Improved catalytic efficiency of Endo-β-1,4-glucanase from Bacillus subtilis BME-15 by directed evolution.

{"article_title":"American Society for Transplantation and Cellular Therapy Series, #6: Management of Invasive Candidiasis in Hematopoietic Cell Transplantation Recipients.","author":"Neofytos D, Steinbach WJ, Hanson K, Carpenter PA, Papanicolaou GA, Slavin MA","journal_title":"Transplantation and cellular therapy","issn":"2666-6367","isbn":"","publication_date":"2009-03-15","volume":"82","issue":"4","first_page":"222","page_count":"","accession_number":"36649748","doi":"","publisher":"Elsevier Inc","doctype":"Practice Guideline","subjects":"United States; Hematopoietic Stem Cell Transplantation adverse effects; Candidiasis, Invasive epidemiology; Candidiasis, Invasive therapy; Adult; Humans; Child; United States epidemiology; Transplant Recipients","interest_area":["Infectious Disease & Vaccines"," Obstetrics & Gynecology"],"abstract":"The Practice Guidelines Committee of the American Society of Transplantation and Cellular Therapy (ASTCT) partnered with its Transplant Infectious Disease Special Interest Group (TID-SIG) to update its 2009 compendium-style infectious disease guidelines for hematopoietic cell transplantation (HCT). A completely new approach was taken with the goal of better serving clinical providers by publishing each standalone topic in the infectious disease series as a concise format of frequently asked questions (FAQ), tables, and figures. Adult and pediatric infectious disease and HCT content experts developed and then answered FAQs and finalized topics with harmonized recommendations made by assigning an A through E strength of recommendation paired with a level of supporting evidence graded I through III. This sixth guideline in the series focuses on invasive candidiasis (IC) with FAQs to address epidemiology, clinical diagnosis, prophylaxis, and treatment of IC, plus special considerations for pediatric, cord blood, haploidentical, and T cell-depleted HCT recipients and chimeric antigen receptor T cell recipients, as well as future research directions. Copyright \u00a9 2023 The American Society for Transplantation and Cellular Therapy. Published by Elsevier Inc. All rights reserved.","url":"https:\/\/search.ebscohost.com\/login.aspx?direct=true&db=mdl&AN=36649748","isPdfLink":true,"isSAML":false,"an":"36649748","number_other":"","type_pub":"","issn_electronic":"2666-6367","languages":"English","language":"eng","date_entry":"","date_update":"","titleSource":"Applied Microbiology & Biotechnology. Mar2009, Vol. 82 Issue 4, p671-679. 9p. 1 Color Photograph, 1 Black and White Photograph, 2 Diagrams, 2 Charts, 2 Graphs.","date_pub_cy":"","type_document":"","contract_publisher":"","authored_on":"2009-03-15","description":"Bacillus subtilis endo-\u03b2-1,4-glucanase (Cel5A) hydrolyzes cellulose by cleavage of the internal bonds in the glucose chains, producing new ends randomly. Using directed evolution techniques of error-prone polymerase chain reaction (PCR) and DNA shuffling, several Cel5A variants with improved catalytic activity had been screened from the mutant library, which contained 71,000 colonies. Compared with the wild-type enzyme, the variants (M44-11, S75 and S78) showed 2.03 to 2.68-fold increased activities toward sodium carboxymethyl cellulose (CMC), while the M44-11 also exhibited a wider pH tolerance and higher thermostability. Structural models of M44-11, S75, S78, and WT proteins revealed that most of the substitutions were not located in the strictly conserved regions, except the mutation V255A of S75, which was closed to the nucleophile Glu257 in the catalytic center of the enzyme. Moreover, V74A and D272G of M44-11, which were not located in the substrate binding sites and the catalytic center, might result in improved stability and catalytic activity. These results provided useful references for directed evolution of the enzymes that belonged to the glycoside hydrolase family 5 (GH5). [ABSTRACT FROM AUTHOR]","upload_link":"https:\/\/search.ebscohost.com\/login.aspx?direct=true&site=eds-live&db=cxh&AN=36649748&authtype=shib&custid=ns346513&group=main&profile=eds","no_of_pages":"","authored_by":"Lin, Ling, Meng, Xin, Liu, Pengfu, Hong, Yuzhi, Wu, Gaobing, Huang, Xiaoluo, Li, Congcong, Dong, Junli, Xiao, Liang, Liu, Ziduo","header":{"DbId":"cxh","DbLabel":"Biomedical Reference Collection: Corporate","An":"36649748","RelevancyScore":"676","PubType":"Academic Journal","PubTypeId":"academicJournal","PreciseRelevancyScore":"675.905517578125"},"plink":"https:\/\/search.ebscohost.com\/login.aspx?direct=true&site=eds-live&db=cxh&AN=36649748&authtype=shib&custid=ns346513&group=main&profile=eds","issn_print":"01757598","physicalDescription":{"Pagination":{"PageCount":"9","StartPage":"671"}},"additionalInfo":{"Authored_By":"Lin, Ling, Meng, Xin, Liu, Pengfu, Hong, Yuzhi, Wu, Gaobing, Huang, Xiaoluo, Li, Congcong, Dong, Junli, Xiao, Liang, Liu, Ziduo","Published_Date":"2009-03-15","Source":"Applied Microbiology & Biotechnology. Mar2009, Vol. 82 Issue 4, p671-679. 9p. 1 Color Photograph, 1 Black and White Photograph, 2 Diagrams, 2 Charts, 2 Graphs.","Languages":"English","Subjects":"BACILLUS subtilis, CATALYSIS, CELLULASE, HYDROGEN-ion concentration, POLYMERASE chain reaction, BINDING sites, DNA, SUBSTRATES (Materials science)","Title_Abbreviations":"Applied Microbiology & Biotechnology","Volume":"82"}}