CN101928689A - Method for industrially preparing Rhodobacter sphaeroides, fermentation liquor of Rhodobacter sphaeroides and application - Google Patents
Method for industrially preparing Rhodobacter sphaeroides, fermentation liquor of Rhodobacter sphaeroides and application Download PDFInfo
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Abstract
The invention relates to bacteria, a production method for preparing a soil water flush microbial fertilizer and a feed additive through fermentation of the bacteria and application of fermentation liquor, in particular to Rhodobacter sphaeroides, a production method for preparing the soil water flush microbial fertilizer and the feed additive through the Rhodobacter sphaeroides and application of the fermentation liquor. The number of the Rhodobacter sphaeroides registered by a China microbial culture collection center is CGMCC NO.3757. Compared with the final products of two preceding patents of the inventor, the fermentation liquor has the advantages of that: the survival rate of the obtained fermentation liquor in soil is increased by 52 to 79 percent compared with that of lighting culture fermentation liquor in soil; the super oxide dlsmutase (SOD) specific activity of extracellular products is increased by 316 percent; the intracellular SOD specific activity is increased by 230 percent; and the fermentation liquor serving as the microbial fertilizer is applied to crop production, so the quality of the crops is effectively improved and the yield is increased. Harmful substances such as nitrate, nitrite and the like in vegetable are degraded. The fermentation liquor has important significance for producing agricultural food safely and guaranteeing human health. The fermentation liquor serving as the feed additive is applied to livestock and bird breed and aquaculture, so growth-promoting, disease-resistant and disease-preventing effects are more obvious.
Description
Technical field
The present invention relates to a kind of bacterium and utilize fermentation using bacteria to prepare the production method of functional fermented liquid and the purposes of fermented liquid, the particularly red bacterium of class ball (Rhodobacter sphaeroides) and utilize its production method for preparing functional fermented liquid, and the purposes of this fermented liquid.
Background technology
At present, because the research of the red bacterium of class ball more and more is subjected to the attention of plant husbandry, aquaculture and field of environment protection, its Application Areas more and more widely.The contriver carries out finding in the suitability for industrialized production application that in former patented technology present technique also should have the extensive space of continuing innovation.The product that the present invention produces more adapts to the field planting survival better under soil and dark condition of the red bacterium of class ball; Bring into play better effect.
Summary of the invention
The purpose of this invention is to provide the red bacterium of a kind ball (Rhodobacter sphaeroides) and utilize it to carry out the fermentation process of the industrially scalable of deep layer, and the fermented liquid that obtains of this method, it is healthy and helpful to human body, can regulate the immunity system of human body, increase resistance of human body.
The present invention relates to the contriver on the basis of its serial patent of invention (Chinese CN1184307C) and (Chinese ZL200510132627.3), according to test and research for many years, bacterial classification is carried out purifying, inducing culture, zymotechnique is transformed; That releases more adapts to as environment-friendly type new fertilizer, novel fodder additive and purifying water body better effects if, application product that cost is lower.In the inventor's series of patents (Chinese CN1184307C), related to similar fermentation process, but through the continuation in several years of contriver test, optimized the composition of fermentation culture, and optimized fermentation condition, its meta-bolites SOD is significantly improved, obtained better effects if, fermented liquid that cost is lower.
With respect to the finished product in two parts of patents before the contriver, the fermented liquid that obtains improves 52%--79% than illumination cultivation liquid in the survival rate of soil, and extracellular products SOD improves 316% than vigor; SOD improves 230% than vigor in the born of the same parents, more helps being applied to plant husbandry and being applied to poultry, fowl breed and aquaculture as fodder additives as microbial fertilizer, and its promotion growth and defending and fighting against diseases effect are more remarkable.
The inventive method is: a kind of utilize the red bacterium of class ball (Rh0d0bacter sphaeroides) Chinese microorganism strain preservation center register on the books the numbering CGMCCNO.3757, after using LB (Luria-Bertani) substratum to add micro-lucifuge inducing culture, mass-producing lucifuge fermentation process is characterized in that: this method is that the red bacterium of class ball (Rh0d0bacter sphaeroides) is passed through 100ml-500ml Erlenmeyer flask lucifuge inducing culture successively, the first class seed pot of 30L-50L, 300L-500L secondary seed jar lucifuge enlarged culturing, the fermentation of 1000L-2000L one-level lucifuge, the fermentation of 3000L-6000L secondary lucifuge, the fermentation of 8000L-16000L terminal lucifuge; Carrying out two-stage seed lucifuge cultivates; The method of three grades of lucifuge fermentations.
The red bacterium of a kind ball of the present invention is for being preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, numbering CGMCC No.3757, preservation time: on April 20th, 2010.
A kind of industrially scalable fermentation process that utilizes the red bacterium of above-mentioned class ball of the present invention, wherein, described bacterial classification is passed through the first class seed pot of 100ml-500ml Erlenmeyer flask, 30L-50L, secondary seed jar, 1000L-2000L one grade fermemtation jar, 3000L-6000L second order fermentation jar and the 8000L-16000L terminal fermentor tank of 300L-500L successively, promptly obtain fermented liquid.
Above-mentioned fermentation condition is respectively:
The 100ml-500ml Erlenmeyer flask, 121 ℃ of sterilizations are cooled to 35 ℃ after the purifying box inoculation, with the tight flask of tinfoil paper paper bag, last shaking table 80-100 commentaries on classics/min; Constant temperature 30-34 ℃; Carry out 36h lucifuge inducing culture;
The first order seed culture tank of 30L-50L; Inoculum size is greater than 10%, PH7.0-7.2, and constant temperature 30-34 ℃, the bottom feeds purified compressed air, opens the tank top exhaust-valve slightly; Make jar internal pressure keep 0.02-0.03Mpa, close the visor lamp, carry out lucifuge and cultivate 21h;
300L-500L secondary seed jar, technological operation such as first class seed pot carry out enlarged culturing and cultivate 21h;
Close 300L-500L secondary seed tank top purging valve; Pressure by purified compressed air is sent into 1000L-2000L one grade fermemtation jar by transport pipe; Inoculum size is greater than 20%; Technological operation such as secondary seed jar carry out one-level lucifuge fermentation 10-12h;
3000L-6000L second order fermentation jar, inoculum size is greater than 25%; Technological operation such as first class seed pot carry out secondary lucifuge fermentation 8h;
8000L-16000L terminal fermentor tank, inoculum size be greater than 30%, and nutrient solution prescription, PH, temperature, venting pressure, the lucifuge operating procedure of its fermentation condition in from the 30L-50L first class seed pot to 8000L-16000L terminal fermentor tank is identical.
The used substratum proportioning of 100ml-500ml Erlenmeyer flask: (every 1000ml substratum, all the other are water)
LB substratum 20-25 gram;
Trace element solution 3-5ml;
Deionized water 950ml;
Adjust PH7.0-7.2 with the 12N sodium hydroxide solution;
Be settled to 1000ml with deionized water.
Wherein said trace element solution proportioning: (1000ml, all the other are water)
Boric acid 300-500 milligram
Ferrous sulfate 200-300 milligram
Manganous sulfate 80-100 milligram
Zinc sulfate 200-300 milligram
Sodium orthomolybdate 100-300 milligram
Copper sulfate 80-100 milligram
Above-mentioned 30L-50L first class seed pot, 300L-500L secondary seed jar, 1000L-2000L one grade fermemtation jar, 3000L-6000L second order fermentation jar, the used fermentation culture proportioning of 8000L-16000L terminal fermentor tank: (1000L, all the other are water):
Ammonium sulfate 1000-1200 gram sal epsom 180-220 gram ferrous sulfate 20-30 gram manganous sulfate 1-2 gram zinc sulfate 20-40 gram copper sulfate 0.1-0.2 gram boric acid 30-50 gram Sodium orthomolybdate 20-30 gram calcium chloride 80-100 gram vitamin H 3-4 milligram disodium ethylene diamine tetraacetate 10-20 gram para-amino benzoic acid 1-2 gram acetate 3.2-4.0 rises VITMAIN B1, Wei ShengsuB2, vitamin B6, each 1 gram sodium hydroxide 1400-1600 gram of nicotinic acid.
The purposes of the red ferment product of class ball of the present invention wherein, is irrigated butt rot place with 20-50 times of fermented liquid and can be treated root rot; Or, with 50-100 doubly fermented liquid spray water fruits and can eat vegetables raw, the SOD of fruit is improved; Or, with 100-150 fermented liquid doubly as the viable bacteria biomaterial; Or, give and to be breeded fish, add 3%-5% in the feed of shrimp or crab, can purify water, reduce disease, improve the speed of growth; Or fermented liquid and water are according to 1: 5-10 adds doubly in the feed that is added to milk cow, pig, chicken or the tap water and feeds.
The SOD enzyme activity of product of the present invention reaches 60.77-119.8u/ml, has improved 230%-316% than illumination fermented liquid.
Product of the present invention sprays vegetables with 50-100 times of lucifuge fermented liquid can degrade nitrate, nitrite, is of value to vegetables safety in production and HUMAN HEALTH.
The bacterial classification source:
In Penglai City, Shandong Province village in the epimere mud of market town Song Jia river.
Separation method:
In getting korneforos mud 500ml the 7-8 month, with prepared culture 2000ml according to the method described above, on average pack in 10 1000ml Erlenmeyer flasks, in the good bottle of bundle tampon plug, external application kraft paper is wrapped, tighten with paper string, place high-temperature sterilization pot internal heating to 121 ℃ to carry out sterilization in 30 minutes after, reduce to 28 ℃-35 ℃.Divide to go into 10 flask culture liquid with mud at leisure respectively fifty-fifty in the stirring of sterilisable chamber inner edge.Regulate pH7.5-8.0 with 10% lactic acid solution then, good with the tampon plug on the former bottle, wrapping kraft paper tightens with paper string, place on 28 ℃ of-35 ℃ of constant temperature shaking tables, shaking speed per minute 120-150 commentaries on classics is shaken, shaking table was cultivated after 40 hours, get the nutrient solution (getting upper strata liquid) of red reaction, move into culture dish (substratum of culture dish is the LB substratum) at the indoor transfer pipet of inoculation, place and cultivate 96h under the above-mentioned culture condition, and then from culture dish, in sterilisable chamber, get the bigger bacterium colony of red point and insert in the LB liquid medium, enter and hold up strong the cultivation, carry out tens of times separation and Culture so repeatedly, hold up strong, the sampling microscopy, reach do not have an assorted bacterium purebred can be produce with kind.
The red bacterium of class ball of the present invention: for being preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, numbering CGMCC No.3757, preservation time: on April 20th, 2010.
Beneficial effect:
With respect to the finished product in two parts of patents before the contriver, the fermented liquid that obtains improves 52%--79% than illumination cultivation liquid in the survival rate of soil, and extracellular products SOD improves 316% than vigor; SOD improves 230% than vigor in the born of the same parents, more helps being applied to plant husbandry and being applied to poultry, fowl breed and aquaculture as fodder additives as microbial fertilizer, and its promotion growth and defending and fighting against diseases effect are more remarkable.
Embodiment 1
Numbering CGMCCNO.3757 registers on the books at Chinese microorganism strain preservation center, described bacterial classification is passed through the first class seed pot of 100ml-500ml Erlenmeyer flask, 30L-50L, secondary seed jar, 1000L-2000L one grade fermemtation jar, 3000L-6000L second order fermentation jar and the 8000L-16000L terminal fermentor tank of 300L-500L successively, promptly obtain fermented liquid.
Above-mentioned fermentation condition is respectively:
The 100ml-500ml Erlenmeyer flask, 121 ℃ of sterilizations are cooled to 35 ℃ after the purifying box inoculation, with the tight flask of tinfoil paper paper bag, last shaking table 100 commentaries on classics/min; 30 ℃ of constant temperature; Carry out 36h lucifuge inducing culture;
The first order seed culture tank of 30L-50L; Inoculum size is greater than 10%, PH7.2, and 34 ℃ of constant temperature, the bottom feeds purified compressed air, opens the tank top exhaust-valve slightly; Make jar internal pressure keep 0.03Mpa, close the visor lamp, carry out lucifuge and cultivate 21h;
300L-500L secondary seed jar, technological operation such as first class seed pot carry out enlarged culturing and cultivate 21h;
Close 300L-500L secondary seed tank top purging valve; Pressure by purified compressed air is sent into 1000L-2000L one grade fermemtation jar by transport pipe; Inoculum size is greater than 20%; Technological operation such as secondary seed jar carry out one-level lucifuge fermentation 12h;
3000L-6000L second order fermentation jar, inoculum size is greater than 25%; Technological operation such as first class seed pot carry out secondary lucifuge fermentation 8h;
8000L-16000L terminal fermentor tank, inoculum size be greater than 30%, and nutrient solution prescription, PH, temperature, venting pressure, the lucifuge operating procedure of its fermentation condition in from the 30L-50L first class seed pot to 8000L-16000L terminal fermentor tank is identical.
The used substratum proportioning of 100ml-500ml Erlenmeyer flask: (every 1000ml substratum)
LB substratum 25 grams;
Trace element solution 5ml;
Deionized water 950ml;
Adjust PH7.2 with the 12N sodium hydroxide solution;
Be settled to 1000ml with deionized water.
Wherein said trace element solution proportioning: (1000ml)
500 milligrams of boric acid
300 milligrams in ferrous sulfate
100 milligrams of manganous sulfates
300 milligrams in zinc sulfate
300 milligrams of Sodium orthomolybdates
100 milligrams in copper sulfate
Above-mentioned 30L-50L first class seed pot, 300L-500L secondary seed jar, 1000L-2000L one grade fermemtation jar, 3000L-6000L second order fermentation jar, the used fermentation culture proportioning of 8000L-16000L terminal fermentor tank: (1000L):
4.0 liters of VITMAIN B1 of ammonium sulfate 1200 gram sal epsom 220 gram ferrous sulfate, 30 gram manganous sulfate 2 gram zinc sulfate, 40 gram copper sulfate 0.2 gram boric acid, 50 gram Sodium orthomolybdate 30 gram calcium chloride, 100 gram vitamin Hs 4 milligrams of disodium ethylene diamine tetraacetate, 20 gram para-amino benzoic acid, 2 gram acetate, Wei ShengsuB2, vitamin B6, each 1 gram sodium hydroxide, 1600 gram of nicotinic acid.
Embodiment 2
Numbering CGMCCNO.3757 registers on the books at Chinese microorganism strain preservation center, described bacterial classification is passed through the first class seed pot of 100ml-500ml Erlenmeyer flask, 30L-50L, secondary seed jar, 1000L-2000L one grade fermemtation jar, 3000L-6000L second order fermentation jar and the 8000L-16000L terminal fermentor tank of 300L-500L successively, promptly obtain fermented liquid.
Above-mentioned fermentation condition is respectively:
The 100ml-500ml Erlenmeyer flask, 121 ℃ of sterilizations are cooled to 35 ℃ after the purifying box inoculation, with the tight flask of tinfoil paper paper bag, last shaking table 80 commentaries on classics/min; 34 ℃ of constant temperature; Carry out 36h lucifuge inducing culture;
The first order seed culture tank of 30L-50L; Inoculum size is greater than 10%, PH7.0, and 30 ℃ of constant temperature, the bottom feeds purified compressed air, opens the tank top exhaust-valve slightly; Make jar internal pressure keep 0.02Mpa, close the visor lamp, carry out lucifuge and cultivate 21h;
300L-500L secondary seed jar, technological operation such as first class seed pot carry out enlarged culturing and cultivate 21h;
Close 300L-500L secondary seed tank top purging valve; Pressure by purified compressed air is sent into 1000L-2000L one grade fermemtation jar by transport pipe; Inoculum size is greater than 20%; Technological operation such as secondary seed jar carry out one-level lucifuge fermentation 10h;
3000L-6000L second order fermentation jar, inoculum size is greater than 25%; Technological operation such as first class seed pot carry out secondary lucifuge fermentation 8h;
8000L-16000L terminal fermentor tank, inoculum size be greater than 30%, and nutrient solution prescription, PH, temperature, venting pressure, the lucifuge operating procedure of its fermentation condition in from the 30L-50L first class seed pot to 8000L-16000L terminal fermentor tank is identical.
The used substratum proportioning of 100ml-500ml Erlenmeyer flask: (every 1000ml substratum)
LB substratum 20 grams;
Trace element solution 3ml;
Deionized water 950ml;
Adjust PH7.0 with the 12N sodium hydroxide solution;
Be settled to 1000ml with deionized water.
Wherein said trace element solution proportioning: (1000ml)
300 milligrams of boric acid
200 milligrams in ferrous sulfate
80 milligrams of manganous sulfates
200 milligrams in zinc sulfate
100 milligrams of Sodium orthomolybdates
80 milligrams in copper sulfate
Above-mentioned 30L-50L first class seed pot, 300L-500L secondary seed jar, 1000L-2000L one grade fermemtation jar, 3000L-6000L second order fermentation jar, the used fermentation culture proportioning of 8000L-16000L terminal fermentor tank: (1000L):
3.2 liters of VITMAIN B1 of ammonium sulfate 1000 gram sal epsom 180 gram ferrous sulfate, 20 gram manganous sulfate 1 gram zinc sulfate, 20 gram copper sulfate 0.1 gram boric acid, 30 gram Sodium orthomolybdate 20 gram calcium chloride, 80 gram vitamin Hs 3 milligrams of disodium ethylene diamine tetraacetate, 10 gram para-amino benzoic acid, 1 gram acetate, Wei ShengsuB2, vitamin B6, each 1 gram sodium hydroxide, 1400 gram of nicotinic acid.
Embodiment 3
Numbering CGMCCNO.3757 registers on the books at Chinese microorganism strain preservation center, described bacterial classification is passed through the first class seed pot of 100ml-500ml Erlenmeyer flask, 30L-50L, secondary seed jar, 1000L-2000L one grade fermemtation jar, 3000L-6000L second order fermentation jar and the 8000L-16000L terminal fermentor tank of 300L-500L successively, promptly obtain fermented liquid.
Above-mentioned fermentation condition is respectively:
The 100ml-500ml Erlenmeyer flask, 121 ℃ of sterilizations are cooled to 35 ℃ after the purifying box inoculation, with the tight flask of tinfoil paper paper bag, last shaking table 90 commentaries on classics/min; 32 ℃ of constant temperature; Carry out 36h lucifuge inducing culture;
The first order seed culture tank of 30L-50L; Inoculum size is greater than 10%, PH7.1, and 32 ℃ of constant temperature, the bottom feeds purified compressed air, opens the tank top exhaust-valve slightly; Make jar internal pressure keep 0.025Mpa, close the visor lamp, carry out lucifuge and cultivate 21h;
300L-500L secondary seed jar, technological operation such as first class seed pot carry out enlarged culturing and cultivate 21h;
Close 300L-500L secondary seed tank top purging valve; Pressure by purified compressed air is sent into 1000L-2000L one grade fermemtation jar by transport pipe; Inoculum size is greater than 20%; Technological operation such as secondary seed jar carry out one-level lucifuge fermentation 11h;
3000L-6000L second order fermentation jar, inoculum size is greater than 25%; Technological operation such as first class seed pot carry out secondary lucifuge fermentation 8h;
8000L-16000L terminal fermentor tank, inoculum size be greater than 30%, and nutrient solution prescription, PH, temperature, venting pressure, the lucifuge operating procedure of its fermentation condition in from the 30L-50L first class seed pot to 8000L-16000L terminal fermentor tank is identical.
The used substratum proportioning of 100ml-500ml Erlenmeyer flask: (every 1000ml substratum)
LB substratum 22 grams;
Trace element solution 4ml;
Deionized water 950ml;
Adjust PH7.1 with the 12N sodium hydroxide solution;
Be settled to 1000ml with deionized water.
Wherein said trace element solution proportioning: (1000ml)
400 milligrams of boric acid
250 milligrams in ferrous sulfate
90 milligrams of manganous sulfates
250 milligrams in zinc sulfate
200 milligrams of Sodium orthomolybdates
90 milligrams in copper sulfate
Above-mentioned 30L-50L first class seed pot, 300L-500L secondary seed jar, 1000L-2000L one grade fermemtation jar, 3000L-6000L level fermentor tank, the used fermentation culture proportioning of 8000L-16000L terminal fermentor tank: (1000L):
3.6 liters of VITMAIN B1 of ammonium sulfate 1100 gram sal epsom 200 gram ferrous sulfate, 25 gram manganous sulfate 1.5 gram zinc sulfate, 30 gram copper sulfate 0.15 gram boric acid, 40 gram Sodium orthomolybdate 25 gram calcium chloride, 90 gram vitamin Hs 3.5 milligrams of disodium ethylene diamine tetraacetate, 15 gram para-amino benzoic acid, 1.5 gram acetate, Wei ShengsuB2, vitamin B6, each 1 gram sodium hydroxide, 1500 gram of nicotinic acid.
Embodiment 4
Numbering CGMCCNO.3757 registers on the books at Chinese microorganism strain preservation center, described bacterial classification is passed through the first class seed pot of 100ml-500ml Erlenmeyer flask, 30L-50L, secondary seed jar, 1000L-2000L one grade fermemtation jar, 3000L-6000L second order fermentation jar and the 8000L-16000L terminal fermentor tank of 300L-500L successively, promptly obtain fermented liquid.
Above-mentioned fermentation condition is respectively:
The 100ml-500ml Erlenmeyer flask, 121 ℃ of sterilizations are cooled to 35 ℃ after the purifying box inoculation, with the tight flask of tinfoil paper paper bag, last shaking table 80 commentaries on classics/min; 30 ℃ of constant temperature; Carry out 36h lucifuge inducing culture;
The first order seed culture tank of 30L-50L; Inoculum size is greater than 10%, PH7.2, and 34 ℃ of constant temperature, the bottom feeds purified compressed air, opens the tank top exhaust-valve slightly; Make jar internal pressure keep 0.03Mpa, close the visor lamp, carry out lucifuge and cultivate 21h;
300L-500L secondary seed jar, technological operation such as first class seed pot carry out enlarged culturing and cultivate 21h;
Close 300L-500L secondary seed tank top purging valve; Pressure by purified compressed air is sent into 1000L-2000L one grade fermemtation jar by transport pipe; Inoculum size is greater than 20%; Technological operation such as secondary seed jar carry out one-level lucifuge fermentation 10h;
3000L-6000L second order fermentation jar, inoculum size is greater than 25%; Technological operation such as first class seed pot carry out secondary lucifuge fermentation 8h;
8000L-16000L terminal fermentor tank, inoculum size be greater than 30%, and nutrient solution prescription, PH, temperature, venting pressure, the lucifuge operating procedure of its fermentation condition in from the 30L-50L first class seed pot to 8000L-16000L terminal fermentor tank is identical.
The used substratum proportioning of 100ml-500ml Erlenmeyer flask: (every 1000ml substratum)
LB substratum 24 grams;
Trace element solution 4ml;
Deionized water 950ml;
Adjust PH7.0 with the 12N sodium hydroxide solution;
Be settled to 1000ml with deionized water.
Wherein said trace element solution proportioning: (1000ml)
500 milligrams of boric acid
200 milligrams in ferrous sulfate
100 milligrams of manganous sulfates
200 milligrams in zinc sulfate
300 milligrams of Sodium orthomolybdates
80 milligrams in copper sulfate
Above-mentioned 30L-50L first class seed pot, 300L-500L secondary seed jar, 1000L-2000L one grade fermemtation jar, 3000L-6000L second order fermentation jar, the used fermentation culture proportioning of 8000L-16000L terminal fermentor tank: (1000L):
4.0 liters of VITMAIN B1 of ammonium sulfate 1200 gram sal epsom 180 gram ferrous sulfate, 30 gram manganous sulfate 1 gram zinc sulfate, 40 gram copper sulfate 0.1 gram boric acid, 50 gram Sodium orthomolybdate 20 gram calcium chloride, 100 gram vitamin Hs 3 milligrams of disodium ethylene diamine tetraacetate, 20 gram para-amino benzoic acid, 1 gram acetate, Wei ShengsuB2, vitamin B6, each 1 gram sodium hydroxide, 1600 gram of nicotinic acid.
Embodiment 5
Numbering CGMCCNO.3757 registers on the books at Chinese microorganism strain preservation center, described bacterial classification is passed through the first class seed pot of 100ml-500ml Erlenmeyer flask, 30L-50L, secondary seed jar, 1000L-2000L one grade fermemtation jar, 3000L-6000L second order fermentation jar and the 8000L-16000L terminal fermentor tank of 300L-500L successively, promptly obtain fermented liquid.
Above-mentioned fermentation condition is respectively:
The 100ml-500ml Erlenmeyer flask, 121 ℃ of sterilizations are cooled to 35 ℃ after the purifying box inoculation, with the tight flask of tinfoil paper paper bag, last shaking table 900 commentaries on classics/min; 30 ℃ of constant temperature; Carry out 36h lucifuge inducing culture;
The first order seed culture tank of 30L-50L; Inoculum size is greater than 10%, PH7.0, and 31 ℃ of constant temperature, the bottom feeds purified compressed air, opens the tank top exhaust-valve slightly; Make jar internal pressure keep 0.028Mpa, close the visor lamp, carry out lucifuge and cultivate 21h;
300L-500L secondary seed jar, technological operation such as first class seed pot carry out enlarged culturing and cultivate 21h;
Close 300L-500L secondary seed tank top purging valve; Pressure by purified compressed air is sent into 1000L-2000L one grade fermemtation jar by transport pipe; Inoculum size is greater than 20%; Technological operation such as secondary seed jar carry out one-level lucifuge fermentation 10h;
3000L-6000L second order fermentation jar, inoculum size is greater than 25%; Technological operation such as first class seed pot carry out secondary lucifuge fermentation 8h;
8000L-16000L terminal fermentor tank, inoculum size be greater than 30%, and nutrient solution prescription, PH, temperature, venting pressure, the lucifuge operating procedure of its fermentation condition in from the 30L-50L first class seed pot to 8000L-16000L terminal fermentor tank is identical.
The used substratum proportioning of 100ml-500ml Erlenmeyer flask: (every 1000ml substratum)
LB substratum 20 grams;
Trace element solution 5ml;
Deionized water 950ml;
Adjust PH7.2 with the 12N sodium hydroxide solution;
Be settled to 1000ml with deionized water.
Wherein said trace element solution proportioning: (1000ml)
500 milligrams of boric acid
250 milligrams in ferrous sulfate
80 milligrams of manganous sulfates
250 milligrams in zinc sulfate
190 milligrams of Sodium orthomolybdates
85 milligrams in copper sulfate
Above-mentioned 30L-50L first class seed pot, 300L-500L secondary seed jar, 1000L-2000L one grade fermemtation jar, 3000L-6000L second order fermentation jar, the used fermentation culture proportioning of 8000L-16000L terminal fermentor tank: (1000L):
3.8 liters of VITMAIN B1 of ammonium sulfate 1050 gram sal epsom 1850 gram ferrous sulfate, 28 gram manganous sulfate 1.8 gram zinc sulfate, 22 gram copper sulfate 0.1 gram boric acid, 35 gram Sodium orthomolybdate 22 gram calcium chloride, 85 gram vitamin Hs 3 milligrams of disodium ethylene diamine tetraacetate, 18 gram para-amino benzoic acid, 1.8 gram acetate, Wei ShengsuB2, vitamin B6, each 1 gram sodium hydroxide, 1450 gram of nicotinic acid.
Concrete application and detected result are as follows:
Detect unit: Ecological Envionment Research Centre, Chinese Academy of Sciences
The people is finished in detection: Bai Zhihui
Class ball red bacterium illumination fermented liquid and lucifuge fermented liquid are to the influence of plant growth and the nitrate nitrite degradation is measured
| Remarks: test period is executed urea 6 days-June 28 May in 2009: imposed in 2 weeks after germination and execute bacterium liquid: be to use at seeding time for the first time, for the second time be to use strain after 1 day at applied nitrogen to weigh: when only executing fermented liquid, shining of the ratio light of lucifuge improves 10%; When applying fermented liquid and urea simultaneously, raising 14% plant height of the ratio illumination of lucifuge: when only executing fermented liquid, the raising 9% of the ratio illumination of lucifuge; When applying fermented liquid and urea simultaneously, the raising 8% of the ratio illumination of lucifuge |
| Nitrate is degraded: with contrasting of use urea, the illumination fermented liquid is 18.8 times to the degraded of nitrate; And the degraded of lucifuge fermented liquid is 28 times.Be better than the illumination fermented liquid.Nitrite degradation: with the do contrast illumination liquid that uses urea be 23 times to the degraded of nitrite; And the degraded of lucifuge fermented liquid is 38 times.Obviously be better than the illumination fermented liquid. |
Detect unit: Ecological Envionment Research Centre, Chinese Academy of Sciences
The people is finished in detection: Zhao Kai
Red bacterium illumination fermentation of class ball and lucifuge fermentation SOD measured value
Deadline: on November 9th, 2008
The red bacterium of class ball promotes the leaf romaine lettuce upgrowth situation that looses
Detect unit: Ecological Envionment Research Centre, Chinese Academy of Sciences
The people is finished in detection: Bai Zhihui Han Qing jasmine
The influence of class ball red bacterium illumination fermented liquid of the present invention and lucifuge fermented liquid survival test and flora in plant rhizosphere district soil:
Under average 22 degrees centigrade of (daytime) conditions of the soil moisture, test and showed in 30 days: the red bacterium of class ball improves 52% at the corn of the ratio illumination fermentation of the survival rate lucifuge fermentation of soil, soybean improves 79%, the probiotic bacterium that vinelandii, actinomycetes, root nodule bacterium etc. are helped plant-growth all has raising in various degree, and the filamentous fungus that causes crop pest has been shown restraining effect in various degree, its mechanism of action remains further to be studied.
More than experiment detects unit:
Ecological Envionment Research Centre, Chinese Academy of Sciences
Finish the people: the auspicious Li Lei of Zhao Zhi
Deadline: 2009.5.9-9.22
Experiment place: east, Wang Tujing village, Haidian District Beijing northeast
Claims (10)
1. the red bacterium of a kind ball is characterized in that: the red bacterium of described class ball is for being preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, numbering CGMCC No.3757, preservation time: on April 20th, 2010.
2. one kind is utilized the industrially scalable fermentation process of the red bacterium of class ball according to claim 1, it is characterized in that: with described preferable methods is the bacterial classification first class seed pot that passes through 100ml-500ml Erlenmeyer flask, 30L-50L successively, secondary seed jar, 1000L-2000L one grade fermemtation jar, 3000L-6000L second order fermentation jar and the 8000L-16000L terminal fermentor tank of 300L-500L, promptly obtains fermented liquid.
3. method as claimed in claim 2: it is characterized in that:
Fermentation condition is respectively:
The 100ml-500ml Erlenmeyer flask, 121 ℃ of sterilizations are cooled to 35 ℃ after the purifying box inoculation, with the tight flask of tinfoil paper paper bag, last shaking table 80-100 commentaries on classics/min; Constant temperature 30-34 ℃; Carry out 36h lucifuge inducing culture; The first order seed culture tank of 30L-50L; Inoculum size is greater than 10%, PH7.0-7.2, and constant temperature 30-34 ℃, the bottom feeds purified compressed air, opens the tank top exhaust-valve slightly; Make jar internal pressure keep 0.02-0.03Mpa, close the visor lamp, carry out lucifuge and cultivate 21h.
300L-500L secondary seed jar, technological operation such as first class seed pot carry out enlarged culturing and cultivate 21h.
Close 300L-500L secondary seed tank top purging valve; Pressure by purified compressed air is by transfer lime
1000L-2000L one grade fermemtation jar is sent in the road; Inoculum size is greater than 20%; Technological operation such as secondary seed jar carry out one-level lucifuge fermentation 10-12h.
3000L-6000L second order fermentation jar, inoculum size is greater than 25%; Technological operation such as first class seed pot carry out secondary lucifuge fermentation 8h.
8000L-16000L terminal fermentor tank, inoculum size be greater than 30%, and nutrient solution prescription, PH, temperature, venting pressure, the lucifuge operating procedure of its fermentation condition in from the 30L-50L first class seed pot to 8000L-16000L terminal fermentor tank is identical.
4. as claim 2 or 3 described methods: it is characterized in that:
The used substratum proportioning of 100ml-500ml Erlenmeyer flask: (every 1000ml substratum, all the other are water)
LB substratum 20-25 gram;
Trace element solution 3-5ml;
Deionized water 950ml;
Adjust PH7.0-7.2 with the 12N sodium hydroxide solution;
Be settled to 1000ml with deionized water.
5. method as claimed in claim 4: it is characterized in that: described trace element solution proportioning: (1000ml, all the other are water)
Boric acid 500-800 milligram
Ferrous sulfate 200-300 milligram
Manganous sulfate 80-100 milligram
Zinc sulfate 200-300 milligram
Sodium orthomolybdate 100-300 milligram
Copper sulfate 80-100 milligram
6. as claim 2 or 3 described methods: it is characterized in that:
30L-50L first class seed pot, 300L-500L secondary seed jar, 1000L-2000L one grade fermemtation jar, 3000L-6000L second order fermentation jar, the used fermentation culture proportioning of 8000L-16000L terminal fermentor tank: (1000L, all the other are water):
Ammonium sulfate 1000-1200 gram sal epsom 180-220 gram ferrous sulfate 20-30 gram manganous sulfate 1-2 gram zinc sulfate 20-40 gram copper sulfate 0.1-0.2 gram boric acid 30-50 gram Sodium orthomolybdate 20-30 gram calcium chloride 80-100 gram vitamin H 3-4 milligram disodium ethylene diamine tetraacetate 10-20 gram para-amino benzoic acid 1-2 gram acetate 3.2-4.0 rises VITMAIN B1, Wei ShengsuB2, vitamin B6, each 1 gram sodium hydroxide 1400-1600 gram of nicotinic acid.
7. the red ferment product of a kind ball is characterized in that: utilize among the claim 2-6 any described method of the red bacterium industrially scalable fermentation of class ball of utilizing to produce.
8. the purposes of fermented liquid described in claim 7 is characterized in that: can treat root rot with butt rot place of 20-50 times of fermented liquid irrigating plant.
9. the purposes of fermented liquid described in claim 7 is characterized in that: with 50-100 doubly fermented liquid spray water fruits and can eat vegetables raw, the SOD of fruit is improved.
10. the purposes of fermented liquid described in claim 7 is characterized in that: use fermented liquid as the viable bacteria biological fodder; Give and to be breeded fish, add 3%-5% in the feed of shrimp or crab, can purify water, reduce disease, improve the speed of growth; Or fermented liquid and water are according to 1: 5-10 adds doubly in the feed that is added to milk cow, pig, chicken or the tap water and feeds.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102352316A (en) * | 2011-09-13 | 2012-02-15 | 无锡中科活力生物技术有限公司 | Composite germ pulp, and production method and application thereof |
| CN103740615A (en) * | 2013-12-26 | 2014-04-23 | 莱州市顺昌水产有限公司 | Photosynthetic bacteria SC01 as well as fast cultivation method and application thereof |
| CN103773714A (en) * | 2013-12-26 | 2014-05-07 | 莱州市顺昌水产有限公司 | Application of photosynthetic bacteria to improving sea farming water quality |
| CN104099395A (en) * | 2014-07-11 | 2014-10-15 | 北京联合大学 | Rhodobacter sphaeroides extracting solution with antioxidant activity and preparation method thereof |
| CN105360609A (en) * | 2015-10-22 | 2016-03-02 | 无锡中科活力生物技术有限公司 | Fodder premix and preparation method thereof |
| CN107400644A (en) * | 2017-07-26 | 2017-11-28 | 沈阳科纳提克生物科技有限公司 | A kind of culture medium and its application in hydrogenlike silicon ion fermentation and a kind of preparation method of sorbic acid |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102352316A (en) * | 2011-09-13 | 2012-02-15 | 无锡中科活力生物技术有限公司 | Composite germ pulp, and production method and application thereof |
| CN102352316B (en) * | 2011-09-13 | 2013-05-29 | 无锡中科活力生物技术有限公司 | Composite germ pulp, and production method and application thereof |
| CN103740615A (en) * | 2013-12-26 | 2014-04-23 | 莱州市顺昌水产有限公司 | Photosynthetic bacteria SC01 as well as fast cultivation method and application thereof |
| CN103773714A (en) * | 2013-12-26 | 2014-05-07 | 莱州市顺昌水产有限公司 | Application of photosynthetic bacteria to improving sea farming water quality |
| CN103773714B (en) * | 2013-12-26 | 2015-12-02 | 莱州市顺昌水产有限公司 | The application of a kind of photosynthetic bacterium in sea farming water correction |
| CN103740615B (en) * | 2013-12-26 | 2016-08-17 | 莱州市顺昌水产有限公司 | Photosynthetic bacteria SC01 and fast culture process thereof and application |
| CN104099395A (en) * | 2014-07-11 | 2014-10-15 | 北京联合大学 | Rhodobacter sphaeroides extracting solution with antioxidant activity and preparation method thereof |
| CN105360609A (en) * | 2015-10-22 | 2016-03-02 | 无锡中科活力生物技术有限公司 | Fodder premix and preparation method thereof |
| CN107400644A (en) * | 2017-07-26 | 2017-11-28 | 沈阳科纳提克生物科技有限公司 | A kind of culture medium and its application in hydrogenlike silicon ion fermentation and a kind of preparation method of sorbic acid |
| CN107400644B (en) * | 2017-07-26 | 2020-12-22 | 宓岚(上海)国际贸易有限公司 | Culture medium, application of culture medium in rhodobacter sphaeroides fermentation and preparation method of sorbic acid |
| CN107663511A (en) * | 2017-11-30 | 2018-02-06 | 沈国青 | A kind of culture medium of nitrite bacteria and preparation method thereof |
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