کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
8360721 1542346 2014 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Expression of soluble recombinant lipoxygenase from Pleurotus sapidus in Pichia pastoris
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Expression of soluble recombinant lipoxygenase from Pleurotus sapidus in Pichia pastoris
چکیده انگلیسی
The first heterologous expression of an iron-containing lipoxygenase from a basidiomycete in Pichia pastoris is reported. Five different expression constructs of the lipoxygenase gene LOX1 from Pleurotus sapidus were cloned and successfully transferred into P. pastoris SMD1168, but only one pPIC9K vector construct was functionally expressed. In this construct the vector-provided α-factor signal sequence was replaced by insertion of a second Kozak sequence between the signal sequence and the LOX1 gene. His+ transformants were screened for their level of resistance to geneticin (G418). Lox1 was expressed under different culture conditions and purified using the N-terminal His-tag. Relative enzyme activity increased significantly 48 h after methanol induction and was highest with 2 ml l−1 inducer. The recombinant enzyme showed an optimal lipoxygenase activity at pH 7 and 30-35 °C and a vmax like the wild-type enzyme.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 95, March 2014, Pages 233-239
نویسندگان
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