title: Assembling a xylanase-lichenase chimera through all-atom molecular dynamics simulations.
abstract: Multifunctional enzyme engineering can improve enzyme cocktails for emerging biofuel technology. Molecular dynamics through structure-based models (SB) is an effective tool for assessing the tridimensional arrangement of chimeric enzymes as well as for inferring the functional practicability before experimental validation. This study describes the computational design of a bifunctional xylanase-lichenase chimera (XylLich) using the xynA and bglS genes from Bacillus subtilis. In silico analysis of the average solvent accessible surface area (SAS) and the root mean square fluctuation (RMSF) predicted a fully functional chimera, with minor fluctuations and variations along the polypeptide chains. Afterwards, the chimeric enzyme was built by fusing the xynA and bglS genes. XylLich was evaluated through small-angle X-ray scattering (SAXS) experiments, resulting in scattering curves with a very accurate fit to the theoretical protein model. The chimera preserved the biochemical characteristics of the parental enzymes, with the exception of a slight variation in the temperature of operation and the catalytic efficiency (kcat/Km). The absence of substantial shifts in the catalytic mode of operation was also verified. Furthermore, the production of chimeric enzymes could be more profitable than producing a single enzyme separately, based on comparing the recombinant protein production yield and the hydrolytic activity achieved for XylLich with that of the parental enzymes.
authors: Laboratório Nacional de Ciência e Tecnologia do Bioetanol, Campinas, SP, Brazil.
authors: Junio
authors: J
authors: Cota
authors: Leandro C
authors: LC
authors: Oliveira
authors: André R L
authors: AR
authors: Damásio
authors: Ana P
authors: AP
authors: Citadini
authors: Zaira B
authors: ZB
authors: Hoffmam
authors: Thabata M
authors: TM
authors: Alvarez
authors: Carla A
authors: CA
authors: Codima
authors: Vitor B P
authors: VB
authors: Leite
authors: Glaucia
authors: G
authors: Pastore
authors: Mario
authors: M
authors: de Oliveira-Neto
authors: Mario T
authors: MT
authors: Murakami
authors: Roberto
authors: R
authors: Ruller
authors: Fabio M
authors: FM
authors: Squina
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology*; D003198:Computer Simulation; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D053838:Scattering, Small Angle
chemical_list: D011993:Recombinant Fusion Proteins; D006026:Glycoside Hydrolases; C028020:licheninase; D043364:Endo-1,4-beta Xylanases