CN108060089A - A kind of Paecilomyces lilacinus solid medium prepares and fermentation method for producing - Google Patents
A kind of Paecilomyces lilacinus solid medium prepares and fermentation method for producing Download PDFInfo
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- 241001465752 Purpureocillium lilacinum Species 0.000 title claims abstract description 85
- 238000000855 fermentation Methods 0.000 title claims abstract description 59
- 230000004151 fermentation Effects 0.000 title claims abstract description 59
- 239000007787 solid Substances 0.000 title claims abstract description 53
- 238000004519 manufacturing process Methods 0.000 title abstract description 11
- 239000002609 medium Substances 0.000 claims abstract description 54
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 38
- 238000002360 preparation method Methods 0.000 claims abstract description 29
- 239000002994 raw material Substances 0.000 claims abstract description 27
- 239000001963 growth medium Substances 0.000 claims abstract description 21
- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 20
- 235000009566 rice Nutrition 0.000 claims abstract description 20
- 239000007788 liquid Substances 0.000 claims abstract description 14
- 240000008042 Zea mays Species 0.000 claims description 33
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 33
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 33
- 235000005822 corn Nutrition 0.000 claims description 33
- 235000013312 flour Nutrition 0.000 claims description 21
- 230000001954 sterilising effect Effects 0.000 claims description 16
- 238000011081 inoculation Methods 0.000 claims description 15
- 239000000843 powder Substances 0.000 claims description 11
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 10
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 10
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 10
- 239000004202 carbamide Substances 0.000 claims description 10
- 239000008103 glucose Substances 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 239000002002 slurry Substances 0.000 claims description 10
- 238000004659 sterilization and disinfection Methods 0.000 claims description 10
- 238000003756 stirring Methods 0.000 claims description 10
- 241000894006 Bacteria Species 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 7
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 5
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 5
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 5
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 claims description 5
- 229910000388 diammonium phosphate Inorganic materials 0.000 claims description 5
- 235000019838 diammonium phosphate Nutrition 0.000 claims description 5
- 241000894007 species Species 0.000 claims description 2
- 240000007594 Oryza sativa Species 0.000 claims 3
- 241000209094 Oryza Species 0.000 abstract description 17
- 229930006000 Sucrose Natural products 0.000 abstract description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 abstract description 4
- 239000005720 sucrose Substances 0.000 abstract description 4
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 abstract 2
- 238000009835 boiling Methods 0.000 abstract 1
- 238000009395 breeding Methods 0.000 abstract 1
- 230000001488 breeding effect Effects 0.000 abstract 1
- 239000002054 inoculum Substances 0.000 abstract 1
- 229920006395 saturated elastomer Polymers 0.000 abstract 1
- 238000011177 media preparation Methods 0.000 description 5
- 241000243785 Meloidogyne javanica Species 0.000 description 3
- 208000035240 Disease Resistance Diseases 0.000 description 2
- 238000010411 cooking Methods 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000012364 cultivation method Methods 0.000 description 2
- 241000609240 Ambelania acida Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 241001236817 Paecilomyces <Clavicipitaceae> Species 0.000 description 1
- 239000010905 bagasse Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000012272 crop production Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 238000010977 unit operation Methods 0.000 description 1
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Abstract
Description
技术领域technical field
本发明涉及微生物技术及生物利用领域，具体涉及一种淡紫拟青霉固体培养基制备及发酵生产方法。The invention relates to the field of microbial technology and biological utilization, in particular to a method for preparing and fermenting a solid culture medium of Paecilomyces lilacinus.
背景技术Background technique
根结线虫已给农作物的生产带来严重的危害。近几年来报道的有效防治根结线虫的真菌很多其中最引人注意的一种是淡紫拟青霉。据文献报道淡紫拟青霉对根结线虫有着明显的抑制效果。针对淡紫拟青霉的大规模发酵生产及培养基制备成为了一个研究热点之一。Root-knot nematodes have caused serious damage to crop production. In recent years, many fungi have been reported to effectively control root-knot nematodes, and one of the most notable ones is Paecilomyces lilacinus. According to literature reports, Paecilomyces lilacinus has obvious inhibitory effect on root-knot nematodes. The large-scale fermentation production and medium preparation of Paecilomyces lilacinus has become one of the research hotspots.
专利201610790557公开了一种淡紫拟青霉发酵生产方法及淡紫拟青霉二级液体培养基的方法，液体培养基采取黄豆粉为主要原料；专利201010209110公开了一种淡紫拟青霉的培养方法，选取麸皮、玉米、甘蔗渣为主要原料；专利201210074067公开了一种淡紫拟青霉及其培养方法和应用，培养基选取米糠、玉米粉为主要原料；专利201611258790公开了淡紫拟青霉及其规模化制备方法，培养基选取玉米秸秆为主要原料。本文提供一种使用廉价原料，分别对淡紫拟青霉固体和液体进行发酵的培养基组成、发酵工艺要点进行研究。Patent 201610790557 discloses a fermentation production method of Paecilomyces lilacinus and a method for the secondary liquid medium of Paecilomyces lilacinus. The liquid medium uses soybean powder as the main raw material; For the cultivation method, bran, corn, and bagasse are selected as main raw materials; patent 201210074067 discloses a Paecilomyces lilacinus and its cultivation method and application, and rice bran and corn flour are selected as the main raw materials for the culture medium; patent 201611258790 discloses Paecilomyces lilacinus Paecilomyces and its large-scale preparation method, the culture medium adopts corn stalks as the main raw material. This article provides a study on the composition of the medium and the key points of the fermentation process for the solid and liquid fermentation of Paecilomyces lilacinus using cheap raw materials.
发明内容Contents of the invention
本发明所要解决的技术问题是提供一种淡紫拟青霉固体培养基制备及发酵生产方法，制备廉价有效的淡紫拟青霉固体培养基，解决了淡紫拟青霉固体发酵的关键操作，扩大了菌体产量，提高了菌体抗病能力。The technical problem to be solved by the present invention is to provide a method for the preparation and fermentation of Paecilomyces lilacinus solid medium, to prepare cheap and effective Paecilomyces lilacinus solid medium, and to solve the key operation of Paecilomyces lilacinus solid fermentation , Expand the production of bacteria and improve the disease resistance of bacteria.
为实现上述目的，本发明采用以下技术方案实现：To achieve the above object, the present invention adopts the following technical solutions:
一种淡紫拟青霉固体培养基制备，具体包括以下步骤：A kind of Paecilomyces lilacinus solid medium preparation specifically comprises the following steps:
1.淡紫拟青霉一级菌种制备1. Preparation of the primary strain of Paecilomyces lilacinus
1)培养基原料：玉米粉5～10g，玉米浆10～15g，脲0.1～0.5g，葡萄糖2～5g，硫酸铵1～2.5g，磷酸氢二铵1～2.5g，酵母粉1～2.5g，水200～500g；1) Culture medium raw materials: corn flour 5-10g, corn steep liquor 10-15g, urea 0.1-0.5g, glucose 2-5g, ammonium sulfate 1-2.5g, diammonium hydrogen phosphate 1-2.5g, yeast powder 1-2.5 g, water 200-500g;
2)配制培养基：取15g～20g玉米粉熬成浆，再将其他原料加入，搅拌，加水，装入1个1000ml的三角瓶；2) Preparation of culture medium: take 15g~20g corn flour and boil it into a slurry, then add other raw materials, stir, add water, and put it into a 1000ml triangular flask;
3)灭菌：把装有培养基的三角瓶放入灭菌锅中，120～125℃，灭菌25～35min；3) Sterilization: Put the triangular flask containing the culture medium into a sterilizing pot, sterilize at 120-125°C for 25-35 minutes;
4)接菌：在无菌室内，把活化后的淡紫拟青霉接种到已灭菌的三角瓶中，放入25～30℃培养箱中培养；4) Inoculation: in the sterile room, inoculate the activated Paecilomyces lilacinus into a sterilized Erlenmeyer flask, and put it into an incubator at 25-30°C for cultivation;
2.淡紫拟青霉二级菌种制备2. Preparation of secondary strains of Paecilomyces lilacinus
1)培养基原料：玉米粉10g～20g，玉米浆20～30g，脲0.5～1g，酵母粉2～5g，葡萄糖10～20g，水800g～1000g；1) Medium raw materials: 10g-20g corn flour, 20-30g corn steep liquor, 0.5-1g urea, 2-5g yeast powder, 10-20g glucose, 800g-1000g water;
2)配制培养基：取30～50g玉米粉熬成浆，再将其他原料加入，搅拌，加水补至1000ml，装入2个1000ml的三角瓶；2) Preparation of medium: Take 30-50g of corn flour and boil it into a slurry, then add other raw materials, stir, add water to make up to 1000ml, and put it into two 1000ml triangular flasks;
3)灭菌：把装有培养基的三角瓶放入灭菌锅中，120～125℃灭菌25～35min；3) Sterilization: put the triangular flask containing the culture medium into a sterilizing pot, and sterilize at 120-125°C for 25-35 minutes;
4)接菌：在无菌室内，把一级菌种接种到已灭菌的三角瓶中，放入25～30℃培养箱中；4) Inoculation: in the sterile room, inoculate the first-grade bacteria into the sterilized Erlenmeyer flask, and put it in the incubator at 25-30°C;
淡紫拟青霉固体发酵方法，具体包括以下步骤：Paecilomyces lilacinus solid fermentation method specifically comprises the following steps:
1.淡紫拟青霉固体发酵培养基制备1. Preparation of Paecilomyces lilacinus solid fermentation medium
将淡紫拟青霉固体发酵培养基陈大米200g～300g倒入盆中，向其注入清水，使大米完全没在清水中,浸泡4～8h，使大米完全吸饱水，同时加入蔗糖0.5～1g，控制蒸汽压力0.12～0.15MPa，温度120～125℃，蒸煮30～35min，降温到25～30℃得淡紫拟青霉固体发酵培养基；Pour 200g~300g of Paecilomyces lilacinus solid fermentation medium old rice into the pot, pour water into it, make the rice completely submerged in the water, soak for 4~8 hours, make the rice fully absorb water, and add 0.5~ 1g, control steam pressure 0.12-0.15MPa, temperature 120-125°C, cook for 30-35min, cool down to 25-30°C to obtain Paecilomyces lilacinus solid fermentation medium;
2.淡紫拟青霉固体发酵培养基接种2. Inoculation of Paecilomyces lilacinus solid fermentation medium
在无菌室内，用无菌量杯接取淡紫拟青霉二级菌种液体，均匀地淋在固体发酵培养基上，混拌，使淡紫拟青霉菌种液体与固体发酵培养基充分混合均匀，放入25～30℃培养箱中里进行发酵培养7～9天，得淡紫拟青霉菌体。In the aseptic room, use a sterile measuring cup to take the liquid of the Paecilomyces lilacinus secondary strain, evenly pour it on the solid fermentation medium, and mix it, so that the Paecilomyces lilacinus liquid and the solid fermentation medium are fully mixed Evenly, put it into an incubator at 25-30° C. to carry out fermentation and culture for 7-9 days to obtain Paecilomyces lilacinus thallus.
淡紫拟青霉菌体孢子数可达8×109cfu/g，纯种率达90％-95％。The number of spores of Paecilomyces lilacinus can reach 8×10 9 cfu/g, and the pure species rate can reach 90%-95%.
与现有的技术相比，本发明的有益效果是：Compared with prior art, the beneficial effect of the present invention is:
1.废弃的陈大米作为发酵培养基原料，适合淡紫拟青霉大量生长繁殖，获得的菌体纯种率高，纯种率达90％-95％，菌体生长能力旺盛，扩大了菌体产量，提高了菌体抗病能力。1. Abandoned old rice is used as the raw material of the fermentation medium, which is suitable for the large-scale growth and reproduction of Paecilomyces lilacinus. Body production, improve the disease resistance of bacteria.
2.本发明通过调整培养基组成、控制发酵生产各参数及操作要点，对发酵生产各单元操作的细分，严格控制反应条件，寻求发酵生产最优化。2. The present invention seeks the optimization of fermentation production by adjusting the composition of the medium, controlling the parameters and operating points of fermentation production, subdividing each unit operation of fermentation production, and strictly controlling the reaction conditions.
3.培养基成本低廉，生产可靠，适合于大规模发酵生产。3. The medium cost is low, the production is reliable, and it is suitable for large-scale fermentation production.
具体实施方式Detailed ways
下面结合实施例对本发明进一步说明：Below in conjunction with embodiment the present invention is further described:
以下实施例对本发明进行详细描述。这些实施例仅是对本发明的最佳实施方案进行描述，并不对本发明的范围进行限制。The following examples describe the present invention in detail. These examples are only to describe the best embodiment of the present invention, and do not limit the scope of the present invention.
实施例1Example 1
一种淡紫拟青霉固体培养基制备，具体包括以下步骤：A kind of Paecilomyces lilacinus solid medium preparation specifically comprises the following steps:
1.淡紫拟青霉一级菌种制备1. Preparation of the primary strain of Paecilomyces lilacinus
1)培养基原料：玉米粉10g，玉米浆12g，脲0.4g，葡萄糖3g，硫酸铵2g，磷酸氢二铵2g，酵母粉2.5g，水500g；1) Medium raw materials: 10g corn flour, 12g corn steep liquor, 0.4g urea, 3g glucose, 2g ammonium sulfate, 2g diammonium hydrogen phosphate, 2.5g yeast powder, 500g water;
2)配制培养基：取18g玉米粉熬成浆，再将其他原料加入，搅拌，加水，装入1个1000ml的三角瓶；2) Preparation of medium: Take 18g of corn flour and boil it into a slurry, then add other raw materials, stir, add water, and put it into a 1000ml triangular flask;
3)灭菌：把装有培养基的三角瓶放入灭菌锅中，121℃，灭菌30min；3) Sterilization: Put the triangular flask containing the culture medium into a sterilizer, sterilize at 121°C for 30 minutes;
4)接菌：在无菌室内，把活化后的淡紫拟青霉接种到已灭菌的三角瓶中，放入28℃培养箱中培养；4) Inoculation: in the sterile room, inoculate the activated Paecilomyces lilacinus into a sterilized Erlenmeyer flask, and put it into a 28°C incubator for cultivation;
2.淡紫拟青霉二级菌种制备2. Preparation of secondary strains of Paecilomyces lilacinus
1)培养基原料：玉米粉15g，玉米浆25g，脲0.6g，酵母粉3g，葡萄糖18g，水900g；1) Medium raw materials: 15g corn flour, 25g corn steep liquor, 0.6g urea, 3g yeast powder, 18g glucose, 900g water;
2)配制培养基：取40g玉米粉熬成浆，再将其他原料加入，搅拌，加水补至1000ml，装入2个1000ml的三角瓶；2) Preparation of culture medium: take 40g of corn flour and boil it into a slurry, then add other raw materials, stir, add water to make up to 1000ml, and put it into two 1000ml triangular flasks;
3)灭菌：把装有培养基的三角瓶放入灭菌锅中，123℃灭菌25min；3) Sterilization: put the triangular flask containing the culture medium into a sterilizer, and sterilize at 123°C for 25 minutes;
4)接菌：在无菌室内，把一级菌种接种到已灭菌的三角瓶中，放入28℃培养箱中；4) Inoculation: in the sterile room, inoculate the first-grade bacteria into a sterilized Erlenmeyer flask, and put it in a 28°C incubator;
淡紫拟青霉固体发酵方法Paecilomyces lilacinus solid fermentation method
1.淡紫拟青霉固体发酵培养基制备1. Preparation of Paecilomyces lilacinus solid fermentation medium
将淡紫拟青霉固体发酵培养基陈大米300g倒入盆中，向其注入清水，使大米完全没在清水中,浸泡6h，使大米完全吸饱水，同时加入蔗糖1g，控制蒸汽压力0.13MPa，温度121℃，蒸煮35min，降温到30℃淡紫拟青霉固体发酵培养基；Pour 300g of Paecilomyces lilacinus solid fermentation medium old rice into the pot, pour clean water into it, make the rice completely submerged in clean water, soak for 6 hours, make the rice fully absorb water, add 1g of sucrose at the same time, control the steam pressure 0.13 MPa, temperature 121°C, cook for 35min, cool down to 30°C Paecilomyces lilacinus solid fermentation medium;
2.淡紫拟青霉固体发酵培养基接种2. Inoculation of Paecilomyces lilacinus solid fermentation medium
在无菌室内，用无菌量杯接取淡紫拟青霉二级菌种液体，均匀地淋在固体发酵培养基上，混拌，使淡紫拟青霉菌种液体与固体发酵培养基充分混合均匀，放入28℃培养箱中里进行发酵培养7～9天，得淡紫拟青霉菌体。。In the aseptic room, use a sterile measuring cup to take the liquid of the Paecilomyces lilacinus secondary strain, evenly pour it on the solid fermentation medium, and mix it, so that the Paecilomyces lilacinus liquid and the solid fermentation medium are fully mixed Evenly, put it into an incubator at 28° C. for fermentation and cultivation for 7 to 9 days to obtain Paecilomyces lilacinus thallus. .
实施例2Example 2
一种淡紫拟青霉固体培养基制备，具体包括以下步骤：A kind of Paecilomyces lilacinus solid medium preparation specifically comprises the following steps:
1.淡紫拟青霉一级菌种制备1. Preparation of the primary strain of Paecilomyces lilacinus
1)培养基原料：玉米粉6g，玉米浆10g，脲0.3g，葡萄糖3g，硫酸铵1.5g，磷酸氢二铵2g，酵母粉2g，水400g；1) Medium raw materials: 6g corn flour, 10g corn steep liquor, 0.3g urea, 3g glucose, 1.5g ammonium sulfate, 2g diammonium hydrogen phosphate, 2g yeast powder, 400g water;
2)配制培养基：取18g玉米粉熬成浆，再将其他原料加入，搅拌，加水，装入1个1000ml的三角瓶；2) Preparation of medium: Take 18g of corn flour and boil it into a slurry, then add other raw materials, stir, add water, and put it into a 1000ml triangular flask;
3)灭菌：把装有培养基的三角瓶放入灭菌锅中，122℃，灭菌25min；3) Sterilization: put the triangular flask containing the culture medium into a sterilizing pot, and sterilize at 122°C for 25 minutes;
4)接菌：在无菌室内，把活化后的淡紫拟青霉接种到已灭菌的三角瓶中，放入28℃培养箱中培养；4) Inoculation: in the sterile room, inoculate the activated Paecilomyces lilacinus into a sterilized Erlenmeyer flask, and put it into a 28°C incubator for cultivation;
2.淡紫拟青霉二级菌种制备2. Preparation of secondary strains of Paecilomyces lilacinus
1)培养基原料：玉米粉10g，玉米浆20g，脲1g，酵母粉4g，葡萄糖15g，水800g；1) Medium raw materials: 10g corn flour, 20g corn steep liquor, 1g urea, 4g yeast powder, 15g glucose, 800g water;
2)配制培养基：取40g玉米粉熬成浆，再将其他原料加入，搅拌，加水补至1000ml，装入2个1000ml的三角瓶；2) Preparation of culture medium: take 40g of corn flour and boil it into a slurry, then add other raw materials, stir, add water to make up to 1000ml, and put it into two 1000ml triangular flasks;
3)灭菌：把装有培养基的三角瓶放入灭菌锅中，120℃灭菌35min；3) Sterilization: put the triangular flask containing the culture medium into a sterilizer, and sterilize at 120°C for 35 minutes;
4)接菌：在无菌室内，把一级菌种接种到已灭菌的三角瓶中，放入28℃培养箱中；4) Inoculation: in the sterile room, inoculate the first-grade bacteria into a sterilized Erlenmeyer flask, and put it in a 28°C incubator;
淡紫拟青霉固体发酵方法Paecilomyces lilacinus solid fermentation method
1.淡紫拟青霉固体发酵培养基制备1. Preparation of Paecilomyces lilacinus solid fermentation medium
将淡紫拟青霉固体发酵培养基陈大米200g倒入盆中，向其注入清水，使大米完全没在清水中,浸泡8h，使大米完全吸饱水，同时加入蔗糖0.8g，控制蒸汽压力0.13MPa，温度125℃，蒸煮30min，降温到25℃得淡紫拟青霉固体发酵培养基；Pour 200g of Paecilomyces lilacinus solid fermentation medium aged rice into the pot, pour clean water into it, make the rice completely submerged in clean water, soak for 8 hours, make the rice fully absorb water, add 0.8g of sucrose at the same time, control the steam pressure 0.13MPa, temperature 125°C, cooking for 30min, cooling to 25°C to obtain Paecilomyces lilacinus solid fermentation medium;
2.淡紫拟青霉固体发酵培养基接种2. Inoculation of Paecilomyces lilacinus solid fermentation medium
在无菌室内，用无菌量杯接取淡紫拟青霉二级菌种液体，均匀地淋在固体发酵培养基上，混拌，使淡紫拟青霉菌种液体与固体发酵培养基充分混合均匀，放入30℃培养箱中里进行发酵培养7～9天，得淡紫拟青霉菌体。In the aseptic room, use a sterile measuring cup to take the liquid of the Paecilomyces lilacinus secondary strain, evenly pour it on the solid fermentation medium, and mix it, so that the Paecilomyces lilacinus liquid and the solid fermentation medium are fully mixed Evenly, put it into an incubator at 30° C. for fermentation and cultivation for 7 to 9 days to obtain Paecilomyces lilacinus thallus.
实施例3Example 3
一种淡紫拟青霉固体培养基制备，具体包括以下步骤：A kind of Paecilomyces lilacinus solid medium preparation specifically comprises the following steps:
1.淡紫拟青霉一级菌种制备1. Preparation of the primary strain of Paecilomyces lilacinus
1)培养基原料：玉米粉8g，玉米浆10g，脲0.5g，葡萄糖3g，硫酸铵2g，磷酸氢二铵2.5g，酵母粉2.5g，水300g；1) Medium raw materials: 8g corn flour, 10g corn steep liquor, 0.5g urea, 3g glucose, 2g ammonium sulfate, 2.5g diammonium hydrogen phosphate, 2.5g yeast powder, 300g water;
2)配制培养基：取20g玉米粉熬成浆，再将其他原料加入，搅拌，加水，装入1个1000ml的三角瓶；2) Preparation of medium: Take 20g of corn flour and boil it into a slurry, then add other raw materials, stir, add water, and put it into a 1000ml triangular flask;
3)灭菌：把装有培养基的三角瓶放入灭菌锅中，123℃，灭菌35min；3) Sterilization: Put the triangular flask containing the culture medium into a sterilizing pot, sterilize at 123°C for 35 minutes;
4)接菌：在无菌室内，把活化后的淡紫拟青霉接种到已灭菌的三角瓶中，放入28℃培养箱中培养；4) Inoculation: in the sterile room, inoculate the activated Paecilomyces lilacinus into a sterilized Erlenmeyer flask, and put it into a 28°C incubator for cultivation;
2.淡紫拟青霉二级菌种制备2. Preparation of secondary strains of Paecilomyces lilacinus
1)培养基原料：玉米粉15g，玉米浆25g，脲0.8g，酵母粉5g，葡萄糖10g，水800g；1) Medium raw materials: 15g corn flour, 25g corn steep liquor, 0.8g urea, 5g yeast powder, 10g glucose, 800g water;
2)配制培养基：取30～50g玉米粉熬成浆，再将其他原料加入，搅拌，加水补至1000ml，装入2个1000ml的三角瓶；2) Preparation of medium: Take 30-50g of corn flour and boil it into a slurry, then add other raw materials, stir, add water to make up to 1000ml, and put it into two 1000ml triangular flasks;
3)灭菌：把装有培养基的三角瓶放入灭菌锅中，120℃灭菌35min；3) Sterilization: put the triangular flask containing the culture medium into a sterilizer, and sterilize at 120°C for 35 minutes;
4)接菌：在无菌室内，把一级菌种接种到已灭菌的三角瓶中，放入28℃培养箱中；4) Inoculation: in the sterile room, inoculate the first-grade bacteria into a sterilized Erlenmeyer flask, and put it in a 28°C incubator;
淡紫拟青霉固体发酵方法Paecilomyces lilacinus solid fermentation method
1.淡紫拟青霉固体发酵培养基制备1. Preparation of Paecilomyces lilacinus solid fermentation medium
将淡紫拟青霉固体发酵培养基陈大米300g倒入盆中，向其注入清水，使大米完全没在清水中,浸泡6h，使大米完全吸饱水，同时加入蔗糖1g，控制蒸汽压力0.15MPa，温度125℃，蒸煮30min，降温到28℃得淡紫拟青霉固体发酵培养基；Pour 300g of Paecilomyces lilacinus solid fermentation medium old rice into the pot, pour clean water into it, make the rice completely submerged in clean water, soak for 6 hours, make the rice fully absorb water, add 1g of sucrose at the same time, control the steam pressure 0.15 MPa, temperature 125°C, cooking for 30min, cooling to 28°C to obtain Paecilomyces lilacinus solid fermentation medium;
2.淡紫拟青霉固体发酵培养基接种2. Inoculation of Paecilomyces lilacinus solid fermentation medium
在无菌室内，用无菌量杯接取淡紫拟青霉二级菌种液体，均匀地淋在固体发酵培养基上，混拌，使淡紫拟青霉菌种液体与固体发酵培养基充分混合均匀，放入28℃培养箱中里进行发酵培养7～9天，得淡紫拟青霉菌体。In the aseptic room, use a sterile measuring cup to take the liquid of the Paecilomyces lilacinus secondary strain, evenly pour it on the solid fermentation medium, and mix it, so that the Paecilomyces lilacinus liquid and the solid fermentation medium are fully mixed Evenly, put it into an incubator at 28° C. for fermentation and cultivation for 7 to 9 days to obtain Paecilomyces lilacinus thallus.
实施1～3淡紫拟青霉固体发酵菌落生长状态见表1；Implementation 1～3 Paecilomyces lilacinus solid fermentation colony growth state is shown in Table 1;
表1：淡紫拟青霉固体发酵菌落生长状态Table 1: Growth status of Paecilomyces lilacinus solid fermentation colonies
。 .
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| CN111454847A (en) * | 2020-04-13 | 2020-07-28 | 陕西赛恩农业科技股份有限公司 | Solid fermentation method and solid fermentation culture medium for paecilomyces lilacinus |
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