillus polymyxa bacteria suspension all is controlled at 10
8Cfu/mL.
Total inoculum size of subtilis and Paenibacillus polymyxa is 10%～14%, and this per-cent is the ratio that the cumulative volume amount of two kinds of bacteria suspensions accounts for fermention medium weight.
The fermentation condition of described solid fermentation is: the initial pH of fermention medium is 4 ~ 5.5,50 ~ 55 ℃ of leavening temperatures.
The zymotechnique of described solid fermentation is: the ratio of bagasse and wheat bran is 55 ℃ of 1.5:1, total inoculum size 12%, the leavening temperatures that mixes bacterium in the initial pH5.5 of fermention medium, the fermention medium.
The present invention is a raw material with sugar refinery bagasse, utilizes subtilis and Paenibacillus polymyxa blended solid fermentation to produce biological humic acids.The ratio of subtilis and Paenibacillus polymyxa is that the various mycetocyte concentration of 2:1(are controlled at 10
8Cfu/mL), two kinds of total inoculum sizes of bacterium are 10%～14%(v/w); Fermentation condition: the ratio of bagasse and wheat bran is 1:1 ~ 1.5:1 in the fermention medium, and the initial pH of substratum is 4 ~ 5.5,50 ~ 55 ℃ of leavening temperatures.The 6d that ferments with this understanding, activated humic acid (xanthohumic acid) increase of production 69.97%.
Remarkable advantage of the present invention: with sugar refinery bagasse is raw material, turns waste into wealth, and realizes the utilization again of ERM, has embodied the development principle of recycling economy, for a new approach has been opened up in the comprehensive utilization technique transformation in the large quantities of big-and-middle-sized sugar refinery of China; Mix the bacteria solid fermentation production technique by seed selection and the good production bacterial classification of exploitation, employing, promote the throughput and the quality level of biological humic product, not only be rich in activated humic acid (xanthohumic acid) in the biological humic product of being produced, and contain profitable probliotics, for the industrialization production that realizes biological humic lays the first stone.
Embodiment:
1, the Screening and Identification of good production bacterial classification
The 7 strain bacteriums (obtaining from sugarcane field soil, paddy soil, rice straw compost, the ground screenings such as tree root, edible fungus culturing matrix of rotting) that obtain from the nature screening are fermented and produce the comparison of humic acids ability, obtain the good production strains A of 4 strains, B, C and D, the rate of increase of xanthohumic acid output is respectively 18.36%, 16.20%, 15.86% and 12.23%.
Get and produce this stronger 4 strain bacterium of humic acids ability, carry out different matched combined (being AB, AC, AD, BC, BD, CD), measure the xanthohumic acid increase of production rate of fermentation back various combination.The result shows that the ferment effect of AB combination is best, and the rate of increase of xanthohumic acid output is a 28.85%(table 1); And when bacterial classification ratio A:B=2:1, the rate of increase of xanthohumic acid output is the highest, is 32.28%(table 2).
The combination of table 1 different strain is to the influence of xanthohumic acid output
Remarks: the xanthohumic acid content of substratum before xanthohumic acid increase of production rate %=(the xanthohumic acid content of substratum before the xanthohumic acid content-fermentation of leavened prod)/fermentation
Table 2 AB bacterial classification different ratios is to the influence of xanthohumic acid output
A, B bacterial classification are carried out 16s rDNA sequential analysis and identify that in conjunction with Physiology and biochemistry determine that A, B bacterial classification are respectively subtilis and Paenibacillus polymyxa, bacterial strain is stored in University of Fuzhou's bio-science and engineering college.
2, fermention medium optimization
In original factory fermention medium, the total content of bagasse and wheat bran accounts for about 95%, is the main component that influences humic acids output.Investigate the influence to xanthohumic acid output of bagasse and wheat bran different ratios (2:1,1.5:1,1:1,1:1.5,1:2), the result shows that when the ratio of bagasse and wheat bran was 1:1, the rate of increase maximum of xanthohumic acid output was 38.13%(table 3).
The different proportionings with wheat bran of table 3 bagasse are to the influence of xanthohumic acid output
Further the initial pH value (pH is respectively 4.0,4.5,5.5,6.5,7.0 and 7.5) to fermention medium is optimized, and when the initial pH value of substratum was 4.0, the rate of increase maximum of xanthohumic acid output was 45.31%(table 4).
Table 4 different fermentations initial pH value of medium is to the influence of xanthohumic acid output
3, the optimization of technological condition for fermentation
Investigate the influence of seed inoculum size (4%, 6%, 8%, 10%, 12%, 14%) to xanthohumic acid output.Experimental result shows that along with the increase of inoculum size, xanthohumic acid output increases gradually; When inoculum size greater than 12%(v/w, cell concn is 10
8Cfu/mL) time,, determine that therefore inoculum size is 12% to xanthohumic acid yield effect little (table 5).
The different seed inoculum sizes of table 5 are to the influence of xanthohumic acid output
Further compare the influence of different fermentations temperature (40 ℃, 45 ℃, 50 ℃, 55 ℃, 60 ℃, 65 ℃) to xanthohumic acid output.The result shows that along with the rising of leavening temperature, xanthohumic acid output increases gradually; When leavening temperature was 55 ℃, the rate of increase maximum of xanthohumic acid output was 52.21%; When temperature during greater than 55 ℃, xanthohumic acid output descend rapidly (table 6).
Table 6 different fermentations temperature is to the influence of xanthohumic acid output
4, orthogonal test
On the basis of above-mentioned experiment of single factor, further adopt orthogonal test to leavening temperature (45 ℃, 50 ℃, 55 ℃), the initial pH(4 of fermention medium, 5.5,6.5), the ratio (1:1,1.5:1,2:1) and the inoculum size (10%, 12%, 14%) of bagasse and wheat bran be optimized.The best zymotechnique that obtains is: 55 ℃ of the ratio 1.5:1 of the initial pH5.5 of fermention medium, bagasse and wheat bran, inoculum size 12%, leavening temperature; The primary and secondary of each factor affecting is in proper order: the ratio＞leavening temperature of bagasse and wheat bran＞fermention medium initial pH value＞inoculum size.
Below be most preferred embodiment of the present invention, further describe the present invention, but the present invention be not limited only to this.
Implement 1
Preparation solid fermentation substratum, wherein the ratio of bagasse and wheat bran is 1.5:1, total content is 94.74%, and adds a spot of urea, groundnut meal, MgSO
4, CaCO
3Deng composition, regulating the initial pH of substratum is 5.5, and during batching, the solid medium moisture content is pinched compound with hand can become piece, and finger tip has water to ooze out but do not drip is advisable; Adopt subtilis and Paenibacillus polymyxa mixed fermentation, the ratio of subtilis and Paenibacillus polymyxa is that the cell concn of two kinds of bacterium of 2:1(is 10
8Cfu/mL), total inoculum size is 12%(v/w); At 55 ℃ of bottom fermentation 6d, the rate of increase of xanthohumic acid output is 69.97%; With compare before the fermentation condition optimization, xanthohumic acid output has improved 41.12%; Compare with utilizing subtilis, the fermentation of Paenibacillus polymyxa list bacterium respectively, xanthohumic acid output has improved 51.61%, 53.77% respectively.
Claims (8)
1. a method of utilizing mixed fungus fermentation to produce biological humic is characterized in that: utilize subtilis and Paenibacillus polymyxa solid fermentation to produce biological humic.
2. the method for utilizing mixed fungus fermentation to produce biological humic according to claim 1, it is characterized in that: the main component of the fermention medium of described solid fermentation is bagasse and wheat bran.
3. the method for utilizing mixed fungus fermentation to produce biological humic according to claim 2 is characterized in that: the ratio of bagasse and wheat bran is 1:1～1.5:1(w/w) in the fermention medium.
4. the method for utilizing mixed fungus fermentation to produce biological humic according to claim 1, it is characterized in that: the ratio of subtilis and Paenibacillus polymyxa is 2:1(v/v).
5. the method for utilizing mixed fungus fermentation to produce biological humic according to claim 1, it is characterized in that: the cell concn of subtilis bacteria suspension and Paenibacillus polymyxa bacteria suspension all is controlled at 10
8Cfu/mL.
6. the method for utilizing mixed fungus fermentation to produce biological humic according to claim 1, it is characterized in that: total inoculum size of subtilis and Paenibacillus polymyxa is 10%～14%, and this per-cent is the ratio that the cumulative volume amount of two kinds of bacteria suspensions accounts for fermention medium weight.
7. the method for utilizing mixed fungus fermentation to produce biological humic according to claim 1, it is characterized in that: the fermentation condition of described solid fermentation is: the initial pH of fermention medium is 4-5.5,50～55 ℃ of leavening temperatures.
8. according to arbitrary described method of utilizing mixed fungus fermentation to produce biological humic among the claim 1-7, it is characterized in that: the zymotechnique of described solid fermentation is: the ratio of bagasse and wheat bran is 55 ℃ of 1.5:1, total inoculum size 12%, the leavening temperatures that mixes bacterium in the initial pH5.5 of fermention medium, the fermention medium.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102517334A (en) * | 2011-12-09 | 2012-06-27 | 宁夏共享集团有限责任公司 | Method for producing biochemical humate by microbially fermented furfural residues |
| CN102557767A (en) * | 2011-12-19 | 2012-07-11 | 西华大学 | Special compound fungus agent for humification of fungus residues of straw mushrooms and manufacturing method thereof |
| CN103467144A (en) * | 2012-06-06 | 2013-12-25 | 福建省诏安县绿洲生化有限公司 | Refined fulvic-acid-containing agricultural microbial inoculum and production method thereof |
| CN110616246A (en) * | 2019-10-11 | 2019-12-27 | 北京大伟嘉生物技术股份有限公司 | Fermentation method for increasing content of sodium fulvate and microbial agent thereof |
| CN111848244A (en) * | 2019-11-03 | 2020-10-30 | 黑龙江丰富农业科技发展有限公司 | Fulvic acid type special fertilizer for industrial hemp |
| CN113233934A (en) * | 2021-04-21 | 2021-08-10 | 陕西省生物农业研究所 | Attapulgite biological organosilicon fertilizer special for saline-alkali soil and preparation method thereof |
-
2011
- 2011-02-22 CN CN2011100420080A patent/CN102174583A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102517334A (en) * | 2011-12-09 | 2012-06-27 | 宁夏共享集团有限责任公司 | Method for producing biochemical humate by microbially fermented furfural residues |
| CN102517334B (en) * | 2011-12-09 | 2014-07-23 | 宁夏共享集团有限责任公司 | Method for producing biochemical humate by microbially fermented furfural residues |
| CN102557767A (en) * | 2011-12-19 | 2012-07-11 | 西华大学 | Special compound fungus agent for humification of fungus residues of straw mushrooms and manufacturing method thereof |
| CN102557767B (en) * | 2011-12-19 | 2014-07-02 | 西华大学 | Special compound fungus agent for humification of fungus residues of straw mushrooms and manufacturing method thereof |
| CN103467144A (en) * | 2012-06-06 | 2013-12-25 | 福建省诏安县绿洲生化有限公司 | Refined fulvic-acid-containing agricultural microbial inoculum and production method thereof |
| CN103467144B (en) * | 2012-06-06 | 2015-01-07 | 福建省诏安县绿洲生化有限公司 | Refined fulvic-acid-containing agricultural microbial inoculum and production method thereof |
| CN110616246A (en) * | 2019-10-11 | 2019-12-27 | 北京大伟嘉生物技术股份有限公司 | Fermentation method for increasing content of sodium fulvate and microbial agent thereof |
| CN111848244A (en) * | 2019-11-03 | 2020-10-30 | 黑龙江丰富农业科技发展有限公司 | Fulvic acid type special fertilizer for industrial hemp |
| CN113233934A (en) * | 2021-04-21 | 2021-08-10 | 陕西省生物农业研究所 | Attapulgite biological organosilicon fertilizer special for saline-alkali soil and preparation method thereof |
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