(A) Construction of complemented and overexpression strains. Lane 1, wild-type (WT); Lane 2, WT:pykF overexpression strain; Lane 3, deletion of cobB gene strain (ΔcobB); Lane 4, ΔcobB:pykF complemented strain; Lane 5, deletion of pykF gene strain (ΔpykF); Lane 6, ΔpykF:pykF complemented strain; Lane 7, ΔpykF:K52R complemented strain; Lane 8, ΔpykF:K68R complemented strain; Lane 9, ΔpykF:K317R complemented strain. Anti-His: His Tag Mouse Monoclonal Antibody. (B) Analysis of pyruvate kinase activity throughout the V. alginolyticus ΔpykF strain and WT:pykF overexpression strain. Pyruvate kinase activity of V. alginolyticus WT was set as 1. (C) Analysis pyruvate kinase activity of site-directed mutagenesis complemented strains. Pyruvate kinase activity of V. alginolyticus ΔpykF:pykF was set as 1. Mean values and standard deviations were calculated based on three replicates (n = 3). Two-tailed P-values were determined by the t-test, and the significance level is 0.05. *p < 0.05.
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