کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
69843 48796 2012 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
The insertion of four residues Isoleucines at the N-terminus of Staphylococcus simulans lipase affects its catalytic and biochemical properties
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی کاتالیزور
پیش نمایش صفحه اول مقاله
The insertion of four residues Isoleucines at the N-terminus of Staphylococcus simulans lipase affects its catalytic and biochemical properties
چکیده انگلیسی

For the sake of evaluating the effect of the hydrophobic residues insertion in the N-terminus of Staphylococcus simulans lipase (SSL), four residues of Isoleucines have been inserted at the N-terminus of this enzyme. The recombinant Staphylococcus simulans lipase (r-SSL) and its constructed mutant (4Ile-SSL) were expressed in Escherichia coli BL21 (DE3) and purified to homogeneity using classical chromatographic techniques. We have performed, then, a comparative study on the biochemical properties of the two enzymes. Due to the insertion of 4Ile at the N-terminus of Staphylococcus simulans lipase (SSL), some important differences in the biochemical properties between r-SSL and 4Ile-SSL have been found. We can essentially notice that, when using short chain triacylglycerols (tributyrin) as substrate, the insertion of four Isoleucines residues (4Ile) was accompanied by an increase in the specific activity (3 fold) as well as in the catalytic efficiency (kcat/KM app.) (2 fold), as compared to the recombinant SSL. Furthermore, our results indicate that the presence of 4Ile at the N-terminus of SSL has greatly affected the pH stability of the enzyme and considerably increased its thermostability.

Surface GRASP representation of closed forms and open forms of SSL and 4Ile-SSL. The catalytic serine is colored in red. The catalytic cavity is shown in yellow. The 4Ile residues are represented in dark blue. The lid domain is colored in green.Figure optionsDownload as PowerPoint slideHighlights
► The insertion of four residues Isoleucines at the N-terminus of SSL.
► Expression and purification of recombinant SSL and its mutant 4Ile-SSL.
► Comparative study on the biochemical properties of r-SSL and 4Ile-SSL.
► The importance of the N-terminus region in the biochemical properties of SSL.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Molecular Catalysis B: Enzymatic - Volume 82, October 2012, Pages 1–7
نویسندگان
, , , , , ,