CN115843936A - Preparation and application of adenosine cyclophosphate-containing pig feed additive - Google Patents
Preparation and application of adenosine cyclophosphate-containing pig feed additive Download PDFInfo
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- IVOMOUWHDPKRLL-KQYNXXCUSA-N Cyclic adenosine monophosphate Chemical compound C([C@H]1O2)OP(O)(=O)O[C@H]1[C@@H](O)[C@@H]2N1C(N=CN=C2N)=C2N=C1 IVOMOUWHDPKRLL-KQYNXXCUSA-N 0.000 title claims abstract description 51
- 239000003674 animal food additive Substances 0.000 title claims abstract description 43
- 238000002360 preparation method Methods 0.000 title abstract description 10
- 238000000855 fermentation Methods 0.000 claims abstract description 36
- 230000004151 fermentation Effects 0.000 claims abstract description 36
- 241000282887 Suidae Species 0.000 claims abstract description 12
- 238000001694 spray drying Methods 0.000 claims abstract description 10
- 241000186073 Arthrobacter sp. Species 0.000 claims abstract description 3
- 238000000034 method Methods 0.000 claims description 12
- 241000186063 Arthrobacter Species 0.000 claims description 7
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 claims description 6
- 229910021591 Copper(I) chloride Inorganic materials 0.000 claims description 3
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- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 claims description 3
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- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 3
- 239000008103 glucose Substances 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
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- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 3
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- HYMXALLEHXXORK-HTDNVCFESA-N (4ar,6r,7r,7as)-6-(6-aminopurin-9-yl)-2-hydroxy-2-oxo-4a,6,7,7a-tetrahydro-4h-furo[3,2-d][1,3,2]dioxaphosphinin-7-ol;(2r,3r,4r,5s)-6-(methylamino)hexane-1,2,3,4,5-pentol Chemical compound CNC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO.C([C@H]1O2)OP(O)(=O)O[C@H]1[C@@H](O)[C@@H]2N1C(N=CN=C2N)=C2N=C1 HYMXALLEHXXORK-HTDNVCFESA-N 0.000 description 4
- YBPWNFMPWVRSOD-QDEZUTFSSA-N 5-[(2S,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]-5-hydroxynonane-4,6-dione Chemical compound C(CCC)(=O)C([C@@H]1[C@H]([C@H]([C@@H](O1)N1C=NC=2C(N)=NC=NC1=2)O)O)(O)C(CCC)=O YBPWNFMPWVRSOD-QDEZUTFSSA-N 0.000 description 4
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- LWFUFLREGJMOIZ-UHFFFAOYSA-N 3,5-dinitrosalicylic acid Chemical compound OC(=O)C1=CC([N+]([O-])=O)=CC([N+]([O-])=O)=C1O LWFUFLREGJMOIZ-UHFFFAOYSA-N 0.000 description 1
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- IVOMOUWHDPKRLL-UHFFFAOYSA-N UNPD107823 Natural products O1C2COP(O)(=O)OC2C(O)C1N1C(N=CN=C2N)=C2N=C1 IVOMOUWHDPKRLL-UHFFFAOYSA-N 0.000 description 1
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- UNXNGGMLCSMSLH-UHFFFAOYSA-N dihydrogen phosphate;triethylazanium Chemical compound OP(O)(O)=O.CCN(CC)CC UNXNGGMLCSMSLH-UHFFFAOYSA-N 0.000 description 1
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- Feed For Specific Animals (AREA)
Abstract
The invention belongs to the field of microbial fermentation and feed additives, and particularly relates to a preparation method and application of a pig feed additive containing adenosine cyclophosphate. The invention uses Arthrobacter sp.A302CGMCC No.3584 as a fermentation strain to prepare adenosine cyclophosphate, and the fermentation broth is directly subjected to spray drying to prepare the pig feed additive, wherein the content of the adenosine cyclophosphate reaches 11.3 percent, and the content of the protein reaches 32.4 percent. The pig feed additive provided by the invention is used for preparing a feed for fattening pigs, can promote the growth of the pigs, increase the slaughter rate, improve the commercial lean meat percentage and improve the meat quality.
Description
Technical Field
The invention belongs to the field of microbial fermentation and feed additives, and particularly relates to a preparation method and application of a pig feed additive containing adenosine cyclophosphate.
Background
With the rapid development of the breeding industry in China and the improvement of the living standard of people, people no longer only meet the requirement of sufficient quantity of meat consumption, but pursue the quality of the meat. Consumers like pork with high lean meat percentage and low fat and cholesterol content, and also demand good color, high tenderness, juiciness, delicious flavor and appropriate content of unsaturated fatty acids. The factors affecting the pork quality include genetics, environment, nutrition, feeding management and the like.
Cyclic amp, a nonspecific small molecule highly bioactive substance, is a second messenger for a variety of hormones, such as growth hormone and epinephrine, and is widely present in the body. The cyclic AMP is stimulated by external signals to activate adenylate cyclase to catalyze ATP cyclization, has important influence on enzyme activity, gene expression, ion channel activity, cell growth and the like, plays a role in regulating extensive nutrient metabolism, and is an animal growth regulator with good application prospect. Adenosine cyclophosphate plays an important role in regulating protein metabolism, lipid metabolism, improving meat quality and the like. As a regulator for animal growth, adenosine cyclophosphate is nontoxic and harmless, and is more and more widely applied to pig production. In addition, compared with the derivatives of adenosine cyclophosphate, such as meglumine adenosine cyclophosphate and dibutyryladenosine cyclophosphate, the adenosine cyclophosphate is easier to be absorbed by organisms, the process of producing the derivatives is omitted, and the production cost is reduced.
However, the adenosine cyclophosphate fermentation yield is low, the separation cost is high, and the application of the product in the feed is not very advantageous. The technology aims to improve the fermentation yield, directly carries bacteria and sprays to obtain the product, saves the separation process, greatly reduces the production cost and makes it possible for adenosine cyclophosphate to enter the feed field.
Disclosure of Invention
The invention aims to solve the technical problem of providing a pig feed additive containing adenosine cyclophosphate aiming at the defects of the prior art.
The invention also aims to solve the technical problem of providing a preparation method of the adenosine cyclophosphate containing pig feed additive.
The invention finally aims to solve the technical problem of providing the application of the adenosine cyclophosphate containing pig feed additive.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows:
a pig feed additive containing adenosine cyclophosphate is prepared by fermenting Arthrobacter to prepare adenosine cyclophosphate, directly spray drying the fermentation liquid,
wherein, the adenosine cyclophosphate content is 2-20wt%, and the protein content is 5-50wt%.
Wherein, the Arthrobacter is Arthrobacter sp.A302CGMCC No.3584, and the detailed construction process of the strain is disclosed in patent CN 102268385B.
A preparation method of a pig feed additive containing adenosine cyclophosphate comprises the following steps:
(1) Inoculating arthrobacterium to a stirring tank containing a fermentation medium for fermentation, stopping fermentation after adenosine cyclophosphate grows stably, and putting the tank to obtain fermentation liquor;
(2) And (2) carrying out spray drying on the fermentation liquor obtained in the step (1) to obtain feed additive powder.
Wherein, in the step (1), the inoculation amount of Arthrobacter is 10% v.
Wherein in the step (1), the fermentation medium comprises the following components: 5-50g/L glucose, 2-20g/L dipotassium hydrogen phosphate, 2-20g/L potassium dihydrogen phosphate, 0.2-2g/L magnesium sulfate, 1-10g/L peptone, 0.001-0.01g/L biotin, 1-10g/L hypoxanthine, feSO 4 0.03g/L，Na 2 MoO 4 0.001g/L，H 3 BO 3 0.002g/L，MnSO 4 0.01g/L，CoCl 2 0.001g/L，CuCl 2 0.005g/L，ZnSO 4 0.01g/L，NaF 0.003g/L，pH 7.0。
Wherein, in the step (1), the cyclic adenosine monophosphate growth is stabilized as follows: the increase amplitude of the adenosine cyclophosphate concentration in 6 hours does not exceed 0.5g/L. Preferably, in example 1 described below, the fermentation is terminated when the increase in adenosine cyclophosphate concentration is stabilized at 16.5. + -. 0.5g/L within 6 hours.
Wherein, in the step (1), the fermentation conditions are as follows: fermentation temperature 30 ℃, initial pH 7.0, aeration ratio 1.
Wherein, in the step (2), the temperature of the spray drying is 150-180 ℃, preferably 170 ℃.
The pig feed additive containing adenosine cyclophosphate is applied to fattening pig feeds.
Wherein the addition amount of the pig feed additive containing adenosine cyclophosphate in the fattening pig feed is 0.01-10 wt% of the dry weight of the whole feed, the preferable range is 0.05-0.5 wt%, and the most preferable range is 0.1wt%.
Wherein, the fattening pig feed is applied to the fattening pig stage, namely used within 70 days before marketing.
Specifically, the production performance (initial weight kg, final weight kg, daily gain g and feed-weight ratio) and the meat quality evaluation (dressing percentage%, eye muscle area cm) of the experimental pig at the fattening stage are evaluated 3 And backfat thickness cm, lean meat percentage%) and the like, to study the influence of the adenosine cyclophosphate-containing pig feed additive on the growth and fattening of the pigs.
Has the advantages that:
1. the adenosine cyclophosphate fermentation liquor produced by the invention is spray-dried to prepare the fattening pig feed additive, the adenosine cyclophosphate content of the fattening pig feed additive reaches 11.3%, and the protein content of the fattening pig feed additive reaches 32.4%.
2. The fattening pig feed additive provided by the invention is used for preparing a fattening pig feed, can promote the growth of pigs, increase the dressing percentage, improve the commercial lean meat percentage and improve the meat quality.
3. Compared with the derivatives of adenosine cyclophosphate, such as meglumine adenosine cyclophosphate and dibutyryladenosine cyclophosphate, the adenosine cyclophosphate is easier to be absorbed by organisms, the process for producing the derivatives is omitted, and the production cost is reduced.
4. The adenosine cyclophosphate fermentation yield is low, the separation cost is high, and the application of the product in feed is not very advantageous. By improving the fermentation yield and then directly carrying out bacteria spray drying to obtain the product, the separation process is omitted, the production cost is greatly reduced, and the adenosine cyclophosphate can enter the field of feeds.
Drawings
The foregoing and/or other advantages of the invention will become further apparent from the following detailed description of the invention when taken in conjunction with the accompanying drawings.
Fig. 1 is a graph of experimental feeding of experimental pigs during the finishing period, wherein the left is experimental group 3: feeding basal diet added with 0.1wt% of feed additive; the right side is control 1: feeding basal diet without feed additive.
Detailed Description
The experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents and materials are commercially available, unless otherwise specified.
The analytical methods used in the following examples are as follows:
the adenosine cyclophosphate product in the fermentation broth is measured by Agilent 1100 High Performance Liquid Chromatography (HPLC). Lichrospher C18 (4.6X250mm, 5 um) of Hanbang science and technology Limited company of Jiangsu Huaiyin of chromatographic column, the column temperature is 30 ℃, the detector is an ultraviolet detector (254 nm), and the mobile phase: v (methanol): v (triethylamine phosphate solution at ph 6.6) =30:70; the flow rate is 0.8mL/min; sample injection amount: 20 μ L. Accurately weighing a adenosine cyclophosphate standard substance, and preparing a standard substance solution with the mass concentration of about 50mg/L by using redistilled triple water; diluting the pretreated sample with deionized water to a proper concentration, filtering with a 0.45 mu m microporous membrane to obtain a sample solution, detecting the sample solution, and calculating the adenosine cyclophosphate yield of the fermented sample according to the peak area.
The protein content in the fermentation liquor is determined by a3,5-dinitrosalicylic acid method.
Example 1: preparation of adenosine cyclophosphate
Preparation of a fermentation medium: 40g/L glucose, 10g/L dipotassium hydrogen phosphate, 10g/L potassium dihydrogen phosphate, 1g/L magnesium sulfate, 5g/L peptone, 0.005g/L biotin, 6g/L hypoxanthine, feSO 4 0.03g/L，Na 2 MoO 4 0.001g/L，H 3 BO 3 0.002g/L，MnSO 4 0.01g/L，CoCl 2 0.001g/L，CuCl 2 0.005g/L，ZnSO 4 0.01g/L, 0.003g/L NaF, pH 7.0, sterilizing at 121 deg.C for 15min, and cooling to about 30 deg.C.
A100L stirring tank is filled with 60L of culture medium, 10% (v: v) of arthrobacter strains are inoculated after sterilization and fermentation culture is carried out under the fermentation conditions of the fermentation temperature of 30 ℃, the initial pH value of 7.0, the aeration ratio of 1. Sampling every 6 hours to determine the concentration of adenosine cyclophosphate, culturing for 72 hours, stabilizing the concentration of adenosine cyclophosphate at about 16.5g/L, and canning after fermentation.
Spray drying the fermentation broth at 150 deg.C, 160 deg.C, 170 deg.C and 180 deg.C respectively to obtain powder product. The adenosine cyclophosphate content is respectively 10.2%,11.0%,11.3% and 10.6% by Agilent 1100 high performance liquid chromatograph, and the protein content is respectively 30.2%,30.5%,32.4% and 29.3% by 3,5-dinitrosalicylic acid method. It was found that the highest adenosine cyclophosphate and protein contents were obtained when the spray drying temperature was 170 ℃.
Example 2: application of fattening pig feed additive
180 Du-growing three-way hybrid pigs with the weight of about 40 kg are selected and divided into 6 groups according to the test requirements, each group has 3 repetitions, each repetition has 10 repetitions, and one repetition is a column. The control group was fed with basal diet, and the test group was fed with basal diet supplemented with feed additive, and the test period was 70 days of fattening period (fig. 1).
Experimental group 1: 0.01wt% of the feed additive of example 1 was added.
Experimental group 2: 0.05wt% of the feed additive of example 1 was added.
Experimental group 3: 0.1wt% of the feed additive of example 1 was added.
Experimental group 4: 0.5wt% of the feed additive of example 1 was added.
Experimental group 5: 1wt% of the feed additive of example 1 was added.
Control group 1: no feed additive is added.
Control group 2: 0.1wt% of commercially available meglumine adenosine cyclophosphate was added.
Control group 3: 0.1wt% of commercially available dibutyryladenosine cyclophosphate was added.
The influence of the pig feed additive on the production performance of the growing and fattening pigs is shown in table 1, and the influence of the pig feed additive on the meat quality of the growing and fattening pigs is shown in table 2.
TABLE 1 influence of pig feed additives on the Productivity of growing finishing pigs
| Treatment of | Initial weight (kg) | Terminal weight (kg) | Daily gain (g) | Material to weight ratio |
| Experimental group 1 | 40.02 | 96.25 | 803.28 | 2.62 |
| Experimental group 2 | 39.85 | 98.53 | 825.42 | 2.51 |
| Experimental group 3 | 40.62 | 101.82 | 874.28 | 2.32 |
| Experimental group 4 | 40.22 | 99.44 | 846.00 | 2.65 |
| Experimental group 5 | 39.69 | 97.15 | 820.85 | 2.72 |
| Control group 1 | 40.26 | 95.12 | 783.71 | 3.12 |
| Control group 2 | 40.23 | 97.53 | 818.57 | 2.61 |
| Control group 3 | 40.53 | 98.48 | 827.85 | 2.50 |
Wherein daily weight gain (g) = (final weight (kg) -initial weight (kg))/day; feed weight ratio = total feed amount (kg)/total weight gain (kg).
TABLE 2 influence of pig feed additives on meat quality of growing-finishing pigs
| Treatment of | Dressing percentage (%) | Eye muscle area (cm) 3 ) | Backfat thickness (cm) | Lean meat percentage (%) |
| Experimental group 1 | 70.43 | 31.38 | 2.65 | 58.36 |
| Experimental group 2 | 72.87 | 32.19 | 2.56 | 62.43 |
| Experimental group 3 | 76.29 | 35.68 | 2.45 | 64.50 |
| Experimental group 4 | 74.16 | 34.87 | 2.52 | 63.54 |
| Experimental group 5 | 71.78 | 32.43 | 2.62 | 60.42 |
| Control group 1 | 65.28 | 27.72 | 2.89 | 54.86 |
| Control group 2 | 73.24 | 32.58 | 2.52 | 60.82 |
| Control group 3 | 74.12 | 33.52 | 2.56 | 61.23 |
In the formula, dressing percentage (%) = net meat (kg)/live pig weight (kg) after slaughter 100%; eye muscle area (cm) 3 ) The measurement and calculation are carried out through a measuring tape; backfat thickness (cm) = measured by a measuring tape; lean meat percentage (%) = lean meat weight (kg)/total dry weight (kg)
Compared with the control group 1 without the added feed additive, the growth performance and the meat quality of the pigs are obviously improved when the product of the embodiment is added into a plurality of groups of pig feeds. The experimental group 3 has the best effect, and compared with the control group 1, the daily gain is increased by 11.56%, the material-weight ratio is reduced by 25.64%, the dressing percentage is increased by 16.87%, the eye muscle area is increased by 28.72%, the back fat thickness is reduced by 15.22%, and the lean meat percentage is increased by 17.57%. Compared with the addition of meglumine adenosine cyclophosphate and dibutyryladenosine cyclophosphate with the same mass percentage, the effect is obviously improved, the daily gain is respectively improved by 6.81 percent and 5.61 percent, the material weight ratio is respectively reduced by 11.11 percent and 7.20 percent, the dressing percentage is respectively improved by 4.16 percent and 2.93 percent, the eye muscle area is respectively increased by 9.52 percent and 6.44 percent, the back fat thickness is respectively reduced by 2.78 percent and 4.30 percent, and the lean meat percentage is respectively improved by 6.05 percent and 5.34 percent.
The invention provides a preparation method of pig feed containing adenosine cyclophosphate and a preparation method and a method for application thereof, and a plurality of methods and ways for realizing the technical scheme are provided, the above description is only a preferred embodiment of the invention, and it should be noted that for a person skilled in the art, a plurality of improvements and decorations can be made without departing from the principle of the invention, and the improvements and decorations are also regarded as the protection scope of the invention. All the components not specified in this embodiment can be implemented by the prior art.
Claims (10)
1. A pig feed additive containing adenosine cyclophosphate is characterized in that adenosine cyclophosphate is prepared by fermenting Arthrobacter, fermentation liquor is directly obtained by spray drying,
wherein, the adenosine cyclophosphate content is 2-20wt%, and the protein content is 5-50wt%.
2. The adenosine cyclophosphate containing pig feed additive according to claim 1, wherein the Arthrobacter is Arthrobacter sp.a302CGMCC No.3584.
3. The method for preparing the pig feed additive containing adenosine cyclophosphate as set forth in claim 1, which comprises the following steps:
(1) Inoculating arthrobacterium to a stirring tank containing a fermentation medium for fermentation, ending the fermentation after adenosine cyclophosphate grows stably, and putting the tank to obtain fermentation liquor;
(2) And (2) carrying out spray drying on the fermentation liquor obtained in the step (1) to obtain feed additive powder.
4. The process of claim 3, wherein in step (1), the inoculation amount of Arthrobacter is 10%.
5. The method according to claim 3, wherein in the step (1), the fermentation medium comprises the following components: 5-50g/L glucose, 2-20g/L dipotassium hydrogen phosphate, 2-20g/L potassium dihydrogen phosphate, 0.2-2g/L magnesium sulfate, 1-10g/L peptone, 0.001-0.01g/L biotin, 1-10g/L hypoxanthine, feSO 4 0.03g/L，Na 2 MoO 4 0.001g/L，H 3 BO 3 0.002g/L，MnSO 4 0.01g/L，CoCl 2 0.001g/L，CuCl 2 0.005g/L，ZnSO 4 0.01g/L，NaF 0.003g/L，pH 7.0。
6. The method according to claim 3, wherein in the step (1), the fermentation is carried out under the following conditions: fermentation temperature 30 ℃, initial pH 7.0, aeration ratio 1.
7. The method according to claim 3, wherein the spray-drying is carried out at a temperature of 150 to 180 ℃ in the step (2).
8. The pig feed additive containing adenosine cyclophosphate according to claim 1, wherein the pig feed additive is applied to fattening pig feed.
9. The use of claim 8, wherein the pig feed additive containing adenosine cyclophosphate is added to the feed for fattening pigs in an amount of 0.01-10% by dry weight of the total feed.
10. The use according to claim 8, wherein the feed is for use during the finishing stage of a growing-finishing pig, i.e. within 70 days before marketing.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102268385A (en) * | 2010-06-04 | 2011-12-07 | 南京工业大学 | Arthrobacter for producing cyclic adenosine monophosphate through fermentation and application thereof |
| CN103571779A (en) * | 2013-11-04 | 2014-02-12 | 河南科技学院 | Fermentation medium, bacterial strain and production method for fermentation production of adenosine cyclophosphate |
| CN115181773A (en) * | 2022-08-24 | 2022-10-14 | 南京工业大学 | Fermentation preparation method of adenosine cyclophosphate |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102268385A (en) * | 2010-06-04 | 2011-12-07 | 南京工业大学 | Arthrobacter for producing cyclic adenosine monophosphate through fermentation and application thereof |
| CN103571779A (en) * | 2013-11-04 | 2014-02-12 | 河南科技学院 | Fermentation medium, bacterial strain and production method for fermentation production of adenosine cyclophosphate |
| CN115181773A (en) * | 2022-08-24 | 2022-10-14 | 南京工业大学 | Fermentation preparation method of adenosine cyclophosphate |
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