Production method of glucose oxidase for feeding

The technology of a glucose oxidase and production method, which is applied in the field of feed processing, can solve problems such as difficult control of fermentation conditions, and achieve the effects of easy control of fermentation conditions, high activity, and less contamination of miscellaneous bacteria

Inactive Publication Date: 2015-03-04

CHANGSHA UNIVERSITY OF SCIENCE AND TECHNOLOGY

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AI-Extracted Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a production method of glucose oxidase for feed, to solve the pr...

Abstract

The invention relates to a production method of glucose oxidase for feeding, and aims at solving the problem of an existing common solid state fermentation technology that the fermentation conditions are difficult to control. The production method of the glucose oxidase for feeding comprises the following steps: culturing activated penicillium notatum for at least one time to obtain penicillium notatum seeds; fermenting the penicillium notatum seeds by adopting a fluidized bed solid state fermentation method; after the fermentation, drying a solid culture medium, a fermentation product and a fermentation strain in a fluidized bed solid state fermentation tank; and then crushing the product to obtain a glucose oxidase finished product. The production method is easy to control, the product is not easily polluted by infectious microbes, and three wastes are not generated so that no pollution to the environment is caused; and after the fermentation, the fermentation product, namely the glucose oxidase, does not need to be purified, and rice dregs and/or rice bran, penicillium notatum strains and the product glucose oxidase can be directly dried to prepare the glucose oxidase for feeding. The production method has a simple process, and the biological cycle utilization of the rice dregs and/or rice bran can be promoted.

Application Domain

Animal feeding stuff

Technology Topic

Penicillium bilaiaeBiotechnology +11

Examples

Experimental program(3)

Example Embodiment

[0030] Example 1

[0031] The production method of feed glucose oxidase is as follows:

[0032] The first step is to activate the strain: take out the slant surface of Penicillium indicum, which produces high glucose oxidase, streak it and inoculate it in a PDA (potato dextrose agar medium, the same below) with a diameter of 10 cm, and incubate at 28°C for 72 hours , And then spread and inoculate it on a PDA solid culture plate with a diameter of 20cm, and culture it at 28°C for 48h to obtain activated specific Penicillium.

[0033] Preparation of potato agar medium: 200g peeled potatoes, cut into small pieces, add water and boil the juice for 30min, filter with gauze, add 15g agar to the filtrate, continue heating and stirring until the agar is dissolved, then add 20g glucose and distilled water to 1000mL, 115 Sterilize at ℃ for 20min, then spread the plate for later use;

[0034] The second step, first-level seed culture: After the first-level liquid seed medium is sterilized at 115°C for 20 minutes, the activated Penicillium indicum is transferred to the first-level seed medium, and the inoculum is a 20cm petri dish to inoculate 5L liquid seed medium After inoculation, cultivate at 32℃ for 72h to obtain the first-level seed liquid.

[0035] The preparation method of the first-level liquid seed culture medium: 2% rice residue, 0.2% NaCl, 0.2% CaCl 2 , 0.2% KH 2 PO 4 , 0.06% MgSO 4 ·7H 2 O, the rest is water;

[0036] The third step, fluidized bed solid-state fermentation: put 45% rice residue and 55% water into a fluidized bed solid-state fermentation tank, mix well, sterilize at 115°C for 20 minutes, and then connect to the first-level seed liquid and mix evenly after cooling. It is 5%, that is, 2000L seed culture medium is inoculated with 40T solid fermentation medium, and sterile air is passed into it, and the pressure is kept at 0.06MPa under 28 conditions. When cultured for 18h, add 0.05% cotton sugar, continue culture for 24h, and dry at 50℃ for 5h to moisture The content is less than or equal to 12%, and it is crushed to a fineness of more than or equal to 85%, which is the finished product of feed glucose oxidase. After testing, the enzyme activity of the product of this example was 30 GOD U/g.

Example Embodiment

[0037] Example 2

[0038] The production method of feed glucose oxidase is as follows:

[0039] The first step, strain activation: take out the preserved slope of high glucose oxidase-producing Penicillium indicum, streak it and inoculate it on a PDA solid culture plate, incubate it at 32℃ for 96 hours and then spread it on a 20cm diameter PDA solid Incubate the plate for 96h at 32°C.

[0040] Preparation of the potato agar medium: 200g of peeled potatoes, cut into small pieces, add water and boil juice for 30min, filter with 8 layers of gauze, add 15g of agar to the filtrate, continue to heat and stir to mix until the agar is dissolved, then add 20g of glucose and make up to 1000mL , Sterilize at 115°C for 20 minutes, then spread the plate for use;

[0041] The second step, first-level seed culture: After the first-level liquid seed medium is sterilized at 115°C for 20 minutes, the activated Penicillium indicum is transferred to the first-level seed medium, and the inoculum is a 20cm petri dish to inoculate 5L liquid seed medium ; After inoculation, cultivate at 32°C for 72 hours to obtain a first-level seed solution.

[0042] Preparation of the first-level liquid seed culture medium: 1% rice residue, 0.05% NaCl, 0.05% CaCl 2 ,0.05%KH 2 PO 4 , 0.01%MgSO 4 ·7H 2 O, 0.001% FeCl 3 , The rest is water;

[0043] The third step, secondary seed culture: After the secondary liquid seed medium is sterilized at 115°C for 20 minutes, inoculate the primary seed liquid with an inoculum amount of 10%, that is, 5L primary seed liquid inoculates 50L secondary seed medium; After incubating at 32°C for 72 hours, a secondary seed liquid is obtained;

[0044] Preparation of the secondary liquid seed culture medium: 3% rice residue, 0.2% NaCl, 0.2% CaCl 2 , 0.2% KH 2 PO 4 , 0.6% MgSO 4 ·7H 2 O, the rest is water;

[0045] The fourth step, seed culture: after the seed medium is sterilized at 115°C for 20 minutes, inoculate the secondary seed liquid with an inoculum amount of 10%, that is, 50L of the primary seed liquid inoculate 500L of the seed medium and cultivate at 32°C for 72 hours to obtain the seed liquid .

[0046] The seed culture medium preparation: 4% rice residue, 0.2% NaCl, 0.2% CaCl 2 , 0.2% KH 2 PO 4 , 0.06% MgSO 4 ·7H 2 O, the rest is water;

[0047] The fifth step, fluidized bed solid-state fermentation: Put 44% rice residue, 11% rice bran and 45% water into a fluidized bed solid-state fermentation tank, mix well, sterilize at 115°C for 20 minutes, cool down and insert seed liquid and mix well. The inoculation amount is 1%, that is, 500L seed medium is used to inoculate 50T solid fermentation medium. Sterile air is passed through, and the pressure is maintained at 32°C at 0.15MPa. When cultured for 24 hours, add 0.1% cotton sugar (mixed with a small amount of aqueous solution and added in spray form) , Continue to cultivate for 12h; after solid-state fermentation, dry for 2h at 60°C until the moisture content is ≤12%, and crush to the fineness ≥85%, which is the finished product of feed glucose oxidase. After testing, the enzyme activity of the product of this example was 33 GOD U/g.

Example Embodiment

[0048] Example 3

[0049] The first step is to activate the strain: take out the preserved slant of Penicillium indicum with high glucose oxidase production, streak it and inoculate it on a PDA solid culture dish with a diameter of 10cm, incubate at 30℃ for 72h, and then spread it on 20cm PDA solid culture plate, culture for 72h at 30℃.

[0050] Preparation of the potato agar medium: 200g of peeled potatoes, cut into small pieces, add water and boil the juice for 30 minutes, filter with 8 layers of gauze, add 15g of agar to the filtrate, continue to heat and stir to mix until the agar is dissolved, then add 20g of glucose, and make up to 1000mL with distilled water , Sterilize at 115°C for 20min, then spread the plate for use

[0051] The second step, first-level seed culture: After the first-level liquid seed medium is sterilized at 115°C for 20 minutes, the activated Penicillium indicum is transferred to the first-level seed medium, and the inoculum is a 20cm petri dish to inoculate 5L liquid seed medium ; Cultivate 60h at 30℃ after inoculation;

[0052] Preparation of the first-level liquid seed culture medium: 1.5% rice residue, 0.05% NaCl, 0.05% CaCl 2 ,0.05%KH 2 PO 4 , 0.03% MgSO 4 ·7H 2 O, 0.025% FeCl 3 , The rest is water;

[0053] The third step, second-level seed culture: After the second-level liquid seed medium is sterilized at 115°C for 20 minutes, inoculate the first-level seed liquid with an inoculum of 8.3% (ie: 5L first-level seed liquid inoculate 60L second-level seed medium) ; Cultivate 60h at 30℃ after inoculation;

[0054] Preparation of the secondary liquid seed culture medium: 1.5% rice residue, 0.05% NaCl, 0.05% CaCl 2 ,0.05%KH 2 PO 4 , 0.01%MgSO 4 ·7H 2 O, 0.020% FeCl 3 , The rest is water;

[0055] The fourth step, seed culture: After the seed medium is sterilized at 115°C for 20 minutes, inoculate the secondary seed liquid with an inoculum amount of 8% (ie: 60L secondary seed liquid inoculate 750L seed medium), cultivate at 30°C for 60 hours;

[0056] The seed culture medium preparation: 2% rice residue, 0.05% NaCl, 0.05% CaCl 2 ,0.05%KH 2 PO 4 , 0.01%MgSO 4 ·7H 2 O, 0.015% FeCl 3 , The rest is water;

[0057] The fifth step, fluidized bed solid-state fermentation: Put 35% rice residue, 15% rice bran and 50% water into a fluidized bed solid-state fermentation tank, mix well, sterilize at 115°C for 20 minutes, cool down and insert seed liquid and mix well. The inoculation amount is 2.5%, that is, 750L seed medium is used to inoculate 30T solid fermentation medium. Sterile air is passed through, and the pressure is maintained at 30°C to 0.10MPa. When cultured for 24 hours, add 0.07% cotton sugar (mixed into a small amount of aqueous solution and added in the form of spray) , Continue to cultivate for 18h; after the solid-state fermentation, dry at 65°C for 3h until the moisture content is ≤12%, and pulverize to the fineness ≥85%, which is the finished product of feed glucose oxidase. After testing, the enzyme activity of the product of this example was 35 GOD U/g.

PUM

Property	Measurement	Unit
Diameter	10.0	cm

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Production method of glucose oxidase for feeding

AI-Extracted Technical Summary

Problems solved by technology

Abstract

Examples

Example Embodiment

Example Embodiment

Example Embodiment

PUM

Description & Claims & Application Information

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