Modification of nitrilase based on computer screening and efficient biosynthesis of 4-cyanobenzoic acid.

4-cyanobenzoic acid serves as a crucial intermediate for the synthesis of various high-value organic compounds. The enzymatic hydrolysis of terephthalonitrile to produce 4-cyanobenzoic acid using nitrilase offers the advantages of a simple reaction pathway, environmental friendliness, and easy product separation. In order to efficiently develop nitrilases that meet industrial production requirements, the virtual screening method used in the study is established and mature. From a total of 371 amino acids in the nitrilase AfNIT, which exhibits activity in terephthalonitrile hydrolysis, three candidate sites (F168, S192, and T201) were identified, and a "small and accurate" mutant library was constructed. The triple mutant F168V/T201N/S192F was screened from this small mutant library with a specific activity of 227.3 U mg-1 , which was 3.8 times higher than that of the wild-type AfNIT. Using the whole-cell biocatalyst containing the mutant F168V/T201N/S192F, terephthalonitrile was successfully hydrolyzed at a concentration of 150 g L-1 to produce 4-cyanobenzoic acid with a final yield of 170.3 g L-1 and a conversion rate of 98.7%. The obtained nitrilase mutant F168V/T201N/S192F in this study can be effectively applied in the biomanufacturing of 4-cyanobenzoic acid using terephthalonitrile as a substrate. Furthermore, the results also demonstrate the significant improvement in predictive accuracy achieved through the latest AI-assisted computer simulation methods. This approach represents a promising and feasible new technological pathway for assisting enzyme engineering research, laying a theoretical foundation for other related studies.

Application of organic polymeric flocculants in centrifugal dewatering of oil refinery sludge.

In order to evaluate the applicability of the organic polymeric flocculants (OPF) in the treatment of oil refinery sludge, experiments were conducted to show that OPF have better performance of flocculation than inorganic flocculants. Both the anionic and cationic OPF have satisfactory flocculation efficiency in oil sludge treatment, but the latter are more cost-efficient. Among the over 20 types of flocculants tested, 2 OPF (CPAM-2 and HPAM-2) were selected as the treatment agents, based on their good treatment performances, oil-resistance and economic feasibility. It was demonstrated in the industrial-scale centrifugal dewatering experiments that the application of either CPAM-2 or HPAM-2 could achieve high treatment efficiency of the oil sludge dewatering and reduce the COD of centrifugal liquid to less than 1000 mg/L.

Micro-electrolysis technology for industrial wastewater treatment.

Experiments were conducted to study the role of micro-electrolysis in removing chromaticity and COD and improving the biodegradability of wastewater from pharmaceutical, dye-printing and papermaking plants. Results showed that the use of micro-electrolysis technology could remove more than 90% of chromaticity and more than 50% of COD and greatly improved the biodegradability of pharmaceutical wastewater. Lower initial pH could be advantageous to the removal of chromaticity. A retention time of 30 minutes was recommended for the process design of micro-electrolysis. For the use of micro-electrolysis in treatment of dye-printing wastewater, the removal rates of both chromaticity and COD were increased from neutral condition to acid condition for disperse blue wastewater; more than 90% of chromaticity and more than 50% of COD could be removed in neutral condition for vital red wastewater.