CN103614315A - Spent mushroom substrate high-temperature decomposition composite bacterial agent and preparation method thereof - Google Patents
Spent mushroom substrate high-temperature decomposition composite bacterial agent and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to a spent mushroom substrate high-temperature decomposition composite bacterial agent, wherein eight thermophilic bacteria such as Aeribacillus pallidus, Geobacillus stearothermophilus, Brevibacillus thermoruber, Clostridium stercorarium subsp. thermolacticum, Clostridium thermocellum, Desulfotomaculum nigrificans and the like are subjected to mixing and mixing culture according to a certain cell number content ratio, composite bacterial agent preparation is performed when decomposition ability is high, different spent mushroom substrates are adopted as absorption carriers, peptone, yeast powder and the like are added, the bacterial liquid is repeatedly sprayed, absorption fermentation is performed, and drying at a temperature of 60 DEG C is performed to obtain the composite bacterial agent. The composite bacterial agent can be used for spent mushroom substrate feeds, spent mushroom substrate fertilizers and energy source conversion and other uses.
Description
Technical field
The present invention relates to decomposition and the comprehensive utilization technique field of agricultural by-products, be specifically related to a kind of bacterium chaff pyrolytic decomposition composite fungus agent.
Background technology
Bacterium chaff is to utilize the lignocellulosic materials such as wood chip, cotton seed hulls, stalk to carry out the substratum residuum after edible mushrooms substituting stuff cultivation results fruit body of edible fungi, is commonly called as edible fungus culturing waste material, bacterium slag etc., is important agricultural by-products resource.In the last few years, mushroom industry develop rapidly, bacterium chaff stock number is increasing, and current most bacterium chaff resources are not used effectively, and cause the wasting of resources and environmental pollution.Therefore, solving bacterium chaff utilizes problem to agricultural resource recycle and curbs environmental pollution of crucial importance.The main component of bacterium chaff is the difficult decomposing lignocellulose component of not utilized by edible mushrooms.At present, the comprehensive utilization of bacterium chaff is broadly divided into the aspects such as development animal-feed, fertilizer, cultivation matrix.The fast decoupled of bacterium chaff is its key link being further developed.Yet, effective report and application of decomposing microbial inoculum fast of also not developing for bacterium chaff specially at present.
Summary of the invention
The present invention is directed to the above-mentioned state of the art, aim to provide a kind of bacterium chaff pyrolytic decomposition composite fungus agent that can effectively decompose fast equal chaff and preparation method thereof.
The implementation of the object of the invention is that a kind of bacterium chaff pyrolytic decomposition composite fungus agent, is comprised of following 8 strain thermophilic bacteriums: jointly by bacterial strain preserving number ATCC51176, DSM3670Aeribacillus pallidus, bacterial strain preserving number DSM2027, ATCC7954Geobacillus stearothermophilus, bacterial strain preserving number DSM7064Brevibacillus thermoruber, bacterial strain preserving number ATCC43739, DSM2910Clostridium stercorarium subsp.thermolacticum, bacterial strain preserving number NCIB10682, ATCC27405Clostridium thermocellum, bacterial strain preserving number DSM574, ATCC19858Desulfotomaculum nigrificans, bacterial strain preserving number DSM21625Geobacillus thermoglucosidasius, bacterial strain preserving number DSM24528Symbiobacterium thermophilum8 strain thermophilic bacterium is respectively 10%-20% by the cell count content in composite fungus agent, 10%-20%, 2%-5%, 10%-15%, 15%-20%, 5%-10%, 10%-15%, the composite fungus agent of the ratio mixed fermentation gained of 15%-20%,
The implication of the cell count content of bacterial strain in mixed fermentation refers to that each strain cell number in composite fungus agent accounts for the percentage composition of the total cell count of composite bacteria.
The method of preparing bacterium chaff pyrolytic decomposition composite fungus agent, 8 strain bacteriums carry out compound bacteria-fermented after cultivating separately; Concrete steps are as follows:
(1) 8 strain bacterium is cultivated separately, and its cultural method is respectively:
Bacterial strain preserving number ATCC51176, the cultivation of DSM3670Aeribacillus pallidus, will be by glucose 1.0g, peptone 7.5g, extractum carnis 5.0g, yeast powder 2.5g, casamino acids 2.5g, sodium-chlor 5.0g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 8.5; The 60 ℃ of static cultivation of lucifuge 48h;
Bacterial strain preserving number DSM2027, the cultivation of ATCC7954Geobacillus stearothermophilus, will be by sodium-chlor 5.0g, extractum carnis 10.0g, peptone 10.0g, the substratum that distilled water 1000.0mL forms, pH value is adjusted: to 7.2; The 60 ℃ of static cultivation of lucifuge 48h;
The cultivation of bacterial strain preserving number DSM7064Brevibacillus thermoruber, will be by glucose 15.0g, yeast powder 5.0g, and Calcium dichloride dihydrate 0.2g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 7.0; The 60 ℃ of static cultivation of lucifuge 48h;
Bacterial strain preserving number ATCC43739, the cultivation of DSM2910Clostridium stercorarium subsp.thermolacticum, will be by dipotassium hydrogen phosphate 0.3g, potassium primary phosphate 0.2g, ammonium chloride 0.5g, magnesium sulfate heptahydrate 0.5g, Calcium dichloride dihydrate 0.25g, sodium-chlor 2.25g, iron vitriol 0.002g, vitamin solution 10.0ml, trace element solution 1.0ml, yeast powder 2.0g, casein peptone 2.0g, resazurin 0.001g, sodium bicarbonate 0.85g, methyl alcohol 10.0ml, Cys hydrochloride monohydrate 0.3g, nine water cure sodium 0.3g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 6.8, at 80%N
2+ 20%CO
2under anaerobic condition, the 60 ℃ of static cultivation of dark 72h,
Described vitamin solution is Biotin2.0mg, Folic acid2.0mg, Pyridoxine-HCl10.0mg, Thiamine-HCl2H
2o5.0mg, Riboflavin5.0mg, Nicotinic acid5.0mg, D-Ca-pantothenate5.0mg, Vitamin B
120.1mg, p-Aminobenzoic acid5.0mg, Lipoic acid5.0mg, adding distil water is settled to the solution of 1000.0ml;
Described trace element solution is 7.7M HCl10.0ml, FeCl
24H
2o1.5g, ZnCl
270.0mg, MnCl
24H
2o100.0mg, H
3bO
36.0mg, CoCl
26H
2o190.0mg, CuCl
22H
2o2.0mg, NiCl
26H
2o24.0mg, Na
2moO
42H
2o36.0mg, adding distil water is settled to the solution of 1000.0ml;
Bacterial strain preserving number NCIB10682, the cultivation of ATCC27405Clostridium thermocellum, will be by ammonium sulfate 1.3g, magnesium chloride hexahydrate 2.6g, potassium primary phosphate 1.43g, dipotassium hydrogen phosphate 5.5g, Calcium dichloride dihydrate 0.13g, β-phospho-glycerol disodium tetrahydrate 6.00g, iron vitriol 1.10mg, reduced glutathion 0.25g, yeast powder 4.5g, resazurin 1.0mg, Microcrystalline Cellulose 10.0g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 7.2; At 80%N
2+ 20%CO
2under anaerobic condition, the 60 ℃ of static cultivation of dark 72h;
Bacterial strain preserving number DSM574, the cultivation of ATCC19858Desulfotomaculum nigrificans, will be by dipotassium hydrogen phosphate 0.5g, ammonium chloride 1.0g, sodium sulfate 1.0g, Calcium dichloride dihydrate 0.1g, magnesium sulfate heptahydrate 2.0g, DL-LACTIC ACID sodium 2.0g, yeast powder 1.0g, resazurin 1.0mg, iron vitriol 0.5g, Thioglycolic acid sodium salt 0.1g, xitix 0.1g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 7.8; At 80%N
2+ 20%CO
2under anaerobic condition, the 60 ℃ of static cultivation of dark 72h;
The cultivation of bacterial strain preserving number DSM21625Geobacillus thermoglucosidasius, will be by casein peptone 15.0g, soy peptone 5.0g, and sodium-chlor 5.0g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 7.3; The 60 ℃ of static cultivation of dark 48h;
The cultivation of bacterial strain preserving number DSM24528Symbiobacterium thermophilum, will be by sodium-chlor 5.0g, SODIUMNITRATE 1.7g, and yeast powder 5.0g, peptone 10.0g, bicarbonate of ammonia 3.2g, the substratum that distilled water 1000.0ml forms,, pH value is adjusted to 7.8; At 80%N
2+ 20%CO
2under anaerobic condition, the 60 ℃ of static cultivation of dark 72h;
(2) compound bacteria-fermented
To after the cultured 8 strain bacteriums of step (1) proportionally mix, obtain mixed strains stoste, the mixed strains stoste that is equivalent to nutrient solution volume 5% is inoculated in fermentation culture, at 60 ℃, dark static cultivation is 5 days, obtains composite bacteria liquid;
Described mixing fermentation culture liquid is: peptone 5.0g, yeast powder 1.0g, calcium carbonate 3g, sodium-chlor 5g, K
2hPO
41g, MgSO
47H
2o0.35g, bacterium chaff powder 10g, distilled water 1000.0mL, pH value is 7.8,121 ℃ of sterilizing 20min;
The blending ratio of described 8 strain bacteriums is: bacterial strain preserving number ATCC51176, DSM3670Aeribacillus pallidus, bacterial strain preserving number DSM2027, ATCC7954Geobacillus stearothermophilus, bacterial strain preserving number DSM7064Brevibacillus thermoruber, bacterial strain preserving number ATCC43739, DSM2910Clostridium stercorarium subsp.thermolacticum, bacterial strain preserving number NCIB10682, ATCC27405Clostridium thermocellum, bacterial strain preserving number DSM574, ATCC19858Desulfotomaculum nigrificans, bacterial strain preserving number DSM21625Geobacillus thermoglucosidasius, bacterial strain preserving number DSM24528Symbiobacterium thermophilum cell count content in mixed strains is respectively 10%-20%, 10%-20%, 2%-5%, 10%-15%, 15%-20%, 5%-10%, 10%-15% and 15%-20%,
(3) preparation of bacterial adsorption carrier: after the bacterium chaff of 80 ℃ of oven dry is pulverized, cross 0.4mm sieve standby;
(4) composite bacteria absorption fermentation: add 0.2% peptone that accounts for bacterium chaff grain weight amount in the bacterium chaff powder of step (3), 0.3% fish meal, 0.25% yeast powder and 50% tap water, sprinkling while stirring accounts for bacterium chaff grain weight 1%, composite bacteria liquid by step (2) gained, be placed in fermentor tank, 60 ℃ of lucifuge heat insulating culture, within every 24 hours, uniform stirring is 1 time, every 48 hours, repeat to spray same bacterium liquid, spray altogether 3 times, the 3rd time is sprayed bacterium liquid measure is 20% of bacterium chaff grain weight, after spraying, lucifuge heat insulating culture is 12 hours, stir and be placed on 60 ℃ of oven dry, when being 20%, overall water content stops being dried, obtain composite fungus agent goods.
Composite fungus agent of the present invention by specific 8 strain bacteriums take bacterium chaff as carrier adsorb fermentation form, 8 strains form bacterium and are thermophilic bacterium, wherein form bacterium Clostridium thermocellum(NCIB10682, ATCC27405) only under strictly anaerobic condition, the lignocellulosic materials such as bacterium chaff are had to faint capacity of decomposition, under the equal conditions that its pure growth is cultivated at composite fungus agent, there is no capacity of decomposition; All the other 7 strains form bacterium and under experiment condition of the present invention, do not find that bacterium chaff is had to capacity of decomposition; But this 8 strain bacterium is carried out to after mixed culture, the lignocellulosic materials such as bacterium chaff are had to remarkable capacity of decomposition according to a certain percentage.This composite fungus agent can be used for the purposes such as edible fungi residue feed, Fertilizer Transformed and bioenergy conversion.
Embodiment
Composite fungus agent of the present invention consists of jointly following 8 strain thermophilic bacteriums: by bacterial strain preserving number ATCC51176, DSM3670Aeribacillus pallidus, bacterial strain preserving number DSM2027, ATCC7954Geobacillus stearothermophilus, bacterial strain preserving number DSM7064Brevibacillus thermoruber, bacterial strain preserving number ATCC43739, DSM2910Clostridium stercorarium subsp.thermolacticum, bacterial strain preserving number NCIB10682, ATCC27405Clostridium thermocellum, bacterial strain preserving number DSM574, ATCC19858Desulfotomaculum nigrificans, bacterial strain preserving number DSM21625Geobacillus thermoglucosidasius, bacterial strain preserving number DSM24528Symbiobacterium thermophilum8 strain thermophilic bacterium is respectively 10%-20% by the cell count content in composite fungus agent, 10%-20%, 2%-5%, 10%-15%, 15%-20%, 5%-10%, 10%-15%, the composite fungus agent of the ratio mixed fermentation gained of 15%-20%.
Preparation method, for above-mentioned 8 strain bacteriums are carried out carrying out mixed culture according to the ratio of appointment, when flora capacity of decomposition is vigorous, carries out composite fungus agent preparation, the different bacterium chaffs of take are absorption carrier, add peptone, yeast powder, fish meal etc., repeatedly spray bacterium liquid, absorption fermentation, 60 ℃ of oven dry and get final product.
Bacterial adsorption carrier is wood chip bacterium chaff, cotton seed hulls bacterium chaff, wood chip cotton seed hulls bacterium chaff, cotton stalk bacterium chaff, corn cob stalk bacterium chaff etc.
Below with example explanation the present invention.
Example 1
(1) 8 strain bacterium is cultivated separately, and its cultural method is respectively:
Bacterial strain preserving number ATCC51176, the cultivation of DSM3670Aeribacillus pallidus, will be by glucose 1.0g, peptone 7.5g, extractum carnis 5.0g, yeast powder 2.5g, casamino acids 2.5g, sodium-chlor 5.0g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 8.5; The 60 ℃ of static cultivation of lucifuge 48h.
Bacterial strain preserving number DSM2027, the cultivation of ATCC7954Geobacillus stearothermophilus, will be by sodium-chlor 5.0g, extractum carnis 10.0g, peptone 10.0g, the substratum that distilled water 1000.0mL forms, pH value is adjusted: to 7.2; The 60 ℃ of static cultivation of lucifuge 48h.
The cultivation of bacterial strain preserving number DSM7064Brevibacillus thermoruber, will be by glucose 15.0g, yeast powder 5.0g, and Calcium dichloride dihydrate 0.2g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 7.0; The 60 ℃ of static cultivation of lucifuge 48h.
Bacterial strain preserving number ATCC43739, the cultivation of DSM2910Clostridium stercorarium subsp.thermolacticum, substratum is: will be by dipotassium hydrogen phosphate 0.3g, potassium primary phosphate 0.2g, ammonium chloride 0.5g, magnesium sulfate heptahydrate 0.5g, Calcium dichloride dihydrate 0.25g, sodium-chlor 2.25g, iron vitriol 0.002g, vitamin solution 10.0ml, trace element solution 1.0ml, yeast powder 2.0g, casein peptone 2.0g, resazurin 0.001g, sodium bicarbonate 0.85g, methyl alcohol 10.0ml, Cys hydrochloride monohydrate 0.3g, nine water cure sodium 0.3g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 6.8, at 80%N
2+ 20%CO
2under anaerobic condition, the 60 ℃ of static cultivation of dark 72h.
Described vitamin solution is Biotin2.0mg, Folic acid2.0mg, Pyridoxine-HCl10.0mg, Thiamine-HCl2H
2o5.0mg, Riboflavin5.0mg, Nicotinic acid5.0mg, D-Ca-pantothenate5.0mg, Vitamin B
120.1mg, p-Aminobenzoic acid5.0mg, Lipoic acid5.0mg, adding distil water is settled to the solution of 1000.0ml.
Described trace element solution is 7.7M HCl10.0ml, FeCl
24H
2o1.5g, ZnCl
270.0mg, MnCl
24H
2o100.0mg, H
3bO
36.0mg, CoCl
26H
2o190.0mg, CuCl
22H
2o2.0mg, NiCl
26H
2o24.0mg, Na
2moO
42H
2o36.0mg, adding distil water is settled to the solution of 1000.0ml.
Bacterial strain preserving number NCIB10682, the cultivation of ATCC27405Clostridium thermocellum will be by ammonium sulfate 1.3g, magnesium chloride hexahydrate 2.6g, potassium primary phosphate 1.43g, dipotassium hydrogen phosphate 5.5g, Calcium dichloride dihydrate 0.13g, β-phospho-glycerol disodium tetrahydrate 6.00g, iron vitriol 1.10mg, reduced glutathion 0.25g, yeast powder 4.5g, resazurin 1.0mg, Microcrystalline Cellulose 10.0g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 7.2; At 80%N
2+ 20%CO
2under anaerobic condition, the 60 ℃ of static cultivation of dark 72h.
Bacterial strain preserving number DSM574, the cultivation of ATCC19858Desulfotomaculum nigrificans will be by dipotassium hydrogen phosphate 0.5g, ammonium chloride 1.0g, sodium sulfate 1.0g, Calcium dichloride dihydrate 0.1g, magnesium sulfate heptahydrate 2.0g, DL-LACTIC ACID sodium 2.0g, yeast powder 1.0g, resazurin 1.0mg, iron vitriol 0.5g, Thioglycolic acid sodium salt 0.1g, xitix 0.1g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 7.8; At 80%N
2+ 20%CO
2under anaerobic condition, the 60 ℃ of static cultivation of dark 72h.
The cultivation of bacterial strain preserving number DSM21625Geobacillus thermoglucosidasius, will be by casein peptone 15.0g, soy peptone 5.0g, and sodium-chlor 5.0g, distilled water 1000.0ml, pH value is adjusted to 7.3; The 60 ℃ of static cultivation of dark 48h.
Preserving number DSM24528Symbiobacterium thermophilum sodium-chlor 5.0g, SODIUMNITRATE 1.7g, yeast powder 5.0g, peptone 10.0g, bicarbonate of ammonia 3.2g, distilled water 1000.0ml, pH value is adjusted to 7.8; (80%N under anaerobic
2+ 20%CO
2), the 60 ℃ of static cultivation of dark 72h.
(2) compound bacteria-fermented
To after the cultured 8 strain bacteriums of step (1) proportionally mix, obtain mixed strains stoste, the mixed strains stoste that is equivalent to nutrient solution volume 5% is inoculated in fermentation culture, at 60 ℃, dark static cultivation is 5 days, obtains composite bacteria liquid;
Described fermentation culture is: peptone 5.0g, yeast powder 1.0g, calcium carbonate 3g, sodium-chlor 5g, K
2hPO
41g, MgSO
47H
2o0.35g, bacterium chaff powder 10g, distilled water 1000.0mL, pH value is 7.8,121 ℃ of sterilizing 20min.
The blending ratio of described 8 strain bacteriums is: bacterial strain preserving number ATCC51176, DSM3670Aeribacillus pallidus, bacterial strain preserving number DSM2027, ATCC7954Geobacillus stearothermophilus, bacterial strain preserving number DSM7064Brevibacillus thermoruber, bacterial strain preserving number ATCC43739, DSM2910Clostridium stercorarium subsp.thermolacticum, bacterial strain preserving number NCIB10682, ATCC27405Clostridium thermocellum, bacterial strain preserving number DSM574, ATCC19858Desulfotomaculum nigrificans, bacterial strain preserving number DSM21625Geobacillus thermoglucosidasius, bacterial strain preserving number DSM24528Symbiobacterium thermophilum cell count content in mixed strains is respectively 20%, 20%, 5%, 10%, 15%, 5%, 10% and 15%,
(3) bacterial adsorption carrier preparation: cross 0.4mm sieve after the wood chip bacterium chaff (in original culture medium of edible fungus, wood chip content accounts for 80%) of 80 ℃ of oven dry is pulverized standby;
(4) composite bacteria absorption fermentation: add 0.2% peptone that accounts for bacterium chaff grain weight amount in above-mentioned wood chip bacterium chaff powder, 0.3% fish meal, 0.25% yeast powder and 50% tap water, spray while stirring and account for the composite bacteria liquid that bacterium chaff grain weight 1% ferments in advance and be placed in fermentor tank, 60 ℃ of lucifuge heat insulating culture, within every 24 hours, uniform stirring is 1 time, every 48 hours, repeat to spray same bacterium liquid, spray altogether 3 times, the 3rd time is sprayed bacterium liquid measure is 20% of bacterium chaff grain weight, after spraying, lucifuge heat insulating culture is 12 hours, stir and be placed on 60 ℃ of oven dry, when being 20%, overall water content stops being dried, obtain composite fungus agent goods, and pack.Composite fungus agent prepared by this example to the capacity of decomposition of different bacterium chaffs in Table 1.
The capacity of decomposition of table 1 composite fungus agent to different bacterium chaffs
Note: wood chip bacterium chaff: in original culture medium of edible fungus, wood chip content accounts for 80%; Cotton seed hulls bacterium chaff: in original culture medium of edible fungus, cotton seed hulls accounts for 80%; Wood chip cotton seed hulls bacterium chaff: in original culture medium of edible fungus, wood chip content accounts for 60%, and cotton seed hulls accounts for 20%; Cotton stalk bacterium chaff: in original culture medium of edible fungus, cotton stalk accounts for 80%; Corn cob stalk bacterium chaff: in original culture medium of edible fungus, corn cob, rice straw and wheat stalk total content are 80%, and wherein corn cob accounts for 30%, and rice straw accounts for 25%, and wheat stalk accounts for 25%.
Example 2, with example 1, different,
(1) the mixed fermentation ratio of described 8 strain bacteriums is: Aeribacillus pallidus(bacterial strain preserving number ATCC51176, DSM3670), Geobacillus stearothermophilus(bacterial strain preserving number DSM2027, ATCC7954), Brevibacillus thermoruber(bacterial strain preserving number DSM7064), Clostridium stercorarium subsp.thermolacticum(bacterial strain preserving number ATCC43739, DSM2910), Clostridium thermocellum(bacterial strain preserving number NCIB10682, ATCC27405), Desulfotomaculum nigrificans(bacterial strain preserving number DSM574, ATCC19858), Geobacillus thermoglucosidasius(bacterial strain preserving number DSM21625), Symbiobacterium thermophilum(bacterial strain preserving number DSM24528) in mixed strains, cell count content is specially 12%, 14%, 3%, 14%, 17%, 10%, 14% and 16%.
Composite fungus agent prepared by this example to the capacity of decomposition of different bacterium chaffs in Table 2.
The capacity of decomposition of table 2 composite fungus agent to different bacterium chaffs
Example 3, with example 1, different,
(1) in described composite bacteria, Aeribacillus pallidus(bacterial strain preserving number ATCC51176, DSM3670), Geobacillus stearothermophilus(bacterial strain preserving number DSM2027, ATCC7954), Brevibacillus thermoruber(bacterial strain preserving number DSM7064), Clostridium stercorarium subsp.thermolacticum(bacterial strain preserving number ATCC43739, DSM2910), Clostridium thermocellum(bacterial strain preserving number NCIB10682, ATCC27405), Desulfotomaculum nigrificans(bacterial strain preserving number DSM574, ATCC19858), Geobacillus thermoglucosidasius(bacterial strain preserving number DSM21625), Symbiobacterium thermophilum(bacterial strain preserving number DSM24528) cell count content is specially 16%, 15%, 4%, 11%, 18%, 7%, 11% and 18%.
Composite fungus agent prepared by this example to the capacity of decomposition of different bacterium chaffs in Table 3.
The capacity of decomposition of table 3 composite fungus agent to different bacterium chaffs
Example 4, with example 1, different,
(1) in described composite bacteria, Aeribacillus pallidus(bacterial strain preserving number ATCC51176, DSM3670), Geobacillus stearothermophilus(bacterial strain preserving number DSM2027, ATCC7954), Brevibacillus thermoruber(bacterial strain preserving number DSM7064), Clostridium stercorarium subsp.thermolacticum(bacterial strain preserving number ATCC43739, DSM2910), Clostridium thermocellum(bacterial strain preserving number NCIB10682, ATCC27405), Desulfotomaculum nigrificans(bacterial strain preserving number DSM574, ATCC19858), Geobacillus thermoglucosidasius(bacterial strain preserving number DSM21625), Symbiobacterium thermophilum(bacterial strain preserving number DSM24528) cell count content is specially 10%, 10%, 2%, 15%, 20%, 8%, 15% and 20%.
Composite fungus agent prepared by this example to the capacity of decomposition of different bacterium chaffs in Table 4.
The capacity of decomposition of table 4 composite fungus agent to different bacterium chaffs
Example 5, with example 4, different,
(3) bacterial adsorption carrier is cotton stalk bacterium chaff (in original culture medium of edible fungus, cotton stalk accounts for 80%), and preparation method is: after the corn cob stalk bacterium chaff of 80 ℃ of oven dry is pulverized, cross 0.4mm sieve standby;
(4) composite bacteria absorption fermentation: add the urea, 0.1% fish meal, 0.1% yeast powder and 45% the tap water that account for bacterium chaff weight 3% in cotton stalk bacterium chaff powder, the composite bacteria liquid fermenting in advance that sprinkling while stirring accounts for rice straw bacterium chaff powder 1% is placed in 50 ℃ of heat insulating culture in jar fermenter, stir 1-2 every day, within every 3-4 days, spray 1 same bacterium liquid, it is 15% of rice straw bacterium chaff grain weight that the 5th is sprayed bacterium liquid measure, 60 ℃ of oven dry immediately after sprinkling, when being 20%, overall water content at once stops being dried, obtain composite fungus agent goods, packing.
Composite fungus agent prepared by this example to the capacity of decomposition of different bacterium chaffs in Table 5.
The capacity of decomposition of table 5 composite fungus agent to different bacterium chaffs
Example 6, with example 4, different is
(3) bacterial adsorption carrier is that (in original culture medium of edible fungus, corn cob, rice straw and wheat stalk total content are 80% to corn cob stalk bacterium chaff, wherein corn cob accounts for 30%, rice straw accounts for 25%, wheat stalk accounts for 25%), preparation method is: after the corn cob stalk bacterium chaff of 80 ℃ of oven dry is pulverized, cross 0.4mm sieve standby;
(4) absorption fermentation: add the urea, 0.1% fish meal, 0.1% yeast powder and 45% the tap water that account for bacterium chaff weight 3% in corn cob stalk bacterium chaff powder, the composite bacteria liquid fermenting in advance that sprinkling while stirring accounts for rice straw bacterium chaff powder 1% is placed in 50 ℃ of heat insulating culture in jar fermenter, stir 1-2 every day, within every 3-4 days, spray 1 same bacterium liquid, it is 15% of rice straw bacterium chaff grain weight that the 5th is sprayed bacterium liquid measure, 60 ℃ of oven dry immediately after sprinkling, when being 20%, overall water content at once stops being dried, obtain composite fungus agent goods, packing.
Composite fungus agent prepared by this example to the capacity of decomposition of different bacterium chaffs in Table 6.
The capacity of decomposition of table 6 composite fungus agent to different bacterium chaffs
For confirming the fermentation capacity of 8 kinds of bacteriums after compound, the applicant has made not bacterium and bacterium Clostridium thermocellum(bacterial strain preserving number NCIB10682, the ATCC27405 of number of the same race) the capacity of decomposition simultaneous test of compound composite fungus agent.In the proportioning of composite fungus agent, to enter be 1: 1 to the proportioning of each bacterium.Each bacterium is decided to be respectively according to the order of sequence: 1. Aeribacillus pallidus(bacterial strain preserving number ATCC51176, DSM3670), 2.: Geobacillus stearothermophilus(bacterial strain preserving number DSM2027, ATCC7954), 3.: Brevibacillus thermoruber(bacterial strain preserving number DSM7064), 4.: Clostridium stercorarium subsp.thermolacticum(bacterial strain preserving number ATCC43739, DSM2910), 5.: Clostridium thermocellum(bacterial strain preserving number NCIB10682, ATCC27405), 6.: Desulfotomaculum nigrificans(bacterial strain preserving number DSM574, ATCC19858), 7.: Geobacillus thermoglucosidasius(bacterial strain preserving number DSM21625), 8.: Symbiobacterium thermophilum(bacterial strain preserving number DSM24528).
Single culture and mixed culture compare in Table 7 the capacity of decomposition of bacterium chaff:
Table 7 bacterium single culture and the mixed culture capacity of decomposition to bacterium chaff
From upper table, bacterium be 5. in 8 strain bacteriums under this experiment condition unique bacterium bacterium chaff to capacity of decomposition, 5. compound bacterium is different that the capacity of decomposition of bacterium chaff is also had to difference from bacterium; 8 kinds of compound rear best capacity of decomposition that just have of bacterium.
A kind of bacterium chaff pyrolytic decomposition composite fungus agent of the present invention is composited by 8 strain thermophilic bacteriums, and composite fungus agent take that to decompose natural wooden fiber's element be principal character.Microorganism adsorption carrier of the present invention is mainly edible fungus bran, and they have good absorption property to the microorganism in this composite fungus agent.Composite fungus agent prepared by the present invention can effectively decompose various edible fungus brans.This composite fungus agent is once made edible fungi residue feed for edible fungus bran being carried out after pyrolytic decomposition is processed, and by Moderate High Temperature, decomposes, and the hemicellulose in bacterium chaff, Mierocrystalline cellulose Partial digestion, increased palatability and the property easy to digest of edible fungi residue feed; This composite fungus agent also can be used for bacterium chaff organic fertilizer production fermenting process, adds after this composite fungus agent, can accelerate bacterium chaff fertilizer maturity, shortens the organic production cycle, improves fertilizer quality.
Claims (3)
1. a bacterium chaff pyrolytic decomposition composite fungus agent fast and effectively, is characterized in that by bacterial strain preserving number ATCC51176, DSM3670Aeribacillus pallidus, bacterial strain preserving number DSM2027, ATCC7954Geobacillus stearothermophilus, bacterial strain preserving number DSM7064Brevibacillus thermoruber, bacterial strain preserving number ATCC43739, DSM2910Clostridium stercorarium subsp.thermolacticum, bacterial strain preserving number NCIB10682, ATCC27405Clostridium thermocellum, bacterial strain preserving number DSM574, ATCC19858Desulfotomaculum nigrificans, bacterial strain preserving number DSM21625Geobacillus thermoglucosidasius, bacterial strain preserving number DSM24528Symbiobacterium thermophilum8 strain thermophilic bacterium is respectively 10%-20% by the cell count content in composite fungus agent, 10%-20%, 2%-5%, 10%-15%, 15%-20%, 5%-10%, 10%-15%, the composite fungus agent of the ratio mixed fermentation gained of 15%-20%,
The implication of the cell count content of bacterial strain in mixed fermentation refers to that each strain cell number in composite fungus agent accounts for the percentage composition of the total cell count of composite bacteria.
2. the method for the preparation a kind of chaff of bacterium fast and effectively pyrolytic decomposition composite fungus agent claimed in claim 1, is characterized in that carrying out compound bacteria-fermented after 8 strain bacteriums are cultivated separately; Concrete steps are as follows:
(1) 8 strain bacterium is cultivated separately, and its cultural method is respectively:
Bacterial strain preserving number ATCC51176, the cultivation of DSM3670Aeribacillus pallidus, will be by glucose 1.0g, peptone 7.5g, extractum carnis 5.0g, yeast powder 2.5g, casamino acids 2.5g, sodium-chlor 5.0g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 8.5; The 60 ℃ of static cultivation of lucifuge 48h;
Bacterial strain preserving number DSM2027, the cultivation of ATCC7954Geobacillus stearothermophilus, will be by sodium-chlor 5.0g, extractum carnis 10.0g, peptone 10.0g, the substratum that distilled water 1000.0mL forms, pH value is adjusted: to 7.2; The 60 ℃ of static cultivation of lucifuge 48h;
The cultivation of bacterial strain preserving number DSM7064Brevibacillus thermoruber, will be by glucose 15.0g, yeast powder 5.0g, and Calcium dichloride dihydrate 0.2g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 7.0; The 60 ℃ of static cultivation of lucifuge 48h;
Bacterial strain preserving number ATCC43739, the cultivation of DSM2910Clostridium stercorarium subsp.thermolacticum, will be by dipotassium hydrogen phosphate 0.3g, potassium primary phosphate 0.2g, ammonium chloride 0.5g, magnesium sulfate heptahydrate 0.5g, Calcium dichloride dihydrate 0.25g, sodium-chlor 2.25g, iron vitriol 0.002g, vitamin solution 10.0ml, trace element solution 1.0ml, yeast powder 2.0g, casein peptone 2.0g, resazurin 0.001g, sodium bicarbonate 0.85g, methyl alcohol 10.0ml, Cys hydrochloride monohydrate 0.3g, nine water cure sodium 0.3g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 6.8, at 80%N
2+ 20%CO
2under anaerobic condition, the 60 ℃ of static cultivation of dark 72h,
Described vitamin solution is Biotin2.0mg, Folic acid2.0mg, Pyridoxine-HCl10.0mg, Thiamine-HCl2H
2o5.0mg, Riboflavin5.0mg, Nicotinic acid5.0mg, D-Ca-pantothenate5.0mg, Vitamin B
120.1mg, p-Aminobenzoic acid5.0mg, Lipoic acid5.0mg, adding distil water is settled to the solution of 1000.0ml;
Described trace element solution is 7.7M HCl10.0ml, FeCl
24H
2o1.5g, ZnCl
270.0mg, MnCl
24H
2o100.0mg, H
3bO
36.0mg, CoCl
26H
2o190.0mg, CuCl
22H
2o2.0mg, NiCl
26H
2o24.0mg, Na
2moO
42H
2o36.0mg, adding distil water is settled to the solution of 1000.0ml;
Bacterial strain preserving number NCIB10682, the cultivation of ATCC27405Clostridium thermocellum, will be by ammonium sulfate 1.3g, magnesium chloride hexahydrate 2.6g, potassium primary phosphate 1.43g, dipotassium hydrogen phosphate 5.5g, Calcium dichloride dihydrate 0.13g, β-phospho-glycerol disodium tetrahydrate 6.00g, iron vitriol 1.10mg, reduced glutathion 0.25g, yeast powder 4.5g, resazurin 1.0mg, Microcrystalline Cellulose 10.0g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 7.2; At 80%N
2+ 20%CO
2under anaerobic condition, the 60 ℃ of static cultivation of dark 72h;
Bacterial strain preserving number DSM574, the cultivation of ATCC19858Desulfotomaculum nigrificans, will be by dipotassium hydrogen phosphate 0.5g, ammonium chloride 1.0g, sodium sulfate 1.0g, Calcium dichloride dihydrate 0.1g, magnesium sulfate heptahydrate 2.0g, DL-LACTIC ACID sodium 2.0g, yeast powder 1.0g, resazurin 1.0mg, iron vitriol 0.5g, Thioglycolic acid sodium salt 0.1g, xitix 0.1g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 7.8; At 80%N
2+ 20%CO
2under anaerobic condition, the 60 ℃ of static cultivation of dark 72h;
The cultivation of bacterial strain preserving number DSM21625Geobacillus thermoglucosidasius, will be by casein peptone 15.0g, soy peptone 5.0g, and sodium-chlor 5.0g, the substratum that distilled water 1000.0ml forms, pH value is adjusted to 7.3; The 60 ℃ of static cultivation of dark 48h;
The cultivation of bacterial strain preserving number DSM24528Symbiobacterium thermophilum, will be by sodium-chlor 5.0g, SODIUMNITRATE 1.7g, and yeast powder 5.0g, peptone 10.0g, bicarbonate of ammonia 3.2g, the substratum that distilled water 1000.0ml forms,, pH value is adjusted to 7.8; At 80%N
2+ 20%CO
2under anaerobic condition, the 60 ℃ of static cultivation of dark 72h;
(2) compound bacteria-fermented
To after the cultured 8 strain bacteriums of step (1) proportionally mix, obtain mixed strains stoste, the mixed strains stoste that is equivalent to nutrient solution volume 5% is inoculated in fermentation culture, at 60 ℃, dark static cultivation is 5 days, obtains composite bacteria liquid;
Described mixing fermentation culture liquid is: peptone 5.0g, yeast powder 1.0g, calcium carbonate 3g, sodium-chlor 5g, K
2hPO
41g, MgSO
47H
2o0.35g, bacterium chaff powder 10g, distilled water 1000.0mL, pH value is 7.8,121 ℃ of sterilizing 20min;
The blending ratio of described 8 strain bacteriums is: bacterial strain preserving number ATCC51176, DSM3670Aeribacillus pallidus, bacterial strain preserving number DSM2027, ATCC7954Geobacillus stearothermophilus, bacterial strain preserving number DSM7064Brevibacillus thermoruber, bacterial strain preserving number ATCC43739, DSM2910Clostridium stercorarium subsp.thermolacticum, bacterial strain preserving number NCIB10682, ATCC27405Clostridium thermocellum, bacterial strain preserving number DSM574, ATCC19858Desulfotomaculum nigrificans, bacterial strain preserving number DSM21625Geobacillus thermoglucosidasius, bacterial strain preserving number DSM24528Symbiobacterium thermophilum cell count content in mixed strains is respectively 10%-20%, 10%-20%, 2%-5%, 10%-15%, 15%-20%, 5%-10%, 10%-15% and 15%-20%,
(3) preparation of bacterial adsorption carrier: after the bacterium chaff of 80 ℃ of oven dry is pulverized, cross 0.4mm sieve standby;
(4) composite bacteria absorption fermentation: add 0.2% peptone that accounts for bacterium chaff grain weight amount in the bacterium chaff powder of step (3), 0.3% fish meal, 0.25% yeast powder and 50% tap water, sprinkling while stirring accounts for bacterium chaff grain weight 1%, composite bacteria liquid by step (2) gained, be placed in fermentor tank, 60 ℃ of lucifuge heat insulating culture, within every 24 hours, uniform stirring is 1 time, every 48 hours, repeat to spray same bacterium liquid, spray altogether 3 times, the 3rd time is sprayed bacterium liquid measure is 20% of bacterium chaff grain weight, after spraying, lucifuge heat insulating culture is 12 hours, stir and be placed on 60 ℃ of oven dry, when being 20%, overall water content stops being dried, obtain composite fungus agent goods.
3. the method for a kind of chaff of bacterium fast and effectively pyrolytic decomposition composite fungus agent of preparation according to claim 2, is characterized in that bacterial adsorption carrier is wood chip bacterium chaff, cotton seed hulls bacterium chaff, wood chip cotton seed hulls bacterium chaff, cotton stalk bacterium chaff, corn cob stalk bacterium chaff.
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