CN102277299B - Preparation method of microbial fermentation medium - Google Patents
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- 238000000855 fermentation Methods 0.000 title claims abstract description 36
- 230000004151 fermentation Effects 0.000 title claims abstract description 36
- 230000000813 microbial effect Effects 0.000 title claims abstract description 29
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 239000002994 raw material Substances 0.000 claims abstract description 25
- 239000010902 straw Substances 0.000 claims abstract description 18
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 14
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000000843 powder Substances 0.000 claims abstract description 9
- 238000000605 extraction Methods 0.000 claims abstract description 5
- 240000008042 Zea mays Species 0.000 claims description 6
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 6
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 6
- 235000005822 corn Nutrition 0.000 claims description 6
- 244000005700 microbiome Species 0.000 claims description 5
- 238000003306 harvesting Methods 0.000 claims description 3
- 230000001954 sterilising effect Effects 0.000 claims description 3
- 238000004659 sterilization and disinfection Methods 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims 1
- 238000010298 pulverizing process Methods 0.000 claims 1
- 238000007873 sieving Methods 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 10
- 241000894006 Bacteria Species 0.000 abstract description 9
- 230000000694 effects Effects 0.000 abstract description 4
- 239000003795 chemical substances by application Substances 0.000 abstract description 2
- 239000012141 concentrate Substances 0.000 abstract description 2
- 239000000126 substance Substances 0.000 abstract description 2
- 239000002609 medium Substances 0.000 description 26
- 238000000034 method Methods 0.000 description 8
- 239000000047 product Substances 0.000 description 7
- 239000001963 growth medium Substances 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 238000011161 development Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 239000003337 fertilizer Substances 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 3
- 238000002386 leaching Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- 150000008163 sugars Chemical class 0.000 description 2
- 238000003809 water extraction Methods 0.000 description 2
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 238000012356 Product development Methods 0.000 description 1
- 241000589180 Rhizobium Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 240000006394 Sorghum bicolor Species 0.000 description 1
- 235000011684 Sorghum saccharatum Nutrition 0.000 description 1
- 235000019764 Soybean Meal Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
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- 210000005069 ears Anatomy 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000004459 forage Substances 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 239000010871 livestock manure Substances 0.000 description 1
- 235000021073 macronutrients Nutrition 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000013586 microbial product Substances 0.000 description 1
- 239000011785 micronutrient Substances 0.000 description 1
- 235000013369 micronutrients Nutrition 0.000 description 1
- 239000002366 mineral element Substances 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
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- -1 yeast powder source Chemical compound 0.000 description 1
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Abstract
本发明公开了一种微生物发酵培养基的制备方法，制备方法包括如下步骤：(a)原料预处理：将作物青秸杆阴干或烘干后切成段并粉碎，过筛；(b)以秸秆粉与水的质量比1：50或1：10加入60～80℃的热水浸提，放置过滤后，在澄清的浸提液中加入碳源，调节pH值为7.0并灭菌后制得微生物发酵培养基或其浓缩液。本发明的微生物发酵培养基的制备方法以作物青秸杆为原料，不仅成本低廉，而且制备工艺简单，发酵效果好，活菌量高，原料经济环保，充足易得，制备的菌剂产品使用时无化学污染，尤其适用于根瘤菌、固氮菌或溶磷菌的发酵生产。The invention discloses a preparation method of a microbial fermentation medium. The preparation method comprises the following steps: (a) raw material pretreatment: dry or oven-dried crop green stalks, cut them into sections, crush them, and sieve them; (b) use The mass ratio of straw powder to water is 1:50 or 1:10, add hot water at 60-80°C for extraction, place and filter, add carbon source to the clarified extraction solution, adjust the pH value to 7.0 and sterilize it. Obtain the microbial fermentation medium or its concentrate. The preparation method of the microbial fermentation medium of the present invention uses crop green stalks as raw materials, not only has low cost, but also has simple preparation process, good fermentation effect, high amount of live bacteria, economical and environmentally friendly raw materials, sufficient and easy to obtain, and the prepared microbial agent products can be used There is no chemical pollution at the time, and it is especially suitable for the fermentation production of rhizobia, nitrogen-fixing bacteria or phosphate-dissolving bacteria.
Description
技术领域 technical field
本发明涉及微生物的发酵技术，尤其涉及一种微生物发酵培养基的制备方法。 The invention relates to microbial fermentation technology, in particular to a method for preparing a microbial fermentation medium.
技术背景 technical background
微生物肥料是目前可持续农业发展的重要组分，由于其无毒无害和不污染环境的特性，目前已广泛应用于作物和饲草生产中，创造了巨大的经济效益。培养基作为微生物菌剂生产的原料，在产品成本和质量的控制中起决定性作用。目前，国内微生物肥料的研制和生产都不同程度存在着资金、原料，技术和设备的缺乏。在产品研发期间采用的实验室培养基配方造价高昂，难以用于大量菌肥产品的工业化生产。目前已有以豆粕，畜粪为原料的培养基质生产技术，降低了部分产品的成本，但由于原料成份的影响因素较多，导致其粗制产品往往质量低劣，精制产品对工艺、设备和技术的要求又较高而影响推广应用。为此，如何在提高培养基质量的同时，最大化降低成本和对周边环境的污染，实现菌肥生产的可持续发展成为亟待解决的问题。另外，我国作物秸秆每年的产量约5亿吨，其中玉米高粱等高杆作物的秸秆产量占相当比例，如何提高秸秆利用率也是可持续农业发展中的重要一环，在玉米、高粱等作物青秸秆中含有大量微生物生长繁殖所需的碳源(包括可直接利用和分解后利用的糖类)、氮源和矿质元素，各成份的自然比例与常用的YEM (酵母-甘露醇)，LB (蛋白胨-酵母)等发酵培养基相近，可为微生物提供几乎完全而可直接利用的营养元素。然而现有技术中却没有以此为原料生产微生物培养培养基的技术方案。 Microbial fertilizer is an important component of sustainable agricultural development. Due to its non-toxic, harmless and non-polluting characteristics, it has been widely used in crop and forage production, creating huge economic benefits. As the raw material for the production of microbial agents, the culture medium plays a decisive role in the control of product cost and quality. At present, the development and production of microbial fertilizers in China are lacking in funds, raw materials, technology and equipment to varying degrees. The laboratory culture medium formula used during product development is expensive and difficult to be used in the industrial production of a large number of bacterial fertilizer products. At present, there is a culture substrate production technology using soybean meal and livestock manure as raw materials, which reduces the cost of some products. However, due to many factors affecting the composition of raw materials, the crude products are often of poor quality, and the refined products have a great impact on the process, equipment and technology. The requirements are higher and affect the popularization and application. For this reason, how to improve the quality of the culture medium while maximizing the reduction of cost and pollution to the surrounding environment and realizing the sustainable development of bacterial fertilizer production has become an urgent problem to be solved. In addition, the annual output of crop straw in my country is about 500 million tons, of which the straw output of high-stalk crops such as corn and sorghum accounts for a considerable proportion. How to improve the utilization rate of straw is also an important part of sustainable agricultural development. Straw contains a large number of carbon sources (including sugars that can be used directly and after decomposition), nitrogen sources and mineral elements required for the growth and reproduction of a large number of microorganisms. The natural ratio of each component is the same as that of commonly used YEM (yeast-mannitol), LB ( Peptone-yeast) and other fermentation media are similar, which can provide almost complete and directly usable nutrients for microorganisms. Yet in the prior art, there is no technical scheme for producing microorganism culture medium with this as raw material.
发明内容 Contents of the invention
本发明旨在克服以上现有技术存在的不足，提供一种制作成本低，发酵效果好，活菌量高的以作物青秸杆为原料制备微生物发酵培养基的方法。 The present invention aims to overcome the deficiencies in the prior art above, and provides a method for preparing microbial fermentation medium with crop green stalks as raw materials, which has low production cost, good fermentation effect and high viable bacteria content.
本发明解决技术问题采用如下技术方案： The present invention solves technical problem and adopts following technical scheme:
一种微生物发酵培养基的制备方法，以作物青秸秆为原料，制备方法包括如下步骤： (a)原料预处理：将作物青秸杆阴干或烘干后切成18～22cm的小段，粉碎后过1.5mm或2mm筛； A method for preparing a microbial fermentation medium, which uses crop green stalks as raw materials. The preparation method includes the following steps: (a) Raw material pretreatment: dry or dry the crop green stalks and cut them into small sections of 18-22 cm. Pass 1.5mm or 2mm sieve;
(b)原料浸提：以秸秆粉与水的质量比1：50或1：10加入60～80℃的热水浸提，放置过滤后，在澄清的浸提液中加入碳源，调节pH值为6.8-7.2并灭菌后制得微生物发酵培养基。 (b) Raw material leaching: the mass ratio of straw powder to water is 1:50 or 1:10, add hot water at 60-80°C for leaching, place and filter, add carbon source to the clarified leaching solution to adjust pH The value is 6.8-7.2 and the microbial fermentation medium is prepared after sterilization.
步骤(b)中秸秆粉与水的质量比1：50时，热水浸提时间为2-12h，加入碳源的质量为使微生物发酵培养基中碳源的浓度为5g/L。 When the mass ratio of straw powder to water in step (b) is 1:50, the hot water extraction time is 2-12 hours, and the quality of carbon source added is such that the concentration of carbon source in the microbial fermentation medium is 5g/L.
步骤(b)中秸秆粉与水的质量比1：10时，热水浸提时间为4-24h，加入碳源的质量为使微生物发酵培养基浓缩液中碳源的浓度为25g/L，该浓缩液经无菌水稀释5倍后制得微生物发酵培养基。 When the mass ratio of straw powder to water in step (b) is 1:10, the hot water extraction time is 4-24h, and the quality of carbon source added is such that the concentration of carbon source in the microbial fermentation medium concentrate is 25g/L, The concentrated solution is diluted 5 times with sterile water to prepare a microbial fermentation medium.
所述作物青秸秆优选果穗收获后48小时内的作物青秸杆。 The crop green straw is preferably the crop green straw within 48 hours after the ears are harvested.
所述作物青秸秆可以是玉米青秸杆。 The crop green stalks may be corn green stalks.
本发明的制备方法所制备的微生物发酵培养基尤其适用于根瘤菌、固氮菌或溶磷菌的发酵生产。 The microbial fermentation medium prepared by the preparation method of the present invention is especially suitable for the fermentation production of rhizobia, nitrogen-fixing bacteria or phosphate-dissolving bacteria.
本发明的有益效果如下： The beneficial effects of the present invention are as follows:
1. 成本低廉：与标准的YEM培养基和LB培养基作为比较，由于该培养基在制备过程中，只需添加相当YEM培养基和LB培养基一半的碳源物质，无需添加酵母粉等氮源，也无需添加额外的大量和微量营养元素，节约了培养基制备中所需的部分试剂。在达到相同或更优发酵效果的前提下，本培养基产品成本约仅为标准YEM培养基的18.58%，为标准LB培养基的24.12%，且成本低于目前最为廉价的豆芽汁培养基。 1. Low cost: Compared with the standard YEM medium and LB medium, since this medium only needs to add half of the carbon source material equivalent to YEM medium and LB medium during the preparation process, there is no need to add nitrogen such as yeast powder source, and there is no need to add additional macronutrients and micronutrients, which saves some of the reagents required in the preparation of the medium. Under the premise of achieving the same or better fermentation effect, the product cost of this medium is only about 18.58% of the standard YEM medium and 24.12% of the standard LB medium, and the cost is lower than the current cheapest bean sprouts juice medium.
2. 制备工艺简单：由于作物青秸秆本身所含的糖类、蛋白、氨基酸和无机盐等的比例和成份符合微生物自然生长的需要，因此除调节pH值和添加适量碳源外无需其他技术要求较高的人工操作，对仪器和生产设备要求不高，便于中小型企业的推广应用。 2. The preparation process is simple: since the ratio and composition of sugars, proteins, amino acids and inorganic salts contained in the crop green straw itself meet the needs of the natural growth of microorganisms, no other technical requirements are required except for adjusting the pH value and adding an appropriate amount of carbon source High manual operation, low requirements for instruments and production equipment, easy to popularize and apply in small and medium-sized enterprises.
3. 发酵效果好，活菌量高：以根瘤菌和固氮溶磷菌RSN19的发酵生产为例，使用本方法制备的培养基产品可获得与标准YEM培养基相同的最大活菌量，而发酵周期缩短6.17%-29.52%。 3. Good fermentation effect and high amount of viable bacteria: Taking the fermentation production of Rhizobium and nitrogen-fixing and phosphorus-fixing bacteria RSN19 as an example, the culture medium prepared by this method can obtain the same maximum viable bacteria as the standard YEM medium, and the fermentation The period is shortened by 6.17%-29.52%.
4. 原料经济环保，充足易得：由于本方法及产品仅需作物青秸秆，不需要额外的无机盐试剂和生物制品作为原料，相对于目前普遍使用的标准发酵培养基而言，不存在原料制备伴随的污染和可能产生的原料紧缺问题，作为一种新的秸秆利用方式，符合目前绿色可持续循环农业的发展要求。 4. The raw materials are economical and environmentally friendly, and they are sufficient and easy to obtain: since this method and products only need crop straw, no additional inorganic salt reagents and biological products are used as raw materials. Compared with the standard fermentation medium commonly used at present, there are no raw materials The pollution associated with the preparation and the possible shortage of raw materials, as a new way of straw utilization, meet the current development requirements of green sustainable circular agriculture.
5. 制备的菌剂产品使用时无化学污染：相对于传统的标准发酵培养基而言，无化学试剂的添加，保证了菌剂在施用于土壤后，不会因化学试剂的存在而对土壤结构和性质产生破坏。 5. The prepared microbial products have no chemical pollution when used: compared with the traditional standard fermentation medium, there is no addition of chemical reagents, which ensures that the bacterial reagents will not affect the soil due to the presence of chemical reagents after they are applied to the soil. structure and properties are destroyed.
具体实施方式 Detailed ways
实施例1Example 1
一种微生物发酵培养基的制备方法，以果实收获后48小时内的玉米青秸杆为原料，制备方法包括如下步骤： (a)原料预处理：将玉米青秸杆阴干或烘干后切成20cm的小段，粉碎后过1.5mm或2mm筛； A method for preparing a microbial fermentation medium, using green corn stalks within 48 hours after fruit harvesting as raw materials, the preparation method comprising the following steps: (a) Raw material pretreatment: dry or dry the green corn stalks and cut them into Small pieces of 20cm, crushed and passed through a 1.5mm or 2mm sieve;
(b)原料浸提：以秸秆粉与水的质量比1：50加入70℃的热水浸提3小时，放置过滤后，在澄清的浸提液中加入碳源，调节碳源浓度5g/L，并调节pH值为7.0，并灭菌后制得微生物发酵培养基。 (b) Extraction of raw materials: The mass ratio of straw powder to water is 1:50, add hot water at 70°C for 3 hours, place and filter, add carbon source to the clarified extract, adjust the carbon source concentration to 5g/ L, and adjust the pH value to 7.0, and prepare the microbial fermentation medium after sterilization.
实施例2Example 2
一种微生物发酵培养基的制备方法，以果穗收获后48小时内的玉米青秸杆为原料，制备方法包括如下步骤： (a)原料预处理：将作物青秸杆阴干或烘干后切成19cm的小段，粉碎后过1.5mm或2mm筛； A method for preparing a microbial fermentation medium, which uses corn green stalks within 48 hours after ear harvesting as a raw material, and the preparation method includes the following steps: (a) Raw material pretreatment: dry or dry the crop green stalks and cut them into Small pieces of 19cm, crushed and passed through a 1.5mm or 2mm sieve;
(b)原料浸提：以秸秆粉与水的质量比1：10加入75℃的热水浸提8小时，放置过滤后，在澄清的浸提液中加入碳源，调节碳源浓度25g/L，并调节pH值为7.0并灭菌后制得微生物发酵培养基浓缩液。 (b) Extraction of raw materials: The mass ratio of straw powder to water is 1:10, adding hot water at 75°C to extract for 8 hours. L, and adjust the pH value to 7.0 and sterilize to obtain the concentrated solution of the microbial fermentation medium.
(c)浓缩液稀释：使用前将培养基浓缩液以无菌水稀释5倍后制得微生物发酵培养基。 (c) Dilution of the concentrated solution: before use, the concentrated solution of the culture medium was diluted 5 times with sterile water to obtain a microbial fermentation medium.
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| CN105754924A (en) * | 2016-04-14 | 2016-07-13 | 上海大学 | Method for improving microbiological cultivation efficiency and fermentation efficiency by aid of straw extract liquid |
| CN108077023A (en) * | 2017-12-29 | 2018-05-29 | 贵州师范学院 | A kind of natural plants soilless culture nutrient fluid based on blue or green stalk leaching liquor |
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