Figure 1 | Scientific Reports

Figure 1

From: Simple and rapid direct cloning and heterologous expression of natural product biosynthetic gene cluster in Bacillus subtilis via Red/ET recombineering

Figure 1

Construction of a heterologous expression host for NPRS/PKS gene cluster based on B. subtilis 1A751.

(A) Detection mutation of the sfp gene in B. subtilis 168 (original strain of B. subtilis 1A751) after alignment with the intact sfp gene from six different B. subtilis strains. (B) Scheme for repairing the insertional mutation of the sfp gene in B. subtilis 1A751. (C) Haemolytic analysis of B. subtilis 1A751 (1A751) and transformants of the sfp-repaired derivative B. subtilis 1A751 + sfp (1A751 + sfp). (D) Analysis of surfactin and fengycin production in B. subtilis 1A751 + sfp. (a) HPLC-MS analysis of surfactin production in B. subtilis 1A751 + sfp. (b) MS spectra of three surfactin peaks(1, 2, and 3). Compound 1 indicates C13-surfactin (m/z 1008.6584 [M + H]+and 1030.6408 [M + Na]+), Compound 2 indicates C15-surfactin (m/z 1036.6897 [M + H]+and 1058.6715 [M + Na]+), Compound 3 indicates C14-surfactin (m/z 1022.6757 [M + H]+and 1044.6576 [M + Na]+), (c) HPLC-MS analysis of fengycin produced from B. subtilis 1A751 + sfp. (d) MS spectra of the three peaks of fengycin produced from B. subtilis 1A751 + sfp. Compound 4 indicates Ala-6-C17 fengycin (m/z 1477.8177 [M + H]+), and compound 5 indicates Val-6-C17 fengycin (m/z 1505.8 [M + H]+). Peak 6 is a mixture of Ala-6-C17 fengycin and Val-6-C17 fengycin.

Back to article page