Increase of d- and l-lactic acids in solution in BG-11 (Fig. S1); schematic representation of the genetic engineering approach at the neutral site (slr0168) and the disruption of the putative d-LDH (slr1556) in the genome of Synechocystis sp. PCC 6803, and colony PCR results for relevant strains showing complete segregation at both sites, confirming the gene insertion in ODSYN, DLEU, and DLEU/ΔDSYN strainsxand the deletion of slr1556 in ΔDSYN and DLEU/ΔDSYN strains (Fig. S2); photoautotropic growth and lactic acid production, shown for Synechocystis wild-type, ΔDSYN, and ODSYN strains (Fig. S3); enzymatic activity assay of purified Slr1556 (Fig. S4); d-lactic acid utilization of Synechocystis wild-type and ΔDSYN strain cell extracts as studied via an NAD(P) reduction assay (Fig. S5); lactic acid detection by HPLC analysis after a prolonged LDH activity assay for wild-type and ΔDSYN strains utilizing NADH and NADPH, respectively, and pyruvate as a starting substrate, and relative NADH- versus NADPH-dependent LDH activity as determined by pyruvate reduction of WT and ODSYN strain cell extracts (Fig. S6): dynamic behavior of growth with d-lactic acid (Fig. S7).
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