Combination of ARTP mutagenesis and color-mediated high-throughput screening to enhance 1-naphthol yield from microbial oxidation of naphthalene in aqueous system

Abstract

Strain QCG of the aerobic bacteria Bacillus cereus is capable of producing 1-naphthol from naphthalene, this strain was first isolated and characterized in this study. Strain QCG was mutagenized to enhance 1-naphthol production, using atmospheric and room temperature plasma (ARTP) technology. Then, a microbial clone screening system was used to accelerate the operation. Meanwhile, a novel color-mediated high-throughput screening using 4-aminoantipyrine was performed to screen mutants. The optimal mutant strain QCG4 produced 19.58±0.34 mg∙L^‒1 1-naphthol from naphthalene that was 47.32% higher than that of the original strain (13.29±0.28 mg∙L^‒1). In addition, the optimal conditions for 1-naphthol production via whole-cell catalysis of strain QCG4 were determined to be an OD₆₀₀ of 40, 150 mg∙L^‒1 naphthalene, and 7.5% dimethyl formamide as a co-solvent at pH 7.5 and 26°C for 3 h, resulting in 41.18±0.12 mg∙L^‒1 1-naphthol, i.e., the mutant strain produces a 2.1-fold higher yield compared to the original strain.

Key words： Bacillus cereus QCG; naphthalene; 1-naphthol; ARTP mutagenesis; high-throughput screening; 4-aminoantipyrine

Acknowledgments

This work was supported by COVESTRO, the National Key Research and Development Program (Grant No. 2016YFA0204300), the Jiangsu Province Natural Science Foundation for Youths (No. BK20170997), and China Postdoctoral Science Foundation (Nos. 2018M642237 and 2017T100359).

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