CN114468121B - Method for fermenting food industry leftovers by using fermentation agent containing Bacillus belgii - Google Patents
Method for fermenting food industry leftovers by using fermentation agent containing Bacillus belgii Download PDFInfo
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Abstract
The invention discloses a method for fermenting food industry leftovers by using a leaven containing Bacillus belgii, which comprises the following steps: (1) inoculating Bacillus velezensis XDY021 bacterial liquid to a sterilized culture medium, and transferring the culture medium into a seeding tank for culture after shaking culture to obtain a starter containing Bacillus velezensis; (2) taking a fermentation substrate consisting of protein-containing leftovers and sugar-containing leftovers, adding water and grinding into slurry; (3) placing the slurry fermentation substrate in a fermentation tank, adjusting pH, adding a fermentation agent containing Bacillus beiLeisi, and fermenting for 24-120 hr. The invention uses the leaven containing the Bacillus beiLeisi to ferment the protein-containing leftovers and the sugar-containing leftovers, the grain size and the content of the water-insoluble substances of the obtained fermented material just meet the requirements of the culture of the water spider, which is beneficial to the feeding of the water spider and is especially suitable for being used as the bait for culturing the water spider. And because the Bacillus belgii XDY021 still exists, the storage is durable, other mixed bacteria cannot breed in the storage process, and the phenomena of flatulence, swelling and the like cannot occur.
Description
Technical Field
The invention relates to application of Bacillus belgii, in particular to a method for fermenting food industry leftovers by using a leavening agent containing the Bacillus belgii.
Background
The process of producing bean curd sticks and soy sauce by bean curd stick factories and soy sauce factories can generate a large amount of waste bean curd stick residues and waste soy sauce residues (hereinafter collectively referred to as protein-containing leftovers), the protein-containing leftovers are always used for supplying fish ponds as auxiliary baits for fish culture, and because the leftovers are not decomposed and can be only used in a small amount, the nutrient utilization rate is low, and the additional value is low; on the other hand, the leftovers are easy to pollute sundry fungus in the processing, transportation and storage processes, and are directly used as feed without being sterilized, fermented and decomposed, so that fish diseases are easily caused; there are many protein-containing leftovers, such as cake and bran of various oil mills.
In the process of producing dried orange peel by using oranges in a dried orange peel factory, only peel is used, pulp (commonly called as dried orange peel slag) is directly discarded, only 30-50 million tons of pulp are generated in the Guangdong Xinhui city every year, and the dried orange peel slag is not basically used all the time, most of the pulp is directly poured into rivers to pollute the surface and underground water body.
In areas such as the Yangtze delta, farmers refer to the fry-opening bait as "water spider", which is a general term for rotifers and cladocera. The water spider is divided into a water spider and a water spider, wherein the water spider mainly refers to rotifers and small branch angles with the size of 0.2-0.5 mm, and the water spider can pass through a 60-mesh screen and is staple food of young fish fries (the first 5 days) which are just hatched. The big water spider mainly refers to cladocera, the size of the big water spider is 0.5-1.0mm, the big water spider is the main food before the conversion of the food in the seedling stage, and the time for taking the big water spider as the main food is about 6-15 days according to the fish species. In aquaculture, particularly in fry culture, the demand of the water spider is large, but the water spider mainly grows naturally all the time, and the artificial culture of the water spider is tried only in recent years.
According to the knowledge, the largest demand of the existing water spiders is a fry field for culturing high-value aquatic animal fries such as micropterus salmoides, mandarin fish, penaeus vannamei boone and other fishes such as four major fish fries, the fry field needs to buy water spiders outside to feed the fries, most of the water spiders are caught in a natural pond or a sea, and the water spiders need to be frozen for preservation after catching, so that the water spiders are dead water spiders which often carry various pathogenic microorganisms or impurities remained in other environments, and the survival rate of fry culture of the fry field is mostly low and is always 20-50 percent or even lower. Therefore, the artificial culture of live water spider to feed the fry is one of the important ways to improve the culture success rate of the fry farm.
The artificial culture of the live water spider needs special bait, the live water spider is mainly cultured by adopting common manure such as chicken manure and the like at present, and feed raw materials such as water spider happy and the like are also adopted for preparation, and the bait has the disadvantages of large pollution to water body, more residual toxin and higher raw material cost, so the leftovers of the food industry are used as the raw materials for producing the special bait for culturing the water spider, and the special bait for developing the water spider is a better choice.
The leftovers containing protein and sugar are fermented and converted into the bait special for culturing the water spider, the special strains suitable for fermenting and decomposing the leftovers are required to be screened, the strains of the conventional fermented feed such as bacillus subtilis, saccharomyces cerevisiae and the like are used for fermenting the leftovers, the strains are often fermented, most of crude protein is fermented into water-soluble amino acid or small peptide which is not suitable for feeding the water spider, and therefore, the leftovers are fermented by screening the strain with the right fermentation degree to convert the leftovers into the bait raw material for the water spider.
Disclosure of Invention
The invention aims to provide a method for fermenting food industry leftovers by using a leavening agent containing bacillus belgii.
The method for fermenting the leftovers of the food industry by using the leavening agent containing the bacillus belgii comprises the following steps:
(1) inoculating Bacillus subtilis XDY021 bacterial liquid into a sterilized culture medium, performing shake flask culture, and transferring into a seeding tank for culture to obtain a starter culture containing Bacillus subtilis;
(2) taking a fermentation substrate consisting of protein-containing leftovers and sugar-containing leftovers, adding water and grinding into slurry;
(3) putting the slurry fermentation substrate into a fermentation tank, adjusting pH to 4-8, adding a fermentation agent containing Bacillus belgii, and fermenting for 24-120 hr.
The medium in step (1) of the present invention comprises: peptone 1-4%, yeast extract 1-4%, glucose 0.5-3%, KH 2 PO 4 0.1-2%, sodium chloride 0.1-1.5%, manganese sulfate 0.01-0.2%, and purified water to 100%.
The shake flask culture in the step (1) of the invention is 1-5 days, and the seeding tank culture is 1-5 days, and anaerobic fermentation or low-oxygen fermentation can be carried out.
The protein-containing leftovers and sugar-containing leftovers obtained in step (2) of the invention are converted according to the carbon-nitrogen ratio of 10-35:1, and then weighed and mixed (the carbon-nitrogen ratio of the invention is calculated according to the organic matter: crude protein, the same applies below). Certainly, in the process of selecting the raw materials, the selection and the dosage are required to be adjusted according to the properties of the raw materials, for example, the salt content of the soy sauce residue is high, the dosage cannot be too large, and the soy sauce residue is required to be matched with the dried beancurd sticks for use.
In the invention, the protein-containing leftovers are the dried bean dregs of a dried bean factory, the soy sauce dregs of a soy sauce factory, the cake pomace of various oil mills and the like; the sugar-containing leftovers can be pericarpium Citri Tangerinae residue from pericarpium Citri Tangerinae factory, waste starch residue from starch factory, medicinal residue from pharmaceutical factory, folium Camelliae sinensis residue from tea beverage factory, and yeast residue from yeast factory.
In the step (2), the fermentation substrate and the water are mixed according to the mass ratio of 1: 0.2-1.5.
The purpose of the invention can be realized by culturing the Bacillus velezensis XDY021 in the step (3) according to the conventional fermentation process and inoculation amount of a 0.1-50 cubic fermentation tank. However, in the present invention, it is preferable that the amount of the Bacillus belvesii-containing fermentation agent to be inoculated in the fermentation tank is in the range of 0.1 to 10% by mass based on the total mass of the fermentation substrate.
In the step (3), the fermentation is low-oxygen fermentation or anaerobic fermentation, the fermentation temperature is 30-45 ℃, and the ventilation volume is 0-5m 3 /H。
A new bacterial strain (Bacillus velezensis) XDY021 of the Bacillus belgii with the deposition number GDMCC No. 62146 can generate a plurality of antibacterial proteins, antibacterial peptides and other substances in fermentation, can inhibit the growth of other mixed bacteria, achieves the effect of inhibiting or killing other mixed bacteria, and is suitable for fermentation substrates polluted by the mixed bacteria. In application, the fermentation degree of the fermentation substrate fermented by the strain, including the substrate particle size, the content of water insoluble substances and the like, just meets the requirements of culture of the water spider, and the phenomenon of over-fermentation or under-fermentation can not occur.
The Bacillus velezensis XDY021 provided by the invention has been deposited in the Guangdong province microorganism collection center at 18.12.2021, and received the collection registration number GDMCC No. 62146.
The invention has the advantages that:
1. the method for fermenting the leftovers in the food industry by using the leavening agent containing the Bacillus belgii is used for fermenting the leftovers containing protein and the leftovers containing sugar, the grain size, the content of water-insoluble substances and the like of the obtained fermented material just meet the requirements of culturing the water spider, and the method is favorable for feeding the water spider and is particularly suitable for culturing the water spider as the water spider bait. And because the Bacillus beleisi XDY021 still exists in the fermentation product, the fermentation product is storable, other mixed bacteria cannot be bred in the storage process, and the phenomena of flatulence, swelling and the like cannot occur.
2. The fermentation process is simple, the used Bacillus belgii XDY021 generates various substances such as bacteriostatic proteins, antibacterial peptides and the like in the fermentation, can inhibit the growth of other mixed bacteria, achieves the effect of inhibiting or killing other mixed bacteria, and is suitable for synchronously realizing sterilization, fermentation and decomposition in the fermentation process.
3. The invention uses protein-containing leftovers and sugar-containing leftovers as fermentation substrates, changes waste into valuable, and recycles the protein-containing leftovers of bean curd stick factories, soy sauce factories, oil mills and the like and the sugar-containing leftovers of dried orange peel factories, yeast factories, tea beverage factories and the like, thereby realizing higher economic value.
Drawings
FIG. 1 shows the sequencing result of Bacillus subtilis XDY 021.
FIG. 2 is a micrograph of a fermentation substrate in test one without fermentation by Bacillus velezensis XDY 021.
FIG. 3 is a micrograph of the fermentation substrate after fermentation with Bacillus velezensis XDY021 in test one.
Detailed Description
Example one
Screening of Bacillus belgii XDY 021: 4 strains of bacillus (XD05B-XD08B) and 1 strain of yeast (XD05J) are purchased from the strain collection center of the institute of microbiology in Guangdong province, 2 strains of yeast (XD06J-XD07J) are purchased from Baijia supermarket in Guangzhou, 4 strains of bacillus (XDY020-XDY023) and 3 strains of yeast (XD05J-XD07J) are separated and screened from the natural world, then the strains are respectively cultured and screened by 3 generations of fermentation substrate consisting of protein-containing leftovers and sugar-containing leftovers, and finally, the strains which can ferment the fermentation substrate into the strains conforming to the culture of the raw materials of the water spider are selected.
The protein-containing leftovers comprise: bean curd stick residues from a bean curd stick factory in south of the city of the Anhui, Guangdong Jiangmen, soy sauce residues from an ancient soy sauce factory in east of the city of the Anhui, Guangdong Jiangmen, peanut bran from a peanut oil factory in Tabangyo, Guangdong Jiangmen, and the like; the sugar-containing leftovers comprise: the tangerine peel residue from a Xinhuotan tangerine peel factory of Guangdong Jiangmen, the tea leaf residue from an Dongpo food and beverage limited company of Guangdong Guangzhou city, the yeast residue from an Angel yeast factory of Guangxi Liuzhou, and the like are prepared according to the carbon nitrogen ratio of 10-35:1 and used as fermentation substrates for screening strains, and the specific formula is shown in a table I, a table five, a table seven, a table nine and a table eleven.
Table one: fermentation substrate formula
The screening method of the Bacillus velezensis comprises the following steps: the first stage is as follows: preparing fermentation substrate according to table I, adding water, grinding, respectively filling into 112 triangular flasks of 500ml, filling 300g per flask, respectively inoculating the above 14 bacteria, respectively inoculating 8 bottles per bacteria, culturing in 35 deg.C incubator for 36 hr, respectively counting total bacteria number, and selecting total bacteria number not less than 10 7 The 4 strains of (1) were XDY020, XDY021, XDY022, and XDY023, which proved to be suitable for growth on this substrate, and further screening was continued. And a second stage: adding water into the fermentation substrate prepared according to the table I, grinding, respectively filling into 32 triangular flasks with 500ml, filling 300g of the fermentation substrate into each flask, respectively inoculating 4 selected bacteria, inoculating 8 bottles of each bacteria, culturing in an incubator with 35 ℃ for 36 hours, and then applying the fermentation product to a culture experiment of the water spider.
Table two: recording the fermentation product detection results of different strains at the seventh day
As can be seen from Table two, the fermentation product of XDY021 strain has the most amount of fermentation products with 1500-1500 meshes and 2200 meshes, and the bacterial amount is moderate.
Culture test:
the fermentation product of XDY020-023 strain fermented rhizoma polygonati residue, soy sauce residue and dried orange peel residue is used for culturing a water spider experiment: in the new biological products limited company's water spider culture test field of the city of the Mount Henshan, set up 12 100L water spider culture jars, add 80L water separately, and inoculate 1L water containing water spider seed source, ferment fermented bamboo dregs + soy sauce dreg + fermented products of the tangerine peel dreg (table one) of 4 kinds of strains that the above-mentioned strain screening method second stage is selected out separately, add the time to be 8 in the morning every day: 30, the addition amount is 10ml per culture tank, 3 culture tanks are added for each strain culture, the abundance of the water spider is observed every day by calculating the number of the water spider in each visual field under a microscope of 160 times (the number of the water spider in each visual field is an integer which is the average number of three culture tanks), the water spider is collected on the seventh day, the water spider is filtered by a 100-mesh sieve and then weighed, and the result is shown in the third table.
Table three: record table of effect of different strains of fermentation products for culturing water spider
| Group of | Day one | The next day | The third day | The fourth day | The fifth day | Day six | Day seven | Weight (D) |
| XDY020 | 5 | 8 | 18 | 38 | 67 | 88 | 107 | 221g |
| XDY021 | 5 | 8 | 20 | 48 | 90 | 124 | 179 | 323g |
| XDY022 | 5 | 7 | 16 | 36 | 83 | 113 | 146 | 240g |
| XDY023 | 5 | 6 | 17 | 37 | 78 | 111 | 155 | 284g |
From Table two and Table three, it can be seen that the amount of the produced water spider is the highest from abundance to yield of XDY021 strain.
After 5 times of repeated experiments, the strain with the highest water spider abundance and the highest water spider yield is selected as XDY021, the strain is sent to molecular genetics laboratory of southern China university of agriculture for DNA sequencing (shown in figure 1), and the result proves that the strain is the strain of Bacillus velezensis.
Test example 1
The screened Bacillus velezensis XDY021 strain (XDY 021 strain for short) is prepared into the feed raw material leavening agent for the water spider:
(1) preparing XDY021 bacterial liquid: inoculating fresh culture of Bacillus subtilis XDY021 into 16 triangular flasks containing culture medium, and culturing in a water bath shaker at 35 deg.C for 48 hr with stirring speed of 20-50 rpm.
(2) Preparation of a culture medium: is prepared from glucose, peptone, yeast extract, KH 2 PO 4 The method comprises the following steps of preparing a culture medium by taking sodium chloride, manganese sulfate and water as main raw materials in percentage by mass:
peptone 2%, yeast extract 3%, glucose 2%, KH 2 PO 4 0.6 percent of sodium chloride, 0.6 percent of manganese sulfate and the balance of water.
A fermentation agent containing Bacillus belgii prepared from Bacillus velezensis XDY021 strain is used for treating the fermented product in the experimental base of traffic stone field in Tanshou village, Tanshan city, a new biological product company Limited in Tanshan city. Preparing a mixture of 500g of the dried bamboo dregs, the soy sauce dregs and the tangerine peel dregs according to the proportion in the table I, adding 400ml of purified water, fully and uniformly mixing, grinding the mixture into slurry through a colloid mill, and detecting the flora condition of the slurry, wherein about 15 strains of yeast, different types of bacillus, escherichia coli, mould and the like are found, and no strain is in a dominant strain (figure 2).
Respectively placing 300g of the three fermentation products into three 500ml triangular flasks, respectively adding 30ml of a fermentation agent containing the bacillus belgii prepared by XDY021 strains, after culturing for 72 hours in a constant-temperature shaking table, stirring at the rotating speed of 20-50rpm, detecting the flora condition again, finding that more than 95 percent of the bacteria in the fermentation products of the three triangular flasks are the bacillus belgii, and other mixed bacteria are less than 5 percent (see Table IV), and basically no escherichia coli, mould and the like exist, wherein the experiment proves that the fermentation agent containing the bacillus belgii can effectively inhibit the growth of other mixed bacteria (figure 3); the Bacillus subtilis (Bacillus velezensis) is proved to generate antibacterial peptide, antibacterial protein and other substances to inhibit the growth of other mixed bacteria in the growth process.
Table four: detection results of proportion of different bacteria before and after fermentation of leftovers
Example two
1. Preparation of a starter culture containing Bacillus belgii: taking XDY021 strain to inoculate into a culture medium which is sterilized at high temperature, and carrying out fermentation culture in a fermentation tank to obtain a starter culture containing Bacillus belgii, namely the fermentation liquor of the feed raw material of the water spider prepared by the Bacillus belgii for standby.
Specifically, the preparation steps are as follows:
(1) preparing XDY021 bacterial liquid: inoculating fresh culture of Bacillus velezensis XDY021 into 16 500ml triangular flasks filled with culture medium, and culturing in a water bath shaker at 35 deg.C for 48h, with stirring speed of 20-50 rpm. The medium in this step is the same as that in step (2).
(2) Preparation of a culture medium: is prepared from glucose, peptone, yeast extract, KH 2 PO 4 Sodium chloride, manganese sulfate and water are used as main raw materials to prepare a culture medium, and the mass percentages of the main raw materials and the water are as follows:
peptone 2%, yeast extract 3%, glucose 2%, KH 2 PO 4 0.6 percent of sodium chloride, 0.6 percent of manganese sulfate and the balance of water.
(3) Inoculating the XDY021 bacterial liquid into the culture medium which is sterilized at high temperature, adjusting the initial pH to 6, and carrying out anaerobic fermentation in a 100L seed tank for 5 days until the final pH is 4.5 to obtain the leaven containing the Bacillus belezii.
2. Preparation of fermentation substrate
Selecting the dried bean curd bamboo residues of south bean curd stick factories of the Anhui city of the Anhui mountain of Guangdong Jiangmen and the soy sauce residues of ancient soy sauce factories of the east China of the Anhui mountain of the Guangdong Jiangmen city as protein-containing leftovers; the tangerine peel residue of the tangerine peel factory of the Xinhui Haoyan tangerine peel of Guangdong Jiangmen is taken as the leftover containing sugar. The raw materials are weighed according to the conversion of the carbon-nitrogen ratio of 26:1 and then mixed, and the specific material formula is shown in the fifth table.
Table five: fermentation substrate formula
The raw materials are uniformly mixed, water is added according to the weight ratio of the mixture to the water of 1:1, and the mixture is ground into slurry by a colloid mill, so that the fermentation substrate is prepared.
3. Placing 70L of the fermentation substrate in 100L fermentation tank, adjusting pH to 6.5, adding 5L of Bacillus beiLeisi-containing leaven, fermenting for 7 days until pH is 5.1 to reach fermentation end point, and controlling ventilation amount to be 0.5m during fermentation process 3 and/H, keeping the low-oxygen fermentation state, and taking 300g of fermentation product for detection, wherein the result is shown in Table six.
Table six: record of fermentation product test results
| Fermentation product (g) | Water-insoluble matters (g) of fermentation broth | 1500-mesh 2200 mesh water-insoluble content (%) | Number of bacteria of fermentation species |
| 300 | 26.22 | 70.56 | 7.1×10 7 |
EXAMPLE III
1. Preparation of a starter culture containing Bacillus belgii: taking XDY021 strain to inoculate into a culture medium which is sterilized at high temperature, and carrying out fermentation culture in a fermentation tank to obtain a starter culture containing Bacillus belezii for later use.
Specifically, the preparation steps are as follows:
(1) preparing XDY021 bacterial liquid: fresh cultures of Bacillus velezensis XDY021 were inoculated into 16 500ml flasks with medium and incubated in a biochemical incubator at 38 ℃ for 48 h. The medium in this step is the same as that in step (2).
(2) Preparation of a culture medium: is prepared from glucose, peptone, yeast extract, KH 2 PO 4 Sodium chloride, manganese sulfate and water are used as main raw materials to prepare a culture medium, and the mass percentages of the main raw materials and the water are as follows:
peptone 3%, yeast extract 2.5%, glucose 2%, KH 2 PO 4 0.5 percent, 0.4 percent of sodium chloride, 0.04 percent of manganese sulfate and the balance of water.
(3) Inoculating the XDY021 bacterial liquid into the culture medium sterilized at high temperature, adjusting initial pH to 6, performing low-oxygen fermentation in 100L seed tank for 5 days with ventilation of 0.3m 3 And/h, until the pH value of the end point is 4.5, and obtaining the leaven containing the Bacillus belgii.
2. Preparation of fermentation substrate
Selecting the dried bean curd bamboo residues of south bean curd stick factories of the Anhui city of the Anhui mountain of Guangdong Jiangmen and the soy sauce residues of ancient soy sauce factories of the east China of the Anhui mountain of the Guangdong Jiangmen city as protein-containing leftovers; tea leaves of Dongpo food and beverage Co., Ltd, Guangdong Guangzhou was used as sugar-containing leftovers. The raw materials are converted and weighed according to the carbon-nitrogen ratio of 24:1 and then mixed, and the specific material formula is shown in Table seven.
TABLE VII: fermentation substrate formula
The raw materials are uniformly mixed, water is added according to the weight ratio of the mixture to the water of 1: 0.8, and the mixture is ground into slurry by a colloid mill, so that the fermentation substrate is prepared.
3. Putting 70L of the fermentation substrate into a 100L fermentation tank, adjusting pH to 6.5, adding 5L of a fermentation agent containing Bacillus belgii, fermenting for 7 days until the pH is 5.0, wherein the fermentation process is anaerobic fermentation, and detecting 300g of fermentation product, and the results are shown in Table eight.
Table eight: record of fermentation product test results
| Fermentation product (g) | Water-insoluble matters (g) of fermentation broth | 1500-mesh 2200 mesh water-insoluble content (%) | Number of bacteria of fermentation species |
| 300 | 28.12 | 69.31 | 6.8×10 7 |
Example four
1. Preparation of a starter culture containing Bacillus belgii: taking XDY021 strain to inoculate into a culture medium which is sterilized at high temperature, and carrying out fermentation culture in a fermentation tank to obtain a starter culture containing Bacillus belezii for later use.
Specifically, the preparation steps are as follows:
(1) preparing XDY021 bacterial liquid: fresh cultures of Bacillus velezensis XDY021 were inoculated into 20 500ml flasks with medium and incubated in a biochemical incubator at 38 ℃ for 48 h. The medium in this step is the same as that in step (2).
(2) Preparation of a culture medium: is prepared from glucose, peptone, yeast extract, KH 2 PO 4 Sodium chloride, manganese sulfate and water are used as main raw materials to prepare a culture medium, and the mass percentages of the main raw materials and the water are as follows:
3.5% of peptone, 2.5% of yeast extract, 3% of glucose and KH 2 PO 4 0.5 percent of sodium chloride, 0.1 percent of manganese sulfate and the balance of water.
(3) Inoculating the XDY021 bacterial liquid into the culture medium which is sterilized at high temperature, adjusting the initial pH to 6, and carrying out anaerobic fermentation in a 100L seeding tank for 5 days until the final pH is 4.7, thus obtaining the leaven containing the Bacillus beleisi.
2. Preparation of fermentation substrate
Taking peanut bran of a peach source peanut oil factory in the Anhushan of Guangdong Jiangmen as protein-containing leftovers; taking tea leaves of Dongpo food and beverage Limited company in Guangdong Guangzhou city as sugar-containing leftovers; the raw materials are converted and weighed according to the carbon-nitrogen ratio of 15:1 and then mixed, and the specific material formula is shown in table nine.
Table nine: fermentation substrate formula
The raw materials are uniformly mixed, water is added according to the weight ratio of the mixture to the water of 1:1.2, and the mixture is ground into slurry by a colloid mill, so that the fermentation substrate is obtained.
4. Putting 70L of the fermentation substrate into a 100L fermentation tank, adjusting pH to 6.5, adding 5L of a fermentation agent containing Bacillus belgii, fermenting for 7 days until the pH is 4.8 to reach the fermentation end point, wherein the fermentation process is anaerobic fermentation, and detecting 300g of fermentation product, and the result is shown in Table ten.
TABLE Ten: record of fermentation product test results
| Fermentation product (g) | Water-insoluble matters (g) of fermentation broth | 1500-mesh 2200 mesh water-insoluble content (%) | Number of bacteria of fermentation species |
| 300 | 26.32 | 67.36 | 5.2×10 7 |
Example five:
1. preparation of a leavening agent containing bacillus belgii: together with the embodiments.
2. Preparation of fermentation substrate
Taking peanut bran of a peach source peanut oil factory in the Anhushan of Guangdong Jiangmen as protein-containing leftovers; taking yeast residue of Angel yeast factory in Guangxi Liuzhou as sugar-containing leftovers. The raw materials are weighed according to the conversion of the carbon-nitrogen ratio of 14:1 and then mixed, and the specific material formula is shown in the eleventh table.
Table eleven: fermentation substrate formula
Mixing the above materials uniformly, adding water at a weight ratio of mixture to water of 1: 0.7, and grinding with colloid mill to obtain slurry.
3. Placing 70L of the fermentation substrate in 100L fermentation tank, adjusting pH to 6.5, adding 5L of Bacillus beiLeisi-containing leaven, fermenting for 7 days until pH is 5.3 to reach fermentation end point, and controlling ventilation amount to be 0.8m during fermentation process 3 And H, keeping the state of low-oxygen fermentation, taking 300g of fermentation products for detection, and obtaining the results shown in the table twelve.
TABLE twelve: recording the fermentation product detection results
| Taking the fermentation product (g) | Water-insoluble (g) of ferment | 1500-mesh 2200 mesh water-insoluble content (%) | Number of bacteria of fermentation species |
| 300 | 28.25 | 66.57 | 2.4×10 7 |
Test example II
In the test field of the culture of the new biological product of the company, new, ltd, in the city of rhizymenia, 12 100 liters of water spider culture pots were set, 80 liters of water was added, 1 liter of water containing water spider seed source was inoculated, and then the fermentation products of the second to fifth examples were added at a time of 8 a.m. each day: 30, the amount added is 10ml per jar per day, 3 jars per example culture were added, the abundance of the water spider was observed per day by counting the number of water spiders in each visual field (the number per day is an average number of three jars taken as an integer) under a 160-fold microscope, and water spiders were collected on the seventh day, and weighed after draining the water with a 100-mesh sieve, the results are shown in table thirteen.
Table thirteen: recording table for effect of different leftover fermentation products for culturing water spider
| Group of | Day one | The next day | The third day | Day four | The fifth day | Day six | The seventh day | Weight (D) |
| Example two | 5 | 8 | 18 | 36 | 73 | 118 | 179 | 323g |
| EXAMPLE III | 5 | 8 | 16 | 39 | 76 | 121 | 170 | 315g |
| Example four | 5 | 7 | 16 | 36 | 80 | 128 | 183 | 326g |
| EXAMPLE five | 5 | 6 | 17 | 38 | 80 | 131 | 188 | 334g |
The results show that the fermentation products fermented by different leftovers have different effects of culturing the water spider, but the difference is not particularly large, which leftovers are used can be determined according to the source condition of raw materials in actual production, and the using effect of the fermentation material can be adjusted by adding different nutrient elements.
Test example III
Taking 10kg of fermentation products in the second to fifth embodiments respectively, vacuumizing by using a vacuum bag, sealing and packaging, placing the fermentation products in a constant-temperature oven with the temperature of 40 ℃ for a storage-resistant experiment, wherein no gas expansion or expansion phenomenon of the products is found after the fermentation products are stored for 15 days, and proving that XDY021 strains in the fermentation products continue to inhibit the growth of other gas-producing mixed bacteria, and the fermentation products have good storage resistance and commodity attributes.
Test example four
10kg of each of the fermentation agents of the bait raw materials of the water spider in the second to the fourth embodiments are taken, vacuum-pumped by a vacuum bag, sealed and packaged, and placed in a constant-temperature oven at 40 ℃ for a storage-resistant experiment, and no product flatulence or swelling phenomenon is found after 15 days of storage, so that the XDY021 bacteria in the fermentation agent continue to inhibit the growth of other aerogenic bacteria, and the fermentation agent has good storage resistance and commodity attributes.
Claims (8)
1. The method for preparing the feed for the water spider from the leftovers of the fermented food industry by using the leavening agent containing the bacillus belgii is characterized by comprising the following steps of:
(1) inoculating Bacillus velezensis XDY021 bacterial liquid to a sterilized culture medium, and transferring the culture medium into a seeding tank for culture after shaking culture to obtain a starter containing Bacillus velezensis for later use;
(2) adding water into a fermentation substrate consisting of protein-containing leftovers and sugar-containing leftovers and grinding the fermentation substrate into slurry;
(3) placing the slurry fermentation substrate into a fermentation tank, adjusting pH to 4-8, adding a fermentation agent containing Bacillus belgii, and fermenting for 24-120 hr to obtain the final product;
the accession number of the Bacillus velezensis XDY021 is GDMCC No. 62146; and (3) converting the protein-containing leftovers and the sugar-containing leftovers in the step (2) according to the carbon-nitrogen ratio of 10-35:1, and weighing and mixing the leftovers and the sugar-containing leftovers.
2. The method for preparing the feed of the water spider by fermenting the leftovers of the food industry with the starter culture containing the bacillus beiLeisi according to claim 1, wherein the culture medium in the step (1) comprises: peptone 1-4%, yeast extract 1-4%, glucose 0.5-3%, KH 2 PO 4 0.1-2%, sodium chloride 0.1-1.5%, manganese sulfate 0.01-0.2%, and purified water to 100%.
3. The method for preparing the feed of the water spider by fermenting the leftovers in the food industry with the starter containing the bacillus beiLeisi according to claim 1, wherein the shake flask culture in the step (1) is performed for 1 to 5 days, and the seeding tank culture is performed for 1 to 5 days.
4. The method for preparing the feed for the spider by fermenting the leftovers in the food industry with the leaven of bacillus beiLeisi according to claim 1, wherein the leftovers containing the protein are the putrid bamboo dregs in a dried bean curd mill, the soy sauce dregs in a soy sauce mill or the cake dregs in an oil mill.
5. The method for preparing the feed for the water spider by fermenting the leftovers in the food industry by the leaven containing the bacillus beilesiensis according to claim 1, wherein the leftovers containing the sugar are tangerine peel dregs in a tangerine peel factory, waste starch dregs in a starch factory, dregs in a pharmaceutical factory, tea dregs in a tea beverage factory or yeast dregs in a yeast factory.
6. The method for preparing the feed for the water spider by fermenting the leftovers in the food industry with the starter culture containing the bacillus beiLeisi according to claim 1, wherein in the step (2), the fermentation substrate and the water are in a mass ratio of 1: 0.2-1.5.
7. The method for preparing the feed for the water spider by fermenting the leftovers in the food industry with the starter containing the Bacillus belgii according to claim 1, wherein in the step (3), the inoculation amount of the starter containing the Bacillus belgii in the fermentation tank is within the range of 0.1-10% of the total mass of the fermentation substrate.
8. The method for preparing feed of Aranea from leftovers of food industry by fermentation with Bacillus belgii as claimed in claim 1, wherein the fermentation in step (3) is low-oxygen fermentation or anaerobic fermentation at 30-45 deg.C and with ventilation of 0-5m 3 /H。
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| CN110551661A (en) * | 2019-09-19 | 2019-12-10 | 岭南师范学院 | Bacillus belgii LfF-1 strain and application thereof in production of protease |
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| CN110438028A (en) * | 2019-06-24 | 2019-11-12 | 东北农业大学 | A kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose |
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