caa a22 4500 60621285X CHVBK 20210128101044.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s10295-015-1635-4 doi (NATIONALLICENCE)springer-10.1007/s10295-015-1635-4 A novel high-throughput and quantitative method based on visible color shifts for screening Bacillus subtilis THY-15 for surfactin production [Elektronische Daten] [Huan Yang, Huimin Yu, Zhongyao Shen] A novel chromatic visible screening method using bromothymol blue (BTB) as a color indicator and cetylpyridinium chloride (CPC) as a mediator was constructed to obtain the high titer surfactin-producing strains. The reliability and quantification accuracy of color shift were also confirmed. Regular chromatic responses from faint yellow-green to dark green and bright blue reflected the different ranges of surfactin concentrations. Moreover, the quantitative accuracy of surfactin quantification in the range of 100-500mg/L was verified by reverse-phase high-performance liquid chromatography (RP-HPLC) using different fermentation supernatant samples. Using this CPC-BTB method, a superior surfactin producer, Bacillus subtilis THY-15, was successfully screened. The producer's surfactin (Srf) titer reached 1240mg/L. RP-HPLC analysis of THY-15 revealed four surfactin isoforms. As identified by amino acid analysis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis, the isoforms of surfactin in fraction 1, 2 and 4 had the same circular peptide sequence of Glu-Leu-Leu-Val-Asp-Leu-Leu but different iso-C13, C14 and C15 fatty acid chains, but the isoform in fraction 3 possessed a special peptide sequence of Glu-Val-Leu-Leu-Asp-Leu-Val. Society for Industrial Microbiology and Biotechnology, 2015 CPC-BTB screening method nationallicence Chromatic responses nationallicence Surfactin nationallicence Bacillus subtilis nationallicence MALDI-TOF MS/MS nationallicence Yang Huan Key Laboratory for Industrial Biocatalysis of the Ministry of Education, Department of Chemical Engineering, Tsinghua University, 100084, Beijing, China aut Yu Huimin Key Laboratory for Industrial Biocatalysis of the Ministry of Education, Department of Chemical Engineering, Tsinghua University, 100084, Beijing, China aut Shen Zhongyao Key Laboratory for Industrial Biocatalysis of the Ministry of Education, Department of Chemical Engineering, Tsinghua University, 100084, Beijing, China aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/8(2015-08-01), 1139-1147 1367-5435 42:8<1139 2015 42 10295 https://doi.org/10.1007/s10295-015-1635-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10295-015-1635-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Yang Huan Key Laboratory for Industrial Biocatalysis of the Ministry of Education, Department of Chemical Engineering, Tsinghua University, 100084, Beijing, China aut NATIONALLICENCE 700 1- Yu Huimin Key Laboratory for Industrial Biocatalysis of the Ministry of Education, Department of Chemical Engineering, Tsinghua University, 100084, Beijing, China aut NATIONALLICENCE 700 1- Shen Zhongyao Key Laboratory for Industrial Biocatalysis of the Ministry of Education, Department of Chemical Engineering, Tsinghua University, 100084, Beijing, China aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/8(2015-08-01), 1139-1147 1367-5435 42:8<1139 2015 42 10295