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Screening of a high-yield strain of avermectin B1a by colony analysis in situ

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Abstract

Avermectin, an agricultural antibiotic, is widely used as an agricultural insecticide and an important lead compound of antibiotics. It is manufactured by Streptomyces avermitilis through fermentation. Manufacturers pay special attention to screening for strains with high fermentation capacity based on morphological properties of the colony and by the result of shake flask fermentation. These traditional screening methods are time-consuming and labor-intensive and require specialized equipment. Moreover, evaluation of colony appearance is highly subjective. To improve and accelerate the screening process, we developed a rapid in situ screening method. Forty-four strains isolated naturally from the spores of industrial high-yielding strains were studied. The data show that the colony fermentation titer is highly correlated with the yield from the shake flask fermentation of avermectin, and the Pearson’s R is 0.990. The total titer of avermectins by shake flask fermentation is also highly correlated with the B1a titer (Pearson’s R is 0.994). This result also shows that strains can be quickly screened by analyzing the colony titer. Pigment rings of the colonies that appeared after growing and maturing on the new medium plate were analyzed. The chosen colonies were directly marked and punched and then extracted with methanol. The fermentation ability can be evaluated by measuring the absorbance at 245 nm. This methodology can be applied in both natural breeding and mutation breeding conditions. By continuously breeding from 2008 to 2020, the flask titer of avermectin B1a increased from 4582 ± 483 to 9197 ± 1134 μg/mL.

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Acknowledgements

The authors are very grateful for the helpful discussion provided by Jianhui Gao and Xi Cheng from Hebei Xingbai Pharmaceutical Group Co. Ltd.

Funding

This study was funded and financially supported by the Funds of Huai’an Municipal Science and Technology Bureau (grant number HAN201610).

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Authors and Affiliations

  1. Jiangsu Food & Pharmaceutical Science College, Huaian, 223003, Jiangsu, China

    Zhongxuan Gou, Feng He, Zhaxi Bamao, Zixuan Li & Tingyu Xu

  2. Hebei Veyong Biochemical Co., LTD, Shijiazhuang City, Hebei, 050011, China

    Zhongxuan Gou, Junhua Li & Tingyu Xu

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  1. Zhongxuan Gou
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  2. Junhua Li
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  5. Zixuan Li
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Contributions

GZX conceived and designed the research and wrote the manuscript. LJH, HF, and ZB conducted the experiments. LZX and XTY contributed analytical tools.

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Correspondence to Zhongxuan Gou.

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Key points

• Avermectin is an agricultural insecticide and a lead compound of antibiotics.

• A high-yielding strain can be identified in situ by analyzing the pigment ring around the colony.

• A high-yielding colony can be screened by measuring the methanol extract of the colony at 245 nm.

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Gou, Z., Li, J., He, F. et al. Screening of a high-yield strain of avermectin B1a by colony analysis in situ. Int Microbiol 26, 123–133 (2023). https://doi.org/10.1007/s10123-022-00279-0

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  • DOI: https://doi.org/10.1007/s10123-022-00279-0

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