Method for reducing fiber binding protein content through microbial fermentation of sesame meal

Abstract

The invention belongs to the technical field of microbial fermentation, and particularly relates to a method for reducing fiber-binding protein content by microbial fermentation of sesame meal, in order to effectively utilize nutritional ingredients of the sesame meal, firstly, aspergillus oryzae yeast and kluyveromyces marxianus are utilized for aerobic fermentation, then enzymolysis is performed, and finally, the content of the fiber-binding protein is reduced. Finally, lactobacillus plantarum and clostridium butyricum are inoculated for anaerobic fermentation, the obtained sesame seed meal fermentation product contains various high-content nutritional ingredients including lactic acid, small peptide and crude protein, the content of fiber binding protein in neutral detergent fibers is reduced, the sesame seed meal fermentation product is used for animal feed, the feed-weight ratio can be effectively reduced, and the feed utilization rate is increased. The diarrhea rate of the piglets is reduced, and the growth performance of the piglets is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial fermentation, and particularly relates to a method for reducing the content of fiber binding protein in sesame meal by microbial fermentation. Background Art

[0002] Sesame meal is the product of sesame oil extraction. It has a crude protein content of 38% to 50%, and the utilization rate of essential amino acids is comparable to that of soybean meal. It is a cheap and high-quality feed protein source worthy of research and development. However, the anti-nutritional factors such as cellulose and tannin contained in sesame meal limit the application of sesame meal. At present, research on dietary fiber is mostly focused on how to degrade non-starch polysaccharides, and how to effectively utilize the nutrients in sesame meal still needs further research. Fibronectin is a glycoprotein connected to fiber in the cell wall and is a cell wall embedded protein. The digestibility of monogastric animals is low, which affects the utilization rate of feed. Studies have found that sesame meal contains a high level of fibronectin. If a method can be found to degrade fibronectin in sesame meal, the digestibility of sesame meal in monogastric animals will be greatly improved, and its utilization rate will be improved. Summary of the invention

[0003] The invention provides a method for reducing the content of fiber binding protein by fermenting sesame meal with microorganisms.

[0004] The technical solution of the present invention is as follows:

[0005] The present invention provides a method for reducing the content of fiber binding protein by fermenting sesame meal with microorganisms, comprising the following steps:

[0006] (1) Mixing sesame meal, bran and hawthorn residue to form a fermentation matrix;

[0007] (2) adding Aspergillus oryzae fermentation product to a fermentation substrate, inoculating Kluyveromyces marxianus, and performing aerobic fermentation to obtain a first fermentation product;

[0008] (3) performing enzymatic hydrolysis on the first fermentation product to obtain a second fermentation product;

[0009] (4) After adding bean dregs to the second fermented product, inoculating Lactobacillus plantarum and Clostridium butyricum to carry out anaerobic fermentation, a sesame meal fermented product is obtained.

[0010] In the step (2), based on the weight of the fermentation substrate, the amount of Aspergillus oryzae fermentation product added is 1%-5%, and 1%-3% of Kluyveromyces marxianus with a viable cell count of more than 1 billion / g is inoculated.

[0011] In the step (4), based on the weight of the fermentation substrate, 2-4% of Lactobacillus plantarum with a live cell count of more than 3 billion / g and 1-3% of Clostridium butyricum with a live cell count of more than 1 billion / g are inoculated.

[0012] In the step (2), in the Aspergillus oryzae fermentation product, the number of Aspergillus oryzae seed spores is higher than 1 billion / g, the activity of neutral protease is not lower than 2000 U / g, the activity of alkaline protease is not lower than 1500 U / g, and the activity of cellulase is not lower than 1000 U / g.

[0013] In the step (1), the fermentation matrix comprises 50%-85% sesame meal, 10%-30% bran, and 5%-20% hawthorn residue.

[0014] After adding dregs of soybeans in step (4), the moisture content of the culture medium is controlled to be 40-43%.

[0015] The step (2) is aerobic fermentation at a temperature of 30-37° C. for 24-48 hours;

[0016] The step (3) of enzymolysis is performed at a temperature of 45-55° C. and a time of 1-6 h;

[0017] The step (4) is anaerobic fermentation at a temperature of 30-37° C. for 72-96 hours.

[0018] Beneficial Effects

[0019] In order to effectively utilize the nutritional components of sesame meal, the invention firstly utilizes Aspergillus oryzae fermentation product and Kluyveromyces marxianus for aerobic fermentation, then performs enzymolysis, and finally inoculates Lactobacillus plantarum and Clostridium butyricum for anaerobic fermentation. The obtained sesame meal fermentation product not only contains a variety of nutritional components with high contents, including lactic acid, small peptides and crude protein, but also reduces the content of fiber binding protein in neutral detergent fiber. The sesame meal fermentation product of the invention is used for animal feed, which can effectively increase the daily feed intake and daily weight gain of piglets, reduce the diarrhea rate of piglets, and improve the growth performance of piglets. DETAILED DESCRIPTION

[0020] The following examples are intended to illustrate the present invention rather than to further limit the present invention.

[0021] The present invention provides a method for reducing the content of fiber binding protein by fermenting sesame meal with microorganisms, comprising the following steps:

[0022] (1) Mix sesame meal, bran and hawthorn residue and use them as a fermentation matrix.

[0023] Hawthorn residue is a byproduct of hawthorn products. It is difficult to store because it contains a lot of water. Adding hawthorn residue to feed can improve the palatability of fermented products and increase the animal's feed intake. The fermented hawthorn residue reduces the cellulose and pectin content and increases the nutritional content.

[0024] Preferably, the fermentation matrix contains 50%-85% sesame meal, 10%-30% bran, and 5%-20% hawthorn residue. The total moisture content of the fermentation matrix is ​​controlled to be about 48%.

[0025] (2) adding Aspergillus oryzae fermentation product to a fermentation substrate, inoculating Kluyveromyces marxianus, and performing aerobic fermentation to obtain a first fermentation product.

[0026] The Aspergillus oryzae fermented product was obtained by inoculating Aspergillus oryzae (CGMCC NO.40471) into a wheat bran solid fermentation medium and fermenting the medium.

[0027] Preferably, in the Aspergillus oryzae fermentation product, the number of Aspergillus oryzae seed spores is higher than 1 billion / g, the activity of neutral protease is not less than 2000 U / g, the activity of alkaline protease is not less than 1500 U / g, and the activity of cellulase is not less than 1000 U / g.

[0028] The Kluyveromyces marxianus was obtained by activating Kluyveromyces marxianus (CICC1275) and inoculating it into a liquid YPD medium and aerobically culturing it at 30° C. for 48 hours.

[0029] Preferably, the amount of Aspergillus oryzae fermentation product added is 1%-5% based on the weight of the fermentation substrate, and 1%-3% of Kluyveromyces marxianus with a viable cell count of more than 1 billion / g is inoculated.

[0030] To ensure the fermentation effect, the aerobic fermentation temperature is 30-37°C and the time is 24-48h.

[0031] (3) The first fermentation product is enzymatically hydrolyzed to obtain a second fermentation product.

[0032] To ensure the fermentation effect, further, the enzymatic hydrolysis temperature is 45-55°C and the time is 1-6h.

[0033] (4) After adding bean dregs to the second fermented product, inoculating Lactobacillus plantarum and Clostridium butyricum to carry out anaerobic fermentation, a sesame meal fermented product is obtained.

[0034] Preferably, after adding dregs in step (4), the moisture content of the matrix is ​​40-43%.

[0035] Regarding Lactobacillus plantarum, Lactobacillus plantarum (CICC6009) was activated and inoculated into liquid MRS culture medium, and statically cultured at 37° C. for 24 hours.

[0036] Regarding Clostridium butyricum, it was obtained by activating Clostridium butyricum (CICC23847) and inoculating it into a seed culture medium, wherein the culture medium components were 10 g / L glucose, 5 g / L whey powder, 10 g / L peptone, 5 g / L beef extract, 2 g K2HPO4, 1.0 g MgSO4·7H2O, 1.0 g NaHCO3, and 0.2 g CaCl2.

[0037] Preferably, based on the weight of the fermentation substrate, 2-4% of Lactobacillus plantarum with a live cell count of more than 3 billion / g and 1-3% of Clostridium butyricum with a live cell count of more than 1 billion / g are inoculated.

[0038] To ensure the fermentation effect, the anaerobic fermentation is further carried out at a temperature of 30-37°C and a time of 72-96 hours.

[0039] In addition, the method further comprises drying the fermented sesame meal at 45-50° C. to make the moisture content of the fermented sesame meal be about 10%.

[0040] In order to effectively utilize the nutritional components of sesame meal, the invention firstly utilizes Aspergillus oryzae fermentation product and Kluyveromyces marxianus for aerobic fermentation, then performs enzymolysis, and finally inoculates Lactobacillus plantarum and Clostridium butyricum for anaerobic fermentation. The obtained sesame meal fermentation product not only contains a variety of nutritional components with high contents, including lactic acid, small peptides and crude protein, but also reduces the content of fiber binding protein in neutral detergent fiber. The sesame meal fermentation product of the invention is used for animal feed, which can effectively reduce the feed-to-weight ratio, reduce the diarrhea rate of piglets, and improve the growth performance of piglets.

[0041] Example 1

[0042] (1) Sesame meal, bran and hawthorn residue are mixed to form a fermentation matrix, wherein the sesame meal accounts for 70%, the bran accounts for 20% and the hawthorn residue accounts for 10%. Water is added to the fermentation matrix to prepare a culture medium, and the moisture content of the culture medium is controlled to be 48%.

[0043] (2) Based on the weight of the fermentation substrate, 1% of Aspergillus oryzae fermentation product was added to the fermentation substrate, and 3% of Kluyveromyces marxianus (CICC1275) with a viable cell count of more than 1 billion / g was inoculated, and aerobic fermentation was carried out at 35° C. for 35 hours to obtain a first fermentation product.

[0044] In the fermentation product of Aspergillus oryzae, the number of Aspergillus oryzae (CGMCC NO.40471) seed spores is higher than 1 billion / g, the activity of neutral protease is not less than 2000U / g, the activity of alkaline protease is not less than 1500U / g, and the activity of cellulase is not less than 1000U / g.

[0045] (3) The first fermentation product was enzymatically hydrolyzed at 50° C. for 4 h to obtain a second fermentation product.

[0046] (4) After adding bean dregs to the second fermented product, the water content of the culture medium is controlled to 42%, and based on the weight of the fermentation substrate, 3% of Lactobacillus plantarum (CICC6009) with a viable cell count of more than 3 billion / g and 2% of Clostridium butyricum (CICC23847) with a viable cell count of more than 1 billion / g are inoculated, and anaerobic fermentation is performed at 35° C. for 80 hours to obtain a sesame meal fermented product.

[0047] Example 2

[0048] (1) Sesame meal, bran and hawthorn residue are mixed to form a fermentation matrix, wherein the sesame meal accounts for 85%, the bran accounts for 10% and the hawthorn residue accounts for 5%. Water is added to the fermentation matrix to prepare a culture medium, and the moisture content of the culture medium is controlled to be 48%.

[0049] (2) Based on the weight of the fermentation substrate, 3% of Aspergillus oryzae fermentation product was added to the fermentation substrate, and 2% of Kluyveromyces marxianus (CICC1275) with a viable cell count of more than 1 billion / g was inoculated, and aerobic fermentation was carried out at 37° C. for 24 hours to obtain a first fermentation product.

[0050] In the fermentation product of Aspergillus oryzae, the number of Aspergillus oryzae (CGMCC NO.40471) seed spores is higher than 1 billion / g, the activity of neutral protease is not less than 2000U / g, the activity of alkaline protease is not less than 1500U / g, and the activity of cellulase is not less than 1000U / g.

[0051] (3) The first fermentation product was enzymatically hydrolyzed at 55° C. for 2 h to obtain a second fermentation product.

[0052] (4) After adding bean dregs to the second fermented product, the water content of the culture medium is controlled to 40%, and based on the weight of the fermentation substrate, 4% of Lactobacillus plantarum (CICC6009) with a viable cell count of more than 3 billion / g and 3% of Clostridium butyricum (CICC23847) with a viable cell count of more than 1 billion / g are inoculated, and anaerobic fermentation is performed at 30° C. for 96 hours to obtain a sesame meal fermented product.

[0053] Example 3

[0054] (1) Sesame meal, bran and hawthorn residue are mixed to form a fermentation matrix, wherein the sesame meal accounts for 60%, the bran accounts for 25% and the hawthorn residue accounts for 15%. Water is added to the fermentation matrix to prepare a culture medium, and the moisture content of the culture medium is controlled to be 48%.

[0055] (2) Based on the weight of the fermentation substrate, 5% of Aspergillus oryzae fermentation product was added to the fermentation substrate, and 1% of Kluyveromyces marxianus (CICC1275) with a viable cell count of more than 1 billion / g was inoculated, and aerobic fermentation was carried out at 30° C. for 48 hours to obtain a first fermentation product.

[0056] In the fermentation product of Aspergillus oryzae, the number of Aspergillus oryzae (CGMCC NO.40471) seed spores is higher than 1 billion / g, the activity of neutral protease is not less than 2000U / g, the activity of alkaline protease is not less than 1500U / g, and the activity of cellulase is not less than 1000U / g.

[0057] (3) The first fermentation product was enzymatically hydrolyzed at 45° C. for 6 h to obtain a second fermentation product.

[0058] (4) After adding bean dregs to the second fermentation product, the water content of the culture medium is controlled to 43%, and based on the weight of the fermentation substrate, 2% of Lactobacillus plantarum (CICC6009) with a viable cell count of more than 3 billion / g and 1% of Clostridium butyricum (CICC23847) with a viable cell count of more than 1 billion / g are inoculated, and anaerobic fermentation is performed at 37° C. for 72 hours to obtain a sesame meal fermentation product.

[0059] Comparative Example 1

[0060] (1) Sesame meal, bran and hawthorn residue are mixed to form a fermentation matrix, wherein the sesame meal accounts for 85%, the bran accounts for 10% and the hawthorn residue accounts for 5%. Water is added to the fermentation matrix to prepare a culture medium, and the moisture content of the culture medium is controlled to be 48%.

[0061] (2) Based on the weight of the fermentation substrate, 3% of Aspergillus oryzae fermentation product is added to the fermentation substrate, 2% of Kluyveromyces marxianus (CICC1275) with a viable cell count of more than 1 billion / g, 4% of Lactobacillus plantarum (CICC6009) with a viable cell count of more than 3 billion / g, and 3% of Clostridium butyricum (CICC23847) with a viable cell count of more than 1 billion / g are inoculated, and enzymatic hydrolysis is carried out at 55° C. for 2 hours to obtain sesame meal fermentation product.

[0062] In the fermentation product of Aspergillus oryzae, the number of Aspergillus oryzae (CGMCC NO.40471) seed spores is higher than 1 billion / g, the activity of neutral protease is not less than 2000U / g, the activity of alkaline protease is not less than 1500U / g, and the activity of cellulase is not less than 1000U / g.

[0063] Comparative Example 2

[0064] (1) Sesame meal, bran and hawthorn residue are mixed to form a fermentation matrix, wherein the sesame meal accounts for 85%, the bran accounts for 10% and the hawthorn residue accounts for 5%. Water is added to the fermentation matrix to prepare a culture medium, and the moisture content of the culture medium is controlled to be 48%.

[0065] (2) Based on the weight of the fermentation substrate, 3% of Aspergillus oryzae fermentation product is added to the fermentation substrate, 2% of Kluyveromyces marxianus (CICC1275) with a viable cell count of more than 1 billion / g, 4% of Lactobacillus plantarum (CICC6009) with a viable cell count of more than 3 billion / g, and 3% of Clostridium butyricum (CICC23847) with a viable cell count of more than 1 billion / g are inoculated, and aerobic fermentation is carried out at 37° C. for 24 hours to obtain a first fermentation product.

[0066] In the fermentation product of Aspergillus oryzae, the number of Aspergillus oryzae (CGMCC NO.40471) seed spores is higher than 1 billion / g, the activity of neutral protease is not less than 2000U / g, the activity of alkaline protease is not less than 1500U / g, and the activity of cellulase is not less than 1000U / g.

[0067] (3) The first fermentation product was enzymatically hydrolyzed at 55° C. for 2 h to obtain a second fermentation product.

[0068] (4) After adding bean dregs to the second fermented product, the water content of the culture medium was controlled to 40%, and anaerobic fermentation was carried out at 30° C. for 96 hours to obtain a sesame meal fermented product.

[0069] Comparative Example 3

[0070] Compared with Example 2, the enzymatic hydrolysis process in step (3) was not performed. Other operations and the amount of bacteria added were the same as those in Example 2.

[0071] Comparative Example 4

[0072] (1) Sesame meal, bran and hawthorn residue are mixed to form a fermentation matrix, wherein the sesame meal accounts for 85%, the bran accounts for 10% and the hawthorn residue accounts for 5%. Water is added to the fermentation matrix to prepare a culture medium, and the moisture content of the culture medium is controlled to be 48%.

[0073] (2) Based on the weight of the fermentation substrate, 3% of Aspergillus oryzae fermentation product is added to the fermentation substrate, 2% of Kluyveromyces marxianus (CICC1275) with a viable cell count of more than 1 billion / g, 4% of Lactobacillus plantarum (CICC6009) with a viable cell count of more than 3 billion / g, and 3% of Clostridium butyricum (CICC23847) with a viable cell count of more than 1 billion / g are inoculated, and anaerobic fermentation is carried out at 30° C. for 96 hours to obtain a sesame meal fermentation product.

[0074] In the fermentation product of Aspergillus oryzae, the number of Aspergillus oryzae (CGMCC NO.40471) seed spores is higher than 1 billion / g, the activity of neutral protease is not less than 2000U / g, the activity of alkaline protease is not less than 1500U / g, and the activity of cellulase is not less than 1000U / g.

[0075] Comparative Example 5

[0076] Compared with Example 2, in step (2), 3% Aspergillus niger fermentation product is added to the fermentation substrate.

[0077] Other operations and the amount of bacteria added are the same as those in Example 2.

[0078] Comparative Example 6

[0079] Compared with Example 2, step (4) inoculates Lactobacillus casei. Other operations and the amount of bacteria added are consistent with those in Example 2.

[0080] Comparative Example 7

[0081] Sesame meal, bran and hawthorn residue were mixed in a ratio of 4:1:1, and the same amount of bacteria as in Example 2 was added and mixed evenly. No fermentation or enzymolysis treatment was performed.

[0082] Experimental results and analysis

[0083] 1. Detection of enzyme activity and bacteria count after aerobic fermentation

[0084] When the target bacteria grow rapidly, the generation of aerobic bacteria and the generation of related enzymes can be effectively inhibited, creating good conditions for the subsequent anaerobic fermentation. The first fermentation products (obtained after aerobic fermentation) of Examples 1-3, Comparative Examples 2, 3, 5, 6, and Comparative Example 7 were tested for related enzyme activities and the number of bacteria. See Table 1 for details.

[0085] Table 1 Detection results of enzyme activity and number of foreign bacteria in different first fermentations

[0086]

[0087] The level of enzyme activity is related to the degree of decomposition and utilization of the solid-state fermentation matrix, and is also related to the subsequent anaerobic fermentation effect. The level of foreign bacteria count is related to the production of toxic and harmful substances during the fermentation process. As shown in Table 1, compared with the comparative example, Examples 1-3 have higher enzyme activity and lower foreign bacteria count, especially the first fermentation product of Example 2 has the highest enzyme activity and the lowest foreign bacteria count. This shows that the first fermentation product obtained by aerobic fermentation according to the method of the present application has higher enzyme activity and lower foreign bacteria count, which is conducive to anaerobic fermentation.

[0088] 2. Nutritional analysis of different sesame meal fermentations

[0089] The number of foreign bacteria in the fermented sesame meal of Examples 1-3 and Comparative Examples 1-7 was detected, and the nutritional components were detected after drying and crushing. See Table 2 for details.

[0090] Table 2 Nutrient composition test results of different sesame meal fermentations

[0091]

[0092]

[0093] Excessive cellulose content in feed not only affects the palatability of feed, but also can cause digestive tract irritation in animals, leading to diseases such as enteritis. Fibrobinding protein is often considered to be a part of cellulose in routine tests. As a cell wall embedded protein, fibrobinding protein has a low digestibility in monogastric animals. This study found that fibrobinding protein in sesame meal can account for more than 20% of crude protein. If fibrobinding protein can be effectively degraded, the utilization rate of sesame meal can be greatly improved.

[0094] As shown in Table 1, compared with Comparative Examples 1-7, the sesame meal fermented products of Examples 1-3 are rich in nutrients and have a higher content. More importantly, the fiber binding protein and crude fiber in the sesame meal neutral detergent fiber are reduced. The crude protein content is increased. This shows that the method of the present application can not only obtain nutrients with a higher content, but also reduce the fiber binding protein content, and effectively utilize the nutrients in the sesame meal.

[0095] 3. Animal testing

[0096] 550 healthy piglets aged 25±2 days were selected and randomly divided into 11 treatments according to the principle of similar weight and consistent gender, with 5 pens in each treatment and 10 pigs in each pen. Among them, the control group was fed with a basic diet, and the experimental group was fed with sesame meal fermentation products of Examples 1-3 and Comparative Examples 1-7 in addition to the basic diet, and the addition amount of sesame meal fermentation products was 10%. The feeding cycle was 20 days. The initial body weight, final body weight, feed intake, and diarrhea were recorded, and the specific results are shown in Table 3.

[0097] Table 3 Effects of different fermented products on growth performance of piglets

[0098]

[0099] As can be seen from Table 3, compared with the control group and comparative examples 1-7 fed with a basic diet, adding the fermented sesame meal of Examples 1-3 to the basic diet can effectively reduce the feed-to-weight ratio and reduce the diarrhea rate of piglets, indicating that the fermented sesame meal obtained by the method of the present application is applied to animal feed to improve the growth performance of piglets.

Claims

1. A method for reducing the content of fiber binding protein by microbial fermentation of sesame meal, characterized in that: The following steps are involved: (1) Mixing sesame meal, bran and hawthorn residue to form a fermentation matrix; (2) adding Aspergillus oryzae fermentation product to a fermentation substrate, inoculating Kluyveromyces marxianus, and performing aerobic fermentation to obtain a first fermentation product; (3) performing enzymatic hydrolysis on the first fermentation product to obtain a second fermentation product; (4) After adding bean dregs to the second fermented product, inoculating Lactobacillus plantarum and Clostridium butyricum to carry out anaerobic fermentation, a sesame meal fermented product is obtained.

2. The method for reducing the content of fiber binding protein by microbial fermentation of sesame meal according to claim 1, characterized in that: In the step (2), based on the weight of the fermentation substrate, the amount of Aspergillus oryzae fermentation product added is 1%-5%, and 1%-3% of Kluyveromyces marxianus with a viable cell count of more than 1 billion / g is inoculated.

3. The method for reducing the content of fiber binding protein by microbial fermentation of sesame meal according to claim 1, characterized in that: In the step (4), based on the weight of the fermentation substrate, 2-4% of Lactobacillus plantarum with a live cell count of more than 3 billion / g and 1-3% of Clostridium butyricum with a live cell count of more than 1 billion / g are inoculated.

4. The method for reducing the content of fiber binding protein by microbial fermentation of sesame meal according to claim 1, characterized in that: In the step (2), in the Aspergillus oryzae fermentation product, the number of Aspergillus oryzae seed spores is higher than 1 billion / g, the activity of neutral protease is not lower than 2000 U / g, the activity of alkaline protease is not lower than 1500 U / g, and the activity of cellulase is not lower than 1000 U / g.

5. The method for reducing the content of fiber binding protein by microbial fermentation of sesame meal according to claim 1, characterized in that: In the step (1), the fermentation matrix comprises 50%-85% sesame meal, 10%-30% bran, and 5%-20% hawthorn residue.

6. The method for reducing the content of fiber binding protein by microbial fermentation of sesame meal according to claim 1, characterized in that: After adding dregs of soybeans in step (4), the moisture content of the culture medium is controlled to be 40-43%.

7. The method for reducing the content of fiber binding protein by microbial fermentation of sesame meal according to claim 1, characterized in that: The step (2) is aerobic fermentation at a temperature of 30-37° C. for 24-48 hours; The step (3) of enzymolysis is performed at a temperature of 45-55° C. and a time of 1-6 h; The step (4) is anaerobic fermentation at a temperature of 30-37° C. for 72-96 hours.

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