CN107058174B - Preparation method of microbial ferment mold liquid with deodorization function - Google Patents
Preparation method of microbial ferment mold liquid with deodorization function Download PDFInfo
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
Abstract
The invention discloses a preparation method of a microorganism enzyme bacterium liquid with a deodorization function, which comprises the following steps: s1: selecting a suitable strain, S2: preparation of medium, S3: method of culturing, S4: and (5) identifying the culture effect. Since most of microbial culture deodorization operations generally require time for contacting and culturing with odor substances, and odor cannot be eliminated instantaneously or in a very short time, the microbial culture solution of the present invention cannot be used as an ideal treatment mode when actually dealing with the current situation that livestock farms discharge a large amount of feces every day.
Description
Technical Field
The invention relates to the technical field of microbial enzymes with deodorization functions, and particularly relates to a preparation method of microbial enzyme bacterial liquid with deodorization functions.
Background
Livestock farms such as swine and cattle and livestock farms are prone to generate odors, and these odors are often caused by spoiled feed, feces and urine collection, manure pushing treatment sites, and the like. The main odorous components are mainly ammonia in pig and cattle farms, and mainly ammonia and hydrogen sulfide in chicken farms. The odor is often strongly irritant and toxic, which not only causes serious harm to health of livestock and poultry and management personnel in the livestock farm, but also causes complaints and objections to the public because the odor is often dissipated and easily drifts with wind, which disturbs the surrounding public, so how to reduce or eliminate the odor in the livestock farm is a problem which needs to be overcome at present.
Various methods for reducing the odor of livestock farms are available on the market, and the methods are summarized to be physical, chemical and biological methods, and a mixed treatment mode of the three methods is also available. The physical method comprises adsorbing odor with charcoal, zeolite, activated carbon, aluminum silicate, silica gel, etc.; the chemical method usually uses ozone or acetic acid deodorant, converts the odorous compound into odorless or low-odor substance through oxidation or pH adjustment reaction, and uses gas-liquid contact to dissolve ozone in liquid, or neutralizes, oxidizes or otherwise chemically reacts the chemical agent and the odorous component in liquid phase to remove the odor; biological methods generally use microorganisms having the ability to eliminate odors to contact odorous substances, and enzymes secreted by the microorganisms with deodorizing function to neutralize, decompose, or convert the odors into odorless or less odorous substances. Such microorganisms include acetic acid-producing bacteria, lactic acid-producing bacteria, alcohol-producing yeasts, and Bacillus subtilis, etc., which decompose ammonia and hydrogen sulfide. In the biological method, microorganisms are usually contacted with odor substances, so that odor-degrading chemical substances or enzymes are secreted when the microorganisms are in a breeding state, thereby achieving the purpose of odor degradation. Studies have shown that various microorganisms such as lactic acid bacteria, bacillus subtilis, enterococcus, etc., all exhibit deodorizing ability when cultured together with odorous substances. However, such an operation of removing odor by microbial culture requires a period of time for contacting and culturing with odor substances, and cannot be completed instantaneously or in a short time, and when applied to the amount of feces discharged in large amounts daily from livestock farms, the problem of deodorization cannot be effectively solved. In addition, the physical method is liable to cause secondary pollution, and the chemical method has a problem of excessively high treatment cost.
Since most of the procedures for removing odor by culturing microorganisms generally require a time for contacting and culturing with an odor substance, and odor cannot be removed instantaneously or in an extremely short time, they cannot be used as an ideal treatment mode in order to cope with the current situation that livestock farms discharge large amounts of feces every day. In view of the above problems, the research and development team of this company has focused on developing a microbial culture solution with a rapid deodorization function, which can reduce or eliminate the odor in a very short time when contacting with the odor substances of livestock and poultry feces and urine by using various organic acids, enzymes, etc. generated and accumulated in the culture solution by the microbes.
Disclosure of Invention
The invention aims to provide a preparation method of a deodorant microbial enzyme bacterial liquid SEM, which comprises the following specific steps:
s1: selecting proper strains, and screening lactobacillus acidophilus, lactobacillus plantarum and beer yeast from lactic acid producing bacteria of various commercially available probiotics as test strains;
s2: the preparation of the culture medium is very suitable for producing culture solution with the deodorization function after a plurality of tests, and the culture medium comprises the following components in percentage by weight: 10% of hydrolyzed soybean protein liquid, 0.5% of monopotassium phosphate, 4% of glucose and the balance of purified water, wherein the hydrolyzed soybean protein liquid is prepared by mixing soybean protein with more than 90% of protein, 10 kg of soybean protein and 100 kg of water are put into a reaction tank, the pH value is adjusted to 5 by using phosphorus, 200 grams of 1200GDU power value unit pineapple ferment is added, stirring and heating are started, heating is stopped when the temperature is slowly increased to 55 ℃, and hydrolysis is finished within about 2 hours to prepare the hydrolyzed soybean protein liquid;
s3: the culture method comprises the steps of firstly, carrying out single pure culture on each strain, then carrying out compound culture, wherein the single pure culture adopts a closed cover, standing for 7 days, slightly opening the cover every day, shaking and mixing for 1-4 times to complete strain culture, during the compound culture, lactobacillus acidophilus, lactobacillus plantarum and beer yeast are inoculated to a fresh culture medium, the inoculation ratio of each strain is 1:100, standing and culturing are carried out at 20-35 ℃, stirring is carried out at least twice every day, each time lasts for 5-15 minutes, the culture solution is completed in about 30 days, the pH value of the culture solution is 3.0-3.5 when the compound culture is completed, and 15% of cane sugar or molasses can be added during the compound culture;
s4: the identification of culture effect, the culture solution contains abundant organic acid and enzyme, have effects of neutralizing ammonia and sulfureted hydrogen, take 100ml container one to put on the electronic scale and return to zero while detecting this number, put 5.0g of standard ammonia liquid with the suction pipe, drip into the sample of the cultured or finished fungus culture solution slowly with the suction pipe, after dripping into 1.5g, slow the speed of dripping into, and shake the cup after adding 1-2 drops each time, mix, use the gaseous detector to measure the ammonia concentration above the mixed solution, stop dripping into the sample when the ammonia concentration measured is zero, record the weight of the sample added, this number value is the ammonia degradation value, according to the size of ammonia degradation value, can reflect the deodorant capacity of the compound culture solution sample.
Preferably, the hydrolyzed soy protein solution that has not been used in step S2 can be stored at 4 ℃ for 1 week, the medium used in less than 10 liters culture is sterilized at 121 ℃ for 15-30 minutes, the medium used in more than 10 liters culture is not sterilized by heating, and the prepared medium in the same day must be inoculated within 1 hour to reduce the proliferation of mixed bacteria.
Preferably, in the step S3, a cover is used for sealing during single strain culture, the strain is left standing for 7 days, the cover is slightly opened every day, and the strain culture is completed by shaking and mixing for 1-4 times.
Preferably, the detection of the ammonia reduction value in step S4 uses a pumping type composite gas detector, which can simultaneously detect ammonia and hydrogen sulfide, and the set upper limit concentrations of ammonia and hydrogen sulfide are 200 ppm and 100ppm, respectively.
Preferably, the standard ammonia solution used in step S4 is a solution prepared by dissolving 6.25g of a solution containing 26% ammonia in 493.75g of pure water.
Compared with the prior art, the invention has the beneficial effects that: since most of microbial culture deodorization operations generally require time for contacting and culturing with odor substances, and odor cannot be eliminated instantaneously or in a very short time, the microbial culture solution of the present invention cannot be used as an ideal treatment mode when actually dealing with the current situation that livestock farms discharge a large amount of feces every day.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1
A preparation method of a deodorant functional microbial enzyme bacterial liquid SEM comprises the following specific steps:
s1: selecting proper strains, and screening lactobacillus acidophilus, lactobacillus plantarum and beer yeast from lactic acid producing bacteria of various commercially available probiotics as test strains;
s2: the preparation of the culture medium is very suitable for producing culture solution with the deodorization function after a plurality of tests, and the culture medium comprises the following components in percentage by weight: 10% of hydrolyzed soybean protein liquid, 0.5% of monopotassium phosphate, 4% of glucose and the balance of purified water, wherein the hydrolyzed soybean protein liquid is prepared by mixing soybean protein with more than 90% of protein, 10 kg of soybean protein and 100 kg of water are put into a reaction tank, the pH value is adjusted to 5 by using phosphoric acid, 200g of pineapple ferment with 1200GDU power value unit is added, stirring and heating are started, heating is stopped when the temperature is slowly increased to 55 ℃, and hydrolysis is finished within about 2 hours to prepare the hydrolyzed soybean protein liquid;
s3: the culture method comprises the steps of firstly, carrying out single pure culture on each strain, then carrying out compound culture, wherein the single pure culture adopts a closed cover, standing for 7 days, slightly opening the cover every day, shaking and mixing for 1-4 times to complete strain culture, during the compound culture, lactobacillus acidophilus, lactobacillus plantarum and beer yeast are inoculated to a fresh culture medium, the inoculation ratio of each strain is 1:100, standing and culturing are carried out at 20-35 ℃, stirring is carried out at least twice every day, each time lasts for 5-15 minutes, the culture solution is completed in about 30 days, the pH value of the culture solution is 3.0-3.5 when the compound culture is completed, and 15% of cane sugar or molasses can be added during the compound culture;
s4: the identification of culture effect, the culture solution contains abundant organic acid and enzyme, have effects of neutralizing ammonia and sulfureted hydrogen, take 100ml container one to put on the electronic scale and return to zero while detecting this number, put 5.0g of standard ammonia liquid with the suction pipe, drip into the sample of the cultured or finished fungus culture solution slowly with the suction pipe, after dripping into 1.5g, slow the speed of dripping into, and shake the cup after adding 1-2 drops each time, mix, use the gaseous detector to measure the ammonia concentration above the mixed solution, stop dripping into the sample when the ammonia concentration measured is zero, record the weight of the sample added, this number value is the ammonia degradation value, according to the size of ammonia degradation value, can reflect the deodorant capacity of the compound culture solution sample.
The hydrolyzed soy protein solution which is not used in step S2 can be stored at 4 ℃ for 1 week, the culture medium with less than 10 liters can be used after being sterilized at 121 ℃ for 30 minutes, the culture medium with more than 10 liters can be used without being heated and sterilized, and the culture medium prepared in the day must be inoculated within 1 hour to reduce the propagation of mixed bacteria. And in the step S3, sealing a cover when a single strain is cultured, standing for 7 days, slightly opening the cover every day, and shaking and mixing for 1-4 times to finish the strain culture. The detection of the ammonia reduction value in step S4 uses a pumping type composite gas detector capable of detecting ammonia and hydrogen sulfide simultaneously, and the set upper limit concentrations of ammonia and hydrogen sulfide are 200 ppm and 100ppm, respectively. The standard ammonia solution used in step S4 was a solution prepared by dissolving 6.25g of a 26% ammonia-containing solution in 493.75g of pure water.
Example 2
A preparation method of a deodorant functional microbial enzyme bacterial liquid SEM comprises the following specific steps:
s1: selecting proper strains, and screening lactobacillus acidophilus, lactobacillus plantarum and beer yeast from lactic acid producing bacteria of various commercially available probiotics as test strains;
s2: the preparation of the culture medium is very suitable for producing culture solution with the deodorization function after a plurality of tests, and the culture medium comprises the following components in percentage by weight: 10% of hydrolyzed soybean protein liquid, 0.5% of monopotassium phosphate, 4% of glucose and the balance of purified water, wherein the hydrolyzed soybean protein liquid is prepared by mixing soybean protein with more than 90% of protein, 10 kg of soybean protein and 100 kg of water are put into a reaction tank, the pH value is adjusted to 5 by using phosphoric acid, 200g of pineapple ferment with 1200GDU power value unit is added, stirring and heating are started, heating is stopped when the temperature is slowly increased to 55 ℃, and hydrolysis is finished within about 2 hours to prepare the hydrolyzed soybean protein liquid;
s3: the culture method comprises the steps of firstly, carrying out single pure culture on each strain, then carrying out compound culture, wherein the single pure culture adopts a closed cover, standing for 7 days, slightly opening the cover every day, shaking and mixing for 1-4 times to complete strain culture, during the compound culture, lactobacillus acidophilus, lactobacillus plantarum and beer yeast are inoculated to a fresh culture medium, the inoculation ratio of each strain is 1:100, standing and culturing are carried out at 20-35 ℃, stirring is carried out at least twice every day, each time lasts for 5-15 minutes, the culture solution is completed in about 30 days, the pH value of the culture solution is 3.0-3.5 when the compound culture is completed, and 15% of cane sugar or molasses can be added during the compound culture;
s4: the identification of culture effect, the culture solution contains abundant organic acid and enzyme, have effects of neutralizing ammonia and sulfureted hydrogen, take 100ml container one to put on the electronic scale and return to zero while detecting this number, put 5.0g of standard ammonia liquid with the suction pipe, drip into the sample of the cultured or finished fungus culture solution slowly with the suction pipe, after dripping into 1.5g, slow the speed of dripping into, and shake the cup after adding 1-2 drops each time, mix, use the gaseous detector to measure the ammonia concentration above the mixed solution, stop dripping into the sample when the ammonia concentration measured is zero, record the weight of the sample added, this number value is the ammonia degradation value, according to the size of ammonia degradation value, can reflect the deodorant capacity of the compound culture solution sample.
The hydrolyzed soy protein solution which is not used in step S2 can be stored at 4 ℃ for 1 week, the culture medium with less than 10 liters can be used after being sterilized at 121 ℃ for 30 minutes, the culture medium with more than 10 liters can be used without being heated and sterilized, and the culture medium prepared in the day must be inoculated within 1 hour to reduce the propagation of mixed bacteria. And in the step S3, sealing a cover when a single strain is cultured, standing for 7 days, slightly opening the cover every day, and shaking and mixing for 1-4 times to finish the strain culture. The detection of the ammonia reduction value in step S4 uses a pumping type composite gas detector capable of detecting ammonia and hydrogen sulfide simultaneously, and the set upper limit concentrations of ammonia and hydrogen sulfide are 200 ppm and 100ppm, respectively. The standard ammonia solution used in step S4 was a solution prepared by dissolving 6.25g of a 26% ammonia-containing solution in 493.75g of pure water.
Example 3
A preparation method of a deodorant functional microbial enzyme bacterial liquid SEM comprises the following specific steps:
s1: selecting proper strains, and screening lactobacillus acidophilus, lactobacillus plantarum and beer yeast from lactic acid producing bacteria of various commercially available probiotics as test strains;
s2: the preparation of the culture medium is very suitable for producing culture solution with the deodorization function after a plurality of tests, and the culture medium comprises the following components in percentage by weight: 10% of hydrolyzed soybean protein liquid, 0.5% of monopotassium phosphate, 4% of glucose and the balance of purified water, wherein the hydrolyzed soybean protein liquid is prepared by mixing soybean protein with more than 90% of protein, 10 kg of soybean protein and 100 kg of water are put into a reaction tank, the pH value is adjusted to 5 by using phosphoric acid, 200g of pineapple ferment with 1200GDU power value unit is added, stirring and heating are started, heating is stopped when the temperature is slowly increased to 55 ℃, and hydrolysis is finished within about 2 hours to prepare the hydrolyzed soybean protein liquid;
s3: the culture method comprises the steps of firstly, carrying out single pure culture on each strain, then carrying out compound culture, wherein the single pure culture adopts a closed cover, standing for 7 days, slightly opening the cover every day, shaking and mixing for 1-4 times to complete strain culture, during the compound culture, lactobacillus acidophilus, lactobacillus plantarum and beer yeast are inoculated to a fresh culture medium, the inoculation ratio of each strain is 1:100, standing and culturing are carried out at 20-35 ℃, stirring is carried out at least twice every day, each time lasts for 5-15 minutes, the culture solution is completed in about 30 days, the pH value of the culture solution is 3.0-3.5 when the compound culture is completed, and 15% of cane sugar or molasses can be added during the compound culture;
s4: the identification of culture effect, the culture solution contains abundant organic acid and enzyme, have effects of neutralizing ammonia and sulfureted hydrogen, take 100ml container one to put on the electronic scale and return to zero while detecting this number, put 5.0g of standard ammonia liquid with the suction pipe, drip into the sample of the cultured or finished fungus culture solution slowly with the suction pipe, after dripping into 1.5g, slow the speed of dripping into, and shake the cup after adding 1-2 drops each time, mix, use the gaseous detector to measure the ammonia concentration above the mixed solution, stop dripping into the sample when the ammonia concentration measured is zero, record the weight of the sample added, this number value is the ammonia degradation value, according to the size of ammonia degradation value, can reflect the deodorant capacity of the compound culture solution sample.
The hydrolyzed soy protein solution which is not used in step S2 can be stored at 4 ℃ for 1 week, the culture medium with less than 10 liters can be used after being sterilized at 121 ℃ for 30 minutes, the culture medium with more than 10 liters can be used without being heated and sterilized, and the culture medium prepared in the day must be inoculated within 1 hour to reduce the propagation of mixed bacteria. And in the step S3, sealing a cover when a single strain is cultured, standing for 7 days, slightly opening the cover every day, and shaking and mixing for 1-4 times to finish the strain culture. The detection of the ammonia reduction value in step S4 uses a pump-suction type composite gas detector capable of detecting ammonia gas and hydrogen sulfide simultaneously, and the set upper limit concentration of ammonia and hydrogen sulfide is 200 ppm and 100 ppm. The standard ammonia solution used in step S4 was a solution prepared by dissolving 6.25g of a 26% ammonia-containing solution in 493.75g of pure water.
After the compound strains are cultured, ammonia reduction values of samples of different culture days can be measured, and the results are shown as the following table:
watch 1
The compound strain culture solution prepared by the invention has the ammonia reduction value less than 5, and when the compound strain culture solution is contacted with livestock manure and urine in equal quantity, ammonia and hydrogen sulfide in the manure can be removed within 5 minutes, and can be respectively reduced to below 1ppm and 0.1ppm, so that when the manure or urine of a farm is treated, odorous manure and urine can be converted into substances with extremely low odor in a moment or extremely short time, the problem of the odor of the farm is solved, a better environment is provided for animals and raising personnel, and the comparison conditions of the ammonia degradation value and the ammonia degradation capability and the odor degradation capability of the livestock manure are as follows:
watch two
Since most of microbial culture deodorization operations generally require time for contacting and culturing with odor substances, and odor cannot be eliminated instantaneously or in a very short time, the microbial culture solution of the present invention cannot be used as an ideal treatment mode when actually dealing with the current situation that livestock farms discharge a large amount of feces every day.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (3)
1. A preparation method of a microorganism ferment bacterial liquid with deodorization function is characterized by comprising the following steps: the preparation method of the deodorant functional microbial ferment bacterial liquid SEM specifically comprises the following steps:
s1: selecting proper strains, and screening lactobacillus acidophilus, lactobacillus plantarum and beer yeast from lactic acid producing bacteria of various commercially available probiotics as test strains;
s2: the preparation of the culture medium is very suitable for producing culture solution with the deodorization function after a plurality of tests, and the culture medium comprises the following components in percentage by weight: 10% of hydrolyzed soybean protein liquid, 0.5% of monopotassium phosphate, 4% of glucose and the balance of purified water, wherein the hydrolyzed soybean protein liquid is prepared by mixing soybean protein with more than 90% of protein, 10 kg of soybean protein and 100 kg of water are put into a reaction tank, the pH value is adjusted to 5 by using phosphoric acid, 200g of pineapple ferment with 1200GDU power value unit is added, stirring and heating are started, heating is stopped when the temperature is slowly increased to 55 ℃, and hydrolysis is finished within about 2 hours to prepare the hydrolyzed soybean protein liquid;
s3: the culture method comprises the steps of firstly, carrying out single pure culture on each strain, then carrying out compound culture, wherein the single pure culture adopts a closed cover, standing for 7 days, slightly opening the cover every day, shaking and mixing for 1-4 times to complete strain culture, during the compound culture, lactobacillus acidophilus, lactobacillus plantarum and beer yeast are inoculated to a fresh culture medium, the inoculation ratio of each strain is 1:100, standing and culturing are carried out at 20-35 ℃, stirring is carried out at least twice every day, each time lasts for 5-15 minutes, the culture solution is completed in about 30 days, the pH value of the culture solution is 3.0-3.5 when the compound culture is completed, and 15% of cane sugar or molasses can be added during the compound culture;
s4: the identification of culture effect, the culture solution contains abundant organic acid and enzyme, have effects of neutralizing ammonia and hydrogen sulfide, measure this number take 100ml container one place return to zero on the electronic scale, put 5.0g of standard ammonia liquid with the suction pipe, the standard ammonia liquid is a solution made by dissolving 6.25g of ammonia liquid containing 26% in 493.75g of pure water, drip into the sample of the cultured or finished fungus culture solution slowly with the suction pipe, after dripping into 1.5g, slow down the speed of dripping into, and shake the cup after adding 1-2 drops each time, mix, measure the ammonia concentration above the mixed solution with the gas detector, stop dripping into the sample when the ammonia concentration measured is zero, record the weight of the sample added, this value is the ammonia degradation value, according to the size of the ammonia degradation value, can reflect the deodorant ability of the culture solution sample.
2. The method for preparing a microorganism ferment bacterial liquid with deodorization function according to claim 1, wherein the method comprises the following steps: the hydrolyzed soy protein solution which is not used in the step S2 can be stored at 4 ℃ for 1 week, the culture medium is used after being sterilized at 121 ℃ for 15-30 minutes when less than 10 liters of the culture medium is cultured, the culture medium which is used more than 10 liters of the culture medium is not heated and sterilized, and the culture medium prepared in the day is required to be inoculated within 1 hour to reduce the propagation of mixed bacteria.
3. The method for preparing a microorganism ferment bacterial liquid with deodorization function according to claim 1, wherein the method comprises the following steps: the detection of the ammonia reduction value in step S4 uses a pumping type composite gas detector capable of detecting ammonia and hydrogen sulfide simultaneously, and the set upper limit concentrations of ammonia and hydrogen sulfide are 200 ppm and 100ppm, respectively.
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