Protein engineering: Methods and applications

Recent Advances in β-Glucosidase Sequence and Structure Engineering: A Brief Review

“…It accelerates the pace of mutagenesis, recombination, and protein selection in an explicit manner to obtain desired properties. The approach typically comprises an iterative cycle of mutagenesis to generate diverse mutants followed by high-throughput screening [ 37 ].…”

“…It entails a thorough comprehension of active sites and functions of enzymes, and specific residues are selected for targeted mutagenesis. The three main methods used to help identify mutation-specific residues are structural analysis, multiple sequence alignment (MSA), and robust computational techniques [ 37 ].…”

“…Guo et al introduced an improved OEP method to increase the amplification efficiency of long-length multisite directed fragments by providing primers continuously [ 70 ]. The whole-plasmid single-round PCR method is involved in PCR replication of double-stranded plasmid DNA template using two complementary primers with the desired mutation for better glucose tolerance [ 37 ].…”

Stabilization challenges and aggregation in protein-based therapeutics in the pharmaceutical industry

“…160 By utilizing these computational tools, researchers can make rational decisions on which modications are the most likely to enhance protein stabilities, leading ultimately to the development of more robust and effective proteins for protein-based therapeutics. 161 Deep learning is applied to unlabelled amino acid sequences to extract essential protein features in rational protein engineering. Alley et al 162 used unsupervised learning with a recurrent articial neural network called UniRep (Unied Representation).…”

“… 160 By utilizing these computational tools, researchers can make rational decisions on which modifications are the most likely to enhance protein stabilities, leading ultimately to the development of more robust and effective proteins for protein-based therapeutics. 161 …”