WO2022012698A1 - 一种烟曲霉菌株及其在降解聚乙烯醇中的应用 - Google Patents

一种烟曲霉菌株及其在降解聚乙烯醇中的应用 Download PDF

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WO2022012698A1
WO2022012698A1 PCT/CN2021/114984 CN2021114984W WO2022012698A1 WO 2022012698 A1 WO2022012698 A1 WO 2022012698A1 CN 2021114984 W CN2021114984 W CN 2021114984W WO 2022012698 A1 WO2022012698 A1 WO 2022012698A1
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aspergillus fumigatus
pva
strain
polyvinyl alcohol
gpva1
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French (fr)
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龙碧波
陈骏佳
谢东
高旭华
杨翀
李圆
王珂
赵阳
刘海露
黄瑶珠
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广东省科学院生物工程研究所
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    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • C12R2001/66—Aspergillus
    • C12R2001/68—Aspergillus fumigatus
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    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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    • 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
    • C12N1/14—Fungi; Culture media therefor
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    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00—Nature of the contaminant
    • C02F2101/30—Organic compounds
    • C02F2101/34—Organic compounds containing oxygen

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  • the invention relates to the technical field of microorganisms, in particular to an Aspergillus fumigatus strain and its application in degrading polyvinyl alcohol.
  • Polyvinyl alcohol is a chemical raw material made from petroleum cracking products. It has good water solubility, adhesion, toughness and abrasion resistance of serosa films, and strong organic solvent resistance. It is widely used in textiles, paper, etc. Product manufacturing, food packaging and medical equipment industries. PVA is the water-soluble polymer with the largest production volume, and its production and consumption are increasing year by year, resulting in a large amount of PVA being discharged into the environment, causing environmental pollution and affecting the ecological balance. PVA has a large surface activity and can form a large number of foams in water, resulting in oxygen-enriched water and increased viscosity, thereby inhibiting or even destroying the respiration of aquatic organisms. Therefore, the degradation of PVA is of great significance to protect the environment.
  • PVA is not easily degraded under natural conditions and needs to be treated by physicochemical or biological methods.
  • the physical and chemical methods of treating PVA include ultrafiltration membrane separation method, radiation method, salt precipitation flocculation method, advanced oxidation method, separate ozone oxidation method, etc., which have problems such as high cost and secondary pollution.
  • biodegrading PVA is more economical and efficient.
  • PVA has poor biodegradability and is difficult to be degraded and utilized by ordinary microorganisms, but it can be degraded by specific functional microorganisms or enzymes. Studies have shown that PVA-degrading bacteria are not widely distributed in nature, and generally only exist in PVA-contaminated environments, such as PVA textile wastewater and papermaking wastewater.
  • the purpose of the present invention is to provide a strain of Aspergillus fumigatus that degrades polyvinyl alcohol, the strain can grow in solid and liquid inorganic salt medium with polyvinyl alcohol (PVA) as the sole carbon source and degrade PVA, and has the advantages of being suitable for environmental application Potential for microbial treatment of PVA contamination.
  • PVA polyvinyl alcohol
  • a strain of Aspergillus fumigatus named Aspergillus fumigatus GPVA1
  • the preservation number is CCTCC NO: M 2020213
  • the preservation date is June 17, 2020.
  • Aspergillus fumigatus (Aspergillus fumigatus) GPVA1 was preserved in the China Typical Culture Collection and Management Center on June 17, 2020, and the preservation number was CCTCC NO: M 2020213, named as Aspergillus fumigatus (Aspergillus fumigatus) GPVA1.
  • the 28S rRNA gene sequence of the above-mentioned Aspergillus fumigatus GPVA1 is as shown in SEQ ID NO:1, the rDNA-ITS gene sequence is as shown in SEQ ID NO:2, and the 18S rRNA gene sequence is as shown in SEQ ID NO:3.
  • the biological characteristics of the above-mentioned Aspergillus fumigatus GPVA1 are: the colony is raised to the surface of the medium, loose, dry, fluffy, with white edges, smoke green in the middle, and a large number of spores are produced, the spores are spherical, and the apical sac In the shape of an inverted flask, no pigment is produced.
  • the present invention provides an inoculum containing the Aspergillus fumigatus strain.
  • the invention provides the application of the Aspergillus fumigatus strain in degrading polyvinyl alcohol.
  • the invention provides the application of the bacterial agent in degrading polyvinyl alcohol.
  • Aspergillus fumigatus GPVA1 is placed in a solid inorganic salt medium and a liquid inorganic salt medium with polyvinyl alcohol (PVA1799) as the sole carbon source for growth and cultivation, and it is found that the Aspergillus fumigatus GPVA1 can be
  • PVA1799 polyvinyl alcohol
  • the PVA concentration in the liquid medium was reduced by 2.72 g/L within 24 hours.
  • the components of the solid inorganic salt medium were K 2 HPO 4 0.7 g, KH 2 PO 4 0.7 g, MgSO 4 ⁇ 7H 2 O 0.7 g, NaCl 0.005 g, FeSO 4 ⁇ 7H 2 O 0.002 g, ZnSO 4 ⁇ 7H 2 O 0.002 g, MnSO 4 ⁇ H 2 O 0.001 g, PVA 10 g, agar 18 g, distilled water 1000 mL, and sterilized.
  • the composition of liquid inorganic salt medium is K 2 HPO 4 0.7g, KH 2 PO 4 0.7g, MgSO 4 ⁇ 7H 2 O 0.7g, NaCl 0.005g, FeSO 4 ⁇ 7H 2 O 0.002g, ZnSO 4 ⁇ 7H 2 O 0.002 g, MnSO 4 ⁇ H 2 O 0.001 g, 10 g of PVA, 1000 mL of distilled water, and sterilized.
  • the application includes: inoculating a strain of Aspergillus fumigatus into wastewater containing polyvinyl alcohol for treatment.
  • the present invention has the following beneficial effects:
  • the present invention provides the Aspergillus fumigatus strain capable of degrading polyvinyl alcohol (PVA) for the first time, and the strain can grow and degrade PVA in a solid inorganic salt medium or a liquid inorganic salt medium with polyvinyl alcohol as the sole carbon source , the concentration of PVA in the liquid medium can be reduced by 2.72g/L within 24 hours, which has the potential to be applied to the treatment of PVA-contaminated microorganisms in the environment.
  • PVA polyvinyl alcohol
  • the Aspergillus fumigatus strain provided by the present invention not only has the ability to degrade polyvinyl alcohol, but also can utilize at least 80 kinds of carbon sources, and has strong carbon source utilization ability.
  • Fig. 1 is the observation picture of the plate colony of Aspergillus fumigatus GPVA1 in Example 2.
  • Figure 2 is an optical microscope (200X) picture of Aspergillus fumigatus GPVA1 in Example 2.
  • Figure 3 is a picture of the degradation of PVA by strain Aspergillus fumigatus GPVA1 in a plate medium.
  • the medium includes the following components: K 2 HPO 4 0.7 g, KH 2 PO 4 0.7 g, MgSO 4 7H 2 O 0.7 g, NaCl 0.005 g, FeSO 4 7H 2 O 0.002 g, ZnSO 4 7H 2 O 0.002 g, MnSO 4 ⁇ H 2 O 0.001 g, PVA 10 g, agar 18 g, and distilled water 1000 mL. Sterilize the culture medium at 121°C for 20 minutes, then cool it down to a point where it is not hot to the touch, and then pour the plate into a sterile operating table.
  • the plastic bag with degradation characteristics was washed in sterile water to obtain a cleaning solution containing microorganisms, which was spread on the inorganic salt medium plate with PVA as the sole carbon source, and cultivated at 30°C to obtain PVA-degrading functional bacteria.
  • strain GPVA1 The detection and identification of strain GPVA1 was entrusted to the China Center for Type Culture Collection. The results are as follows:
  • the strain grows well on PDA medium. After culturing at 28°C for 3 days, the colony is raised to the surface of the medium, loose, dry, fluffy, with white edges and smoke green in the middle ( Figure 1). A large number of spores are produced, and the spores are Spherical, the apical capsule is inverted flask-shaped, and no pigment is produced ( Figure 2).
  • the biolog FF plate was used to detect the utilization of 95 carbon sources by GPVA1, and the results showed that it could utilize 80 of them.
  • the total DNA of GPVA1 was extracted, and the target gene rDNA-ITS (primer ITS1: 5'-TCCGTAGGTGAACCTGCGG-3'; ITS4: 5'-TCCTCCGCTTATTGATATGC-3'), 28S rRNA gene (primer NL1: 5'-GCATATCAATAAGCGGAGGAAAAG- 3'; NL4: 5'-GGTCCGTGTTTCAAGACGG-3'), 18S rRNA gene (primers NS1: 5'-GTAGTCATATGCTTGTCTC-3'; NS8: 5'-TCCGCAGGTTCACCTACGGA-3').
  • the sequence of the amplified product is as follows:
  • the rDNA-ITS gene, 28S rRNA gene, and 18S rRNA gene were compared by blast in the NCBI database, and the results showed that the bacterial strain and some strains of Aspergillus fumigatus had a common origin, so the bacterial strain was identified as: Aspergillus fumigatus, Named Aspergillus fumigatus GPVA1.
  • the medium included the following components: K 2 HPO 4 0.7 g, KH 2 PO 4 0.7 g, MgSO 4 7H 2 O 0.7 g, NaCl 0.005 g, FeSO 4 7H 2 O 0.002 g, ZnSO 4 7H 2 O 0.002 g, MnSO 4 ⁇ H 2 O 0.001 g, PVA 10 g, agar 18 g, and distilled water 1000 mL. Sterilize the culture medium at 121°C for 20 minutes, then cool it down to a point where it is not hot to the touch, and then pour the plate into a sterile operating table.
  • a liquid inorganic salt medium containing PVA as the sole carbon source was prepared.
  • the medium included the following components: K 2 HPO 4 0.7 g, KH 2 PO 4 0.7 g, MgSO 4 7H 2 O 0.7 g, NaCl 0.005 g, FeSO 4 7H 2 O 0.002 g, ZnSO 4 7H 2 O 0.002 g , MnSO 4 ⁇ H 2 O 0.001 g, PVA 10 g, and distilled water 1000 mL.
  • the medium was sterilized at 121 °C for 20 min, and then inoculated with GPVA1 after cooling. One day later, PVA in the medium was detected by boric acid-iodine spectrophotometry at a wavelength of 645 nm.

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Abstract

一种烟曲霉菌株及其在降解聚乙烯醇中的应用，其特征在于，命名为烟曲霉（ Aspergillus fumigillus）GPVA1，保藏编号为CCTCC NO：M2020213，保藏日期为2020年6月17日。该菌株能在以聚乙烯醇为唯一碳源的固体无机盐培养基或液体无机盐培养基中生长并降解聚乙烯醇。

Description

一种烟曲霉菌株及其在降解聚乙烯醇中的应用 技术领域
本发明涉及微生物技术领域，尤其涉及一种烟曲霉菌株及其在降解聚乙烯醇中的应用。
背景技术
聚乙烯醇(PVA)是以石油裂解产物为原料制作的化工原料，具有良好的水溶性、黏附性、浆膜强韧性和耐磨性、强有机溶剂耐受性，被广泛应用于纺织、纸制品制造、食品包装和医疗器械等行业。PVA是生产量最大的水溶性聚合物，且其生产和消费量逐年递增，导致大量PVA排入环境，造成了环境污染，影响生态平衡。PVA具有较大的表面活性，能在水中形成大量泡沫，导致水体富氧，黏度增大，从而抑制甚至破坏水生生物的呼吸活动。因此，PVA的降解处理对保护环境具有重要的意义。
PVA在自然条件下不易被降解，需要通过物理化学或生物的方法处理。处理PVA的物理化学方法包括超滤膜分离法、辐射法、盐析絮凝法、高级氧化法、单独臭氧氧化法等，其存在成本高和二次污染等问题。相比之下，生物降解PVA更加经济和高效。
PVA可生化性差，难以被普通微生物降解利用，但是可以被特定的功能微生物或酶降解。研究表明，PVA降解菌在自然界中的分布并不广泛，一般仅存在于被PVA污染的环境中，如PVA纺织废水和造纸废水。自1936年Nord首次报道了镰刀菌可以降解PVA以来，已有部分PVA降解菌得到分离纯化，如Pseudomonas O-3、Pseudomonas vesiculars PD、Pseudomonas putida、Bacillus magaterium、Acinetiobactor sp.、Xanthamonas sp.等，但是，绝大部分菌株因为效率低等原因，仍无法满足实际应用的需要。筛选分离高效菌株是研发PVA微生物降解技术的关键。
因此，有必要筛选能满足实际应用要求的新微生物资源，为PVA污染微生物修复提供支撑。
发明内容
本发明提供了的目的在于提供一株降解聚乙烯醇的烟曲霉，该菌株能在以聚乙烯醇(PVA)为唯一碳源的固体和液体无机盐培养基生长并降解PVA，具有应用于环境中PVA污染微生物处理的潜力。
具体技术方案如下：
一种烟曲霉菌株，命名为烟曲霉(Aspergillus fumigatus)GPVA1，保藏编号为CCTCC NO:M 2020213，保藏日期为2020年6月17日。
上述烟曲霉(Aspergillus fumigatus)GPVA1于2020年6月17日保藏于中国典型培 养物菌种保藏管理中心，保藏编号为CCTCC NO:M 2020213，命名为烟曲霉(Aspergillus fumigatus)GPVA1。
上述烟曲霉(Aspergillus fumigatus)GPVA1的28S rRNA基因序列如SEQ ID NO:1所示，rDNA-ITS基因序列如SEQ ID NO:2所示，18S rRNA基因序列如SEQ ID NO:3所示。
上述烟曲霉(Aspergillus fumigatus)GPVA1的生物学特征为：菌落向培养基表面隆起，疏松、干燥、呈绒毛状、边缘为白色，中间为烟绿色，有大量孢子产生，孢子为球型，顶囊为倒烧瓶状，无色素产生。
本发明提供了一种包含所述的烟曲霉菌株的菌剂。
本发明提供了所述的烟曲霉菌株在降解聚乙烯醇中的应用。
本发明提供了所述的菌剂在降解聚乙烯醇中的应用。
本发明将烟曲霉(Aspergillus fumigatus)GPVA1置于以聚乙烯醇(PVA1799)为唯一碳源的固体无机盐培养基和液体无机盐培养基进行生长和培养，发现该烟曲霉(Aspergillus fumigatus)GPVA1能够24小时内将液体培养基中PVA浓度减少2.72g/L。
其中，固体无机盐培养基的成分为K 2HPO 4 0.7g、KH 2PO 4 0.7g、MgSO 4·7H 2O 0.7g、NaCl 0.005g、FeSO 4·7H 2O 0.002g、ZnSO 4·7H 2O 0.002g、MnSO 4·H 2O 0.001g、PVA 10g、琼脂18g、蒸馏水1000m L，灭菌。
液体无机盐培养基的成分为K 2HPO 4 0.7g、KH 2PO 4 0.7g、MgSO 4·7H 2O 0.7g、NaCl 0.005g、FeSO 4·7H 2O 0.002g、ZnSO 4·7H 2O 0.002g、MnSO 4·H 2O 0.001g、PVA 10g、蒸馏水1000m L，灭菌。
进一步地，所述的应用，包括：将烟曲霉菌株接种至含聚乙烯醇的废水中进行处理。
与现有技术相比，本发明具有以下有益效果：
(1)本发明首次提供了能够降解聚乙烯醇(PVA)的烟曲霉菌株，该菌株能在以聚乙烯醇为唯一碳源的固体无机盐培养基或液体无机盐培养基中生长并降解PVA，在24小时内能将液体培养基中PVA的浓度减少2.72g/L，具有应用于环境中PVA污染微生物处理的潜力。
(2)本发明提供的烟曲霉菌株不仅具有聚乙烯醇降解能力，而且能够利用至少80种碳源，碳源利用能力强。
附图说明
图1为实施例2中烟曲霉(Aspergillus fumigatus)GPVA1的平板菌落观察图片。
图2为实施例2中烟曲霉(Aspergillus fumigatus)GPVA1的光学显微镜镜检(200X)图片。
图3为菌株烟曲霉(Aspergillus fumigatus)GPVA1在平板培养基中降解PVA的图片。
具体实施方式
下面结合具体实施例对本发明作进一步描述，以下列举的仅是本发明的具体实施例，但本发明的保护范围不仅限于此。
实施例1一株PVA降解功能菌株的筛选
1.1制作含PVA为唯一碳源的无机盐培养基平板：
培养基包括以下成分：K 2HPO 4 0.7g、KH 2PO 4 0.7g、Mg SO 4·7H 2O 0.7g、NaCl0.005g、FeSO 4·7H 2O 0.002g、ZnSO 4·7H 2O 0.002g、MnSO 4·H 2O 0.001g、PVA 10g、琼脂18g、蒸馏水1000m L。将培养基在121℃灭菌20min，然后冷却至不烫手后于无菌操作台内倒平板。
1.2培养PVA降解功能菌：
将具有降解特征的塑料袋在无菌水中清洗，得到含有微生物的清洗液，并将其涂在以PVA为唯一碳源的无机盐培养基平板上，于30℃培养，获得PVA降解功能菌。
1.3分离PVA降解功能菌株单菌落：
用无菌水冲洗长有PVA降解功能菌的平板，配制含菌悬浮液。按10 -1、10 -2、10 -3、10 -4、10 -5稀释含菌悬浮液，各吸取0.1ml涂在以PVA为唯一碳源的无机盐培养基平板上，30℃培养箱中倒置培养，获得长势良好、可降解PVA功能菌株单菌落。
1.4纯化PVA降解功能菌：
在以PVA为唯一碳源的无机盐培养基平板上挑取长势良好的单菌落接种到新的平板上，在30℃培养箱中倒置培养，待其长出孢子，用无菌水冲洗平板，配制浓度约为10 6个/mL的孢子悬浮液，按10 -1、10 -2、10 -3、10 -4、10 -5稀释孢子液，各吸取0.1ml涂在以PVA为唯一碳源的无机盐培养基平板上，30℃培养箱中倒置培养，确保得到纯净的PVA降解功能菌株，命名为GPVA1。
实施例2菌株GPVA1的形态、碳源利用、分子生物学鉴定
菌株GPVA1的检测鉴定委托中国典型培养物保藏中心实施，结果如下：
2.1形态学特征
菌株在PDA培养基上生长良好，28℃培养3天后，菌落向培养基表面隆起，疏松、干燥、呈绒毛状、边缘为白色，中间为烟绿色(图1)，有大量孢子产生，孢子为球型，顶囊为倒烧瓶状，无色素产生(图2)。
2.2碳源利用特征
用biolog FF板检测GPVA1对95种碳源的利用情况，结果表明其能利用其中的80种。
表1 GPVA1菌株的碳源利用检测
  检测项目 结果   检测项目 结果
A1 水 - B7 D-半乳糖 +
A2 吐温80 + B8 D-半乳糖醛酸 +
A3 N-乙酰-半乳糖胺 - B9 龙胆二糖 +
A4 N-乙酰-葡萄糖胺 + B10 D-葡萄糖酸 +
A5 N-乙酰-甘露糖胺 - B11 D-葡萄糖胺 +
A6 核糖醇 - B12 α-D-葡萄糖 +
A7 苦杏仁甙 + C1 1-磷酸-葡萄糖 +
A8 D-阿拉伯糖 - C2 葡糖醛酰胺 +
A9 L-阿拉伯糖 + C3 D-葡萄糖醛酸 +
A10 D-阿拉伯糖醇 + C4 甘油 +
A11 熊果苷 + C5 糖原 +
A12 D-纤维二糖 + C6 M-肌醇 +
B1 α-环糊精 + C7 2-酮基-D-葡萄糖酸 +
B2 β-环糊精 + C8 α-D-乳糖 +
B3 葡聚糖 + C9 乳果糖 -
B4 I-赤丝藻醇 + C10 麦芽糖醇 +
B5 D-果糖 + C11 麦芽糖 +
B6 L-岩藻糖 + C12 麦芽三糖 +
D1 D-甘露醇 + F7 P-邻羟基苯乙酸 +
D2 D-甘露糖 + F8 α-酮戊二酸 +
D3 D-松三糖 + F9 D-乳酸甲酯 +
D4 D-蜜二糖 + F10 L-乳酸 +
D5 α-甲基-D-半乳糖苷 + F11 D-苹果酸 +
D6 β-甲基-D-半乳糖苷 + F12 L-苹果酸 +
D7 α-甲基-D-葡萄糖苷 + G1 D-葡糖二酸 -
D8 β-甲基-D-葡萄糖苷 + G2 癸二酸 +
D9 异麦芽酮糖 + G3 琥珀酰胺酸 -
D10 D-阿洛酮糖 + G4 琥珀酸 -
D11 D-棉子糖 + G5 琥珀酸单甲酯 +
D12 L-鼠李糖 + G6 N-乙酰-L-谷氨酸 -
E1 D-核糖 + G7 丙酸胺 +
E2 水杨苷 + G8 L-丙氨酸 +
E3 景天庚酫聚糖 + G9 L-丙氨酰甘氨酸 +
E4 D-山梨醇 + G10 L-天冬酰胺 +
E5 L-山梨糖 + G11 L-天(门)冬氨酸 +
E6 水苏糖 + G12 L-谷氨基酸 +
E7 蔗糖 + H1 L-甘氨酰谷氨酸 -
E8 D-塔格糖 - H2 L-鸟氨酸 +
E9 D-海藻糖 + H3 L-苯基丙氨酸 +
E10 松二糖 + H4 L-脯氨酸 +
E11 木糖醇 + H5 L-焦谷氨酸 +
E12 D-木糖 + H6 L-丝氨酸 +
F1 γ-丁胺酸 + H7 L-苏氨酸 +
F2 溴代丁二酸 - H8 2-乙醇胺 -
F3 反丁烯二酸 - H9 腐胺 +
F4 β-羟基异丁酸 - H10 腺苷 +
F5 γ-羟基异丁酸 + H11 尿苷 +
F6 L-鼠李糖 + H12 腺嘌呤核苷酸 +
+：阳性反应；-：阴性反应；
2.3分子生物学特征
提取GPVA1的总DNA，用PCR扩增目的基因rDNA-ITS(引物ITS1:5‘-TCCGTAGGTGAACCTGCGG-3’；ITS4:5‘-TCCTCCGCTTATTGATATGC-3’)、28S  rRNA基因(引物NL1:5’-GCATATCAATAAGCGGAGGAAAAG-3’；NL4:5’-GGTCCGTGTTTCAAGACGG-3’)、18S rRNA基因(引物NS1:5‘-GTAGTCATATGCTTGTCTC-3’；NS8:5‘-TCCGCAGGTTCACCTACGGA-3’)。扩增产物序列如下：
1)28S rRNA的基因序列如SEQ ID NO:1所示；
2)rDNA-ITS的基因序列如SEQ ID NO:2所示；
3)18SrRNA的基因序列如SEQ ID NO:3所示。
将rDNA-ITS基因、28S rRNA基因、18S rRNA基因在NCBI数据库中作blast比对，结果显示该菌株与Aspergillus fumigatus某些菌株具有共同的起源，故将菌株鉴定为:烟曲霉(Aspergillus fumigatus)，命名为Aspergillus fumigatus GPVA1。
该菌株已于2020年6月17日保存于中国典型培养物菌种保藏管理中心，保存号为CCTCC M 2020213，保藏地址为中国，武汉，武汉大学。
实施例3菌株GPVA1在平板培养基中降解PVA
制作含PVA为唯一碳源的无机盐培养基平板。培养基包括以下成分：K 2HPO 4 0.7g、KH 2PO 4 0.7g、Mg SO 4·7H 2O 0.7g、NaCl 0.005g、FeSO 4·7H 2O 0.002g、ZnSO 4·7H 2O 0.002g、MnSO 4·H 2O 0.001g、PVA 10g、琼脂18g、蒸馏水1000m L。将培养基在121℃灭菌20min，然后冷却至不烫手后于无菌操作台内倒平板。在平板上接种GPVA1后让其在30℃培养。菌落铺满平板后取小块有菌丝的培养基小块，转接到新的培养基平板上，于30℃培养2天。用喷壶往培养基中喷硼酸和碘化钾-碘溶液，让其与培养基中的PVA生成蓝绿色络合物，以证实其中PVA的存在。
结果显示(图3)，在GPVA1菌斑的周围形成了一个透明圈，与周围的蓝绿色(图中体现为灰色)形成鲜明的对比，表明菌株GPVA1降解了平板培养基中的PVA。
实施例4 GPVA1对PVA的降解
制作含PVA为唯一碳源的液体无机盐培养基。培养基包括以下成分：K 2HPO 4 0.7g、KH 2PO 4 0.7g、MgSO 4·7H 2O 0.7g、NaCl 0.005g、FeSO 4·7H 2O 0.002g、ZnSO 4·7H 2O 0.002g、MnSO 4·H 2O 0.001g、PVA 10g、蒸馏水1000m L。将培养基在121℃灭菌20min，冷却后接种GPVA1，1天后用硼酸-碘分光光度法在645nm波长检测培养基中的PVA。
结果显示，GPVA1在24小时降解了PVA2.72g/L。
Figure PCTCN2021114984-appb-000001
Figure PCTCN2021114984-appb-000002
Figure PCTCN2021114984-appb-000003
Figure PCTCN2021114984-appb-000004
Figure PCTCN2021114984-appb-000005

Claims (7)

  1. 一种烟曲霉菌株，其特征在于，命名为烟曲霉(Aspergillus fumigatus)GPVA1，保藏编号为CCTCC NO:M 2020213，保藏日期为2020年6月17日。
  2. 如权利要求1所述的烟曲霉菌株，其特征在于，所述烟曲霉(Aspergillus fumigatus)GPVA1的28S rRNA基因序列如SEQ ID NO:1所示，rDNA-ITS基因序列如SEQ ID NO:2所示，18S rRNA基因序列如SEQ ID NO:3所示。
  3. 一种包含权利要求1所述的烟曲霉菌株的菌剂。
  4. 如权利要求1所述的烟曲霉菌株在降解聚乙烯醇中的应用。
  5. 如权利要求4所述的应用，其他特征在于，将烟曲霉菌株接种至含聚乙烯醇的废水中进行处理。
  6. 如权利要求3所述的菌剂在降解聚乙烯醇中的应用。
  7. 如权利要求6所述的应用，其他特征在于，将烟曲霉菌株接种至含聚乙烯醇的废水中进行处理。
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