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Acidification of the Culture Medium by Products of Glucose Metabolism Inhibits the Synthesis of Heterologous Extracellular α-amylase by Bacillus subtilis 168

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Abstract

The synthesis of α-amylase AmyM3 from B. flexus 406 in the recombinant strain B. subtilis 168-28 was significantly repressed during submerged cultivation in nutrient medium supplemented with 1% glucose. Repression of α-amylase synthesis by glucose in B. subtilis cells is observed at the level of transcription and is mediated by the catabolite control protein CcpA. The effect of catabolite repression was eliminated after the deletion of a putative CRE site in the amyM3 gene, but the amount of extracellular α-amylase still significantly decreased in the presence of glucose. It was shown that, despite the high level of amyM3 gene activity, a shift in the cultivation medium pH to 5.8–6.0 due to glucose-overflow metabolites excreted by B. subtilis interferes with the synthesis of active extracellular α-amylase, which probably affects postsecretory folding of the enzyme. This observation is a newly demonstrated facet of the CcpA-mediated effect of preferentially utilized sugars on extracellular α-amylase production in Bacillus subtilis.

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Funding

The work was carried out within the framework of the assignment of the state program of scientific research “Biotechnology” for 2018–2020.

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  1. Belarusian State University, 220030, Minsk, Belarus

    A. V. Kachan & A. N. Evtushenkov

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  1. A. V. Kachan
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  2. A. N. Evtushenkov
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Correspondence to A. V. Kachan.

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The authors declare that they have no conflict of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.

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Kachan, A.V., Evtushenkov, A.N. Acidification of the Culture Medium by Products of Glucose Metabolism Inhibits the Synthesis of Heterologous Extracellular α-amylase by Bacillus subtilis 168 . Appl Biochem Microbiol 57, 443–451 (2021). https://doi.org/10.1134/S0003683821040062

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