کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2020965 1069221 2009 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Cloning, expression and characterization of recombinant elastase from Pseudomonas aeruginosa in Picha pastoris
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Cloning, expression and characterization of recombinant elastase from Pseudomonas aeruginosa in Picha pastoris
چکیده انگلیسی

The gene lasB from Pseudomonas aeruginosa, which encoded elastase, was cloned and firstly successfully expressed in Pichia pastoris stain KM71 under the control of AOX promoter. The effects on the recombinant elastase activities of different pH, different temperatures and different metal ions were assayed. The full-length gene (1497 bp) encodes a preproenzyme including an N-terminal signal peptide (23 aa), a propeptide (197 aa) and mature elastase (301 aa). The recombinant elastase was secreted into culture supernatants using signal sequence from lasB and showed a single band at about 34 kDa by SDS–PAGE. The recombinant elastase expression hit the highest level of approximately 450 mg/L and the specific elastolytic activity of the recombinant elastase was 130 U/ml, which was approximately 26-fold higher than that of elastase obtained from P. aeruginosa. The optimal temperature and pH of the recombinant elastase was 28 °C and 7.4, respectively. The enzyme possessed high resistance to heat, and can be activated by Ca2+. These enzyme properties suggested that it could be produced in an industrial scale and has the potential to be a commercial enzyme.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 63, Issue 2, February 2009, Pages 69–74
نویسندگان
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