CN101554485A - Air purifying material of microorganism absorption degradation, preparation method and applications thereof - Google Patents
Air purifying material of microorganism absorption degradation, preparation method and applications thereof Download PDFInfo
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Abstract
本发明公开了一种微生物吸附降解的空气净化材料及制备方法和用途，微生物吸附降解的空气净化材料是海藻酸钙凝胶包埋着经交叉培养驯化后的假单胞菌属或不动杆菌属的粒径在2～4mm的生物颗粒。微生物吸附降解的空气净化材料可用于室内甲醛和苯系物污染的长期净化，净化效果显著。该产品具有固定化细胞活性高，稳定性好，能够持久有效地降低室内甲醛与苯系物等有害化学污染物的含量，利于实现生物催化剂的回收、再生和重复使用的优点，并严格避免了微生物对室内环境的污染。原材料价格低廉，制作工艺简便易行，制成品可在多种反应器中应用。The invention discloses an air purification material for microbial adsorption and degradation, a preparation method and application thereof. The air purification material for microbial adsorption and degradation is calcium alginate gel embedded with Pseudomonas or Acinetobacter after cross-cultivation and domestication It is a biological particle with a particle size of 2 to 4 mm. The air purification material degraded by microbial adsorption can be used for long-term purification of indoor formaldehyde and benzene series pollution, and the purification effect is remarkable. This product has the advantages of high activity of immobilized cells and good stability, and can effectively and permanently reduce the content of harmful chemical pollutants such as formaldehyde and benzene series in the room, which is beneficial to the recovery, regeneration and reuse of biocatalysts, and strictly avoids Microbial contamination of the indoor environment. The price of raw materials is low, the manufacturing process is simple and easy, and the finished product can be applied in various reactors.
Description
技术领域 technical field
本发明属于空气净化领域，涉及一种微生物吸附降解的空气净化材料及制备方法和用途。The invention belongs to the field of air purification, and relates to an air purification material for microbial adsorption and degradation, a preparation method and an application.
背景技术 Background technique
随着科技的进步，人们对室内空气品质的要求越来越高，化学污染物作为室内空气品质的重要部分，越来越受到人们的关注。随着人们对室内装修的追求，大量装饰材料进入到室内环境中，装饰材料中的胶粘剂，合成木板以及油漆涂料等都会释放出大量挥发性有机化合物。其中，甲醛和苯系物是室内装修引发空气污染中最常见且危害最严重的污染物，也是最迫切需要去除的化学污染物。With the advancement of science and technology, people have higher and higher requirements for indoor air quality. As an important part of indoor air quality, chemical pollutants have attracted more and more attention. With people's pursuit of interior decoration, a large number of decorative materials have entered the indoor environment. Adhesives in decorative materials, synthetic wood boards, and paints and coatings will release a large amount of volatile organic compounds. Among them, formaldehyde and benzene series are the most common and most harmful pollutants in air pollution caused by interior decoration, and they are also the most urgent chemical pollutants that need to be removed.
甲醛(HCHO)，又名“蚁醛”，在常温下是一种无色、有强烈刺激性气味的气体，易溶于水、醇和醚，它有凝固蛋白质的作用，具有活泼的化学性质和生物学性质。甲醛是一种活泼的分子，有一对羰基，会与胺类(还有蛋白质中的氨基酸)、氨基化合物、硫化物以及有机物的嘌呤发生剧烈反应。它还会与微生物组织形成亲核加合物，引起DNA-DNA和DNA-蛋白质的交叉连接键，会对DNA、微生物的突变以及哺乳动物细胞造成伤害，且致癌。甲醛沸点为19.5℃，故在室温时极易挥发，随着温度的上升挥发速度加快。目前已被世界卫生组织确定为致癌和致畸形物质，是公认的变态反应源，也是潜在的强致突变物之一。苯系物(benzene series)在环境污染监测与控制中，通常是指苯、甲苯、邻二甲苯、问二甲苯、对二甲苯、乙苯、苯乙烯、异丙苯等8种化合物，是无色具有特殊芳香气味的气体，经皮肤接触和吸收引起中毒，会造成嗜睡、头痛、呕吐等。苯系物已被国际癌症研究机构确认为有毒致癌物质。苯及苯化合物主要来自于合成纤维、塑料、燃料、橡胶等，隐藏在油漆、各种涂料的添加剂以及各种胶粘剂、防水材料中，还可来自燃料和烟叶的燃烧。在家庭装修中，胶带、粘合剂、墙纸、油漆和家具表面都是苯和苯的同系物超标的来源。统计显示，12％的装修出现了苯超标，甲苯超标率为14.4％，二甲苯则达到了21.3％。同时，甲醛、苯等污染物挥发期可长达十几二十年，居室中长期存在的低浓度污染物对人体健康的潜在威胁不容小视。Formaldehyde (HCHO), also known as "formaldehyde", is a colorless gas with a strong pungent odor at room temperature. It is easily soluble in water, alcohol and ether. It has the effect of coagulating proteins and has active chemical properties and biological properties. Formaldehyde is a lively molecule with a pair of carbonyl groups that can react violently with amines (and amino acids in proteins), amino compounds, sulfides, and purines in organic matter. It will also form nucleophilic adducts with microbial tissues, causing DNA-DNA and DNA-protein cross-links, causing damage to DNA, microbial mutations, and mammalian cells, and causing cancer. The boiling point of formaldehyde is 19.5°C, so it is very volatile at room temperature, and the volatilization speed is accelerated as the temperature rises. It has been determined by the World Health Organization as a carcinogenic and teratogenic substance, a recognized source of allergies, and one of the potential strong mutagens. Benzene series (benzene series) in environmental pollution monitoring and control, usually refers to 8 kinds of compounds such as benzene, toluene, o-xylene, p-xylene, p-xylene, ethylbenzene, styrene, cumene, etc. It is a gas with a special aromatic odor, which can cause poisoning through skin contact and absorption, and can cause drowsiness, headache, and vomiting. Benzene series have been identified as toxic carcinogens by the International Agency for Research on Cancer. Benzene and benzene compounds mainly come from synthetic fibers, plastics, fuel, rubber, etc., and are hidden in paint, various coating additives, various adhesives, and waterproof materials, and can also come from the burning of fuel and tobacco leaves. In home improvement, tapes, adhesives, wallpapers, paints, and furniture surfaces are all sources of excessive levels of benzene and benzene homologues. Statistics show that 12% of the decorations have exceeded the standard for benzene, 14.4% for toluene, and 21.3% for xylene. At the same time, the volatilization period of formaldehyde, benzene and other pollutants can be as long as ten or twenty years, and the potential threat to human health caused by long-term low-concentration pollutants in the living room cannot be underestimated.
甲醛对人体健康的影响主要表现在嗅觉异常、刺激、过敏、肺功能异常、肝功能异常和免疫功能异常等方面。长期接触低浓度的甲醛引起的主要症状是流泪、打喷嚏、咳嗽，甚至出现眼结膜炎以及支气管炎等，而且，甲醛还具有遗传毒性，在职业接触甲醛的人群中，外周血白细胞的DNA-蛋白交联物质DPC(DNA-protein Cross-links，DPC)水平将增高，DPC是化学物质毒作用的分子生物标志。与此同时，苯在1993年就被世界卫生组织确定为强致癌物，长期接触苯会引起骨髓与遗传损害，造成白细胞、血小板减少，导致再生障碍性贫血，甚至发生白血病，近些年来，很多劳动卫生学资料表明，长期接触苯系混合物的工人患再生障碍性贫血的比率较高，另外，苯还可导致胎儿的先天性缺陷。在室内空气中甲醛污染受到普遍重视的情况下，室内苯污染的预防与治理将是改善室内空气品质的又一重点问题。The lot of documents record, formaldehyde mainly shows aspects such as allotriosmia, stimulation, allergy, abnormal pulmonary function, dysfunction of liver and immunologic dysfunction to health effects. The main symptoms caused by long-term exposure to low concentrations of formaldehyde are tearing, sneezing, coughing, and even conjunctivitis and bronchitis. Moreover, formaldehyde is also genetically toxic. In people who are occupationally exposed to formaldehyde, the DNA of peripheral blood leukocytes- The level of protein cross-linking substance DPC (DNA-protein Cross-links, DPC) will increase, and DPC is a molecular biomarker of chemical substance toxicity. At the same time, benzene was identified as a strong carcinogen by the World Health Organization in 1993. Long-term exposure to benzene can cause bone marrow and genetic damage, reduce white blood cells and platelets, lead to aplastic anemia, and even leukemia. In recent years, many Occupational hygiene data show that workers who have been exposed to benzene-based mixtures for a long time have a higher rate of aplastic anemia. In addition, benzene can also cause congenital defects in fetuses. In the case that formaldehyde pollution in indoor air is generally taken seriously, the prevention and control of indoor benzene pollution will be another key issue to improve indoor air quality.
目前在室内空气品质研究领域，能够有效去除室内化学污染物的方法相对匮乏，而且均在不同程度上存在着各种不可逾越的缺陷：At present, in the field of indoor air quality research, methods that can effectively remove indoor chemical pollutants are relatively scarce, and all of them have various insurmountable defects to varying degrees:
活性碳过滤：这种方法虽然能够消除某些化学气体和异味，但活性炭属于消耗性产品，需经常更换。且对某些化学性污染效果很有限(例如甲醛)。Activated carbon filtration: Although this method can eliminate some chemical gases and odors, activated carbon is a consumable product and needs to be replaced frequently. And it has very limited effect on some chemical pollution (such as formaldehyde).
等离子：等离子空气净化系统是利用氧分子带电特点，运用阴极管等电极射出电子，将空气中的氧分子离子化，它能杀灭及中和分解空气中的细菌和化学污染物。缺点是有氮氧化合物污染，需高新风量加以淡化。其设备体积较大，耗电和价格都较高。Plasma: The plasma air purification system utilizes the charging characteristics of oxygen molecules, uses electrodes such as cathode tubes to emit electrons, and ionizes oxygen molecules in the air. It can kill and neutralize bacteria and chemical pollutants in the air. The disadvantage is that there is nitrogen oxide pollution, which requires high fresh air volume to desalinate. Its equipment is larger in size, power consumption and price are higher.
静电除尘：主要原理是产生高压静电，将有尘埃等可吸入粒子的空气通过电极时，被吸引和收集。对生物性和化学性污染效果欠佳。我国很多发电厂都有高压静电除尘器。近年，国内外此类产品加了紫外管及过滤网，开发成中央空调空气净化设备，但从体积、安装、价格，以及效果上都有局限性。Electrostatic dust removal: The main principle is to generate high-voltage static electricity, and when the air with dust and other respirable particles passes through the electrodes, it is attracted and collected. Ineffective against biological and chemical pollution. Many power plants in my country have high-voltage electrostatic precipitators. In recent years, such products at home and abroad have added ultraviolet tubes and filters to develop into central air-conditioning air purification equipment, but there are limitations in terms of volume, installation, price, and effect.
光触媒：常见的是把光触媒剂喷在室内四壁，利用室内灯光中400nm光波或窗外自然光中的紫外光催化，以产生羟基和负离子来进行净化。其净化强度和稳定度的弱点不言而喻，所以，尽管有净化效果，但要达到卫生部的消毒、净化标准则很难。Photocatalyst: It is common to spray the photocatalyst on the four walls of the room, and use the 400nm light wave in the indoor light or the ultraviolet photocatalysis in the natural light outside the window to produce hydroxyl and negative ions for purification. The weakness of its purification strength and stability is self-evident. Therefore, despite the purification effect, it is difficult to meet the disinfection and purification standards of the Ministry of Health.
植物法处理：许多现有的研究成果表明绿色植物对室内的空气污染有一定的净化作用。绿色植物能通过将空气中的化学物质转化为生长所需养料，从而有效地降低室内空气污染，但此类净化作用非常有限，其原因如下：1、植物进行的是光合作用，一般在白天吸入部分有害物质，而在夜晚不具备这个功能。而人的生活规律一般都是白天室外，夜晚在室内，在人最需要空气净化时，植物没有办法起到相应的作用；2、植物可吸收的有害物质量极为有限。国家标准对甲醛的释放值要求低于0.08mg/m3，一般刚装修完的房子，甲醛不低于0.2mg/m3，而作为吸收甲醛效率较高的波斯顿蕨，每小时吸收的甲醛仅为20μg。以一个面积30m2，高3m，甲醛值为0.2mg/m3的房间为例，需要在房间内放置540盆波斯顿蕨，才能在1小时后将有害物质降为0.08mg/m3，在净化过程中需保证房间内光线充足，且不再有甲醛释放。因此，植物也仅能对有害物质起到辅助的治理作用。Plant method treatment: Many existing research results show that green plants have a certain purification effect on indoor air pollution. Green plants can effectively reduce indoor air pollution by converting chemical substances in the air into nutrients needed for growth, but this kind of purification effect is very limited. The reasons are as follows: 1. Plants perform photosynthesis and generally inhale during the day Some harmful substances, but do not have this function at night. And the law of people's life is generally outside during the day and indoors at night. When people need air purification most, plants can't play a corresponding role; 2. The amount of harmful substances that plants can absorb is extremely limited. The national standard requires that the formaldehyde release value be less than 0.08mg/m 3 . Generally, for houses that have just been renovated, the formaldehyde is not less than 0.2mg/m 3 . As Boston fern, which has a high formaldehyde absorption efficiency, the formaldehyde absorbed per hour Only 20μg. Taking a room with an area of 30m 2 , a height of 3m, and a formaldehyde value of 0.2mg/m 3 as an example, 540 pots of Boston ferns need to be placed in the room to reduce the harmful substances to 0.08mg/m 3 after 1 hour. During the purification process, it is necessary to ensure that there is sufficient light in the room and that no formaldehyde is released. Therefore, plants can only play an auxiliary role in controlling harmful substances.
生物法去除空气中挥发性有机物的实质即通过微生物生长过程中的新陈代谢活动，以有机物作为生长所需的养分而分解转化的过程。在这一点上与生物法处理废水的原理是一致的，但由于大气并非微生物理想的生长环境，因此微生物通常不能在空气环境中正常生长，这使得生物法在空气净化中的应用受到限制。尽管目前生物法在工业废水处理和工业废气治理方面均有不同程度的应用，但关于微生物法降解室内空气化学污染的相关资料和研究在国内外还很少见，尚处前沿技术。The biological method to remove the essence of volatile organic compounds in the air is the process of decomposing and transforming organic matter as the nutrients required for growth through the metabolic activities in the growth process of microorganisms. At this point, it is consistent with the principle of biological treatment of wastewater, but because the atmosphere is not an ideal growth environment for microorganisms, microorganisms usually cannot grow normally in the air environment, which limits the application of biological methods in air purification. Although biological methods are currently applied to varying degrees in industrial wastewater treatment and industrial waste gas treatment, relevant information and research on the degradation of indoor air chemical pollution by microbial methods are still rare at home and abroad, and are still at the forefront of technology.
本发明利用假单胞菌属菌种体内含有的甲醛脱氢酶和甲醛歧化酶可对甲醛进行有效分解(N.Adroer，“Mechanism of formaldehyde biodegradation by pseudomonas putida”)，最终产物为水和二氧化碳，对室内环境不会产生负面影响，安全可靠。此外，假单胞菌属和不动杆菌属可对室内如苯、甲苯、二甲苯等的苯系物同样具有高效的降解作用(MarceloHenrique Otenio，“Benzene，toluene and xylene biodegradation by Pseudomonas putida CCMI852”)，在国内外文献中对此已有详细阐述。The present invention utilizes the formaldehyde dehydrogenase and formaldehyde dismutase contained in the Pseudomonas strains to effectively decompose formaldehyde (N. Adroer, "Mechanism of formaldehyde biodegradation by pseudomonas putida"), and the final products are water and carbon dioxide. No negative impact on the indoor environment, safe and reliable. In addition, Pseudomonas and Acinetobacter can also efficiently degrade indoor benzene series such as benzene, toluene and xylene (MarceloHenrique Otenio, "Benzene, toluene and xylene biodegradation by Pseudomonas putida CCMI852") , which has been elaborated in domestic and foreign literature.
发明内容 Contents of the invention
本发明的目的是克服现有技术中的不足，提供一种微生物吸附降解的空气净化材料。The purpose of the present invention is to overcome the deficiencies in the prior art, and provide an air purification material degraded by microbial adsorption.
本发明的第二个目的是提供一种微生物吸附降解的空气净化材料的制备方法。The second object of the present invention is to provide a method for preparing an air purification material degraded by microbial adsorption.
本发明的第三个目的是提供一种微生物吸附降解的空气净化材料的用途。The third object of the present invention is to provide the use of an air purification material degraded by microbial adsorption.
本发明的技术方案概述如下：Technical scheme of the present invention is summarized as follows:
一种微生物吸附降解的空气净化材料，是海藻酸钙凝胶包埋着经交叉培养驯化后的假单胞菌属或不动杆菌属的粒径在2～4mm的生物颗粒。An air purification material for microbial adsorption and degradation is calcium alginate gel embedded with cross-cultured and acclimated biological particles of the genus Pseudomonas or Acinetobacter with a particle size of 2 to 4 mm.
所述假单胞菌属菌体为类产碱假单胞菌Pseudomonaspseudoalcaligenes(CGMCC1.31)、铜绿假单胞菌Pseudomonas.Aeruginosa(CGMCC1.1129)、恶臭假单胞菌Pseudomonasputida(CGMCC1.1130)、荧光假单胞菌Pseudomonas fluorescens(CICC10271)或睾丸酮假单胞菌Pseudomonas testosteroni(CICC20542)。The bacteria of the genus Pseudomonas are Pseudomonas pseudoalcaligenes (CGMCC1.31), Pseudomonas.Aeruginosa (CGMCC1.1129), Pseudomonasputida (CGMCC1.1130), Pseudomonas fluorescens (CICC10271) or Pseudomonas testosteroni (CICC20542).
所述不动杆菌属菌体为乙酸钙不动杆菌Acinetobacter calcoaceticus(CGMCC 1.2004)。The Acinetobacter thallus is Acinetobacter calcoaceticus (CGMCC 1.2004).
一种微生物吸附降解的空气净化材料的制备方法，包括如下步骤：A preparation method of an air purification material degraded by microbial adsorption, comprising the steps of:
(1)对假单胞菌属或不动杆菌属进行活化和纯化培养：(1) Activation and purification of Pseudomonas or Acinetobacter:
(2)配制A培养基、B培养基和C培养基：(2) Prepare A medium, B medium and C medium:
所述A培养基为全营养培养基，pH为7.0，组成为：The A medium is a complete nutrient medium with a pH of 7.0 and consists of:
所述B培养基为无糖矿物盐培养基，浓度为g/L，组成为：Described B culture medium is sugar-free mineral salt culture medium, and concentration is g/L, consists of:
溶剂为蒸馏水；The solvent is distilled water;
所述C培养基为加糖矿物盐培养基，是在B培养基中加入葡萄糖配制而成，所述葡萄糖的浓度为0.37g/L；The C medium is a sugar-added mineral salt medium, which is prepared by adding glucose to the B medium, and the concentration of the glucose is 0.37g/L;
(3)对假单胞菌属或不动杆菌属进行交叉培养驯化：(3) Cross-cultivation and domestication of Pseudomonas or Acinetobacter:
将步骤(2)制备的A培养基、B培养基和C培养基分别放入培养皿中，使15-30mL/皿；Put the A medium, B medium and C medium prepared in step (2) into the culture dish respectively, so that 15-30mL/dish;
将质量浓度为0.01％～0.03％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含低浓度甲醛的B培养基和含低浓度甲醛的C培养基，每皿加1mL；Sterilize formaldehyde aqueous solution with a mass concentration of 0.01% to 0.03% and add them to Petri dish B and Petri dish C respectively to make B medium containing low concentration formaldehyde and C medium containing low concentration formaldehyde, add 1mL to each dish ;
将质量浓度为0.02％～0.06％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含中浓度甲醛的B培养基和含中浓度甲醛的C培养基，每皿加1mL；Sterilize formaldehyde aqueous solution with a mass concentration of 0.02% to 0.06% and add it to Petri dish B and Petri dish C respectively to make B medium containing medium-concentration formaldehyde and C medium containing medium-concentration formaldehyde, add 1mL to each dish ;
将质量浓度为0.04～0.12％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含高浓度甲醛的B培养基和含高浓度甲醛的C培养基，每皿加1mL；Sterilize formaldehyde aqueous solution with a mass concentration of 0.04-0.12% and add it to Petri dish B and Petri dish C respectively to make medium B with high concentration of formaldehyde and medium C with high concentration of formaldehyde, add 1 mL to each dish;
将所述步骤(1)制备的纯化菌种接种到所述含低浓度甲醛的B培养基中25～35℃培养2～3天后，转接到所述含低浓度甲醛的C培养基中25～35℃培养2～3天；接着转接到所述含中浓度甲醛的B培养基中25～35℃培养2～3天后，转接到所述含中浓度甲醛的C培养基中25～35℃培养2～3天；之后转接到所述含高浓度甲醛的B培养基中25～35℃培养2～3天，再转接到所述含高浓度甲醛的C培养基中25～35℃培养2～3天，最后转接到所述A培养基中25～35℃培养1～2天，达到对数生长期或稳定期，取菌落制成固体琼脂斜面保藏菌种，25℃～35℃培养1～2天后，得到已驯化的菌体，4℃冰箱内保存待用；Inoculate the purified strains prepared in the step (1) into the B medium containing low-concentration formaldehyde at 25 to 35°C for 2 to 3 days, then transfer to the C medium containing low-concentration formaldehyde for 25 Cultivate at ~35°C for 2 to 3 days; then transfer to the B medium containing medium concentration of formaldehyde and culture at 25 to 35°C for 2 to 3 days, then transfer to the C medium containing medium concentration of formaldehyde for 25 ~ Cultivate at 35°C for 2-3 days; then transfer to the B medium containing high-concentration formaldehyde and culture at 25-35°C for 2-3 days, then transfer to the C medium containing high-concentration formaldehyde for 25-3 days Cultivate at 35°C for 2 to 3 days, and finally transfer to the A medium for 1 to 2 days at 25 to 35°C to reach the logarithmic growth phase or stable phase, take the colonies to make solid agar slant to preserve the strains, 25°C After culturing at ~35°C for 1-2 days, the acclimatized bacteria are obtained and stored in a refrigerator at 4°C until use;
(4)用海藻酸钙包埋已驯化的菌体：(4) Embedding the domesticated bacteria with calcium alginate:
将步骤(3)制备的已驯化的菌体接种到装有500mL已灭菌的营养肉汤液体培养基的三角瓶中，25℃～35℃下在摇床上恒温培养1～2天，菌体密度达到109-1010个/mL，得菌悬液，取50mL菌悬液注入灭菌的离心管中，在4000rpm下分离15-30分钟，分离所得菌体细胞经灭菌生理盐水洗涤2～3次，加蒸馏水至50mL，使菌体细胞悬浮于蒸馏水中，得加水菌悬液；Inoculate the acclimatized thalline prepared in step (3) into a conical flask equipped with 500 mL of sterilized nutrient broth liquid medium, and cultivate it on a shaker at a constant temperature of 1 to 2 days at 25°C to 35°C, and the thalline When the density reaches 10 9 -10 10 cells/mL, the bacterial suspension is obtained, and 50 mL of the bacterial suspension is injected into a sterilized centrifuge tube, separated at 4000 rpm for 15-30 minutes, and the separated bacterial cells are washed with sterilized normal saline for 2 ~3 times, add distilled water to 50mL, suspend the bacterial cells in distilled water, and add water bacterial suspension;
按体积比为100∶0.5～1的比例将质量浓度为2％-5％的海藻酸钠水溶液与所述加水菌悬液混匀后制成混合液体，用针形管将所述混合液体滴入质量浓度为5％-10％的CaCl2水溶液中，静置7～8小时，得到2～5mm的颗粒，滤出颗粒后用生理盐水和蒸馏水洗净，制成一种生物吸附空气净化材料。Mix the sodium alginate aqueous solution with a mass concentration of 2%-5% and the water-added bacteria suspension at a volume ratio of 100:0.5 to 1 to make a mixed liquid, and use a needle-shaped tube to drop the mixed liquid Put it into a CaCl2 aqueous solution with a mass concentration of 5%-10%, let it stand for 7-8 hours to obtain particles of 2-5mm, filter out the particles and wash them with physiological saline and distilled water to make a bioabsorption air purification material.
所述步骤(1)优选的是：The step (1) is preferably:
取冷冻真空干燥管内假单胞菌属或不动杆菌属菌体，在无菌环境下注入80-120mL的营养肉汤液体培养基，在摇床25～35℃下连续培养1～2天活化菌种，利用平板划线分离法分离出单个菌落，在25～35℃恒温培养箱内连续培养1～2天，转移至固体琼脂斜面培养基培养1～2天，于4℃冰箱内保存待用，所述营养肉汤液体培养基的组成为：牛肉膏1.5g，蛋白胨1g，NaCl1.5g，蒸馏水100mL；所述固体琼脂斜面培养基的组成为：营养肉汤液体培养基100mL，琼脂4-5g，调节pH为7-7.5。Take Pseudomonas or Acinetobacter bacteria in the freeze-drying tube, inject 80-120mL nutrient broth liquid culture medium in a sterile environment, and continuously cultivate for 1-2 days on a shaker at 25-35°C to activate Bacterial strains, a single colony was isolated by the plate streak separation method, continuously cultured in a constant temperature incubator at 25-35°C for 1-2 days, transferred to a solid agar slant medium for 1-2 days, and stored in a refrigerator at 4°C until Use, the composition of described nutrient broth liquid medium is: beef extract 1.5g, peptone 1g, NaCl1.5g, distilled water 100mL; The composition of described solid agar slant medium is: nutrient broth liquid medium 100mL, agar 4 -5g, adjust the pH to 7-7.5.
所述假单胞菌属菌体为类产碱假单胞菌Pseudomonas pseudoalcaligenes(CGMCC1.31)、铜绿假单胞菌Pseudomonas.Aeruginosa(CGMCC1.1129)、恶臭假单胞菌Pseudomonasputida(CGMCC1.1130)、荧光假单胞菌Pseudomonas fluorescens(CICC10271)或睾丸酮假单胞菌Pseudomonas testosteroni(CICC20542)。The bacterium of the genus Pseudomonas is Pseudomonas pseudoalcaligenes (CGMCC1.31), Pseudomonas.Aeruginosa (CGMCC1.1129), Pseudomonasputida (CGMCC1.1130) , Pseudomonas fluorescens (CICC10271) or Pseudomonas testosteroni (CICC20542).
所述不动杆菌属菌体为乙酸钙不动杆菌Acinetobacter calcoaceticus(CGMCC 1.2004)。The Acinetobacter thallus is Acinetobacter calcoaceticus (CGMCC 1.2004).
微生物吸附降解的空气净化材料在吸附降解空气化学污染物中的应用。The application of microbial adsorption and degradation air purification materials in the adsorption and degradation of air chemical pollutants.
本发明的优点是：The advantages of the present invention are:
微生物吸附降解的空气净化材料可用于室内甲醛和苯系物污染的长期净化，净化效果显著。该产品具有固定化细胞活性高，稳定性好，能够持久有效地降低室内甲醛与苯系物等有害化学污染物的含量，利于实现生物催化剂的回收、再生和重复使用的优点，并严格避免了微生物对室内环境的污染。原材料价格低廉，制作工艺简便易行，制成品可在多种反应器中应用，在改善室内空气品质和去除化学污染物等净化领域有着广阔的市场前景。The air purification material degraded by microbial adsorption can be used for long-term purification of indoor formaldehyde and benzene series pollution, and the purification effect is remarkable. This product has the advantages of high activity of immobilized cells and good stability, and can effectively and permanently reduce the content of harmful chemical pollutants such as formaldehyde and benzene series in the room, which is beneficial to the recovery, regeneration and reuse of biocatalysts, and strictly avoids Microbial contamination of the indoor environment. The price of raw materials is low, the manufacturing process is simple and easy, and the finished product can be applied in various reactors, and has broad market prospects in the purification fields such as improving indoor air quality and removing chemical pollutants.
附图说明 Description of drawings
图1为本发明一种微生物吸附降解的空气净化材料的外观扫描电镜照片。Fig. 1 is a scanning electron micrograph of the appearance of an air purification material degraded by microbial adsorption of the present invention.
图2为本发明一种微生物吸附降解的空气净化材料的内部网孔结构扫描电镜照片。Fig. 2 is a scanning electron micrograph of the internal mesh structure of an air purification material degraded by microbial adsorption of the present invention.
图3为本发明一种微生物吸附降解的空气净化材料的外观。Fig. 3 is the appearance of an air purification material degraded by microbial adsorption of the present invention.
图4为本发明微生物吸附降解的空气净化材料降解室内甲醛的效果。Fig. 4 is the effect of degrading indoor formaldehyde by the air purification material degraded by microbial adsorption of the present invention.
具体实施方式 Detailed ways
下面结合具体实施例对本发明作进一步的说明。The present invention will be further described below in conjunction with specific examples.
实施例1Example 1
一种微生物吸附降解的空气净化材料，是海藻酸钙凝胶包埋着经交叉培养驯化后的恶臭假单胞菌Pseudomonas putida(CGMCC 1.1130)的粒径在2～4mm的生物颗粒。An air purification material for microbial adsorption and degradation, which is a biological particle with a particle size of 2-4mm of Pseudomonas putida (CGMCC 1.1130) after cross-cultivation and domestication embedded in calcium alginate gel.
利用海藻酸钙凝胶包埋恶臭假单胞菌制成粒径在2～4mm的生物颗粒，将微生物包埋于海藻酸钙高分子链骨架形成的孔径约为100～500μm的较大圆孔内，有效防止了微生物细胞的向外渗漏而对室内环境造成污染。海藻酸钙凝胶颗粒是一种三维梯状结构体系，结构疏松，利于物质传递，表面的微孔结构(孔径约为20-50μm)有利于水分和VOCs等小分子进入，内部结构疏松，非常有利于物质传递，进入的水分和VOCs分子很容易由海藻酸钙凝胶表面向其体系内部进行扩散，进而有利于凝胶内部的微生物降解化学污染物的深度进行，因此可高效去除室内化学污染物。Use calcium alginate gel to embed Pseudomonas putida to make biological particles with a particle size of 2-4 mm, and embed the microorganisms in larger circular holes with a pore size of about 100-500 μm formed by the calcium alginate polymer chain skeleton , Effectively prevent the outward leakage of microbial cells and cause pollution to the indoor environment. Calcium alginate gel particles are a three-dimensional ladder-like structure system with a loose structure that is conducive to material transfer. The microporous structure on the surface (pore size is about 20-50 μm) is conducive to the entry of small molecules such as water and VOCs. The internal structure is loose and very It is conducive to material transfer, and the entering water and VOCs molecules are easy to diffuse from the surface of calcium alginate gel to the interior of the system, which is conducive to the depth of microbial degradation of chemical pollutants inside the gel, so it can efficiently remove indoor chemical pollution thing.
海藻酸钙凝胶珠是一种密度分布不均匀的体系，由外向内可分为致密层，较致密层和疏松内核，本发明中用于包埋恶臭假单胞菌的海藻酸钙凝胶表面微孔孔径约为20-50μm，内部网孔孔径约为100-500μm，细菌在海藻酸钙凝胶中所占体积约为0.88％，生物颗粒粒径约为2-4mm。见图1、图2和图3。Calcium alginate gel beads are a system with uneven density distribution, which can be divided into dense layer, denser layer and loose inner core from outside to inside. The calcium alginate gel used for embedding Pseudomonas putida in the present invention The surface micropore diameter is about 20-50 μm, the internal mesh aperture is about 100-500 μm, the volume of bacteria in the calcium alginate gel is about 0.88%, and the particle size of biological particles is about 2-4mm. See Figure 1, Figure 2 and Figure 3.
实施例2Example 2
一种微生物吸附降解的空气净化材料，是海藻酸钙凝胶包埋着经交叉培养驯化后的类产碱假单胞菌Pseudomonas pseudoalcaligenes(CGMCC 1.31)的粒径在2～4mm的生物颗粒。An air purification material for microbial adsorption and degradation, which is a calcium alginate gel embedded with cross-cultivated Pseudomonas pseudoalcaligenes (CGMCC 1.31) biological particles with a particle size of 2-4mm.
实施例3Example 3
一种微生物吸附降解的空气净化材料，是海藻酸钙凝胶包埋着经交叉培养驯化后的铜绿假单胞菌Pseudomonas.Aeruginosa(CGMCC1.1129)的粒径在2～4mm的生物颗粒。An air purification material for microbial adsorption and degradation is calcium alginate gel embedded with biological particles of Pseudomonas.
实施例4Example 4
一种微生物吸附降解的空气净化材料，是海藻酸钙凝胶包埋着经交叉培养驯化后的荧光假单胞菌Pseudomonas fluorescens(CICC10271)的粒径在2～4mm的生物颗粒。An air purification material for microbial adsorption and degradation, which is a biological particle with a diameter of 2-4 mm of Pseudomonas fluorescens (CICC10271) that has been domesticated by cross-cultivation and embedded in calcium alginate gel.
实施例5Example 5
一种微生物吸附降解的空气净化材料，是海藻酸钙凝胶包埋着经交叉培养驯化后的睾丸酮假单胞菌Pseudomonas testosteroni(CICC20542)的粒径在2～4mm的生物颗粒。An air purification material for microbial adsorption and degradation is calcium alginate gel embedded with biological particles of Pseudomonas testosteroni (CICC20542) with a particle size of 2-4mm after cross-cultivation and domestication.
实施例6Example 6
一种微生物吸附降解的空气净化材料，是海藻酸钙凝胶包埋着经交叉培养驯化后的乙酸钙不动杆菌Acinetobacter calcoaceticus：(CGMCC 1.2004)的粒径在2～4mm的生物颗粒。An air purification material for microbial adsorption and degradation, which is a calcium alginate gel embedded with cross-cultivated Acinetobacter calcoaceticus: (CGMCC 1.2004) biological particles with a particle size of 2 to 4 mm.
实施例7Example 7
一种微生物吸附降解的空气净化材料的制备方法，包括如下步骤：A preparation method of an air purification material degraded by microbial adsorption, comprising the steps of:
(1)对恶臭假单胞菌进行活化和纯化培养：(1) Activate and purify Pseudomonas putida:
取冷冻真空干燥管内恶臭假单胞菌Pseudomonasputida(CGMCC1.1130)菌体，在无菌环境下注入100mL的营养肉汤液体培养基，在摇床30℃下连续培养1.5天活化菌种，利用平板划线分离法分离出单个菌落，在30℃恒温培养箱内连续培养1.5天，转移至固体琼脂斜面培养基培养1.5天，于4℃冰箱内保存待用，所述营养肉汤液体培养基的组成为：牛肉膏1.5g，蛋白胨1g，NaCl1.5g，蒸馏水100mL；所述固体琼脂斜面培养基的组成为：营养肉汤液体培养基100mL，琼脂5g，调节pH为7；Take Pseudomonasputida (CGMCC1.1130) cells in the freeze-drying tube, inject 100mL of nutrient broth liquid medium in a sterile environment, and continuously cultivate the activated strains for 1.5 days on a shaker at 30°C. A single colony was isolated by the streak separation method, continuously cultured in a 30°C constant temperature incubator for 1.5 days, transferred to a solid agar slant medium for 1.5 days, and stored in a 4°C refrigerator for use. The nutrient broth liquid medium It consists of: 1.5g of beef extract, 1g of peptone, 1.5g of NaCl, and 100mL of distilled water; the composition of the solid agar slant medium is: 100mL of nutrient broth liquid medium, 5g of agar, and adjust the pH to 7;
(2)配制A培养基、B培养基和C培养基：(2) Prepare A medium, B medium and C medium:
所述A培养基为全营养培养基，pH为7.0，组成为：The A medium is a complete nutrient medium with a pH of 7.0 and consists of:
所述B培养基为无糖矿物盐培养基，浓度为g/L，组成为：Described B culture medium is sugar-free mineral salt culture medium, and concentration is g/L, consists of:
溶剂为蒸馏水；The solvent is distilled water;
所述C培养基为加糖矿物盐培养基，是在B培养基中加入葡萄糖配制而成，所述葡萄糖的浓度为0.37g/L；The C medium is a sugar-added mineral salt medium, which is prepared by adding glucose to the B medium, and the concentration of the glucose is 0.37g/L;
(3)对恶臭假单胞菌进行交叉培养驯化：(3) Pseudomonas putida is cross-cultured and domesticated:
将步骤(2)制备的A培养基、B培养基和C培养基分别放入培养皿中，使20mL/皿；Put the A medium, B medium and C medium prepared in step (2) into the culture dish respectively to make 20mL/dish;
将质量浓度为0.02％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含低浓度甲醛的B培养基和含低浓度甲醛的C培养基，每皿加1mL；Add 0.02% formaldehyde aqueous solution to Petri dish B and Petri dish C respectively after sterilizing the bacteria to make medium B with low concentration of formaldehyde and medium C with low concentration of formaldehyde, add 1 mL to each dish;
将质量浓度为0.04％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含中浓度甲醛的B培养基和含中浓度甲醛的C培养基，每皿加1mL；Sterilize 0.04% formaldehyde aqueous solution and add them to Petri dish B and Petri dish C respectively to make B medium containing medium-concentration formaldehyde and C medium containing medium-concentration formaldehyde, add 1 mL to each dish;
将质量浓度为0.08％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含高浓度甲醛的B培养基和含高浓度甲醛的C培养基，每皿加1mL；Add 0.08% formaldehyde aqueous solution to Petri dish B and Petri dish C respectively after sterilizing the bacteria to make medium B with high concentration of formaldehyde and medium C with high concentration of formaldehyde, add 1 mL to each dish;
将所述步骤(1)制备的纯化菌种接种到所述含低浓度甲醛的B培养基中30℃培养3天，转接到所述含低浓度甲醛的C培养基中30℃培养3天，再转接到所述含中浓度甲醛的B培养基中30℃培养3天，再转接到所述含中浓度甲醛的C培养基中30℃培养3天，再转接到所述含高浓度甲醛的B培养基中30℃培养3天，再转接到所述含高浓度甲醛的C培养基中30℃培养3天，再转接到所述A培养基中30℃培养2天，达到对数生长期，取菌落制成固体琼脂斜面保藏菌种，30℃培养2天后，得到已驯化的菌体，4℃冰箱内保存待用；Inoculate the purified strains prepared in the step (1) into the B medium containing low-concentration formaldehyde and cultivate at 30°C for 3 days, then transfer to the C medium containing low-concentration formaldehyde and cultivate at 30°C for 3 days , then transferred to the B medium containing medium concentration formaldehyde for 3 days at 30°C, then transferred to the medium C medium containing formaldehyde for 3 days at 30°C, and then transferred to the medium containing formaldehyde Cultivate in medium B with high concentration of formaldehyde at 30°C for 3 days, then transfer to medium C containing high concentration of formaldehyde and culture at 30°C for 3 days, then transfer to medium A and culture at 30°C for 2 days , to reach the logarithmic growth phase, take the colony and make it into a solid agar slant to preserve the strain, cultivate it at 30°C for 2 days, and obtain the domesticated bacteria, and store it in a refrigerator at 4°C until use;
(4)用海藻酸钙包埋已驯化的菌体：(4) Embedding the domesticated bacteria with calcium alginate:
将步骤(3)制备的已驯化的菌体接种到装有500mL已灭菌的营养肉汤液体培养基的三角瓶中，30℃下在摇床上恒温培养2天，菌体密度达到1010个/mL，得菌悬液，取50mL菌悬液注入灭菌的离心管中，在4000rpm下分离20分钟，分离所得菌体细胞经灭菌生理盐水洗涤3次，加蒸馏水至50mL，使菌体细胞悬浮于蒸馏水中，得加水菌悬液；Inoculate the acclimatized thalline prepared in step (3) into a conical flask equipped with 500 mL of sterilized nutrient broth liquid medium, and culture it on a shaker at a constant temperature for 2 days at 30° C., and the thalline density reaches 10 10 /mL, to obtain the bacterial suspension, take 50mL of the bacterial suspension and inject it into a sterilized centrifuge tube, separate it at 4000rpm for 20 minutes, wash the separated bacterial cells with sterilized normal saline for 3 times, add distilled water to 50mL, and make the bacterial cells The cells are suspended in distilled water, and the bacterial suspension in water is added;
按体积比为100∶1的比例将质量浓度为2％的海藻酸钠水溶液与所述加水菌悬液混匀后制成混合液体，用针形管将所述混合液体滴入质量浓度为8％的CaCl2水溶液中，静置8小时，得到2～5mm的颗粒，滤出颗粒后用生理盐水和蒸馏水洗净，制成一种生物吸附空气净化材料。According to the volume ratio of 100:1, the sodium alginate aqueous solution with a mass concentration of 2% is mixed with the water-added bacterial suspension to make a mixed liquid, and the mixed liquid is dripped into the mixed liquid with a mass concentration of 8 with a needle tube. % CaCl 2 aqueous solution, let it stand for 8 hours to obtain particles of 2-5mm, filter out the particles and wash them with physiological saline and distilled water to make a biological adsorption air purification material.
实施例8Example 8
一种微生物吸附降解的空气净化材料的制备方法，包括如下步骤：A preparation method of an air purification material degraded by microbial adsorption, comprising the steps of:
(1)对类产碱假单胞菌进行活化和纯化培养：(1) Activate and purify Pseudomonas pseudoalcaligenes:
取冷冻真空干燥管内类产碱假单胞菌Pseudomonas pseudoalcaligenes(CGMCC 1.31)菌体，在无菌环境下注入80mL的营养肉汤液体培养基，在摇床25℃下连续培养2天活化菌种，利用平板划线分离法分离出单个菌落，在25℃恒温培养箱内连续培养2天，转移至固体琼脂斜面培养基培养1天，于4℃冰箱内保存待用，营养肉汤液体培养基的组成与实施例7相同；固体琼脂斜面培养基的组成为：营养肉汤液体培养基100mL，琼脂4g，调节pH为7.5；Take the Pseudomonas pseudoalcaligenes (CGMCC 1.31) cells in the freeze-drying tube, inject 80 mL of nutrient broth liquid medium in a sterile environment, and continuously cultivate the bacteria in a shaker at 25 °C for 2 days to activate the strains. A single colony was isolated by the plate streak separation method, continuously cultured in a constant temperature incubator at 25°C for 2 days, transferred to a solid agar slant medium for 1 day, and stored in a refrigerator at 4°C until use. The composition is the same as in Example 7; the composition of the solid agar slant medium is: 100 mL of nutrient broth liquid medium, 4 g of agar, and the pH is adjusted to 7.5;
(2)配制A培养基、B培养基和C培养基：同实施例7；(2) preparation of A medium, B medium and C medium: with embodiment 7;
(3)对类产碱假单胞菌进行交叉培养驯化：(3) Carry out cross-cultivation domestication to Pseudomonas pseudoalcaligenes:
将步骤(2)制备的A培养基、B培养基和C培养基分别放入培养皿中，使25mL/皿；Put the A medium, B medium and C medium prepared in step (2) into the culture dish respectively to make 25mL/dish;
将质量浓度为0.01％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含低浓度甲醛的B培养基和含低浓度甲醛的C培养基，每皿加1mL；Add 0.01% formaldehyde aqueous solution to Petri dish B and Petri dish C respectively after sterilizing the bacteria to make medium B with low concentration of formaldehyde and medium C with low concentration of formaldehyde, add 1 mL to each dish;
将质量浓度为0.02％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含中浓度甲醛的B培养基和含中浓度甲醛的C培养基，每皿加1mL；Sterilize 0.02% formaldehyde aqueous solution and add them to Petri dish B and Petri dish C respectively to make B medium containing medium-concentration formaldehyde and C medium containing medium-concentration formaldehyde, add 1 mL to each dish;
将质量浓度为0.04的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含高浓度甲醛的B培养基和含高浓度甲醛的C培养基，每皿加1mL；After sterilizing the formaldehyde solution with a mass concentration of 0.04, add it to Petri dish B and Petri dish C respectively to make medium B with high concentration of formaldehyde and medium C with high concentration of formaldehyde, add 1 mL to each dish;
将所述步骤(1)制备的纯化菌种接种到所述含低浓度甲醛的B培养基中35℃培养2天，转接到所述含低浓度甲醛的C培养基中35℃培养2天，再转接到所述含中浓度甲醛的B培养基中35℃培养2天，再转接到所述含中浓度甲醛的C培养基中35℃培养2天，再转接到所述含高浓度甲醛的B培养基中35℃培养2天，再转接到所述含高浓度甲醛的C培养基中35℃培养2天，再转接到所述A培养基中35℃培养1天，达到对数生长期，取菌落制成固体琼脂斜面保藏菌种，35℃培养1天后，得到已驯化的菌体，4℃冰箱内保存待用；Inoculate the purified strain prepared in the step (1) into the B medium containing low-concentration formaldehyde and cultivate at 35°C for 2 days, then transfer to the C medium containing low-concentration formaldehyde and cultivate at 35°C for 2 days , then transferred to the B medium containing medium concentration formaldehyde for 2 days at 35°C, then transferred to the medium C medium containing formaldehyde for 2 days at 35°C, and then transferred to the medium containing formaldehyde Culture in medium B with high concentration of formaldehyde at 35°C for 2 days, then transfer to medium C containing high concentration of formaldehyde and culture at 35°C for 2 days, then transfer to medium A and culture at 35°C for 1 day , to reach the logarithmic growth phase, take the colony and make it into a solid agar slant to preserve the strain, cultivate it at 35°C for 1 day, and obtain the domesticated bacteria, and store it in the refrigerator at 4°C until use;
(4)用海藻酸钙包埋已驯化的菌体：(4) Embedding the domesticated bacteria with calcium alginate:
将步骤(3)制备的已驯化的菌体接种到装有500mL已灭菌的营养肉汤液体培养基的三角瓶中，35℃下在摇床上恒温培养1天，菌体密度达到109个/mL，得菌悬液，取50mL菌悬液注入灭菌的离心管中，在4000rpm下分离30分钟，分离所得菌体细胞经灭菌生理盐水洗涤2次，加蒸馏水至50mL，使菌体细胞悬浮于蒸馏水中，得加水菌悬液；Inoculate the domesticated thallus prepared in step (3) into a conical flask containing 500 mL of sterilized nutrient broth liquid medium, and culture it on a shaker at a constant temperature at 35°C for 1 day, and the thallus density reaches 109 /mL, to obtain the bacterial suspension, take 50mL of the bacterial suspension and inject it into a sterilized centrifuge tube, separate it at 4000rpm for 30 minutes, wash the separated bacterial cells twice with sterile normal saline, add distilled water to 50mL, and make the bacterial The cells are suspended in distilled water, and the bacterial suspension in water is added;
按体积比为100∶1的比例将质量浓度为3％的海藻酸钠水溶液与所述加水菌悬液混匀后制成混合液体，用针形管将所述混合液体滴入质量浓度为7％的CaCl2水溶液中，静置7小时，得到2～5mm的颗粒，滤出颗粒后用生理盐水和蒸馏水洗净，制成一种生物吸附空气净化材料。According to the volume ratio of 100:1, the sodium alginate aqueous solution with a mass concentration of 3% is mixed with the water-added bacterial suspension to make a mixed liquid, and the mixed liquid is dripped into the mixed liquid with a mass concentration of 7% with a needle tube. % CaCl 2 aqueous solution, let it stand for 7 hours to obtain particles of 2-5mm, filter out the particles and wash them with physiological saline and distilled water to make a biological adsorption air purification material.
实施例9Example 9
一种微生物吸附降解的空气净化材料的制备方法，包括如下步骤：A preparation method of an air purification material degraded by microbial adsorption, comprising the steps of:
(1)对铜绿假单胞菌进行活化和纯化培养：(1) Pseudomonas aeruginosa is activated and purified:
取冷冻真空干燥管内铜绿假单胞菌Pseudomonas.Aeruginosa(CGMCC 1.1129)菌体，在无菌环境下注入90mL的营养肉汤液体培养基，在摇床35℃下连续培养1天活化菌种，利用平板划线分离法分离出单个菌落，在35℃恒温培养箱内连续培养1天，转移至固体琼脂斜面培养基培养2天，于4℃冰箱内保存待用，营养肉汤液体培养基的组成同实施例7；固体琼脂斜面培养基的组成同实施例7；Take Pseudomonas.Aeruginosa (CGMCC 1.1129) cells in the freeze-drying tube, inject 90 mL of nutrient broth liquid medium in a sterile environment, and continuously cultivate for 1 day on a shaker at 35°C to activate the strains. A single colony was isolated by streaking on a flat plate, continuously cultured in a constant temperature incubator at 35°C for 1 day, transferred to a solid agar slant medium for 2 days, and stored in a refrigerator at 4°C until use. The composition of the nutrient broth liquid medium With embodiment 7; The composition of solid agar slant medium is with embodiment 7;
(2)配制A培养基、B培养基和C培养基：同实施例7；(2) preparation of A medium, B medium and C medium: with embodiment 7;
(3)对铜绿假单胞菌进行交叉培养驯化：(3) Pseudomonas aeruginosa is cross-cultured and domesticated:
将步骤(2)制备的A培养基、B培养基和C培养基分别放入培养皿中，使30mL/皿；Put the A medium, B medium and C medium prepared in step (2) into the culture dish respectively to make 30mL/dish;
将质量浓度为0.03％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含低浓度甲醛的B培养基和含低浓度甲醛的C培养基，每皿加1mL；Add 0.03% formaldehyde aqueous solution to Petri dish B and Petri dish C respectively after sterilizing the bacteria to make medium B with low concentration of formaldehyde and medium C with low concentration of formaldehyde, add 1 mL to each dish;
将质量浓度为0.06％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含中浓度甲醛的B培养基和含中浓度甲醛的C培养基，每皿加1mL；Add 0.06% formaldehyde aqueous solution to Petri dish B and Petri dish C respectively after sterilizing the bacteria to make B medium containing medium-concentration formaldehyde and C medium containing medium-concentration formaldehyde, add 1 mL to each dish;
将质量浓度为0.12％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含高浓度甲醛的B培养基和含高浓度甲醛的C培养基，每皿加1mL；Sterilize 0.12% formaldehyde aqueous solution and add them to Petri dish B and Petri dish C respectively to make medium B with high concentration of formaldehyde and medium C with high concentration of formaldehyde, add 1 mL to each dish;
将所述步骤(1)制备的纯化菌种接种到所述含低浓度甲醛的B培养基中25℃培养3天，转接到所述含低浓度甲醛的C培养基中25℃培养3天，再转接到所述含中浓度甲醛的B培养基中25℃培养3天，再转接到所述含中浓度甲醛的C培养基中25℃培养3天，再转接到所述含高浓度甲醛的B培养基中25℃培养3天，再转接到所述含高浓度甲醛的C培养基中25℃培养3天，再转接到所述A培养基中25℃培养1天，达到稳定期，取菌落制成固体琼脂斜面保藏菌种，25℃培养2天后，得到已驯化的菌体，4℃冰箱内保存待用；Inoculate the purified strains prepared in the step (1) into the B medium containing low-concentration formaldehyde and cultivate at 25°C for 3 days, then transfer to the C medium containing low-concentration formaldehyde and cultivate at 25°C for 3 days , then transferred to the B medium containing medium concentration formaldehyde for 3 days at 25°C, then transferred to the medium C medium containing formaldehyde for 3 days at 25°C, and then transferred to the medium containing formaldehyde Cultivate in medium B with high concentration of formaldehyde at 25°C for 3 days, then transfer to medium C containing high concentration of formaldehyde and culture at 25°C for 3 days, then transfer to medium A and culture at 25°C for 1 day , reach the stable period, take the colony to make a solid agar slant to preserve the strain, cultivate it at 25°C for 2 days, and obtain the acclimatized bacteria, and store it in the refrigerator at 4°C until use;
(4)用海藻酸钙包埋已驯化的菌体：(4) Embedding the domesticated bacteria with calcium alginate:
将步骤(3)制备的已驯化的菌体接种到装有500mL已灭菌的营养肉汤液体培养基的三角瓶中，25℃下在摇床上恒温培养2天，菌体密度达到109个/mL，得菌悬液，取50mL菌悬液注入灭菌的离心管中，在4000rpm下分离15分钟，分离所得菌体细胞经灭菌生理盐水洗涤3次，加蒸馏水至50mL，使菌体细胞悬浮于蒸馏水中，得加水菌悬液；Inoculate the acclimatized thalline prepared in step (3) into a conical flask equipped with 500 mL of sterilized nutrient broth liquid medium, and cultivate it at a constant temperature on a shaker for 2 days at 25 ° C, and the thalline density reaches 10 9 /mL, to obtain the bacterial suspension, take 50mL of the bacterial suspension and inject it into a sterilized centrifuge tube, separate it at 4000rpm for 15 minutes, wash the separated bacterial cells with sterilized normal saline for 3 times, add distilled water to 50mL, and make the bacterial cells The cells are suspended in distilled water, and the bacterial suspension in water is added;
按体积比为100∶0.5的比例将质量浓度为2％的海藻酸钠水溶液与所述加水菌悬液混匀后制成混合液体，用针形管将所述混合液体滴入质量浓度为5％的CaCl2水溶液中，静置7小时，得到2～5mm的颗粒，滤出颗粒后用生理盐水和蒸馏水洗净，制成一种生物吸附空气净化材料。According to the volume ratio of 100:0.5, the sodium alginate aqueous solution with a mass concentration of 2% is mixed with the water-added bacterial suspension to make a mixed liquid, and the mixed liquid is dripped into the mixed liquid with a mass concentration of 5% with a needle tube. % CaCl 2 aqueous solution, stand still for 7 hours to obtain 2-5mm particles, filter out the particles and wash them with physiological saline and distilled water to make a bioabsorption air purification material.
实施例10Example 10
一种微生物吸附降解的空气净化材料的制备方法，包括如下步骤：A preparation method of an air purification material degraded by microbial adsorption, comprising the steps of:
(1)对荧光假单胞菌进行活化和纯化培养：(1) Activate and purify Pseudomonas fluorescens:
取冷冻真空干燥管内荧光假单胞菌Pseudomonas fluorescens(CICC 10271)菌体，在无菌环境下注入120mL的营养肉汤液体培养基，在摇床25℃下连续培养1天活化菌种，利用平板划线分离法分离出单个菌落，在25℃恒温培养箱内连续培养1天，转移至固体琼脂斜面培养基培养2天，于4℃冰箱内保存待用，营养肉汤液体培养基的组成同实施例7；固体琼脂斜面培养基的组成同实施例8；Take Pseudomonas fluorescens (CICC 10271) cells in the freeze-drying tube, inject 120 mL of nutrient broth liquid medium in a sterile environment, and continuously cultivate the strains on a shaker at 25°C for 1 day to activate the strains. A single colony was isolated by the streak separation method, continuously cultured in a constant temperature incubator at 25°C for 1 day, transferred to a solid agar slant medium for 2 days, and stored in a 4°C refrigerator for later use. The composition of the nutrient broth liquid medium was the same as Embodiment 7; The composition of solid agar slant medium is with
(2)配制A培养基、B培养基和C培养基：同实施例7；(2) preparation of A medium, B medium and C medium: with embodiment 7;
(3)对荧光假单胞菌进行交叉培养驯化：(3) Pseudomonas fluorescens is cross-cultured and domesticated:
将步骤(2)制备的A培养基、B培养基和C培养基分别放入培养皿中，使15mL/皿；Put the A medium, B medium and C medium prepared in step (2) into the culture dish respectively to make 15mL/dish;
将质量浓度为0.02％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含低浓度甲醛的B培养基和含低浓度甲醛的C培养基，每皿加1mL；Add 0.02% formaldehyde aqueous solution to Petri dish B and Petri dish C respectively after sterilizing the bacteria to make medium B with low concentration of formaldehyde and medium C with low concentration of formaldehyde, add 1 mL to each dish;
将质量浓度为0.04％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含中浓度甲醛的B培养基和含中浓度甲醛的C培养基，每皿加1mL；Sterilize 0.04% formaldehyde aqueous solution and add them to Petri dish B and Petri dish C respectively to make B medium containing medium-concentration formaldehyde and C medium containing medium-concentration formaldehyde, add 1 mL to each dish;
将质量浓度为0.08％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含高浓度甲醛的B培养基和含高浓度甲醛的C培养基，每皿加1mL；Add 0.08% formaldehyde aqueous solution to Petri dish B and Petri dish C respectively after sterilizing the bacteria to make medium B with high concentration of formaldehyde and medium C with high concentration of formaldehyde, add 1 mL to each dish;
将所述步骤(1)制备的纯化菌种接种到所述含低浓度甲醛的B培养基中28℃培养3天，转接到所述含低浓度甲醛的C培养基中28℃培养3天，再转接到所述含中浓度甲醛的B培养基中28℃培养3天，再转接到所述含中浓度甲醛的C培养基中28℃培养3天，再转接到所述含高浓度甲醛的B培养基中28℃培养3天，再转接到所述含高浓度甲醛的C培养基中28℃培养3天，再转接到所述A培养基中28℃培养2天，达到对数生长期，取菌落制成固体琼脂斜面保藏菌种，28℃培养2天后，得到已驯化的菌体，4℃冰箱内保存待用；Inoculate the purified strain prepared in the step (1) into the B medium containing low concentration of formaldehyde and cultivate it at 28°C for 3 days, then transfer it to the C medium containing low concentration of formaldehyde and cultivate it at 28°C for 3 days , then transferred to the B medium containing medium concentration formaldehyde for 3 days at 28°C, then transferred to the medium C medium containing formaldehyde for 3 days at 28°C, and then transferred to the medium containing formaldehyde Cultivate in medium B with high concentration of formaldehyde at 28°C for 3 days, then transfer to medium C containing high concentration of formaldehyde and culture at 28°C for 3 days, then transfer to medium A and culture at 28°C for 2 days , to reach the logarithmic growth phase, take the colony and make it into a solid agar slant to preserve the strain, cultivate it at 28°C for 2 days, and obtain the domesticated bacteria, and store it in the refrigerator at 4°C until use;
(4)用海藻酸钙包埋已驯化的菌体：(4) Embedding the domesticated bacteria with calcium alginate:
将步骤(3)制备的已驯化的菌体接种到装有500mL已灭菌的营养肉汤液体培养基的三角瓶中，28℃下在摇床上恒温培养2天，菌体密度达到1010个/mL，得菌悬液，取50mL菌悬液注入灭菌的离心管中，在4000rpm下分离30分钟，分离所得菌体细胞经灭菌生理盐水洗涤3次，加蒸馏水至50mL，使菌体细胞悬浮于蒸馏水中，得加水菌悬液；Inoculate the domesticated thalline prepared in step (3) into a conical flask equipped with 500 mL of sterilized nutrient broth liquid medium, and culture it on a shaker at a constant temperature at 28 ° C for 2 days, and the thalline density reaches 10 10 /mL, to obtain the bacterial suspension, take 50mL of the bacterial suspension and inject it into a sterilized centrifuge tube, separate it at 4000rpm for 30 minutes, wash the separated bacterial cells with sterilized normal saline for 3 times, add distilled water to 50mL, and make the bacterial cells The cells are suspended in distilled water, and the bacterial suspension in water is added;
按体积比为100∶0.8的比例将质量浓度为5％的海藻酸钠水溶液与所述加水菌悬液混匀后制成混合液体，用针形管将所述混合液体滴入质量浓度为10％的CaCl2水溶液中，静置8小时，得到2～5mm的颗粒，滤出颗粒后用生理盐水和蒸馏水洗净，制成一种生物吸附空气净化材料。According to the volume ratio of 100:0.8, the sodium alginate aqueous solution with a mass concentration of 5% is mixed with the water-added bacterial suspension to make a mixed liquid, and the mixed liquid is dripped into the mixed liquid with a mass concentration of 10 with a needle tube. % CaCl 2 aqueous solution, let it stand for 8 hours to obtain particles of 2-5mm, filter out the particles and wash them with physiological saline and distilled water to make a biological adsorption air purification material.
实施例11Example 11
一种微生物吸附降解的空气净化材料的制备方法，包括如下步骤：A preparation method of an air purification material degraded by microbial adsorption, comprising the steps of:
(1)对睾丸酮假单胞菌进行活化和纯化培养：(1) Pseudomonas testosterone is activated and purified:
取冷冻真空干燥管内睾丸酮假单胞菌Pseudomonas testosteroni(CICC20542)菌体，在无菌环境下注入110mL的营养肉汤液体培养基，在摇床35℃下连续培养2天活化菌种，利用平板划线分离法分离出单个菌落，在35℃恒温培养箱内连续培养2天，转移至固体琼脂斜面培养基培养1天，于4℃冰箱内保存待用，营养肉汤液体培养基的组成同实施例7；固体琼脂斜面培养基的组成同实施例7；Take Pseudomonas testosteroni (CICC20542) cells in the freeze vacuum drying tube, inject 110mL of nutrient broth liquid medium in a sterile environment, and continuously cultivate the activated strains for 2 days at 35°C on a shaker, and use a plate to draw A single colony was isolated by the line separation method, continuously cultured in a constant temperature incubator at 35°C for 2 days, transferred to a solid agar slant medium for 1 day, and stored in a refrigerator at 4°C until use. The composition of the nutrient broth liquid medium was the same as that of the implementation Example 7: The composition of solid agar slant medium is the same as in Example 7;
(2)配制A培养基、B培养基和C培养基：同实施例7；(2) preparation of A medium, B medium and C medium: with embodiment 7;
(3)对睾丸酮假单胞菌进行交叉培养驯化：(3) Pseudomonas testosterone is cross-cultured and domesticated:
将步骤(2)制备的A培养基、B培养基和C培养基分别放入培养皿中，使20mL/皿；Put the A medium, B medium and C medium prepared in step (2) into the culture dish respectively to make 20mL/dish;
将质量浓度为0.02％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含低浓度甲醛的B培养基和含低浓度甲醛的C培养基，每皿加1mL；Add 0.02% formaldehyde aqueous solution to Petri dish B and Petri dish C respectively after sterilizing the bacteria to make medium B with low concentration of formaldehyde and medium C with low concentration of formaldehyde, add 1 mL to each dish;
将质量浓度为0.05的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含中浓度甲醛的B培养基和含中浓度甲醛的C培养基，每皿加1mL；Sterilize formaldehyde aqueous solution with a mass concentration of 0.05 and add it to Petri dish B and Petri dish C respectively to make B medium containing medium-concentration formaldehyde and C medium containing medium-concentration formaldehyde, add 1 mL to each dish;
将质量浓度为0.10％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含高浓度甲醛的B培养基和含高浓度甲醛的C培养基，每皿加1mL；Sterilize 0.10% formaldehyde aqueous solution and add them to Petri dish B and Petri dish C respectively to make medium B with high concentration of formaldehyde and medium C with high concentration of formaldehyde, add 1 mL to each dish;
将所述步骤(1)制备的纯化菌种接种到所述含低浓度甲醛的B培养基中28℃培养3天，转接到所述含低浓度甲醛的C培养基中28℃培养3天，再转接到所述含中浓度甲醛的B培养基中28℃培养3天，再转接到所述含中浓度甲醛的C培养基中28℃培养3天，再转接到所述含高浓度甲醛的B培养基中28℃培养3天，再转接到所述含高浓度甲醛的C培养基中28℃培养3天，再转接到所述A培养基中28℃培养2天，达到对数生长期，取菌落制成固体琼脂斜面保藏菌种，28℃培养2天后，得到已驯化的菌体，4℃冰箱内保存待用；Inoculate the purified strain prepared in the step (1) into the B medium containing low concentration of formaldehyde and cultivate it at 28°C for 3 days, then transfer it to the C medium containing low concentration of formaldehyde and cultivate it at 28°C for 3 days , then transferred to the B medium containing medium concentration formaldehyde for 3 days at 28°C, then transferred to the medium C medium containing formaldehyde for 3 days at 28°C, and then transferred to the medium containing formaldehyde Cultivate in medium B with high concentration of formaldehyde at 28°C for 3 days, then transfer to medium C containing high concentration of formaldehyde and culture at 28°C for 3 days, then transfer to medium A and culture at 28°C for 2 days , to reach the logarithmic growth phase, take the colony and make it into a solid agar slant to preserve the strain, cultivate it at 28°C for 2 days, and obtain the domesticated bacteria, and store it in the refrigerator at 4°C until use;
(4)用海藻酸钙包埋已驯化的菌体：同实施例7。(4) Embedding the domesticated bacteria with calcium alginate: the same as in Example 7.
实施例12Example 12
一种微生物吸附降解的空气净化材料的制备方法，包括如下步骤：A preparation method of an air purification material degraded by microbial adsorption, comprising the steps of:
(1)对乙酸钙不动杆菌进行活化和纯化培养：(1) Activate and purify Acinetobacter calcoacetate:
取冷冻真空干燥管内乙酸钙不动杆菌Acinetobacter calcoaceticus：(CGMCC 1.2004)菌体，在无菌环境下注入100mL的营养肉汤液体培养基，在摇床30℃下连续培养1天活化菌种，利用平板划线分离法分离出单个菌落，在30℃恒温培养箱内连续培养1天，转移至固体琼脂斜面培养基培养1天，于4℃冰箱内保存待用，营养肉汤液体培养基的组成同实施例7；固体琼脂斜面培养基的组成同实施例8Take Acinetobacter calcoaceticus: (CGMCC 1.2004) cells in the freeze-drying tube, inject 100mL of nutrient broth liquid medium in a sterile environment, and continuously cultivate for 1 day on a shaker at 30°C to activate the strains. A single colony was isolated by streaking on a flat plate, continuously cultured in a constant temperature incubator at 30°C for 1 day, transferred to a solid agar slant medium for 1 day, and stored in a refrigerator at 4°C until use. The composition of the nutrient broth liquid medium With embodiment 7; The composition of solid agar slant medium is with
(2)配制A培养基、B培养基和C培养基：(同实施例7)(2) preparation of A medium, B medium and C medium: (with embodiment 7)
(3)对乙酸钙不动杆菌进行交叉培养驯化：(3) Acinetobacter calcoacetate is cross-cultured and domesticated:
将步骤(2)制备的A培养基、B培养基和C培养基分别放入培养皿中，使25mL/皿；Put the A medium, B medium and C medium prepared in step (2) into the culture dish respectively to make 25mL/dish;
将质量浓度为0.03％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含低浓度甲醛的B培养基和含低浓度甲醛的C培养基，每皿加1mL；Add 0.03% formaldehyde aqueous solution to Petri dish B and Petri dish C respectively after sterilizing the bacteria to make medium B with low concentration of formaldehyde and medium C with low concentration of formaldehyde, add 1 mL to each dish;
将质量浓度为0.06％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含中浓度甲醛的B培养基和含中浓度甲醛的C培养基，每皿加1mL；Add 0.06% formaldehyde aqueous solution to Petri dish B and Petri dish C respectively after sterilizing the bacteria to make B medium containing medium-concentration formaldehyde and C medium containing medium-concentration formaldehyde, add 1 mL to each dish;
将质量浓度为0.12％的甲醛水溶液除菌后分别加入到B培养皿和C培养皿中制成含高浓度甲醛的B培养基和含高浓度甲醛的C培养基，每皿加1mL；Sterilize 0.12% formaldehyde aqueous solution and add them to Petri dish B and Petri dish C respectively to make medium B with high concentration of formaldehyde and medium C with high concentration of formaldehyde, add 1 mL to each dish;
将所述步骤(1)制备的纯化菌种接种到所述含低浓度甲醛的B培养基中28℃培养3天，转接到所述含低浓度甲醛的C培养基中28℃培养3天，再转接到所述含中浓度甲醛的B培养基中28℃培养3天，再转接到所述含中浓度甲醛的C培养基中28℃培养3天，再转接到所述含高浓度甲醛的B培养基中28℃培养3天，再转接到所述含高浓度甲醛的C培养基中28℃培养3天，再转接到所述A培养基中28℃培养2天，达到对数生长期，取菌落制成固体琼脂斜面保藏菌种，28℃培养2天后，得到已驯化的菌体，4℃冰箱内保存待用；Inoculate the purified strain prepared in the step (1) into the B medium containing low concentration of formaldehyde and cultivate it at 28°C for 3 days, then transfer it to the C medium containing low concentration of formaldehyde and cultivate it at 28°C for 3 days , then transferred to the B medium containing medium concentration formaldehyde for 3 days at 28°C, then transferred to the medium C medium containing formaldehyde for 3 days at 28°C, and then transferred to the medium containing formaldehyde Cultivate in medium B with high concentration of formaldehyde at 28°C for 3 days, then transfer to medium C containing high concentration of formaldehyde and culture at 28°C for 3 days, then transfer to medium A and culture at 28°C for 2 days , to reach the logarithmic growth phase, take the colony and make it into a solid agar slant to preserve the strain, cultivate it at 28°C for 2 days, and obtain the domesticated bacteria, and store it in the refrigerator at 4°C until use;
(4)用海藻酸钙包埋已驯化的菌体：同实施例7。(4) Embedding the domesticated bacteria with calcium alginate: the same as in Example 7.
实施例13Example 13
净化效果：取实施例7-12制备的微生物吸附降解的空气净化材料装载入底部带网孔的容器内，并可在容器底部放置粒径为18-25mm的陶粒，将容器置于空调系统加湿段后，可使加湿后的气流强制通过容器，从而制成气体强制流动容器。当加湿后的空气经底部网孔均匀流过装有微生物吸附降解的空气净化材料的气体强制流动容器时，污染物将被生物颗粒吸附并降解，其净化效果如下所述：Purification effect: take the air purification material prepared by microbial adsorption and degradation prepared in Examples 7-12 and load it into a container with a mesh at the bottom, and place ceramsite with a particle size of 18-25mm at the bottom of the container, and place the container in an air conditioner After the humidification section of the system, the humidified airflow can be forced through the container to make a gas forced flow container. When the humidified air evenly flows through the gas forced flow container equipped with microbial adsorption and degradation air purification materials through the bottom mesh, the pollutants will be adsorbed and degraded by biological particles, and the purification effect is as follows:
在一个体积0.8×0.8×1设有循环送排风装置的实验舱中，舱内甲醛和苯系物的初始浓度分别为0.7mg/m3和0.8g/m3。当未添加微生物吸附降解的空气净化材料时，实验舱内污染物的浓度衰减缓慢，在12小时内仍未降至0.1mg/m3和0.11mg/m3(0.1mg/m3和0.11mg/m3分别为国家标准对室内甲醛和苯系物释放值规定的上限浓度)。而将填充有实施例7-12制备的微生物吸附降解的空气净化材料的气体强制流动容器置于实验舱内之后，舱内污染物的浓度衰减非常迅速，在9小时内舱内甲醛浓度既可降低至0.1mg/m3以下，同时10h后苯系物浓度可降至0.11mg/m3以下。对于实际建筑中甲醛和苯系物浓度超标100％的房间(即房间内甲醛和苯系物浓度分别为0.2mg/m3和0.22～0.25mg/m3)，利用微生物吸附降解的空气净化材料去除居室内甲醛和苯系物至健康浓度范围仅需4-5h。图4表征了实验舱内甲醛浓度衰减的情况。In an experimental cabin with a volume of 0.8×0.8×1 equipped with a circulating air supply and exhaust device, the initial concentrations of formaldehyde and benzene series in the cabin were 0.7mg/m 3 and 0.8g/m 3 respectively. When no air purification material for microbial adsorption and degradation was added, the concentration of pollutants in the experimental cabin decayed slowly, and did not drop to 0.1mg/ m3 and 0.11mg/ m3 (0.1mg/ m3 and 0.11mg/m3) within 12 hours /m 3 are the upper limit concentrations stipulated by national standards for indoor formaldehyde and benzene series release values). After the gas forced flow container filled with the air-purifying material for microbial adsorption degradation prepared in Examples 7-12 is placed in the experimental cabin, the concentration of the pollutants in the cabin decays very rapidly, and the formaldehyde concentration in the cabin can be reached within 9 hours. Reduced to below 0.1mg/m 3 , and the concentration of benzene series can be reduced to below 0.11mg/m 3 after 10 hours. For rooms where the concentration of formaldehyde and benzene series exceeds the standard by 100% in the actual building (that is, the concentration of formaldehyde and benzene series in the room is 0.2mg/m 3 and 0.22-0.25mg/m 3 respectively), air purification materials that use microbial adsorption and degradation It only takes 4-5 hours to remove formaldehyde and benzene series in the living room to a healthy concentration range. Figure 4 characterizes the attenuation of formaldehyde concentration in the experimental cabin.
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