کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2021189 | 1069234 | 2008 | 11 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Characterization and cloning of chitin deacetylases from Rhizopus circinans
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Characterization and cloning of chitin deacetylases from Rhizopus circinans Characterization and cloning of chitin deacetylases from Rhizopus circinans](/preview/png/2021189.png)
چکیده انگلیسی
Chitin deacetylase catalyzes hydrolysis of the acetamido groups of N-acetylglucosamine of chitin in fungal cell walls. Here a chitin deacetylase secreted by Rhizopus circinans was purified to homogeneity and partially characterized. The enzyme exhibits an apparent molecular weight of â¼75 kDa. At 37 °C it shows optimal activity at pH 5.5-6. Its pH stability and thermal stability are good. Mn2+ and Mg2+ slightly enhance the activity of the enzyme and Cu2+ strongly inhibits it. An R. circinans cDNA library was constructed and screened with a homologous probe synthesized by RT-PCR or with synthetic primers derived from the N-terminal amino-acid sequence of the native purified chitin deacetylase. Three chitin deacetylase cDNAs (RC, D2, and I3/2) were isolated from the cDNA library and sequenced. These cDNAs exhibit features characteristic of chitin deacetylase sequences: the presence of a polysaccharide deacetylase domain, a metal-binding triad, the conserved catalytic residues, and high homology with various chitin deacetylase genes. The cDNAs were cloned in a Pichia pastoris expression system and produced as polyhistidine-tagged proteins. Only one recombinant enzyme (called RC) was active under the tested conditions. It was purified to homogeneity in a single step and further characterized. The protein showed an apparent molecular mass of â¼75 kDa and, like the native enzyme, showed optimal activity at pH 5.5-6 at 37 °C. It was strongly inhibited by Cu2+. The isolation of several chitin deacetylase cDNAs from the same microorganism is discussed.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 59, Issue 1, May 2008, Pages 127-137
Journal: Protein Expression and Purification - Volume 59, Issue 1, May 2008, Pages 127-137
نویسندگان
Carole Gauthier, Fabienne Clerisse, Jacques Dommes, Marie-France Jaspar-Versali,