WO2019061961A1 - 一种利用城市污泥制备的超高温好氧发酵菌剂及其制备方法 - Google Patents

一种利用城市污泥制备的超高温好氧发酵菌剂及其制备方法 Download PDF

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WO2019061961A1
WO2019061961A1 PCT/CN2018/073426 CN2018073426W WO2019061961A1 WO 2019061961 A1 WO2019061961 A1 WO 2019061961A1 CN 2018073426 W CN2018073426 W CN 2018073426W WO 2019061961 A1 WO2019061961 A1 WO 2019061961A1
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preparation
fermentation
sludge
ultra
temperature aerobic
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周顺桂
陈志�
廖汉鹏
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福建农林大学
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Priority to US16/826,261 priority Critical patent/US11898140B2/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/04—Preserving or maintaining viable microorganisms
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F7/00—Fertilisers from waste water, sewage sludge, sea slime, ooze or similar masses
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/34—Sugars
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales

Definitions

  • the invention relates to the field of microbial culture and application, in particular to a preparation method of an ultra-high temperature aerobic fermentation microbial preparation prepared by using urban sludge and a preparation method thereof.
  • High temperature composting is an important means for the harmless treatment and resource utilization of organic solid waste.
  • the essence of organic matter is the catabolism of organic matter under the action of microorganisms to generate heat energy, which promotes the conversion of organic matter to stable humus.
  • High temperature is a double-edged sword for aerobic composting.
  • the rapid decomposing of materials and the harmlessness of hygiene are inseparable from high temperatures, which are conducive to the formation of humus and the killing of pathogenic bacteria;
  • important constituents of microbial organisms such as proteins and nucleic acids are sensitive to temperature. It will irreversibly destroy or even die with the increase of temperature, thereby inhibiting the activity of the microbial population, which makes the subsequent composting process difficult to operate stably.
  • thermophilic microorganisms In traditional aerobic composting, the optimum growth temperature of most thermophilic microorganisms is 50-60 °C, which makes the traditional compost have low stack temperature, slow degradation rate of organic matter, incomplete killing of pathogenic microorganisms, long fermentation cycle and so on. .
  • extreme thermophilic bacteria have harsh growth conditions and low survival rate in the natural environment, which makes them easily inactivated by other indigenous dominant microorganisms in practical applications, thus affecting the treatment effect of ultra-high temperature aerobic fermentation technology.
  • Urban sludge is an inevitable deposit in the process of urban domestic sewage treatment. China's annual sludge production is about 8 million tons of dry weight (32 million tons of wet mud cake), and is increasing at a rate of 10% to 15% per year. The problem of environmental pollution caused by sludge has become increasingly prominent, which has caused great potential safety hazards, environmental pressures and economic burdens. At the same time, municipal sludge contains nutrients such as carbon, nitrogen, phosphorus and trace elements that can be used by microorganisms. The use of sludge as a raw material for the fermentative production of microbial inoculum not only provides a new resource for urban sludge, but also greatly reduces the cost of industrial fermentation of microorganisms.
  • the object of the present invention is to disclose an ultra-high temperature aerobic fermentation microbial preparation prepared by using municipal sludge and a preparation method thereof, so as to improve the fermentation production efficiency of the extreme thermophilic bacteria and the activity and preservation period of the ultra-high temperature fermentation microbial agent.
  • the preparation method of the ultra-high temperature aerobic fermenting agent prepared by using municipal sludge is prepared by the following steps: after the activation of the ultra-high temperature aerobic bacteria, the fermentation is carried out, the supernatant is removed by the fermentation liquid, and the protective agent is added and stirred. Uniform, dried, ground by a pulverizer, and sieved.
  • the medium used for the fermentation is prepared from municipal sludge.
  • the preparation method of the culture medium used for the fermentation is: taking urban sludge and water to adjust the solid content of the sludge, adding the enemy, yeast powder, ammonium chloride, sucrose, soluble starch, potassium dihydrogen phosphate, adjusting the pH, adding The composite carrier is stirred evenly and sterilized and cooled to standby.
  • the protective agent consists of skimmed milk powder, soluble starch, glycerin, and water.
  • the composite carrier consists of at least one of kaolin, biochar, and wheat bran.
  • the composite carrier content is from 50 to 100 g/L.
  • the sludge has a solid content of 2 to 3%.
  • the hyperthermia aerobic bacteria is at least one of Thermusthermophilus, Calditerricolayamamurae, Calditerricolasatsumensis, Thermaerobactercomposti, Geobacillus thermocatenulatus or Thermaerobacter subterraneus.
  • the invention adopts the seasoning and optimization preparation of the sludge culture medium, not only reduces the fermentation cost of the ultra-high temperature aerobic bacteria, but also improves the fermentation efficiency, and solves the problem that the super-high temperature aerobic fermentation production is difficult;
  • the addition of the composite carrier and the protective agent in the invention can effectively improve the survival rate and the preservation period of the ultra-high temperature aerobic fermentation agent, and is suitable for long-term preservation;
  • the ultra-high temperature fermentation microbial agent is produced by simple fermentation, which reduces the cost of the application, and is suitable for large-scale industrial production and application.
  • the components of the CYS medium were: yeast extract 4.0 g, hydrolyzed tyrosin 6.0 g, soluble starch 3.0 g, NaCl 30.0 g, MgCl 2 ⁇ 6H 2 O 0.27 g, CaCl 2 0.025 g, FeSO 4 ⁇ 7H 2 O 0.01 g, trace element 100 ⁇ L (Na 2 MoO 4 ⁇ 2H 2 O 12.0g / L, VOSO 4 ⁇ xH 2 O 1.0g / L, MnCl 2 5.0g / L, ZnSO 4 ⁇ 7H 2 O 0.6g / L, CuSO 4 5H 2 O 0.15 g/L, CoCl 2 ⁇ 6H 2 O 8.0 g/L, NiCl 2 ⁇ 6H 2 O 0.2 g/L), initial pH 7.5, 0.15 MPa, and sterilization at 121 ° C for 30 min.
  • the shake flask solution was inoculated to a seed tank containing a sludge fermentation medium at 2%, and the aeration rate was 100 m 3 /h at 75 ° C, the stirring speed was 180 rpm, and the fermentation time was 16 hours.
  • Calditerricolasatsumensis FAFU012 seed solution was inoculated into the fermentor at a seeding rate of 2%, at 75 ° C, aeration of 100 m 3 /h, agitation speed of 180 rpm, and fermentation time of 24 hours.
  • the fermentation broth is subjected to plate and frame filtration to remove the supernatant, and 1% of a protective agent is added.
  • the protective agent is composed of skim milk powder, soluble starch, glycerin and water.
  • the ultra-high temperature aerobic bacteria produced by fermentation of the sludge fermentation medium were randomly sampled and tested according to the National Standard of the People's Republic of China, Agricultural Microbial Inoculants (GB 20287-2006). After testing, the number of viable cells was 2.8 ⁇ 10 9 /g.
  • the components of the CYS medium were: yeast extract 4.0 g, hydrolyzed tyrosin 6.0 g, soluble starch 3.0 g, NaCl 30.0 g, MgCl 2 ⁇ 6H 2 O 0.27 g, CaCl 2 0.025 g, FeSO 4 ⁇ 7H 2 O 0.01 g, trace element 100 ⁇ L (Na 2 MoO 4 ⁇ 2H 2 O 12.0g / L, VOSO 4 ⁇ xH 2 O 1.0g / L, MnCl 2 5.0g / L, ZnSO 4 ⁇ 7H 2 O 0.6g / L, CuSO 4 5H 2 O 0.15 g/L, CoCl 2 ⁇ 6H 2 O 8.0 g/L, NiCl 2 ⁇ 6H 2 O 0.2 g/L), initial pH 7.5, 0.15 MPa, and sterilization at 121 ° C for 30 min.
  • the shake flask solution was inoculated to a seed tank containing a sludge fermentation medium at 2%, 80 ° C, aeration rate of 100 m 3 /h, stirring speed of 180 rpm, and fermentation time of 16 hours.
  • Calditerricolayamamurae UTM801 seed solution was inoculated into the fermentor at a seeding rate of 2%, at 85 ° C, aeration of 100 m 3 /h, agitation speed of 180 rpm, and fermentation time of 24 hours.
  • the fermentation broth is subjected to plate and frame filtration to remove the supernatant, and 1% of a protective agent is added.
  • the protective agent is composed of skim milk powder, soluble starch, glycerin and water.
  • the ultra-high temperature aerobic bacteria produced by fermentation of the sludge fermentation medium were randomly sampled and tested according to the National Standard of the People's Republic of China, Agricultural Microbial Inoculants (GB 20287-2006). After testing, the number of viable cells was 4.5 ⁇ 10 9 /g.
  • the components of the CYS medium were: yeast extract 4.0 g, hydrolyzed tyrosin 6.0 g, soluble starch 3.0 g, NaCl 30.0 g, MgCl 2 ⁇ 6H 2 O 0.27 g, CaCl 2 0.025 g, FeSO 4 ⁇ 7H 2 O 0.01 g, trace element 100 ⁇ L (Na 2 MoO 4 ⁇ 2H 2 O 12.0g / L, VOSO 4 ⁇ xH 2 O 1.0g / L, MnCl 2 5.0g / L, ZnSO 4 ⁇ 7H 2 O 0.6g / L, CuSO 4 5H 2 O 0.15 g/L, CoCl 2 ⁇ 6H 2 O 8.0 g/L, NiCl 2 ⁇ 6H 2 O 0.2 g/L), initial pH 7.5, 0.15 MPa, and sterilization at 121 ° C for 30 min.
  • the shake flask solution was inoculated to a seed tank containing a sludge fermentation medium at 2%, and the aeration rate was 100 m 3 /h at 75 ° C, the stirring speed was 180 rpm, and the fermentation time was 16 hours.
  • Thermusthermophilus HB8 seed solution was inoculated into the fermentor at a seeding rate of 2%, at 75 ° C, aeration of 100 m 3 /h, agitation speed of 180 rpm, and fermentation time of 24 hours.
  • the fermentation broth is subjected to plate and frame filtration to remove the supernatant, and 1% of a protective agent is added.
  • the protective agent is composed of skim milk powder, soluble starch, glycerin and water.
  • the ultra-high temperature aerobic bacteria produced by fermentation of the sludge fermentation medium were randomly sampled and tested according to the national standard of the People's Republic of China, Agricultural Microbial Inoculants (GB 20287-2006). After testing, the number of viable cells was 2.3 ⁇ 10 9 /g. Effect of different treatments of Example 4 on Calditerricola fermentation and microbial quality
  • the activated Calditerricolasatsumensis FAFU012 was subjected to seed tank fermentation, fermentor fermentation and microbial preparation, respectively, according to Table 5.
  • the seed tank fermentation was inoculated with 2% of the activated bacterial solution in a seed tank containing the sludge fermentation medium, 75 ° C, aeration rate of 100 m 3 /h, stirring speed of 180 rpm, fermentation time of 16 hours; fermentation
  • the can fermentation was carried out by inoculating the seed solution to a fermenter at a rate of 2%, at 75 ° C, a can pressure of 0.06 MPa, aeration of 100 m 3 /h, a stirring speed of 180 rpm, and a fermentation time of 24 hours.
  • the fermentation broth removes the supernatant by plate and frame filtration, adding the corresponding protective agent, mixing at high speed, drying at 90 ° C to constant weight, grinding through a pulverizer, passing through a 80 mesh sieve, and dispensing .

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Abstract

一种利用城市污泥制备的超高温好氧发酵菌剂，其制备方法是：将超高温好氧菌活化后，进行发酵，发酵液去除上清液，加入保护剂搅拌混匀，烘干，经粉磨机粉磨，并筛分装。发酵所用的培养基是用城市污泥制备的。

Description

一种利用城市污泥制备的超高温好氧发酵菌剂及其制备方法 技术领域
本发明涉及微生物培养与应用领域，特别涉及到一种利用城市污泥制备的超高温好氧发酵菌剂的制备方法及其制备方法。
背景技术
高温堆肥是有机固体废物无害化处理与资源化利用的重要手段，其实质是有机物在微生物作用下分解代谢产生热能，促使有机物向稳定的腐殖质转化过程。高温对于好氧堆肥是一把双刃剑。一方面，物料的快速腐熟和卫生学无害化离不开高温，高温有利于促进腐殖质的形成和病原菌的杀灭；而另一方面，蛋白质、核酸等微生物有机体的重要组成物质对温度敏感，将随着温度的升高遭受不可逆的破坏甚至死亡，从而抑制微生物种群活性，导致后续堆肥过程难以稳定运行。在传统好氧堆肥中，大多数嗜热微生物的最适生长温度为50-60℃，这使得传统堆肥存在堆体温度低，有机物降解速率慢，病原微生物杀灭不彻底，发酵周期长等缺点。
2008年，日本学者Y.Oshima将一株极端嗜热菌Calditerricolasatsumensis应用于有机固废的堆肥中，有效提高堆肥的温度和病原物的杀灭效率。2012年，周顺桂等研究人员将另一株极端嗜热菌Calditerricolayamamurae应用于污泥堆肥中(一种极端嗜热菌UTM801及其应用，专利申请号：201210343222.4)，并发明了超高温好氧堆肥技术。与传统堆肥相比，超高温好氧堆肥技术可有效提高堆肥温度，促进有机物降解，有效杀灭病原微生物，缩短发酵周期，降低有机固体废弃物堆肥处理的成本。然而，极端嗜热细菌生长条件严苛，在自然环境中存活率低，使其在实际应用中容易被其他本土优势微生物所替代而失活，从而影响超高温好氧发酵技术的处理效果。基于此，在实际应用中需要定期加入相应极端嗜热菌剂，以维持堆肥中的极端嗜热菌的菌群数量，保持超高温好氧发酵的技术优势。因此，利用低成本的有机废弃物，通过简单的发酵生产超高温发酵菌剂，提高极端嗜热菌的活性和保藏期，对于降低超高温好氧堆肥技术大规模工业化应用的成本具有重要的实践意义。
城市污泥是城市生活污水处理过程中必然产生的沉淀物。我国污泥年产生量约800万吨干重(折成湿泥饼3200万吨)，并且以每年10％～15％的速度递增。污泥产生的环境污染问题日益突出，已造成极大的安全隐患、环境压力和经济负担。同时，城市污泥中含有可被微生物利用的碳、氮、磷以及微量元素等营养物质。利用污泥为原料进行微生物菌剂的发酵生产， 不仅能够为城市污泥提供新的资源化途径，而且可大幅度降低微生物的工业发酵的成本。国内外已有多个专利报道采用污泥作为主要原料发酵生产微生物菌剂。然而，极端嗜热菌生长条件严苛，难以保存，且城市污泥中养分含量有限，成份不稳定，直接以污泥为唯一原料进行发酵培养，菌体生长缓慢，发酵水平低，又不稳定。因此，在现有技术中还未发现利用污泥制备超高温发酵菌剂的报道。
发明内容
本发明的目的在于公开了一种利用城市污泥制备的超高温好氧发酵菌剂及其制备方法，以提高极端嗜热菌的发酵生产效率以及超高温发酵菌剂的活性和保藏期。
本发明所采取的技术方案是：
一种利用城市污泥制备的超高温好氧发酵菌剂的制备方法，由以下步骤制备而成：将超高温好氧菌活化后，进行发酵，发酵液去除上清液，加入保护剂搅拌混匀，烘干，经粉磨机粉磨，并筛分装。
优选的，发酵所用的培养基是用城市污泥制备的。
优选的，发酵所用的培养基的制备方法是：取城市污泥加水调节污泥含固率，加入泡敌、酵母粉、氯化铵、蔗糖、可溶性淀粉、磷酸二氢钾，调节pH，加入复合载体，搅拌均匀，灭菌冷却至备用。
优选的，保护剂由脱脂奶粉、可溶性淀粉、甘油、水组成。
进一步优选的，保护剂中，脱脂奶粉∶可溶性淀粉∶甘油∶水＝(2～3)∶(1～2)∶10∶(85～87)，其中脱脂奶粉和可溶性淀粉按质量计，单位g；甘油和水按体积计，单位mL。
优选的，复合载体由高岭土，生物炭，麦麸中的至少一种组成。
进一步优选的，复合载体含量为50～100g/L。
进一步优选的，复合载体由生物炭，麦麸组成，其中按质量比计生物炭∶麦麸＝1.5～3∶1。
优选的，污泥含固率为2～3％。
优选的，超高温好氧菌是Thermusthermophilus、Calditerricolayamamurae、Calditerricolasatsumensis、Thermaerobactercomposti、Geobacillusthermocatenulatus或Thermaerobactersubterraneus中的至少一种。
本发明的有益效果是：
1、利用城市污泥为主要原料进行发酵，降低了发酵成本，同时为解决城市污泥的污染问题提供了一种思路，有助于保护和改善生态环境；
2、本发明通过污泥培养基的调料和优化配制，不但降低了超高温好氧菌的发酵成本，而 且提高了发酵效率，解决了超高温好氧菌发酵生产困难的问题；
3、相较于常规方法制备的发酵菌剂，本发明中复合载体和保护剂的添加可有效提高超高温好氧发酵菌剂的存活率和保藏期，适合长期的保存；
4、通过简单的发酵生产超高温发酵菌剂，降低了应用的成本，从而适合大规模工业化的生产和应用。
具体实施方式
实施例1生产Calditerricolasatsumensis超高温好氧发酵菌剂
(1)城市污泥样品
取北京市某污水处理厂的浓缩污泥，该污泥的基本性质见表1.
表1.供试污泥基本性质
Figure PCTCN2018073426-appb-000001
(2)污泥发酵培养基配置
加水调节污泥含固率至2％，加入0.2％的泡敌、1g/L酵母粉、0.5g/L氯化铵、0.3g/L蔗糖、0.2g/L可溶性淀粉、0.3g/L磷酸二氢钾，调节pH至7.0，加入50g/L生物炭/麦麸(w/w＝2/1)复合载体，搅拌均匀，100℃灭菌45分钟，冷却至70-80℃备用。
(3)摇瓶培养从保藏正常的CalditerricolasatsumensisFAFU012平板培养基上挑取菌落，接种到装有100mL CYS液体培养基的300mL三角瓶中，于80℃、180转/分钟，培养12小时。CYS培养基的成分为：酵母提取物4.0g、水解酪合蛋白6.0g、可溶性淀粉3.0g、NaCl 30.0g、MgCl 2·6H 2O 0.27g、CaCl 2 0.025g、FeSO 4·7H 2O 0.01g、微量元素100μL(Na 2MoO 4·2H 2O 12.0g/L、VOSO 4·xH 2O 1.0g/L、MnCl 2 5.0g/L、ZnSO 4·7H 2O 0.6g/L、CuSO 4·5H 2O 0.15g/L、CoCl 2·6H 2O 8.0g/L、NiCl 2·6H 2O 0.2g/L)、初始pH 7.5，0.15MPa，121℃灭菌30min。
(4)种子罐发酵
按2％将上述摇瓶菌液接种于装有污泥发酵培养基的种子罐中，75℃、通气量为100m 3/h、搅拌速度180转/分、发酵时间16小时。
(5)发酵罐发酵
按2％的接种量将Calditerricolasatsumensis FAFU012种子液接种到发酵罐中，于75℃、通气量为100m 3/h、搅拌速度180转/分、发酵时间24小时。
(6)菌剂制备
发酵液经板框压滤法去除上清液，加入1％的保护剂，保护剂由脱脂奶粉，可溶性淀粉，甘油和水组成。其中，脱脂奶粉∶可溶性淀粉∶甘油∶水＝3∶2∶10∶85(w/w/v/v)，质量单位为g，体积单位为mL。高速搅拌混匀，90℃烘干，经粉磨机粉磨，并过80目筛，分装。
(7)成品质量检验
经过污泥发酵培养基发酵生产的超高温好氧菌剂通过随机抽样，并按按中华人民共和国国家标准《农用微生物菌剂》(GB 20287-2006)进行检测。经过检测，活菌数为2.8×10 9个/g。
实施例2生产Calditerricolayamamurae超高温好氧发酵菌剂
(1)城市污泥样品
取河南省郑州市的浓缩污泥，该污泥的基本性质见表2.
表2供试污泥基本性质
Figure PCTCN2018073426-appb-000002
(2)污泥发酵培养基配置
加水调节污泥含固率至3％，加入0.2％的泡敌、1.5g/L酵母粉、1.0g/L氯化铵、0.5g/L蔗糖、0.5g/L可溶性淀粉、0.5g/L磷酸二氢钾，用调节pH至7.0，加入80g/L生物炭/麦麸(w/w＝1.5/1)复合载体，搅拌均匀，100℃灭菌45分钟，冷却至70-80℃备用。
(3)摇瓶培养从保藏正常的CalditerricolayamamuraeUTM801平板培养基上挑取菌落，接种到装有100mL CYS液体培养基的300mL三角瓶中，于80℃、180转/分钟，培养12小时。CYS培养基的成分为：酵母提取物4.0g、水解酪合蛋白6.0g、可溶性淀粉3.0g、NaCl 30.0g、MgCl 2·6H 2O 0.27g、CaCl 2 0.025g、FeSO 4·7H 2O 0.01g、微量元素100μL(Na 2MoO 4·2H 2O 12.0g/L、VOSO 4·xH 2O 1.0g/L、MnCl 2 5.0g/L、ZnSO 4·7H 2O 0.6g/L、CuSO 4·5H 2O 0.15g/L、CoCl 2·6H 2O 8.0g/L、NiCl 2·6H 2O 0.2g/L)、初始pH 7.5，0.15MPa，121℃灭菌30min。
(4)种子罐发酵
按2％将上述摇瓶菌液接种于装有污泥发酵培养基的种子罐中，80℃、通气量为100m 3/h、搅拌速度180转/分、发酵时间16小时。
(5)发酵罐发酵
按2％的接种量将Calditerricolayamamurae UTM801种子液接种到发酵罐中，于85℃、通气量为100m 3/h、搅拌速度180转/分、发酵时间24小时。
(6)菌剂制备
发酵液经板框压滤法去除上清液，加入1％的保护剂，保护剂由脱脂奶粉，可溶性淀粉，甘油和水组成。其中，脱脂奶粉∶可溶性淀粉∶甘油∶水＝3∶2∶10∶85(w/w/v/v)，质量单位为g，体积单位为mL。高速搅拌混匀，90℃烘干，经粉磨机粉磨，并过80目筛，分装。
(7)成品质量检验
经过污泥发酵培养基发酵生产的超高温好氧菌剂通过随机抽样，并按按中华人民共和国国家标准《农用微生物菌剂》(GB 20287-2006)进行检测。经过检测，活菌数为4.5×10 9个/g。
实施例3生产Thermusthermophilus超高温好氧发酵菌剂
(1)城市污泥样品
取福建省福州市某污水处理厂的浓缩污泥，该污泥的基本性质见表3.
表3.供试污泥基本性质
Figure PCTCN2018073426-appb-000003
(2)污泥发酵培养基配置
加水调节污泥含固率至3％，加入0.2％的泡敌、1.5g/L酵母粉、0.5g/L氯化铵、0.5g/L蔗糖、0.5g/L可溶性淀粉、1g/L磷酸二氢钾，调节pH至7.0，加入100g/L生物炭/麦麸(1.5/1)复合载体，搅拌均匀，100℃灭菌45分钟，冷却至70-80℃备用。
(3)摇瓶培养从保藏正常的ThermusthermophilusHB8平板培养基上挑取菌落，接种到装有100mL CYS液体培养基的300mL三角瓶中，于75℃、180转/分钟，培养12小时。CYS培养基的成分为：酵母提取物4.0g、水解酪合蛋白6.0g、可溶性淀粉3.0g、NaCl 30.0g、MgCl 2·6H 2O 0.27g、CaCl 2 0.025g、FeSO 4·7H 2O 0.01g、微量元素100μL(Na 2MoO 4·2H 2O 12.0g/L、VOSO 4·xH 2O 1.0g/L、MnCl 2 5.0g/L、ZnSO 4·7H 2O 0.6g/L、CuSO 4·5H 2O 0.15g/L、CoCl 2·6H 2O 8.0g/L、NiCl 2·6H 2O 0.2g/L)、初始pH 7.5，0.15MPa，121℃灭菌30min。
(4)种子罐发酵
按2％将上述摇瓶菌液接种于装有污泥发酵培养基的种子罐中，75℃、通气量为100m 3/h、搅拌速度180转/分、发酵时间16小时。
(5)发酵罐发酵
按2％的接种量将ThermusthermophilusHB8种子液接种到发酵罐中，于75℃、通气量为100m 3/h、搅拌速度180转/分、发酵时间24小时。
(6)菌剂制备
发酵液经板框压滤法去除上清液，加入1％的保护剂，保护剂由脱脂奶粉，可溶性淀粉，甘油和水组成。其中，脱脂奶粉∶可溶性淀粉∶甘油∶水＝(2～3)∶(1～2)∶10∶(85～87)(w/w/v/v)，质量单位为g，体积单位为mL。高速搅拌混匀，90℃烘干，经粉磨机粉磨，并过80目筛，分装。
(7)成品质量检验
经过污泥发酵培养基发酵生产的超高温好氧菌剂通过随机抽样，并按中华人民共和国国家标准《农用微生物菌剂》(GB 20287-2006)进行检测。经过检测，活菌数为2.3×10 9个/g。实施例4不同处理对Calditerricola发酵和菌剂质量的影响
(1)城市污泥样品
取福建省福州市某污水处理厂的浓缩污泥，该污泥的基本性质见表4.
表4.供试污泥基本性质
Figure PCTCN2018073426-appb-000004
(2)发酵菌株的活化
摇瓶培养从保藏正常的CalditerricolasatsumensisFAFU012平板培养基上挑取菌落，接种到CYS液体培养基中，于75℃、180转/分钟，培养16小时。
(3)菌株发酵和菌剂制备
按表5不同处理分别对活化的CalditerricolasatsumensisFAFU012进行种子罐发酵、发酵罐发酵和菌剂制备。其中，种子罐发酵按2％将活化菌液接种于装有污泥发酵培养基的种子罐中，75℃、通气量为100m 3/h、搅拌速度180转/分、发酵时间16小时；发酵罐发酵按2％的接种量将种子液接种到发酵罐中，于75℃、罐压0.06MPa、通气量为100m 3/h、搅拌速度180转/分、发酵时间24小时。发酵后，发酵液经板框压滤法去除上清液，加入相应的保护剂，高速搅拌混匀，90℃烘干至恒重，经粉磨机粉磨，并过80目筛，分装。
表5不同处理的CalditerricolasatsumensisFAFU012发酵和菌剂制备
Figure PCTCN2018073426-appb-000005
Figure PCTCN2018073426-appb-000006
(4)发酵效果和菌剂质量检验
表6不同处理对CalditerricolasatsumensisFAFU012发酵和菌剂质量的影响
Figure PCTCN2018073426-appb-000007
上述结果表明，与其他方案相比，按照本发明方案制备的Calditerricola菌剂，有效菌活性高，保存效果好。

Claims (10)

  1. 一种利用城市污泥制备的超高温好氧发酵菌剂，其制备方法是：将超高温好氧菌活化后，进行发酵，发酵液去除上清液，加入保护剂搅拌混匀，烘干，经粉磨机粉磨，并筛分装。
  2. 根据权利要求1所述的制备方法，其特征在于，发酵所用的培养基是用城市污泥制备的。
  3. 根据权利要求1所述的制备方法，其特征在于，发酵所用的培养基的制备方法是：取城市污泥加水调节污泥含固率，加入泡敌、酵母粉、氯化铵、蔗糖、可溶性淀粉、磷酸二氢钾，调节pH，加入复合载体，搅拌均匀，灭菌冷却至备用。
  4. 根据权利要求1所述的制备方法，其特征在于，所述保护剂由脱脂奶粉、可溶性淀粉、甘油、水组成。
  5. 根据权利要求1所述的制备方法，其特征在于，保护剂中，脱脂奶粉∶可溶性淀粉∶甘油∶水＝(2～3)∶(1～2)∶10∶(85～87)，其中脱脂奶粉和可溶性淀粉按质量计，单位g；甘油和水按体积计，单位mL。
  6. 根据权利要求3所述的制备方法，其特征在于，所述复合载体由高岭土，生物炭，麦麸中的至少一种组成。
  7. 根据权利要求3所述的制备方法，其特征在于，所述复合载体含量为50～100g/L。
  8. 根据权利要求3所述的制备方法，其特征在于，污泥含固率为2～3％。
  9. 根据权利要求3所述的制备方法，其特征在于，所述复合载体由生物炭，麦麸组成，按质量比计，生物炭∶麦麸＝1.5～3∶1。
  10. 根据权利要求1～9任一项所述的制备方法，其特征在于，所述超高温好氧菌是Thermusthermophilus、Calditerricolayamamurae、Calditerricolasatsumensis、Thermaerobactercomposti、Geobacillusthermocatenulatus或Thermaerobactersubterraneus中的至少一种。
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113214999A (zh) * 2021-03-24 2021-08-06 中国科学院微生物研究所 一株地霉tn42及其在污水处理中的应用

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108033817B (zh) * 2017-11-27 2019-11-26 福建农林大学 一种快速消减有机固体废物中抗生素及抗性基因的方法
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CN112725229B (zh) * 2020-12-31 2022-05-17 福建农林大学 一种减少氧化亚氮排放的复合菌剂及其应用
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CN113652371A (zh) * 2021-08-17 2021-11-16 湖南泰洁环保科技有限公司 一种超高温餐厨垃圾处理菌剂及其制备方法
CN114480184B (zh) * 2022-01-13 2023-07-18 中农创达(北京)环保科技有限公司 超嗜热菌的培养和冻干菌粉的制备方法
CN114606163A (zh) * 2022-03-16 2022-06-10 中南大学 一种有机废物高温好氧功能强化菌剂及其制备方法与应用
CN115215432B (zh) * 2022-07-09 2023-07-21 同济大学 一种适用于城镇污水处理的颗粒污泥及其快速生成方法
CN116144573A (zh) * 2022-10-26 2023-05-23 中建能源管理(辽宁)有限公司 一种利用城市污泥制备的超高温好氧发酵菌剂的制备方法

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS51115988A (en) * 1975-03-31 1976-10-13 Mitsubishi Petrochem Co Ltd Process for rreparing thermophilic l-asparaginase
CN1793330A (zh) * 2005-11-07 2006-06-28 浙江工商大学 一种新型高活性乳酸菌制品的制备方法
CN101186889A (zh) * 2007-12-14 2008-05-28 华南理工大学 一种处理污水污泥的嗜热微生物制剂及其制备方法和应用
CN101338295A (zh) * 2008-08-09 2009-01-07 甘肃省科学院生物研究所 双歧杆菌冷冻浓缩发酵剂及其制备方法
CN102660487A (zh) * 2012-01-21 2012-09-12 大地绿源环保科技(北京)有限公司 一株嗜热地芽孢杆菌utm02及其应用
CN102703351A (zh) * 2012-01-21 2012-10-03 大地绿源环保科技(北京)有限公司 一株芽孢杆菌utm03及其应用
CN102851247A (zh) * 2012-09-14 2013-01-02 大地绿源环保科技(北京)有限公司 一种极端嗜热菌utm801及其应用
CN107937303A (zh) * 2017-11-10 2018-04-20 福建农林大学 一种极端嗜热菌协同发酵生产复合超高温堆肥菌剂的方法

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TW200301303A (en) * 2001-12-25 2003-07-01 Sanyu Co Ltd Novel microorganism
US6966983B1 (en) * 2004-10-01 2005-11-22 Mixing And Mass Transfer Technologies, Llc Continuous multistage thermophilic aerobic sludge digestion system
CN101273757B (zh) * 2008-03-31 2011-02-09 北京市农林科学院 罗伊氏乳酸杆菌冻干制剂及其制备方法
DK2342346T3 (da) * 2008-09-24 2013-11-04 Hyperthermics Holding As Thermotoga til behandling af biomasse
JP6376798B2 (ja) * 2014-03-27 2018-08-22 三菱重工業株式会社 洗濯排水生物処理製剤を用いた洗濯排水処理装置及び方法
CN105060954A (zh) * 2015-08-30 2015-11-18 常州亚环环保科技有限公司 一种活性污泥生产颗粒有机肥的方法

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS51115988A (en) * 1975-03-31 1976-10-13 Mitsubishi Petrochem Co Ltd Process for rreparing thermophilic l-asparaginase
CN1793330A (zh) * 2005-11-07 2006-06-28 浙江工商大学 一种新型高活性乳酸菌制品的制备方法
CN101186889A (zh) * 2007-12-14 2008-05-28 华南理工大学 一种处理污水污泥的嗜热微生物制剂及其制备方法和应用
CN101338295A (zh) * 2008-08-09 2009-01-07 甘肃省科学院生物研究所 双歧杆菌冷冻浓缩发酵剂及其制备方法
CN102660487A (zh) * 2012-01-21 2012-09-12 大地绿源环保科技(北京)有限公司 一株嗜热地芽孢杆菌utm02及其应用
CN102703351A (zh) * 2012-01-21 2012-10-03 大地绿源环保科技(北京)有限公司 一株芽孢杆菌utm03及其应用
CN102851247A (zh) * 2012-09-14 2013-01-02 大地绿源环保科技(北京)有限公司 一种极端嗜热菌utm801及其应用
CN107937303A (zh) * 2017-11-10 2018-04-20 福建农林大学 一种极端嗜热菌协同发酵生产复合超高温堆肥菌剂的方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
LIAO HANPENG ET AL: "Development of Hperthermophinic Aerobic Composting and Its Engineering Applications in Organic Solid Wastes", JOURNAL OF FUJIAN AGRICULTURE AND FORESTRY UNIVERSITY ( NATURAL SCIENCE EDITION), vol. 46, no. 4, 31 July 2017 (2017-07-31), pages 439 - 444 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113214999A (zh) * 2021-03-24 2021-08-06 中国科学院微生物研究所 一株地霉tn42及其在污水处理中的应用

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