Enhanced catalytic activity and stability of lactate dehydrogenase for cascade catalysis of D-PLA by rational design.

Serving as a vital medical intermediate and an environmentally-friendly preservative, D-PLA exhibits substantial potential across various industries. In this report, the urgent need for efficient production motivated us to achieve the rational design of lactate dehydrogenase and enhance catalytic efficiency. Surprisingly, the enzymatic properties revealed that a mutant enzyme, LrLDHT247I/D249A/F306W/A214Y (LrLDH-M1), had a viable catalytic advantage. It demonstrated a 3.3-fold increase in specific enzyme activity and approximately a 2.08-fold improvement of Kcat. Correspondingly, molecular docking analysis provided a supporting explanation for the lower Km and higher Kcat/Km of the mutant enzyme. Thermostability analysis exhibited increased half-lives and the deactivation rate constants decreased at different temperatures (1.47-2.26-fold). In addition, the mutant showed excellent resistance abilities in harsh environments, particularly under acidic conditions. Then, a two-bacterium (E. coli/pET28a-lrldh-M1 and E. coli/pET28a-ladd) coupled catalytic system was developed and realized a significant conversion rate (77.7%) of D-phenyllactic acid, using 10 g/L L-phenylalanine as the substrate in a two-step cascade reaction.

Lactate dehydrogenase encapsulated in a metal-organic framework: A novel stable and reusable biocatalyst for the synthesis of D-phenyllactic acid.

In this study, we synthesized a novel biocatalyst by encapsulating lactate dehydrogenase (LDH) in the metal-organic framework ZIF-90 by one-pot embedding. It showed strong biological activity for efficient synthesis of D-phenyllactic acid (D-PLA). The morphology and structure of LDH@ZIF-90 was systematically characterized by powder X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy, confocal laser scanning microscopy (CLSM) and gas sorption. According to thermogravimetric analysis (TGA), the enzyme loading of the biocatalyst was 3 %. The Michaelis-Menten constant (Km) and maximal reaction rate (Vmax) of LDH@ZIF-90 were similar to those of free LDH, which proved that ZIF-90 had good biocompatibility to encapsulate LDH. At the same time, LDH@ZIF-90 exhibited enhanced tolerance to temperature, pH and organic solvents, and its reusability was greatly improved with 68 % of initial enzyme activity remaining after 7 rounds of recylcing. Overall, LDH encapsulated in ZIF-90 may be an economically competitive and environmentally friendly novel biocatalyst for the synthesis of D-PLA.

Magnetic NiFe2O4/MWCNTs functionalized cellulose bioadsorbent with enhanced adsorption property and rapid separation.

Magnetic NiFe2O4 nanoparticles and multi-walled carbon nanotubes functionalized cellulose composite (m-NiFe2O4/MWCNTs@cellulose) as a magnetic bioadsorbent was prepared and used for effectively removing Congo Red (CR) from aqueous solution. The chemical and physical properties of the prepared m-NiFe2O4/MWCNTs@cellulose were characterized by XRD, TGA, FT-IR, VSM, SEM and TEM. Batch experiments were carried out to investigate the adsorption capacity and mechanisms. Effects of different adsorption parameters such as initial CR concentration, adsorbent dosage and temperature were studied. Results demonstrated that m-NiFe2O4/MWCNTs@cellulose had high adsorption capacity for CR from aqueous solution. The obtained experimental data fitted well with the pseudo-second-order equation and followed the Langmuir isotherm model with a maximum adsorption capacity of 95.70 mg g-1 for CR. The m-NiFe2O4/MWCNTs@cellulose with rapid magnetic separation and high adsorption capacity can be a promising and recyclable engineering biomaterials for purification and treatment of practical wastewater.