Isolation and identification of an erythromycin degradation bacterium strain and its biodegradation characteristics

doi:10.3969/j.issn.1004-1524.2021.01.16

Acta Agriculturae Zhejiangensis ›› 2021, Vol. 33 ›› Issue (1): 131-141.DOI: 10.3969/j.issn.1004-1524.2021.01.16

• Environmental Science • Previous Articles Next Articles

Isolation and identification of an erythromycin degradation bacterium strain and its biodegradation characteristics

XU Shuangyan¹(), ZHANG Tao¹, ZHANG Cheng¹^,², LIN Hui³, SHUI Xianlei¹, ZHENG Huabao¹^,²^,^*()

1. College of Environmental and Resource Sciences, Zhejiang A & F University, Hangzhou 311300, China
2. Key Laboratory of Soil Contamination Bioremediation of Zhejiang Province, Hangzhou 311300, China
3. Institute of Environmental Resources and Soil Fertilizer, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021,China

Received:2020-07-03 Online:2021-01-25 Published:2021-01-25
Contact: ZHENG Huabao

Abstract

Abstract:

In order to obtain efficient erythromycin degradation bacteria, erythromycin was used as the sole carbon source, and gradient domestication method was used to isolate and screen erythromycin degradation bacteria. It was shown that an erythromycin degradation bacterium strain Ery-6 was isolated from the soil of an organic fertilizer production workshop where chicken manure was stacked for a long time. The Ery-6 strain was identified as Methylobacillus sp. through colony morphology identification and 16S rDNA sequence analysis. Further, high performance liquid chromatography (HPLC) was used to measure the degradation rate of erythromycin, and the degradation conditions of erythromycin by Ery-6 strain were optimized, including different erythromycin concentration, culture temperature, rotation speed, initial pH value, exogenous carbon and nitrogen sources and metal ions. The Ery-6 strain could reach a degradation rate of 88.68% after 48 h in inorganic salt medium containing 100 mg·L^-1 erythromycin under temperature of 35 ℃, rotation speed of 120 r·min^-1, initial pH value of 7.0, and 50 mg·L^-1 sucrose. Meanwhile, the Ery-6 strain could tolerate 1 000 mg·L^-1 high-concentration erythromycin, and exhibited a degradation rate of 31.95% after 48 h under temperature of 35 ℃, rotation speed of 120 r·min^-1, initial pH value of 7.0. The Ery-6 strain had a good tolerance to various metal ions. Cu²⁺ could inhibit its growth, and pose direct effect on the degradation of erythromycin by Ery-6. The present study for the first time reported that the Methylobacillus sp. strain had the ability to degrade erythromycin, and it could tolerate high concentration of erythromycin, and maintain a certain degradation rate, which provided new microorganism for biodegradation of erythromycin in the environment.

Key words: erythromycin, degrading bacteria, isolation, degradation characteristics

CLC Number:

S154.39

XU Shuangyan, ZHANG Tao, ZHANG Cheng, LIN Hui, SHUI Xianlei, ZHENG Huabao. Isolation and identification of an erythromycin degradation bacterium strain and its biodegradation characteristics[J]. Acta Agriculturae Zhejiangensis, 2021, 33(1): 131-141.

Figures/Tables 10

Table 1 Gradient elution procedure for erythromycin determination

t/min	流动相比例Mobile phase ratio/%
t/min	A	B
0	90.0	10.0
1.0	90.0	10.0
2.0	80.0	20.0
3.5	77.0	23.0
5.0	76.0	24.0
8.0	76.5	23.5
9.0	70.0	30.0
16	70.0	30.0
20	50.0	50.0
24	90.0	10.0
25	90.0	10.0

Fig.1 Colony morphology (A), transmission electron microscopy (B) and phylogenetic tree (C) of Ery-6 strain

Fig.2 Growth curve (A) and biodegradation kinetic curve (B) of Ery-6 strain Ct/C0 was the ratio of erythromycin concentration at t time to the initial concentrution.CK, Control withow Ery-6 strain inoculation.

Table 2 Biodegradation kinetic equation and kinetic parameter of erythromycin

处理Treatment	一级反应方程First order reaction equation	k/h^-1	t_1/2/h	R²
不接菌Not inoculated	C_t=92.14e^-0.00049^t	0.000 49	88.42	0.928 4
接菌Inoculated	C_t=57.69e^-0.00753^t	0.007 53	30.65	0.925 9

Fig.3 Effect of initial erythromycin concentration on erythromycin degradation by Ery-6 strain

Fig.4 Effect of temperature on erythromycin degradation by Ery-6 strain

Fig.5 Effect of rotation speed on erythromycin degradation by Ery-6 strain

Fig.6 Effect of initial pH on erythromycin degradation by Ery-6 strain

Fig.7 Effect of exogenous carbon or nitrogen source on erythromycin degradation by Ery-6 strain Bars marked without the same letters indicated significant difference at P<0.05. The same as below. No exogenous carbon or nitrogen source was added in the control.

Fig.8 Effect of exogenous metal ions on erythromycin degradation by Ery-6 strain No exogenous metal ion was added in the control.

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Isolation and identification of an erythromycin degradation bacterium strain and its biodegradation characteristics

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