CN115039834A - Method for improving polypeptide content of egg powder by bacterial enzyme synergistic treatment - Google Patents
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- 239000000843 powder Substances 0.000 title claims abstract description 60
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 35
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 35
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 35
- 238000000034 method Methods 0.000 title claims abstract description 26
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 25
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 25
- 230000002195 synergetic effect Effects 0.000 title claims abstract description 24
- 230000001580 bacterial effect Effects 0.000 title claims abstract description 20
- 238000000855 fermentation Methods 0.000 claims abstract description 67
- 230000004151 fermentation Effects 0.000 claims abstract description 67
- 239000000463 material Substances 0.000 claims abstract description 30
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 24
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 19
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 19
- 241000894006 Bacteria Species 0.000 claims abstract description 19
- 239000004365 Protease Substances 0.000 claims abstract description 19
- 108091005804 Peptidases Proteins 0.000 claims abstract description 16
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 16
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 16
- 238000002360 preparation method Methods 0.000 claims abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 13
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 12
- 239000004310 lactic acid Substances 0.000 claims abstract description 12
- 238000002156 mixing Methods 0.000 claims abstract description 11
- 238000001035 drying Methods 0.000 claims abstract description 9
- 241000186660 Lactobacillus Species 0.000 claims abstract description 7
- 229940039696 lactobacillus Drugs 0.000 claims abstract description 7
- 230000001954 sterilising effect Effects 0.000 claims abstract description 7
- 235000013601 eggs Nutrition 0.000 claims description 48
- 229940088598 enzyme Drugs 0.000 claims description 32
- 235000019419 proteases Nutrition 0.000 claims description 14
- 230000000694 effects Effects 0.000 claims description 7
- 108091005508 Acid proteases Proteins 0.000 claims description 3
- 108090000145 Bacillolysin Proteins 0.000 claims description 3
- 102000002322 Egg Proteins Human genes 0.000 claims description 3
- 108010000912 Egg Proteins Proteins 0.000 claims description 3
- 108091005507 Neutral proteases Proteins 0.000 claims description 3
- 102000035092 Neutral proteases Human genes 0.000 claims description 3
- 108090000526 Papain Proteins 0.000 claims description 3
- 235000013345 egg yolk Nutrition 0.000 claims description 3
- 210000002969 egg yolk Anatomy 0.000 claims description 3
- 229940055729 papain Drugs 0.000 claims description 3
- 235000019834 papain Nutrition 0.000 claims description 3
- 235000013305 food Nutrition 0.000 claims description 2
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims 1
- 238000009472 formulation Methods 0.000 claims 1
- 239000000203 mixture Substances 0.000 claims 1
- 102000004169 proteins and genes Human genes 0.000 abstract description 10
- 108090000623 proteins and genes Proteins 0.000 abstract description 10
- 239000002994 raw material Substances 0.000 abstract description 4
- 241000235342 Saccharomycetes Species 0.000 abstract description 3
- 239000002131 composite material Substances 0.000 abstract description 3
- 238000011161 development Methods 0.000 abstract description 3
- 238000004659 sterilization and disinfection Methods 0.000 abstract description 2
- 238000010521 absorption reaction Methods 0.000 description 9
- 238000013461 design Methods 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 235000019629 palatability Nutrition 0.000 description 4
- 238000007696 Kjeldahl method Methods 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000015816 nutrient absorption Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000013777 protein digestion Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/14—Pretreatment of feeding-stuffs with enzymes
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L15/00—Egg products; Preparation or treatment thereof
- A23L15/25—Addition or treatment with microorganisms or enzymes
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
The invention discloses a method for improving the polypeptide content of egg powder by bacterial enzyme synergistic treatment, which comprises the following steps: step 1: preparing a fermentation base material: and (3) carrying out high-pressure steam sterilization on the egg powder to obtain a fermentation base material. Step 2: preparing a fermentation preparation: mixing Bacillus subtilis, lactobacillus, yeast and protease at a certain ratio. And step 3: inoculating and fermenting: and (3) mixing the fermentation base material prepared in the step (1) with the fermentation preparation prepared in the step (2), adjusting the material-water ratio to obtain a fermentation mixed material, then fermenting, and drying after fermentation to obtain the bacterium-enzyme synergistic fermentation egg powder. According to the invention, a brand-new method for fermenting the egg powder by the aid of the cooperation of multiple bacterial enzymes is designed by optimizing the cooperation fermentation of the composite bacterial enzymes and the proportion and fermentation conditions of raw materials, the egg powder is fermented by a fermentation system of bacillus subtilis, lactic acid bacteria, saccharomycetes and protease, the polypeptide content of the fermented egg powder is detected, and a basis is provided for scientific development and utilization of protein materials.
Description
Technical Field
The invention relates to the technical field of fermentation, in particular to a method for improving the polypeptide content of egg powder by bacterial enzyme synergistic treatment.
Background
The egg powder is a high-quality protein material, has the characteristics of high nutritive value, rich immune globulin, lysozyme and the like, and is an ideal substitute for eggs.
At present, protein resources are in short supply worldwide, and the reduction of the cost of protein materials and the improvement of the protein digestion and absorption rate are the key points of nutrition research.
The absorption of the polypeptide has the characteristics of high speed, low energy consumption, high absorption rate and the like, and the absorption of the polypeptide in the animal body has an absorption mechanism which is independent from the amino acid, so that the absorption inhibition caused by mutual competition of free amino acid and common absorption sites can be effectively reduced, and the utilization of protein is facilitated. In addition, the polypeptide can obviously improve the nutrient absorption of animals, improve the production performance of the animals and reduce the diarrhea rate.
However, in the prior art, the method for increasing the polypeptide content of the egg powder is mainly an enzymolysis method, but the enzymolysis method has limited capability of increasing the polypeptide, the enzymolysis can be inhibited by an enzymolysis product after a period of time, the polypeptide content cannot be further effectively increased, and the enzymolysis method is easy to generate bitter taste and affects the palatability of raw materials.
Disclosure of Invention
1. Technical problem to be solved
The invention aims to solve the problems that in the prior art, the method for improving the polypeptide content of egg powder mainly adopts an enzymolysis method, but the enzymolysis method has limited capability of improving the polypeptide, enzymolysis can be inhibited by an enzymolysis product after a period of time, the polypeptide content cannot be further effectively improved, and the palatability is influenced.
2. Technical scheme
In order to achieve the purpose, the invention adopts the following technical scheme:
the method for improving the content of the polypeptide in the egg powder by the fungus-enzyme synergistic treatment comprises the following steps:
step 1: preparing a fermentation base material: sterilizing the egg powder by high-pressure steam to obtain a fermentation base material;
step 2: preparing a fermentation preparation: mixing Bacillus subtilis, lactobacillus, yeast and protease at a certain ratio;
and step 3: inoculating and fermenting: and (3) mixing the fermentation base material prepared in the step (1) with the fermentation preparation prepared in the step (2), adjusting the material-water ratio to obtain a fermentation mixed material, then fermenting, and drying after fermentation to obtain the bacterium-enzyme synergistic fermentation egg powder.
Preferably, the egg powder in step 1 comprises whole egg powder, albumen powder and egg yolk powder.
Preferably, the mixed bacterial species include lactic acid bacteria, bacillus subtilis and yeast.
Preferably, the protease in step 1 comprises one or more of neutral protease, acid protease and papain, and the addition amount is 0.4%.
Preferably, in the step 2, the mass ratio of the lactic acid bacteria to the bacillus subtilis to the yeast is 2:1.05:1, and the concentration of the bacteria is as follows: the concentration of the lactic acid bacteria is as follows: 3X 10 9 cfu/g, Bacillus subtilis 1.15 × 10 11 cfu/g, concentration of yeast 2X 10 10 cfu/g。
Preferably, the enzyme activity of the protease in the step 2 is 1 × 10 5 U/g。
Preferably, in the step 3, the water content is 80%, the fermentation temperature is 35-37 ℃, the fermentation is carried out for 72 hours, and the drying is carried out at the low temperature of 60 ℃ until the water content is 10%.
The invention also provides the bacterial enzyme synergistic fermented egg powder prepared by the preparation method.
The invention also provides application of the bacterial enzyme synergistic fermented egg powder in food and feed.
3. Advantageous effects
Compared with the prior art, the invention has the advantages that:
(1) according to the invention, a brand-new method for fermenting the egg powder by the cooperation of multiple bacterial enzymes is designed by optimizing the cooperation fermentation of the composite bacterial enzymes and the proportion and fermentation conditions of raw materials, the egg powder is fermented by a fermentation system of bacillus subtilis, lactic acid bacteria, saccharomycetes and protease, the polypeptide content of the fermented egg powder is detected, and a basis is provided for scientific development and utilization of protein materials.
(2) According to the invention, the polypeptide content of the fermented egg powder is increased, the nutritional level is obviously increased, the smell and palatability of the egg powder are also obviously improved, and the digestion and absorption of protein can be promoted after the egg powder is fermented by the aid of the bacteria and the enzyme, so that the utilization rate is increased.
Drawings
FIG. 1 is a graph showing the effect of enzyme addition on polypeptide production;
FIG. 2 is a graph comparing the effect of enzymatic pH on polypeptide production.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.
Example 1:
the method for improving the content of the polypeptide in the egg powder by the fungus-enzyme synergistic treatment comprises the following steps:
step 1: preparing a fermentation base material: and (3) carrying out high-pressure steam sterilization on the egg powder to obtain a fermentation base material.
Step 2: preparing a fermentation preparation: mixing Bacillus subtilis, lactobacillus, yeast and protease at a certain ratio.
And step 3: inoculating and fermenting: mixing the fermentation base material prepared in the step 1 and the fermentation preparation prepared in the step 2, adjusting the material-water ratio to obtain a fermentation mixed material, then fermenting, drying after fermentation to obtain the bacterial enzyme synergistic fermentation egg powder, wherein the water content is 80%, the fermentation temperature is 35-37 ℃, the fermentation is 72 hours, and the drying is carried out at the low temperature of 60 ℃ until the water content is 10%.
According to the invention, a brand-new method for fermenting the egg powder by the cooperation of multiple bacterial enzymes is designed by optimizing the cooperation fermentation of the composite bacterial enzymes and the proportion and fermentation conditions of raw materials, the egg powder is fermented by a fermentation system of bacillus subtilis, lactic acid bacteria, saccharomycetes and protease, the polypeptide content of the fermented egg powder is detected, and a basis is provided for scientific development and utilization of protein materials.
According to the invention, the polypeptide content of the fermented egg powder is increased, the nutritional level is obviously increased, the smell and palatability of the egg powder are also obviously improved, and the digestion and absorption of protein can be promoted after the egg powder is fermented by the aid of the bacteria and the enzyme, so that the utilization rate is increased.
Example 2:
the implementation contents of the above embodiments can be referred to the above description, and the embodiments herein are not repeated in detail; in the embodiment of the present application, the difference from the above embodiment is:
in the invention, the egg powder in the step 1 comprises whole egg powder, protein powder and egg yolk powder, the mixed bacteria comprise lactic acid bacteria, bacillus subtilis and yeast, the protease comprises one or more of neutral protease, acid protease and papain, and the addition amount is 0.4%.
In the invention, in the step 2, the mass ratio of the lactic acid bacteria, the bacillus subtilis and the yeast is 2:1.05:1, and the concentration of the bacteria is as follows: the concentration of the lactic acid bacteria is as follows: 3X 10 9 cfu/g, Bacillus subtilis 1.15 × 10 11 cfu/g, concentration of yeast 2X 10 10 cfu/g, enzyme activity of protease is 1 x 10 5 U/g。
Example 3:
the implementation contents of the above embodiments can be referred to the above description, and the embodiments herein are not repeated in detail; in the embodiment of the present application, the difference from the above embodiment is:
in the invention, the pH value of the protease is optimized, and the method comprises the following steps:
(1) preparing a base material: sterilizing the egg powder by high-pressure steam to obtain a fermentation base material;
(2) protease is added according to 0.2%, and enzymolysis pH is adjusted to 5.5, 6.0, 6.5, 7.0, 7.5, and enzymolysis is carried out for 2 hours.
(3) And (3) detecting the content of the polypeptide by using a trichloroacetic acid-Kjeldahl method.
As shown in FIG. 2, the optimum enzymatic pH was 6.5.
Example 4:
the implementation contents of the above embodiments can be referred to the above description, and the embodiments herein are not repeated in detail; in the embodiment of the present application, the difference from the above embodiment is:
the optimal three-bacterium inoculation ratio is designed and screened by using Plakett-Burman (PB), and the method comprises the following steps:
(1) preparing a fermentation base material: sterilizing the egg powder by high-pressure steam to obtain a fermentation base material;
(2) preparing a fermentation preparation: mixing Bacillus subtilis, lactobacillus and yeast at different mass ratio according to Plackett-Burman design table 1.
(3) Inoculating and fermenting: and (3) mixing the fermentation base material prepared in the step (1) with the fermentation preparation prepared in the step (2), adjusting the material-water ratio to 50% to obtain a fermentation mixed material, then fermenting for 72 hours, and drying at 60 ℃ after fermentation to obtain the bacterium-enzyme synergistic fermentation egg powder.
(4) And (3) detecting the content of the polypeptide by using a trichloroacetic acid-Kjeldahl method.
TABLE 1 Effect of inoculation with bacteria at different ratios on polypeptide production
According to table 1Y-2.97 a-0.89B-2.48C +7.56, lactobacillus optimalis: b, bacillus subtilis: the ratio of yeast is 2:1.05: 1.
Example 5:
the implementation contents of the above embodiments can be referred to the above description, and the embodiments herein are not repeated in detail; in the embodiment of the present application, the difference from the above embodiment is:
the method for designing and screening the optimal fermentation conditions of the strains by using the Plakett-Burman (PB) comprises the following steps:
(1) preparing a fermentation base material: sterilizing the egg powder by high-pressure steam to obtain a fermentation base material;
(2) preparing a fermentation preparation: the mixed strains were designed according to the different conditions according to Plackett-Burman design Table 2.
(3) Inoculating and fermenting: mixing the fermentation base material prepared in the step (1) and the fermentation preparation prepared in the step (2), adjusting the material-water ratio according to the design table 2, then fermenting for different times at different temperatures according to the design table 2, and drying at 60 ℃ after fermentation to obtain the bacterial enzyme synergistic fermented egg powder.
(4) And (3) detecting the content of the polypeptide by using a trichloroacetic acid-Kjeldahl method.
TABLE 2
From table 2, it can be seen:
Y＝14.92+8.14*A+1.37*B+0.0998*C+0.7076*D+1.12*AB+0.0638*
AC+1.22*AD+0.0638*AC+1.22*AD+0.0777*BC+0.0594*BD+0.0729
*CD-4.34*A 2 -0.0031*B 2 -1.12*C 2 -1.03*D 2 。
further analysis of Table 2 revealed that the optimum fermentation material-water ratio was 94.50%, the ratio of fungal enzyme was 9.52, the optimum fermentation time was 76.09 hours, the optimum fermentation temperature was 41.53 ℃, and the maximum value of the obtained polypeptide was 21.50%
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 able to cover the technical scope of the present invention and the equivalent alternatives or modifications according to the technical solution and the inventive concept of the present invention within the technical scope of the present invention.
Claims (10)
1. The method for improving the polypeptide content of the egg powder by the bacterial enzyme synergistic treatment is characterized by comprising the following steps:
step 1: preparing a fermentation base material: sterilizing the egg powder by high-pressure steam to obtain a fermentation base material;
step 2: preparing a fermentation preparation: mixing Bacillus subtilis, lactobacillus, yeast and protease at a certain ratio;
and step 3: inoculating and fermenting: and (3) mixing the fermentation base material prepared in the step (1) with the fermentation preparation prepared in the step (2), adjusting the material-water ratio to obtain a fermentation mixed material, then fermenting, and drying after fermentation to obtain the bacterium-enzyme synergistic fermentation egg powder.
2. The method for increasing polypeptide content of egg powder by bacterial-enzyme synergistic treatment according to claim 1, wherein the egg powder in the step 1 comprises whole egg powder, albumen powder and egg yolk powder.
3. The method for increasing polypeptide content in egg powder by bacterial-enzyme synergistic treatment according to claim 1, wherein the mixed bacterial strain comprises lactic acid bacteria, bacillus subtilis and yeast.
4. The method for increasing the content of polypeptides in egg powder by using bacterial-enzyme synergistic treatment according to claim 1, wherein the protease in the step 1 comprises one or more of neutral protease, acid protease and papain, and the addition amount is 0.4%.
5. The method for improving the content of the polypeptide in the egg powder through the bacterial-enzyme synergistic treatment according to claim 1, wherein the mass ratio of the lactic acid bacteria to the bacillus subtilis to the yeast in the step 2 is 2:1.05:1, and the concentration of the bacteria is as follows: the concentration of the lactic acid bacteria is as follows: 3X 10 9 cfu/g, Bacillus subtilis 1.15 × 10 11 cfu/g, concentration of yeast 2X 10 10 cfu/g。
6. The method for increasing the content of polypeptides in egg powder by bacterial-enzyme synergistic treatment according to claim 1, wherein the enzyme activity of said protease in step 2 is 1 x 10 5 U/g。
7. The method for increasing polypeptide content in egg powder by bacterial-enzyme synergistic treatment according to claim 1, wherein the water content in the step 3 is 80%, the fermentation temperature is 35-37 ℃, the fermentation is carried out for 72 hours, and the drying is carried out at a low temperature of 60 ℃ until the water content is 10%.
8. The bacterial enzyme synergistic fermented egg powder prepared by the preparation method of any one of claims 1 to 7.
9. A bacterial enzyme synergistic preparation for increasing polypeptide content of egg powder comprises lactobacillus, Bacillus subtilis, yeast and protease.
10. Use of a bacterial enzyme synergistic formulation according to claim 9 in food and feed.
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|---|---|---|---|---|
| CN104585505A (en) * | 2015-01-21 | 2015-05-06 | 东北农业大学 | Method for synergistic fermentation of soybean meal by employing bacillus subtilis and neutral protease |
| CN110115347A (en) * | 2019-05-13 | 2019-08-13 | 扬州大学 | A method of the modified preparation egg white powder with high foamability of enzymatic hydrolysis cooperative fermentation |
| CN110447759A (en) * | 2019-08-29 | 2019-11-15 | 段青文 | A kind of additive for farm animal feed and preparation method thereof and animal meat products |
| CN111748598A (en) * | 2020-06-16 | 2020-10-09 | 李忠军 | Small molecule peptide protein powder and preparation method thereof |
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- 2022-06-28 CN CN202210748746.5A patent/CN115039834A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104585505A (en) * | 2015-01-21 | 2015-05-06 | 东北农业大学 | Method for synergistic fermentation of soybean meal by employing bacillus subtilis and neutral protease |
| CN110115347A (en) * | 2019-05-13 | 2019-08-13 | 扬州大学 | A method of the modified preparation egg white powder with high foamability of enzymatic hydrolysis cooperative fermentation |
| CN110447759A (en) * | 2019-08-29 | 2019-11-15 | 段青文 | A kind of additive for farm animal feed and preparation method thereof and animal meat products |
| CN111748598A (en) * | 2020-06-16 | 2020-10-09 | 李忠军 | Small molecule peptide protein powder and preparation method thereof |
Non-Patent Citations (1)
| Title |
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| 孙郡: "蛋体水解液的益生菌发酵酶解工艺及产品制备研究", 粮油食品科技, no. 03, pages 122 - 128 * |
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