Improved catalytic activity of a novel aspartate kinase by site-directed saturation mutagenesis

Characterization of aspartokinase double mutants using a combination of experiments and simulations

“…The single mutants A380I, T379L, and A380 M and double mutant T379 N/A380C were obtained by inducing saturation mutations and high-throughput screening, and the enzyme activity was improved; however, enzymatic properties were not improved. Our previous studies confirmed that sites 379 and 380 are important sites affecting enzyme activity [ 12 , 13 ]. In this study, we used sites 379 and 380 as saturated mutation sites ( Fig.…”

Process Cascade for the Production of Green Polymers from CO2 and Electric Energy

“…These traits are advantageous for using M. extorquens as a producer of l-lysine because the precursor oxaloacetate is supplied with increased metabolic flux [29]. From recent studies it is known that the enzyme aspartate kinase (LysC, phosphorylating aspartate to aspartyl phosphate) plays a central role in the biosynthesis of l-lysine and catalyzes the rate-limiting step in the corresponding enzymatic reaction cascade [49,50]. Moreover, it is known that activity of LysC is sensitive against l-lysine, depicting a feedback inhibition and limiting l-lysine biosynthesis upon certain intracellular levels [51].…”

Enabling technology and core theory of synthetic biology

“…Considerable progress has been made with the creation of "smart libraries" (Qu et al, 2020). These approaches have been reported to successfully improve enzyme properties such as thermostability, catalytic activity, and enantioselectivity (Fan et al, 2022;Tan et al, 2022;Tong et al, 2022). The combination of enzyme engineering and systems metabolic engineering has also significantly increased the metabolic flux of target products (Qian et al, 2019;Yang et al, 2020).…”