{"doi":"10.1371/journal.pone.0108292","title":"Combining Rational and Random Strategies in β-Glucosidase Zm-p60.1 Protein Library Construction","abstract":null,"journal":"PLoS ONE","year":2014,"id":648198,"datarank":0.2813160602931734,"base_score":1.3862943611198906,"endowment":1.3862943611198906,"self_citation_contribution":0.20794415416798362,"citation_network_contribution":0.07337190612518979,"self_endowment_contribution":0.20794415416798362,"citer_contribution":0.07337190612518979,"corpus_percentile":null,"corpus_rank":null,"citation_count":3,"citer_count":3,"citers_with_citation_signal":3,"citers_with_endowment":3,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1689088,"name":"Pavel Klimeš","orcid":null,"position":1,"is_corresponding":false},{"id":1689090,"name":"Pavel Mazura","orcid":null,"position":2,"is_corresponding":false},{"id":254551,"name":"Břetislav Brzobohatý","orcid":"0000-0002-1552-1314","position":3,"is_corresponding":false},{"id":1689087,"name":"Dušan Turek","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Combining Rational and Random Strategies in β-Glucosidase Zm-p60.1 Protein Library Construction","abstract":"Saturation mutagenesis is a cornerstone technique in protein engineering because of its utility (in conjunction with appropriate analytical techniques) for assessing effects of varying residues at selected positions on proteins' structures and functions. Site-directed mutagenesis with degenerate primers is the simplest and most rapid saturation mutagenesis technique. Thus, it is highly appropriate for assessing whether or not variation at certain sites is permissible, but not necessarily the most time- and cost-effective technique for detailed assessment of variations' effects. Thus, in the presented study we applied the technique to randomize position W373 in β-glucosidase Zm-p60.1, which is highly conserved among β-glucosidases. Unexpectedly, β-glucosidase activity screening of the generated variants showed that most variants were active, although they generally had significantly lower activity than the wild type enzyme. Further characterization of the library led us to conclude that a carefully selected combination of randomized codon-based saturation mutagenesis and site-directed mutagenesis may be most efficient, particularly when constructing and investigating randomized libraries with high fractions of positive hits.","is_dataset_classified":null,"base_score":1.3862943611198906,"endowment":1.3862943611198906,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"25260034","pmcid":null,"openalex_id":"https://openalex.org/W1964597525","authors":[],"funders":[],"total_grants":0,"fwci":0.3105,"citation_percentile":0.63712655,"influential_citations":0,"citation_trend":[{"year":2016,"count":1},{"year":2018,"count":1},{"year":2023,"count":1}],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://doi.org/10.1371/journal.pone.0108292","host_type":"journal"},{"url":"https://doi.org/10.1371/journal.pone.0108292","host_type":"publisher"},{"url":"http://dx.plos.org/10.1371/journal.pone.0108292","host_type":"publisher"},{"url":"https://pubmed.ncbi.nlm.nih.gov/25260034","host_type":"repository"},{"url":"https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0108292","host_type":"repository"},{"url":"http://hdl.handle.net/11104/0239818","host_type":"repository"},{"url":"https://doaj.org/article/13e17aed09cd4655b62d24bcfcbab278","host_type":"repository"},{"url":"https://figshare.com/articles/dataset/_Combining_Rational_and_Random_Strategies_in_946_Glucosidase_Zm_p60_1_Protein_Library_Construction_/1184972","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/4178128","host_type":"repository"}],"fields_of_study":["Enzyme Production and Characterization","CRISPR and Genetic Engineering","Transgenic Plants and Applications"],"mesh_terms":["beta-Glucosidase","Codon","Zea mays","Enzyme Activation","Hydrolysis","Plant Proteins","Substrate Specificity","Protein Engineering","Gene Library","Mutagenesis","Computational Biology","Databases, Protein"],"keywords":["Saturated mutagenesis","Mutagenesis","Protein engineering","Directed evolution","Computational biology","Directed Molecular Evolution","Site-directed mutagenesis","Biology","Genetics","DNA shuffling","Biochemistry","Mutation","Mutant","Enzyme","Gene"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-10T02:21:36.474236Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}