title: Preparation and properties of Clostridium thermocellum lichenase deletion variants and their use for construction of bifunctional hybrid proteins.
abstract: Major properties (pH and temperature optimum, stability) of lichenase (beta-1,3-1,4-glucanase) deletion variants from Clostridium thermocellum were comparatively studied. The deletion variant LicBM2 was used to create hybrid bifunctional proteins by fusion with sequences of the green fluorescent protein (GFP) from Aequorea victoria. The data show that in hybrid proteins both GFP and lichenase retain their major properties, namely, GFP remains a fluorescent protein and the lichenase retains activity and high thermostability. Based on the results of this investigation and results that have been obtained earlier, the use of the deletion variants of lichenase and the bifunctional hybrid proteins as reporter proteins is suggested.
authors.affiliation: Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, 117809 Russia. kostyam@vigg.ru
authors.forename: K A
authors.initials: KA
authors.lastname: Musiychuk
authors.forename: I V
authors.initials: IV
authors.lastname: Goldenkova
authors.forename: R M
authors.initials: RM
authors.lastname: Abdeev
authors.forename: N S
authors.initials: NS
authors.lastname: Kobets
authors.forename: E S
authors.initials: ES
authors.lastname: Piruzian
mesh_terms: D003013:Clostridium / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D017353:Gene Deletion; D017930:Genes, Reporter; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics*; D049452:Green Fluorescent Proteins; D006863:Hydrogen-Ion Concentration; D008164:Luminescent Proteins / Q000378:metabolism; D008967:Molecular Biology / Q000379:methods; D016296:Mutagenesis; D010957:Plasmids / Q000378:metabolism; D011489:Protein Denaturation; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism*; D013696:Temperature; D013997:Time Factors; D014466:Ultraviolet Rays
chemical_list: D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D006026:Glycoside Hydrolases; C028020:licheninase