CN109234186B - novel thermophilic lactic acid bacteria and application thereof in wet corn soaking - Google Patents
novel thermophilic lactic acid bacteria and application thereof in wet corn soaking Download PDFInfo
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- C08B30/00—Preparation of starch, degraded or non-chemically modified starch, amylose, or amylopectin
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- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention relates to a novel thermophilic lactic acid bacterium and application thereof in wet corn soaking, wherein the strain preservation information of the thermophilic lactic acid bacterium is as follows: the name of the depository: china general microbiological culture Collection center; the address of the depository: western road No.1, north west city of township, beijing, institute of microbiology, china academy of sciences; the preservation date is as follows: 18 months 4 in 2018; the preservation number is: CGMCC No. 15623; and (3) classification and naming: bacillus coagulans Bacillus coagulons.
Description
Technical Field
The invention belongs to the field of microbial agents, and particularly relates to novel thermophilic lactic acid bacteria and application thereof in wet corn soaking.
background
the traditional technology uses old pulp and new acid (0.2% sulfurous acid solution) to dip in a soaking tank, the whole soaking process comprises adding old pulp in a new tank, adding new acid to corn with long soaking time, and the tank-to-tank circulation is regular. The circulation process can maintain a certain concentration difference between the soluble substances in the corn and the soaking solution, so as to soak out the soluble substances in the corn. However, sulfur dioxide generated in the traditional soaking process is a main component of acid rain, so that environmental pollution is caused, and meanwhile, sulfurous acid can corrode equipment and pipelines, so that the production cost is increased. In the final product, the residue of sulfur dioxide can affect the quality of the starch product and the utilization of byproducts, reducing economic benefits. Therefore, it is urgent to find a new soaking condition which can replace sulfurous acid. The traditional tank-inverting soaking process needs a plurality of tanks connected in series, is not suitable for the condition of low corn starch demand, and has complicated tank-inverting process and increased production and equipment cost. In the corn wet-process soaking process, the abundant thermophilic lactic acid bacteria in the soaking solution play an important role. The lactobacillus has the capability of degrading protein, so that insoluble protein is converted into soluble protein, and simultaneously, the lactic acid and sulfurous acid generated by lactobacillus fermentation can cause the cell wall to be in a porous state, so that macromolecular protease and water molecules which cannot pass through the corn epidermis originally can enter the corn kernel, the water absorption speed of the corn kernel is accelerated, the separation of starch and protein in the corn is promoted, and the purpose of shortening the soaking time is achieved. The thermophilic lactobacillus bacterial liquid is added in the corn soaking process, so that the content of lactobacillus in the soaking liquid can be increased, the function of the lactobacillus is fully exerted, and the soaking effect is improved. However, the lactobacillus bacterial liquid has the limitations of short storage life, low stability, difficult transportation and the like. Therefore, the development of a microbial agent which can avoid the use of sulfurous acid and can obviously improve the wet-process soaking effect of the corn is an effective way for solving the defects.
Disclosure of Invention
The invention provides thermophilic lactic acid bacteria LAB 0#, which are separated from corn steep water (old pulp). The strain preservation information of thermophilic lactic acid bacteria LAB 0# of the invention is as follows: the name of the depository: china general microbiological culture Collection center; the address of the depository: western road No.1, north west city of township, beijing, institute of microbiology, china academy of sciences; the preservation date is as follows: 18 months 4 in 2018; the preservation number is: CGMCC No. 15623; and (3) classification and naming: bacillus coagulans Bacillus coagulons.
another embodiment of the present invention provides a thermophilic lactic acid bacterium LAB # 1 isolated from corn steep water (old pulp). The strain preservation information of thermophilic lactic acid bacteria LAB 1# of the invention is as follows: the name of the depository: china general microbiological culture Collection center; the address of the depository: western road No.1, north west city of township, beijing, institute of microbiology, china academy of sciences; the preservation date is as follows: 18 months 4 in 2018; the preservation number is: CGMCC No. 15624; and (3) classification and naming: staphylococcus succinogenes succinicus.
Another embodiment of the present invention provides a microbial agent, characterized in that the functional microorganism in the microbial agent is selected from one or two of thermophilic lactic acid bacteria LAB 0#, thermophilic lactic acid bacteria LAB 1 #.
the microbial agent also comprises a freeze-drying protective agent, wherein the freeze-drying protective agent consists of the following freeze-drying protective components: 100-300mg/L of rhamnolipid, 80-120g/L of glucose, 120-150g/L of skim milk and 12-15mL/L of glycerol.
Another embodiment of the invention provides a microbial agent a, which is characterized in that the microbial agent a consists of thermophilic lactic acid bacteria LAB 0# and a freeze-drying protective agent, wherein the viable count of the thermophilic lactic acid bacteria LAB 0# is more than or equal to 3.6 × 109/g, and the freeze-drying protective agent consists of the following freeze-drying protective components: 200-300mg/L of rhamnolipid, 80-120g/L of glucose, 120-150g/L of skim milk and 12-15mL/L of glycerol.
Another embodiment of the present invention provides a method for producing microbial agent a, which comprises the steps of:
(1) Preparing a seed solution: inoculating thermophilic lactobacillus LAB 0# into MRS liquid culture medium, and performing shake culture at 50-55 deg.C for 24 hr to obtain seed solution;
(2) Preparation of bacterial sludge: inoculating the seed solution obtained in the step (1) into a fermentation culture medium, performing shake culture at 50-55 ℃ for 24 hours, centrifuging the fermentation product, removing the supernatant, collecting the thallus, and washing with normal saline to obtain bacterial sludge;
(3) Preparation of the microbial agent: and (3) uniformly mixing the bacterial sludge obtained in the step (2) with a freeze-drying protective agent to obtain a bacterial suspension, pre-freezing the bacterial suspension at a temperature of between 70 ℃ below zero and 80 ℃ for 3 to 4 hours, transferring the bacterial suspension to a freeze dryer, freeze-drying the bacterial suspension for 20 to 24 hours to obtain bacterial powder, and packaging the bacterial powder to obtain the microbial agent A.
The formula of the MRS liquid culture medium in the step (1) is that each liter of MRS liquid culture medium contains 10.0g of peptone, 5.0g of beef extract, 4.0g of yeast extract, 20.0g of glucose, Tween-801.0 mL, 2.0g of dipotassium hydrogen phosphate, 5.0g of sodium acetate, 2.0g of triammonium citrate, 0.2g of magnesium sulfate, 0.05g of manganese sulfate and the balance of distilled water; the rotating speed of the shaking table is 100-;
The formula of the fermentation medium in the step (2) is that each liter of fermentation medium contains 10g of glucose, 10g of yeast extract, 10g of peptone and the balance of distilled water; the rotating speed of the shaking table is 100-; the inoculation amount of the seed liquid inoculated into the fermentation medium is 5 percent; the centrifugal speed is 5000r/min, and the centrifugal time is 10-15 min;
the volume ratio of the bacterial sludge to the freeze-drying protective agent in the step (3) is 1:1.5-2.0, and the freeze-drying protective agent consists of the following freeze-drying protective components: 200-300mg/L of rhamnolipid, 80-120g/L of glucose, 120-150g/L of skim milk and 12-15mL/L of glycerol.
Another embodiment of the invention provides a microbial agent B, which is characterized in that the microbial agent B consists of thermophilic lactic acid bacteria LAB 1# and a freeze-drying protective agent, wherein the viable count of the thermophilic lactic acid bacteria LAB 1# is more than or equal to 3.6 × 109/g, and the freeze-drying protective agent consists of the following freeze-drying protective components: 200mg/L of rhamnolipid 100-.
Another embodiment of the present invention provides a method for producing the microbial agent B, which comprises the steps of:
(1) preparing a seed solution: inoculating thermophilic lactobacillus LAB 1# into an MRS liquid culture medium, and performing shake culture at 50-55 ℃ for 24 hours to obtain seed liquid;
(2) preparation of bacterial sludge: inoculating the seed solution obtained in the step (1) into a fermentation culture medium, performing shake culture at 50-55 ℃ for 24 hours, centrifuging the fermentation product, removing the supernatant, collecting the thallus, and washing with normal saline to obtain bacterial sludge;
(3) Preparation of the microbial agent: and (3) uniformly mixing the bacterial sludge obtained in the step (2) with a freeze-drying protective agent to obtain a bacterial suspension, pre-freezing the bacterial suspension at a temperature of between 70 ℃ below zero and 80 ℃ for 3 to 4 hours, transferring the bacterial suspension to a freeze dryer, freeze-drying the bacterial suspension for 20 to 24 hours to obtain bacterial powder, and packaging the bacterial powder to obtain the microbial agent B.
the formula of the MRS liquid culture medium in the step (1) is that each liter of MRS liquid culture medium contains 10.0g of peptone, 5.0g of beef extract, 4.0g of yeast extract, 20.0g of glucose, Tween-801.0 mL, 2.0g of dipotassium hydrogen phosphate, 5.0g of sodium acetate, 2.0g of triammonium citrate, 0.2g of magnesium sulfate, 0.05g of manganese sulfate and the balance of distilled water; the rotating speed of the shaking table is 100-;
The formula of the fermentation medium in the step (2) is that each liter of fermentation medium contains 10g of glucose, 10g of yeast extract, 10g of peptone and the balance of distilled water; the rotating speed of the shaking table is 100-; the inoculation amount of the seed liquid inoculated into the fermentation medium is 5 percent; the centrifugal speed is 5000r/min, and the centrifugal time is 10-15 min;
The volume ratio of the bacterial sludge to the freeze-drying protective agent in the step (3) is 1:1.5-2.0, and the freeze-drying protective agent consists of the following freeze-drying protective components: 200mg/L of rhamnolipid 100-.
A microbial agent composition, characterized in that the composition consists of the microbial agent a and the microbial agent B; wherein the mass ratio of the two is 1: 1.0-1.5.
Another embodiment of the invention provides application of the microbial agent, the microbial agent A and/or B and the microbial agent composition in wet corn soaking.
Another embodiment of the invention provides application of the microbial agent, the microbial agent A and/or B and the microbial agent composition in corn starch extraction.
The invention provides a process for extracting starch by wet corn soaking, which is characterized by comprising the following steps:
(1) mixing corn, microbial agent and water, and soaking at 50-55 deg.C for 12-20 hr to obtain soaking mixture;
(2) and (2) transferring the soaking mixture obtained in the step (1) into a grinder to carry out coarse grinding, supplementing a microbial agent to continue soaking for 8-12h, carrying out fine grinding, sequentially sieving by a 50-250 mesh sieve, standing the filtrate overnight, carrying out centrifugal separation, and drying to obtain the corn starch.
In the step (1), the mass ratio of the corn to the microbial agent to the water is 1: 0.01-0.03: 3-5; the coarse crushing is used for crushing the corn particles into 5-8 pieces;
The dosage of the microbial inoculum in the step (2) is 0.5 time of that in the step (1).
Compared with the prior art, the invention has the advantages that: (1) the invention provides 2 novel thermophilic lactic acid bacteria, and the thermophilic lactic acid bacteria are prepared into microbial agents which can be used as direct-vat-set biological agents to be directly used for soaking and extracting starch in wet corn, so that artificial addition of lactic acid and sulfurous acid is avoided; (2) the microbial agent A and the microbial agent B are used in combination, so that the corn starch yield can be improved, the soaking time is reduced, and the corn soaking efficiency by a wet method is greatly improved; (3) when the microbial agent is prepared, rhamnolipid with a strong protection effect on LAB 0# and LAB 1# is selected as one of the components of the protective agent, so that the freeze-drying survival rate can be remarkably enhanced and is over 90 percent.
Detailed Description
In order to facilitate a further understanding of the invention, the following examples are provided to illustrate it in more detail. However, these examples are only for better understanding of the present invention and are not intended to limit the scope or the principle of the present invention, and the embodiments of the present invention are not limited to the following.
example 1 screening of LAB 0#, LAB 1#
10mL of corn soaking water (old pulp) is taken, diluted to 10-1, 10-2, 10-3, 10-4, 10-5, 10-6 and 10-7 times of the original concentration respectively, and then 0.3mL of corn soaking water (old pulp) is taken and spread on an MC plate (50 +/-2) DEG C for 24 hours. Picking single colony with calcium dissolving ring, streaking, separating and purifying repeatedly. Separating out bacterial colonies with large calcium-dissolving rings and unique bacterial colonies, and inoculating the bacterial colonies to an inclined plane for preservation, wherein the bacterial colonies are respectively numbered as LAB 0# and LAB 1 #. The corn steep water (old pulp) is provided by a maize starch production workshop of the diamond flower group company Limited (Jining, Shandong).
The MC culture medium comprises soybean peptone 5.0g/L, beef extract 3.0g/L, yeast extract 3.0g/L, glucose 20.0g/L, lactose 20.0g/L, calcium carbonate 10.0g/L, agar 15.0g/L, neutral red 0.05g/L, and pH value of 6.0 + -0.1 at 25 deg.C.
Example 2 preparation of microbial inoculum A
(1) preparing a seed solution: inoculating thermophilic lactobacillus LAB 0# into MRS liquid culture medium, and performing shake culture at 50-55 deg.C for 24 hr at 100r/min to obtain seed solution; wherein the formula of the MRS liquid culture medium is that each liter of MRS liquid culture medium contains 10.0g of peptone, 5.0g of beef extract, 4.0g of yeast extract, 20.0g of glucose, 801.0 mL of tween-sodium, 2.0g of dipotassium hydrogen phosphate, 5.0g of sodium acetate, 2.0g of triammonium citrate, 0.2g of magnesium sulfate, 0.05g of manganese sulfate and the balance of distilled water;
(2) Preparation of bacterial sludge: inoculating the seed solution obtained in the step (1) into a fermentation culture medium according to the inoculation amount of 5%, carrying out shaking table culture at the temperature of 50-55 ℃ for 24 hours at 100r/min, centrifuging the fermentation product (the centrifugal rotation speed is 5000r/min, the centrifugation time is 10min), discarding supernatant, collecting thalli, and washing with physiological saline to obtain bacterial sludge; wherein the formula of the fermentation medium is that each liter of fermentation medium contains 10g of glucose, 10g of yeast extract, 10g of peptone and the balance of distilled water;
(3) Preparation of the microbial agent: uniformly mixing the bacterial sludge obtained in the step (2) with a freeze-drying protective agent according to the volume ratio of 1:1.5 to obtain a bacterial suspension, pre-freezing the bacterial suspension at-80 ℃ for 3 hours, transferring the bacterial suspension to a freeze dryer for freeze drying for 20 hours to obtain bacterial powder, and packaging the bacterial powder to obtain the microbial agent A (hereinafter referred to as a product a 1); the freeze-drying protective agent consists of the following freeze-drying protective components: 200mg/L of rhamnolipid, 120g/L of glucose, 120g/L of skim milk and 15mL/L of glycerol. The viable count of the product a1 is 3.6 multiplied by 109/g, and the freezing survival rate is 92.0%.
uniformly mixing the bacterial sludge obtained in the step (2) with a freeze-drying protective agent according to the volume ratio of 1:1.5 to obtain a bacterial suspension, pre-freezing the bacterial suspension at-80 ℃ for 3 hours, transferring the bacterial suspension to a freeze dryer for freeze drying for 20 hours to obtain bacterial powder, and packaging the bacterial powder to obtain the microbial agent (hereinafter referred to as product a 1-1); the freeze-drying protective agent consists of the following freeze-drying protective components: 120g/L glucose, 120g/L skim milk and 15mL/L glycerol. The freezing survival rate of the product a1-1 is 62.3%.
Uniformly mixing the bacterial sludge obtained in the step (2) with a freeze-drying protective agent according to the volume ratio of 1:1.5 to obtain a bacterial suspension, pre-freezing the bacterial suspension at-80 ℃ for 3 hours, transferring the bacterial suspension to a freeze dryer for freeze drying for 20 hours to obtain bacterial powder, and packaging the bacterial powder to obtain the microbial agent (hereinafter referred to as product a 1-2); the freeze-drying protective agent consists of the following freeze-drying protective components: rhamnolipid 50mg/L, glucose 120g/L, skim milk 120g/L, glycerol 15 mL/L. The freezing survival rate of the product a1-2 is 76.8%.
Example 3 preparation of microbial inoculum A
(1) preparing a seed solution: inoculating thermophilic lactobacillus LAB 0# into MRS liquid culture medium, and performing shake culture at 50-55 deg.C for 24 hr at 120r/min to obtain seed solution; wherein the formula of the MRS liquid culture medium is that each liter of MRS liquid culture medium contains 10.0g of peptone, 5.0g of beef extract, 4.0g of yeast extract, 20.0g of glucose, 801.0 mL of tween-sodium, 2.0g of dipotassium hydrogen phosphate, 5.0g of sodium acetate, 2.0g of triammonium citrate, 0.2g of magnesium sulfate, 0.05g of manganese sulfate and the balance of distilled water;
(2) Preparation of bacterial sludge: inoculating the seed solution obtained in the step (1) into a fermentation culture medium according to the inoculation amount of 5%, carrying out shaking table culture at the temperature of 50-55 ℃ for 24 hours at 120r/min, centrifuging the fermentation product (the centrifugal rotation speed is 5000r/min, the centrifugal time is 15min), discarding supernatant, collecting thalli, and washing with normal saline to obtain bacterial sludge; wherein the formula of the fermentation medium is that each liter of fermentation medium contains 10g of glucose, 10g of yeast extract, 10g of peptone and the balance of distilled water;
(3) preparation of the microbial agent: uniformly mixing the bacterial sludge obtained in the step (2) with a freeze-drying protective agent according to the volume ratio of 1:2.0 to obtain a bacterial suspension, pre-freezing the bacterial suspension at-70 ℃ for 4 hours, transferring the bacterial suspension to a freeze dryer for freeze drying for 24 hours to obtain bacterial powder, and packaging the bacterial powder to obtain the microbial agent A (hereinafter referred to as a product a 2); the freeze-drying protective agent consists of the following freeze-drying protective components: 300mg/L of rhamnolipid, 80g/L of glucose, 150g/L of skim milk and 12mL/L of glycerol. The viable count of the product a2 is 3.7 multiplied by 109/g, and the freezing survival rate is 94.5%.
Example 4 preparation of microbial inoculum B
(1) preparing a seed solution: inoculating thermophilic lactobacillus LAB 1# into an MRS liquid culture medium, and performing shake culture at 50-55 ℃ at 100r/min for 24 hours to obtain seed liquid; wherein the formula of the MRS liquid culture medium is that each liter of MRS liquid culture medium contains 10.0g of peptone, 5.0g of beef extract, 4.0g of yeast extract, 20.0g of glucose, 801.0 mL of tween-sodium, 2.0g of dipotassium hydrogen phosphate, 5.0g of sodium acetate, 2.0g of triammonium citrate, 0.2g of magnesium sulfate, 0.05g of manganese sulfate and the balance of distilled water;
(2) Preparation of bacterial sludge: inoculating the seed solution obtained in the step (1) into a fermentation culture medium according to the inoculation amount of 5%, carrying out shaking table culture at the temperature of 50-55 ℃ for 24 hours at 100r/min, centrifuging the fermentation product (the centrifugal rotation speed is 5000r/min, the centrifugation time is 10min), discarding supernatant, collecting thalli, and washing with physiological saline to obtain bacterial sludge; wherein the formula of the fermentation medium is that each liter of fermentation medium contains 10g of glucose, 10g of yeast extract, 10g of peptone and the balance of distilled water;
(3) preparation of the microbial agent: uniformly mixing the bacterial sludge obtained in the step (2) with a freeze-drying protective agent according to the volume ratio of 1:1.5 to obtain a bacterial suspension, pre-freezing the bacterial suspension at-80 ℃ for 3 hours, transferring the bacterial suspension to a freeze dryer for freeze drying for 20 hours to obtain bacterial powder, and packaging the bacterial powder to obtain the microbial agent B (hereinafter referred to as product B1); the freeze-drying protective agent consists of the following freeze-drying protective components: 100mg/L of rhamnolipid, 120g/L of glucose, 120g/L of skim milk and 15mL/L of glycerol. The viable count of the product b1 is 4.0 multiplied by 109/g, and the freezing survival rate is 91.2%.
Uniformly mixing the bacterial sludge obtained in the step (2) with a freeze-drying protective agent according to the volume ratio of 1:1.5 to obtain a bacterial suspension, pre-freezing the bacterial suspension at-80 ℃ for 3 hours, transferring the bacterial suspension to a freeze dryer for freeze drying for 20 hours to obtain bacterial powder, and packaging the bacterial powder to obtain the microbial agent (hereinafter referred to as product b 1-1); the freeze-drying protective agent consists of the following freeze-drying protective components: 120g/L glucose, 120g/L skim milk and 15mL/L glycerol. The freezing survival rate of the product b1-1 is 63.1%.
Uniformly mixing the bacterial sludge obtained in the step (2) with a freezing protective agent according to the volume ratio of 1:1.5 to obtain bacterial suspension, pre-freezing the bacterial suspension for 3 hours at the temperature of minus 80 ℃, transferring the bacterial suspension to a freeze dryer for freeze drying for 20 hours to obtain bacterial powder, and packaging the bacterial powder to obtain the microbial agent (hereinafter referred to as product b 1-2); the freeze-drying protective agent consists of the following freeze-drying protective components: rhamnolipid 50mg/L, glucose 120g/L, skim milk 120g/L, glycerol 15 mL/L. The freezing survival rate of the product b1-2 is 79.3%.
Example 5 preparation of microbial inoculum B
(1) preparing a seed solution: inoculating thermophilic lactobacillus LAB 1# into an MRS liquid culture medium, and performing shake culture at 50-55 ℃ at 120r/min for 24 hours to obtain seed liquid; wherein the formula of the MRS liquid culture medium is that each liter of MRS liquid culture medium contains 10.0g of peptone, 5.0g of beef extract, 4.0g of yeast extract, 20.0g of glucose, 801.0 mL of tween-sodium, 2.0g of dipotassium hydrogen phosphate, 5.0g of sodium acetate, 2.0g of triammonium citrate, 0.2g of magnesium sulfate, 0.05g of manganese sulfate and the balance of distilled water;
(2) Preparation of bacterial sludge: inoculating the seed solution obtained in the step (1) into a fermentation culture medium according to the inoculation amount of 5%, carrying out shaking table culture at the temperature of 50-55 ℃ for 24 hours at 120r/min, centrifuging the fermentation product (the centrifugal rotation speed is 5000r/min, the centrifugal time is 15min), discarding supernatant, collecting thalli, and washing with normal saline to obtain bacterial sludge; wherein the formula of the fermentation medium is that each liter of fermentation medium contains 10g of glucose, 10g of yeast extract, 10g of peptone and the balance of distilled water;
(3) preparation of the microbial agent: uniformly mixing the bacterial sludge obtained in the step (2) with a freeze-drying protective agent according to the volume ratio of 1:2.0 to obtain a bacterial suspension, pre-freezing the bacterial suspension at-70 ℃ for 4 hours, transferring the bacterial suspension to a freeze dryer for freeze drying for 24 hours to obtain bacterial powder, and packaging the bacterial powder to obtain the microbial agent B (hereinafter referred to as product B2); the freeze-drying protective agent consists of the following freeze-drying protective components: 200mg/L of rhamnolipid, 80g/L of glucose, 150g/L of skim milk and 12mL/L of glycerol. The freezing survival rate of the product b2 is 93.6%.
example 6
(1) Mixing semen Maydis (10kg), product a1(300g), and water (30kg), soaking at 50-55 deg.C for 20 hr to obtain soaking mixture;
(2) and (2) transferring the soaking mixture obtained in the step (1) into a grinder to carry out coarse grinding and crushing to 5-8 petals, adding a product a1(150g), continuing soaking for 12h, carrying out fine grinding, sequentially sieving by a 50-250 mesh sieve and standing the filtrate overnight, carrying out centrifugal separation and drying to obtain the corn starch, wherein the starch yield is 70.2%.
Example 7
(1) mixing semen Maydis (10kg), product a1(300g), and water (30kg), soaking at 50-55 deg.C for 12 hr to obtain soaking mixture;
(2) and (2) transferring the soaking mixture obtained in the step (1) into a grinder to carry out coarse grinding and crushing to 5-8 petals, adding a product a1(150g), continuing soaking for 8 hours, carrying out fine grinding, sequentially sieving by a 50-250 mesh sieve and standing the filtrate overnight, carrying out centrifugal separation and drying to obtain the corn starch, wherein the starch yield is 62.3%.
example 8
(1) Mixing semen Maydis (10kg), product a1(100g), and water (30kg), soaking at 50-55 deg.C for 12 hr to obtain soaking mixture;
(2) And (2) transferring the soaking mixture obtained in the step (1) into a grinder to carry out coarse grinding and crushing to 5-8 petals, adding a product a1(50g), continuing soaking for 8 hours, carrying out fine grinding, sequentially sieving by a 50-250 mesh sieve, standing the filtrate overnight, carrying out centrifugal separation, and drying to obtain the corn starch, wherein the starch yield is 54.7%.
example 9
(1) Mixing semen Maydis (10kg), product a2(300g), and water (50kg), soaking at 50-55 deg.C for 20 hr to obtain soaking mixture;
(2) And (2) transferring the soaking mixture obtained in the step (1) into a grinder to carry out coarse grinding and crushing to 5-8 petals, adding a product a2(150g), continuing soaking for 12h, carrying out fine grinding, sequentially sieving by a 50-250 mesh sieve and standing the filtrate overnight, carrying out centrifugal separation and drying to obtain the corn starch, wherein the starch yield is 70.5%.
Example 10
(1) mixing semen Maydis (10kg), product b1(300g), and water (30kg), soaking at 50-55 deg.C for 20 hr to obtain soaking mixture;
(2) And (2) transferring the soaking mixture obtained in the step (1) into a grinder to carry out coarse grinding and crushing to 5-8 petals, adding a product b1(150g), continuing soaking for 12h, carrying out fine grinding, sequentially sieving by a 50-250 mesh sieve and standing the filtrate overnight, carrying out centrifugal separation and drying to obtain the corn starch, wherein the starch yield is 70.7%.
example 11
(1) Mixing corn (10kg), product b1(300g) and water (30kg), and soaking at 50-55 deg.C for 12 hr to obtain soaking mixture;
(2) and (2) transferring the soaking mixture obtained in the step (1) into a grinder to carry out coarse grinding and crushing to 5-8 petals, adding a product b1(150g), continuing soaking for 8 hours, carrying out fine grinding, sequentially sieving by a 50-250 mesh sieve and standing the filtrate overnight, carrying out centrifugal separation and drying to obtain the corn starch, wherein the starch yield is 63.5%.
Example 12
(1) Mixing semen Maydis (10kg), product b1(100g), and water (30kg), soaking at 50-55 deg.C for 12 hr to obtain soaking mixture;
(2) And (2) transferring the soaking mixture obtained in the step (1) into a grinder to carry out coarse grinding and crushing to 5-8 petals, adding a product b1(50g), continuing soaking for 8 hours, carrying out fine grinding, sequentially sieving by a 50-250-mesh sieve, standing the filtrate overnight, carrying out centrifugal separation, and drying to obtain the corn starch, wherein the starch yield is 55.2%.
Example 13
(1) Mixing semen Maydis (10kg), product b2(300g), and water (50kg), soaking at 50-55 deg.C for 20 hr to obtain soaking mixture;
(2) and (2) transferring the soaking mixture obtained in the step (1) into a grinder to carry out coarse grinding and crushing to 5-8 petals, adding a product b2(150g), continuing soaking for 12h, carrying out fine grinding, sequentially sieving by a 50-250 mesh sieve and standing the filtrate overnight, carrying out centrifugal separation and drying to obtain the corn starch, wherein the starch yield is 70.6%.
Example 14
(1) mixing corn (10kg), product a1(50g), product b1(50g) and water (30kg), and soaking at 50-55 deg.C for 12 hr to obtain soaking mixture;
(2) And (2) transferring the soaking mixture obtained in the step (1) into a grinder to carry out coarse grinding and crushing to 5-8 petals, adding a product a1(25g) and a product b1(25g), continuing to soak for 8 hours, carrying out fine grinding, sequentially sieving by a 50-250 mesh sieve, standing the filtrate overnight, carrying out centrifugal separation, and drying to obtain the corn starch, wherein the starch yield is 72.1%.
Example 15
(1) Mixing corn (10kg), product a1(40g), product b2(60g) and water (30kg), and soaking at 50-55 deg.C for 12 hr to obtain soaking mixture;
(2) And (2) transferring the soaking mixture obtained in the step (1) into a grinder to carry out coarse grinding and crushing to 5-8 petals, adding a product a1(20g) and a product b2(30g), continuing to soak for 8 hours, carrying out fine grinding, sequentially sieving by a 50-250 mesh sieve, standing the filtrate overnight, carrying out centrifugal separation, and drying to obtain the corn starch, wherein the starch yield is 71.9%.
Example 16
(1) mixing corn (10kg) with water (30kg), inoculating LAB 0# with 5% amount, soaking at 50-55 deg.C for 20 to obtain soaking mixture;
(2) and (2) transferring the soaking mixture obtained in the step (1) into a grinder to carry out coarse grinding and crushing to obtain 5-8 petals, then adding half of LAB 0# (namely 2.5%) to continue soaking for 12h, carrying out fine grinding, sequentially sieving by a sieve of 50 meshes and a sieve of 250 meshes, standing the filtrate overnight, carrying out centrifugal separation, and drying to obtain the corn starch, wherein the starch yield is 23.5%.
example 17
(1) mixing corn (10kg) and water (30kg), inoculating LAB 1# with 5% amount, soaking at 50-55 deg.C for 20 to obtain soaking mixture;
(2) And (2) transferring the soaking mixture obtained in the step (1) into a grinder to carry out coarse grinding and crushing to obtain 5-8 petals, then adding half of LAB 1# (namely 2.5%) to continue soaking for 12h, carrying out fine grinding, sequentially sieving by a sieve of 50 meshes and a sieve of 250 meshes, standing the filtrate overnight, carrying out centrifugal separation, and drying to obtain the corn starch, wherein the starch yield is 24.7%.
Example 18
(1) Mixing corn (10kg) and water (30kg), sequentially inoculating LAB 0# with the amount of 2.5% and LAB 1# with the amount of 2.5%, and soaking at 50-55 deg.C for 20 to obtain a soaking mixture;
(2) And (2) transferring the soaking mixture obtained in the step (1) into a grinder to carry out coarse grinding and crushing to obtain 5-8 petals, then adding half of LAB 0# and LAB 1# (namely 1.25 percent respectively) to continue soaking for 12 hours, carrying out fine grinding, sequentially sieving by using a 50-250-mesh sieve, standing the filtrate overnight, carrying out centrifugal separation, and drying to obtain the corn starch, wherein the starch yield is 30.2 percent.
Claims (5)
1. A microbial agent A is characterized in that the microbial agent A consists of thermophilic lactic acid bacteria LAB 0# and a freeze-drying protective agent; the freeze-drying protective agent consists of the following freeze-drying protective components: 200-300mg/L of rhamnolipid, 80-120g/L of glucose, 120-150g/L of skim milk and 12-15mL/L of glycerol;
the strain preservation information of the thermophilic lactic acid bacteria LAB 0 #: the name of the depository: china general microbiological culture Collection center; the address of the depository: western road No.1, north west city of township, beijing, institute of microbiology, china academy of sciences; the preservation date is as follows: 18 months 4 in 2018; the preservation number is: CGMCC No. 15623; and (3) classification and naming: bacillus coagulans Bacillus coagulons.
2. The process for preparing microbial preparation A according to claim 1, comprising the steps of:
(1) Preparing a seed solution: inoculating thermophilic lactobacillus LAB 0# into MRS liquid culture medium, and performing shake culture at 50-55 deg.C for 24 hr to obtain seed solution;
(2) Preparation of bacterial sludge: inoculating the seed solution obtained in the step (1) into a fermentation culture medium, performing shake culture at 50-55 ℃ for 24 hours, centrifuging the fermentation product, removing the supernatant, collecting the thallus, and washing with normal saline to obtain bacterial sludge;
(3) preparation of the microbial agent: uniformly mixing the bacterial sludge obtained in the step (2) with a freeze-drying protective agent to obtain a bacterial suspension, pre-freezing the bacterial suspension at a temperature of between 70 ℃ below zero and 80 ℃ for 3 to 4 hours, transferring the bacterial suspension to a freeze dryer for freeze drying for 20 to 24 hours to obtain bacterial powder, and packaging the bacterial powder to obtain the microbial agent A;
the formula of the MRS liquid culture medium in the step (1) is that each liter of MRS liquid culture medium contains 10.0g of peptone, 5.0g of beef extract, 4.0g of yeast extract, 20.0g of glucose, Tween-801.0 mL, 2.0g of dipotassium hydrogen phosphate, 5.0g of sodium acetate, 2.0g of triammonium citrate, 0.2g of magnesium sulfate, 0.05g of manganese sulfate and the balance of distilled water;
The formula of the fermentation medium in the step (2) is that each liter of fermentation medium contains 10g of glucose, 10g of yeast extract, 10g of peptone and the balance of distilled water; the inoculation amount of the seed liquid inoculated into the fermentation medium is 5 percent; the centrifugal speed is 5000r/min, and the centrifugal time is 10-15 min;
The volume ratio of the bacterial sludge to the freeze-drying protective agent in the step (3) is 1:1.5-2.0, and the freeze-drying protective agent consists of the following freeze-drying protective components: 200-300mg/L of rhamnolipid, 80-120g/L of glucose, 120-150g/L of skim milk and 12-15mL/L of glycerol.
3. The process according to claim 2, wherein the rotational speed of the rocking platforms in steps (1) and (2) is 100-120 r/min.
4. the use of microbial inoculant a according to claim 1 in wet corn steeping.
5. The use of microbial inoculant a according to claim 1 in the extraction of corn starch.
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| CN102077904A (en) * | 2010-03-11 | 2011-06-01 | 哈尔滨龙猪科技开发有限公司 | Method for preparing biological feed additive of lactobacillus |
| CN102839140A (en) * | 2012-09-07 | 2012-12-26 | 吉林中粮生化科技有限公司 | L-lactic acid producing strain separated and screened out of corn soaking water |
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