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Figure S1. Effects of 1,4-BL concentration and addition time on S. hygroscopicus 5008 cell growth (A and C) and VAL-A production (B and D). Values are mean ± SEM (standard error of mean) of four replications. Data points with asterisk are significantly different at the level of P < 0.05.

Figure S2. Amino acid sequence alignment of ShbR1. ShbR2 and ShbR3. ShbR1 have 40% and 29% identity to ShbR2 and ShbR3, respectively. ShbR2 has 45% identity to ShbR3.

Figure S3. Inactivation of shbR1, shbR2, and shbR3 in S. hygroscopicus 5008. A: Schematic deletion of shbR1 in S. hygroscopicus 5008 and PCR analysis of S. hygroscopicus5008 and the shbR1mutant. Lane 1, S. hygroscopicus5008; lane 2, positive control; lane 3, 1 kb marker; lanes 4 and 5, shbR1 mutants. B: Schematic deletion of shbR2 in S. hygroscopicus 5008 and PCR analysis of S. hygroscopicus 5008 and the shbR2 mutant. Lane 1, S. hygroscopicus 5008; lane 2, positive control; lane 3, 1 kb marker; lanes 4 and 5, shbR2 mutants. C: Schematic deletion of shbR3 in S. hygroscopicus 5008 and PCR analysis of S. hygroscopicus 5008 and the shbR3 mutant. Lane 1, positive control; lane 2, S. hygroscopicus 5008; lane 3, 1 kb marker; lanes 4 and 5, shbR3 mutants.

Figure S4. SDS–PAGE analysis of the purified ShbR1, ShbR2, ShbR3, and AdpA–H.

Figure S5. Schematic deletion of adpA–H in S. hygroscopicus 5008 and PCR analysis of S. hygroscopicus 5008 and the adpA–Hmutant. Lane 1, S. hygroscopicus 5008; lane 2, positive control; lane 3, 1 kb marker; lanes 4 and 5, adpA–H mutants.

Table SI. Sequences of primer pairs for quantitative real-time PCR (qRT-PCR) assay.

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