Protein Engineering for Improving and Diversifying Natural Product Biosynthesis

Innovations in CAZyme gene diversity and its modification for biorefinery applications

“…Protein engineering is the espoused method for modifying CAZyme for improved activity and stability together with augmentation of the products biosynthesis. It has also helped in improved screening and regulation of gene expression by engineering the sensors for enzymes [ 73 ]. The two general approaches for protein engineering involves directed evolution and rational design ( Fig.…”

“…Although, the technique of directed evolution has helped in the blooming of various industrially valuable CAZymes, it doesn’t always necessarily generate mutant with increased activity but reverse can also happen. Moreover, labor intensive screening procedure in absence of high throughput screening techniques makes rational design of CAZymes a preferred option for those enzymes with adequate structural information [ 73 ]. Here site directed mutagenesis is applied to achieve preconceived amino acid mutation based on the structural and catalytic information of the enzyme of interest [ 103 ].…”

A roadmap to engineering antiviral natural products synthesis in microbes

“…Bioprocess control with feedback cascades and feeding pattern will be critical to improve the titer and productivity [ 27 ]. The commonly-adopted metabolic engineering strategies are outlined in Figure 2 , which have been extensively covered in previous reviews [ 16 , 17 , 28 , 29 , 30 ]. We will highlight the detailed engineering strategies for the microbial production of specific antiviral natural products in the following section.…”

Genetically Encodable Scaffolds for Optimizing Enzyme Function

“…The exponential increase in genomic and protein sequences made freely available has aided the search for enzymes to carry out chemical transformations that might find use in the areas of applications mentioned, among others [ 10 ]. Common bottlenecks in the translation of this wealth of genomic enzymological information into biotechnological applications include enzyme instability, lower activity when expressed in heterologous hosts, interference from host metabolism, and suboptimal metabolic flux [ 11 ]. Two established strategies to overcome these limitations are (i) rational design and (ii) directed evolution of enzymes to identify sequence variants that confer desired properties.…”

“…Such properties include stability against denaturation induced by elevated temperatures, organic solvents or non-physiological pH, higher catalytic efficiency, and utilizing or generating noncanonical metabolites. The considerable list of achievements in using sequence space exploration for improving enzyme function and properties has been documented in other reviews [ 11 , 12 , 13 ].…”