A fermentation extraction method for oligosaccharides and its application

Abstract

The present invention relates to the technical field of oligosaccharide preparation, and discloses a fermentation extraction method of oligosaccharides and its application. The present invention develops soybean oligosaccharides with good physiological functions and high added value, improves yield and purity, reduces losses, protects the structure and activity of soybean oligosaccharides to the maximum extent, has good safety, low preparation cost, and short production cycle. The present invention uses defatted soybean meal as raw material, which can not only improve resource utilization and solve environmental protection problems, but also can produce good economic and social benefits, and has broad application prospects.

Description

Technical Field

[0001] The invention relates to the technical field of oligosaccharide preparation, and in particular to a fermentation extraction method for oligosaccharides and application thereof. Background Art

[0002] my country is the birthplace of soybeans, and its soybean production has always been among the highest in the world. Soybean processing enterprises mainly produce oils and soybean protein, and more than 95% of the soybean meal produced by soybean oil soaking plants is high-temperature soybean meal. Highly denatured defatted soybean meal is obtained by high-temperature desolventizing after soybean oil soaking. Since the protein in highly denatured soybean meal is almost completely denatured, it is almost impossible to separate edible products from it by ordinary physical methods. At present, high-temperature soybean meal is mainly used as feed, and is rarely used in food, and is limited to brewing food. Soybean meal contains about 45% to 50% protein, 10% to 15% oligosaccharides, 20% to 25% polysaccharides and cellulose, and has extremely high development and utilization value. Soybean oligosaccharides are the general term for soluble carbohydrates in soybeans. Its main components are stachyose, raffinose and sucrose, among which raffinose and stachyose are the functional factors with physiological activity of oligosaccharides. Soybean oligosaccharides have excellent physical and chemical properties such as high stability, good safety, low sweetness and low caloric value. They also have the physiological effects of promoting the proliferation of beneficial bacteria such as bifidobacteria in the human intestine and inhibiting the growth and reproduction of harmful bacteria in the human intestine.

[0003] However, most of the existing efficient purification technologies for various oligosaccharides are still in the laboratory stage, with low extraction rates and are not suitable for industrial production; the removal rate of impurities by a single method is low and the product loss rate is high; and because the properties of the three components of soybean oligosaccharides are relatively similar, it is difficult to obtain a single component with high purity, resulting in a higher cost for high-purity components.

[0004] Therefore, how to obtain a fermentation extraction method for oligosaccharides and its application is a problem that those skilled in the art need to solve urgently. Summary of the invention

[0005] In view of this, the present invention provides a fermentation extraction method of oligosaccharides and application thereof.

[0006] In order to achieve the above object, the present invention adopts the following technical solution:

[0007] A fermentation extraction method for oligosaccharides comprises the following steps:

[0008] (1) Putting defatted soybean meal into a grinder, crushing it, sieving it to obtain powder, and soaking it with purified water to obtain a mixed liquid;

[0009] (2) After soaking, adjust the pH of the mixed solution with NaOH solution, then add the complex enzyme, turn on the microwave-assisted extraction, stir, and then filter to obtain a filtrate;

[0010] (3) The filtrate is mixed evenly with yeast for inoculation, added to a fermentation tank, air is introduced, stirred and fermented, and centrifuged after fermentation is completed to obtain the supernatant;

[0011] (4) Add activated carbon to the supernatant, stir, let stand, filter, and concentrate under reduced pressure to obtain a concentrated solution;

[0012] (5) The concentrated solution is decolorized by adsorption on a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components are collected and freeze-dried to obtain soybean oligosaccharides.

[0013] Preferably, after the powder is pulverized in step (1), it is sieved through a 80-100 mesh sieve, and the ratio of powder to purified water is 1:10-15.

[0014] Preferably, the complex enzyme in step (2) is cellulase, hemicellulase and papain in a dosage ratio of 3:1:5, the added amount of the complex enzyme accounts for 1-1.5% of the powder mass, the enzyme activity of cellulase is 10000-100000 U / g, the enzyme activity of hemicellulase is 10000-100000 U / g, and the enzyme activity of papain is 10000-100000 U / g.

[0015] Preferably, in step (2), the pH is adjusted to 8-8.5, the extraction temperature is 50-60° C., and the extraction time is 2-3 h.

[0016] Preferably, in step (2), the microwave power is 800-1200 W, and the stirring speed is 300-400 r / min.

[0017] Preferably, in step (3), the yeast accounts for 1-3% of the mass of the filtrate, and the live bacteria parameter in the yeast is 10 8 ~10 9 CFU / mL, the fermentation temperature is 36-38℃, the stirring speed is 50-100r / min, and the fermentation time is 16-20h.

[0018] Preferably, the centrifugal speed in step (3) is 3000-3500 r / min, and the time is 15-20 min.

[0019] Preferably, in step (4), the mass of activated carbon accounts for 3-8% of the supernatant, and the supernatant is concentrated under reduced pressure to 1 / 4-1 / 3 of the original supernatant volume.

[0020] Preferably, in step (5), the macroporous adsorption resin is selected from one of D-101, HP-20, and AB-8.

[0021] As an inventive concept identical to the above technical solution, the present invention also requests protection for the use of the oligosaccharides obtained by the above method in food and health products.

[0022] It can be seen from the above technical solution that, compared with the prior art, the present invention provides a fermentation extraction method for oligosaccharides and its application, which has the following beneficial effects:

[0023] (1) The present invention develops soybean oligosaccharides with good physiological functions and high added value, improves yield and purity, reduces losses, and protects the structure and activity of soybean oligosaccharides to the maximum extent. It has good safety, low preparation cost, and short production cycle.

[0024] (2) The present invention uses defatted soybean meal as raw material, which can not only improve resource utilization and solve environmental protection problems, but also produce good economic and social benefits and has broad application prospects. DETAILED DESCRIPTION

[0025] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0026] Example 1

[0027] A fermentation extraction method for oligosaccharides comprises the following steps:

[0028] (1) Putting defatted soybean meal into a grinder, crushing it, sieving it to obtain powder, and soaking it with purified water to obtain a mixed liquid;

[0029] (2) After soaking, adjust the pH of the mixed solution with NaOH solution, then add the complex enzyme, turn on the microwave-assisted extraction, stir, and then filter to obtain a filtrate;

[0030] (3) The filtrate is mixed evenly with yeast for inoculation, added to a fermentation tank, air is introduced, stirred and fermented, and centrifuged after fermentation is completed to obtain the supernatant;

[0031] (4) Add activated carbon to the supernatant, stir, let stand, filter, and concentrate under reduced pressure to obtain a concentrated solution;

[0032] (5) The concentrated solution is decolorized by adsorption on a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components are collected and freeze-dried to obtain soybean oligosaccharides.

[0033] After the powder is pulverized in step (1), it is passed through an 80-mesh sieve, and the ratio of powder to purified water is 1:10.

[0034] The compound enzyme in step (2) is cellulase, hemicellulase and papain in a dosage ratio of 3:1:5, the added amount of the compound enzyme accounts for 1% of the powder mass, the enzyme activity of cellulase is 20000U / g, the enzyme activity of hemicellulase is 20000U / g, and the enzyme activity of papain is 20000U / g.

[0035] In step (2), the pH is adjusted to 8, the extraction temperature is 50° C., and the extraction time is 2 h.

[0036] In step (2), the microwave power is 800 W and the stirring speed is 300 r / min.

[0037] In step (3), the yeast accounts for 1% of the mass of the filtrate, and the parameter of live bacteria in the yeast is 10 9 CFU / mL, the fermentation temperature was 36°C, the stirring speed was 50r / min, and the fermentation time was 16h.

[0038] The centrifugal speed in step (3) is 3000 r / min and the time is 15 min.

[0039] In step (4), the mass of activated carbon accounts for 3% of the supernatant, and the supernatant is concentrated under reduced pressure to 1 / 4 of the original supernatant volume.

[0040] In step (5), the macroporous adsorption resin is selected from D-101.

[0041] Example 2

[0042] A fermentation extraction method for oligosaccharides comprises the following steps:

[0043] (1) Putting defatted soybean meal into a grinder, crushing it, sieving it to obtain powder, and soaking it with purified water to obtain a mixed liquid;

[0044] (2) After soaking, adjust the pH of the mixed solution with NaOH solution, then add the complex enzyme, turn on the microwave-assisted extraction, stir, and then filter to obtain a filtrate;

[0045] (3) The filtrate is mixed evenly with yeast for inoculation, added to a fermentation tank, air is introduced, stirred and fermented, and centrifuged after fermentation is completed to obtain the supernatant;

[0046] (4) Add activated carbon to the supernatant, stir, let stand, filter, and concentrate under reduced pressure to obtain a concentrated solution;

[0047] (5) The concentrated solution is decolorized by adsorption on a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components are collected and freeze-dried to obtain soybean oligosaccharides.

[0048] After the powder is pulverized in step (1), it is passed through a 80-100 mesh sieve, and the mass ratio of powder to purified water is 1:15.

[0049] The complex enzyme in step (2) is cellulase, hemicellulase and papain in a dosage ratio of 3:1:5, the added amount of the complex enzyme accounts for 1.5% of the powder mass, the enzyme activity of cellulase is 20000 U / g, the enzyme activity of hemicellulase is 20000 U / g, and the enzyme activity of papain is 20000 U / g.

[0050] In step (2), the pH is adjusted to 8.5, the extraction temperature is 60° C., and the extraction time is 3 h.

[0051] In step (2), the microwave power is 1200 W and the stirring speed is 400 r / min.

[0052] In step (3), the yeast powder accounts for 3% of the filtrate mass, and the live bacteria parameter in the yeast is 10 9 CFU / mL, the fermentation temperature was 38°C, the stirring speed was 100r / min, and the fermentation time was 20h.

[0053] In step (3), the pH is adjusted to 5.5, the centrifugal speed is 3500 r / min, and the time is 20 min.

[0054] In step (4), the mass of activated carbon accounts for 8% of the supernatant, and the supernatant is concentrated under reduced pressure to 1 / 3 of the original supernatant volume.

[0055] In step (5), the macroporous adsorption resin is selected from AB-8.

[0056] Example 3

[0057] A fermentation extraction method for oligosaccharides comprises the following steps:

[0058] (1) Putting defatted soybean meal into a grinder, crushing it, sieving it to obtain powder, and soaking it with purified water to obtain a mixed liquid;

[0059] (2) After soaking, adjust the pH of the mixed solution with NaOH solution, then add the complex enzyme, turn on the microwave-assisted extraction, stir, and then filter to obtain a filtrate;

[0060] (3) The filtrate is mixed evenly with yeast for inoculation, added to a fermentation tank, air is introduced, stirred and fermented, and centrifuged after fermentation is completed to obtain the supernatant;

[0061] (4) Add activated carbon to the supernatant, stir, let stand, filter, and concentrate under reduced pressure to obtain a concentrated solution;

[0062] (5) The concentrated solution is decolorized by adsorption on a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components are collected and freeze-dried to obtain soybean oligosaccharides.

[0063] After the powder is pulverized in step (1), it is passed through a 100-mesh sieve, and the mass ratio of the powder to purified water is 1:15.

[0064] The complex enzyme in step (2) is cellulase, hemicellulase and papain in a dosage ratio of 3:1:5, the added amount of the complex enzyme accounts for 1.2% of the powder mass, the enzyme activity of cellulase is 20000 U / g, the enzyme activity of hemicellulase is 20000 U / g, and the enzyme activity of papain is 20000 U / g.

[0065] In step (2), the pH value is adjusted to 8.5, the extraction temperature is 55° C., and the extraction time is 2.5 h.

[0066] In step (2), the microwave power is 1000 W and the stirring speed is 350 r / min.

[0067] In step (3), the yeast accounts for 2% of the mass of the filtrate, and the parameter of live bacteria in the yeast is 10 9 CFU / mL, the fermentation temperature was 37°C, the stirring speed was 80r / min, and the fermentation time was 18h.

[0068] In step (3), the pH is adjusted to 5, the centrifugal speed is 3200 r / min, and the time is 18 min.

[0069] In step (4), the mass of activated carbon accounts for 6% of the supernatant, and the supernatant is concentrated under reduced pressure to 1 / 3 of the original supernatant volume.

[0070] In step (5), the macroporous adsorption resin is selected from AB-8.

[0071] Comparative Example 1

[0072] A method for extracting oligosaccharides by fermentation is different from Example 3 in that no complex enzyme is added in step (2), and the rest is the same.

[0073] Comparative Example 2

[0074] A method for extracting oligosaccharides by fermentation, which differs from Example 3 in that the composite enzyme in step (2) is cellulase and hemicellulase in a dosage ratio of 3:1, and the rest is the same.

[0075] Comparative Example 3

[0076] A method for extracting oligosaccharides by fermentation, which differs from Example 3 in that the complex enzyme in step (2) is replaced by papain, and the rest is the same.

[0077] Comparative Example 4

[0078] A method for extracting oligosaccharides by fermentation, which differs from Example 3 in that in step (3), the yeast accounts for 0.5% of the mass of the filtrate, and the parameter of live bacteria in the yeast is 109 CFU / mL, and the rest were the same.

[0079] The stachyose and raffinose contents of the products of each embodiment and comparative example are shown in Table 1. It can be seen that the extraction method of the present invention greatly improves the purity of oligosaccharides and reduces losses. Both complex enzymes and yeast have an important influence on the process. After changing the parameters of the comparative example, the effect is significantly reduced. In particular, cellulase and hemicellulase react with papain, which has a synergistic effect, enhances the enzymolysis effect, and can improve the yield and purity of oligosaccharides. The test results are shown in Table 1.

[0080] Table 1: Test results

[0081] Group Stachyose (%) Raffinose (%) Example 1 64.3 25.4 Example 2 65.7 28.1 Example 3 69.1 27.8 Comparative Example 1 40.8 11.5 Comparative Example 2 60.5 21.3 Comparative Example 3 56.4 18.6 Comparative Example 4 57.2 19.7

[0082] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0083] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. Application of a fermentation extraction method in improving the purity of stachyose and raffinose, characterized in that: The steps of the fermentation extraction method are as follows: (1) Putting defatted soybean meal into a grinder, crushing it, sieving it to obtain powder, and soaking it with purified water to obtain a mixed liquid; (2) After soaking, adjust the pH of the mixed solution with NaOH solution, then add the complex enzyme, turn on the microwave-assisted extraction, stir, and then filter to obtain a filtrate; (3) The filtrate is mixed evenly with yeast for inoculation, added to a fermentation tank, air is introduced, stirred and fermented, and centrifuged after fermentation is completed to obtain the supernatant; (4) Add activated carbon to the supernatant, stir, let stand, filter, and concentrate under reduced pressure to obtain a concentrated solution; (5) Decolorizing the concentrated solution by adsorption on a macroporous adsorption resin column, eluting with deionized water, collecting the sugar-containing components, and freeze-drying to obtain soybean oligosaccharides; The compound enzyme in step (2) is cellulase, hemicellulase and papain in a dosage ratio of 3:1:5, and the added amount of the compound enzyme accounts for 1-1.5% of the powder mass; In step (3), the yeast accounts for 1-3% of the mass of the filtrate, and the parameter of live bacteria in the yeast is 10 8 ~10 9 CFU / mL, fermentation temperature is 36-38°C, stirring speed is 50-100r / min, and fermentation time is 16-20h; In step (2), the pH is adjusted to 8-8.5, the extraction temperature is 50-60°C, and the extraction time is 2-3 hours.

2. The use according to claim 1, characterized in that: After the powder is pulverized in step (1), it is passed through a 80-100 mesh sieve, and the ratio of powder to purified water is 1:10-15.

3. The use according to claim 1, characterized in that: In step (2), the microwave power is 800-1200 W, and the stirring speed is 300-400 r / min.

4. The use according to claim 1, characterized in that: The centrifugal speed in step (3) is 3000-3500 r / min, and the time is 15-20 min.

5. The use according to claim 1, characterized in that: In step (4), the activated carbon mass accounts for 3-8% of the supernatant, and the supernatant is concentrated under reduced pressure to 1 / 4-1 / 3 of the original supernatant volume.

6. The use according to claim 1, characterized in that: In step (5), the macroporous adsorption resin is selected from one of D-101, HP-20, and AB-8.