Screening and Identification of Aflatoxin B1 Degradation Strains and Preliminary Exploration of Effective Degradation Components
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摘要: 黄曲霉毒素B1（AFB1）是具有强毒性、强致癌性的一种真菌毒素。为了筛选出AFB1降解菌，将实验室保藏的真菌菌株与AFB1共培养，选取降解率最高的菌株，经形态学和ITS rDNA分析确定其种属，分离菌株不同组分（菌悬液、菌体、孢子液、发酵液）后探究其降解AFB1的有效组分，并将有效组分菌体破碎后作进一步探究。结果表明：培养72 h后的郝克氏青霉MD1菌悬液和未破碎菌体对AFB1降解率分别为98.15%和28.20%，菌体破碎后对AFB1的降解效果更好，培养24 h后对AFB1的降解率为55.40%。因此，郝克氏青霉MD1的菌悬液和菌体内的活性组分能有效降解AFB1。本研究筛选出的郝克氏青霉MD1扩大了AFB1降解菌的菌种库，为AFB1的生物降解提供了一定的参考价值。Abstract: Aflatoxin B1 (AFB1) is a type of fungaltoxin with strong toxicity and strong carcinogenicity. To screen degrading bacteria of AFB1, co-culture of fungi strains which were stored in laboratory and AFB1 was carried out and the strains with the highest degradation rate were chosen. Species of the chosen strains were determined through morphology and ITS rDNA analysis. After separation of different components (bacterial suspension, thalli, spore suspension and fermentation liquor) in strains, the effective components for AFB1 degradation were explored, and the effective components thalli was broken for further investigation. Results showed that the AFB1 degrading rates by Penicillium herquei MD1 bacterial suspension cultured by 72 h and the unbroken thalli were 98.15% and 28.20%, respectively. The degradation of AFB1 was better after the fragmentation of the thalli, and the degradation rate of AFB1 was 55.40% after 24 h of incubation. Therefore, the active components in MD1 bacterial suspension and thalli can degrade AFB1 effectively. The screened MD1 expands the degrading bacterial library of AFB1 and provide some references to biological degradation of AFB1.
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Key words:
- aflatoxin B1 /
- Penicillium herquei /
- biodegradation /
- bioactive ingredients /
- identification
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图 1 菌株MD1的菌落形态（A）和400×显微镜（B）、1800×扫描电镜（C）下的特征
Figure 1. MD1 colony morphology (A) and characteristics under 400 times microscope (B) and 1800 times scanning electron microscope (C)
图 2 菌株MD1的ITS rDNA基因PCR产物电泳图谱
注：M：DNA Maker D5000；1：菌株MD1的ITS rDNA基因。
Figure 2. Electrophoretic profile of ITS rDNA gene from srain MD1 by PCR
图 4 菌悬液（A）、菌体（B）、孢子液（C）、发酵液（D）对AFB1的降解效果
注：不同小写字母表示具有显著性差异，P<0.05；图5同。
Figure 4. Degradation of AFB1 by fungal suspension (A), mycelium (B), spores (C) and culture filtrate (D)
表 1 流动相梯度洗脱程序
Table 1. Gradient elution program of mobile phase
时间（min） 流速（mL/min） A（%） B（%） 0.0~3.0 0.4 90 10 3.0~3.5 0.4 40 60 3.5~4.6 0.4 10 90 4.6~6.0 0.4 90 10 
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表 2 AFB1的保留时间、监测离子及质谱采集参数
Table 2. Retention time, monitoring ion and mass spectrum acquisition parameters of AFB1
目标物 保留时间（min） 监测离子对 锥孔电压（V） 碰撞能量（V） 驻留时间（s） AFB1 3.37 313.2>241.1 40 36 0.017 313.2>285.1* 40 24 0.017 注：*为定量离子对。
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表 3 降解黄曲霉毒素 B1 菌株的筛选结果
Table 3. Screening results of the strains degrading aflatoxin B1
菌株编号 AFB1降解率（%） MD1 99.20±1.30 S1 73.17±0.87 25P11 42.00±2.37 30P2 39.55±3.00 25P14 33.87±1.77 25P3 30.20±2.96 35P5 27.90±6.66 30P6 24.76±3.30 25M5 15.07±4.88 S2 12.44±0.79 25P5 3.82±3.87
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