کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8328579 | 1540206 | 2018 | 35 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Secretory expression of recombinant small laccase from Streptomyces coelicolor A3(2) in Pichia pastoris
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
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چکیده انگلیسی
This work reports for the first time the secretory expression of the small laccase (SLAC) from Streptomyces coelicolor A3(2) in Pichia pastoris. Using an AOX1 promoter and α factor as a secretion signal, the recombinant P. pastoris harbouring the laccase gene (rSLAC) produced high titres of extracellular laccase (500â¯Â±â¯10â¯U/l), which were further increased seven fold by pre-incubation at 80â¯Â°C for 30â¯min. The enzyme (â¼38â¯kDa) had an optimum activity at 80â¯Â°C, but optimum pH varied with substrate used. Km values for ABTS, SGZ and 2,6-DMP were 142.85â¯Î¼M, 10â¯Î¼M and 54.55â¯Î¼M and the corresponding kcat values were 60.6â¯sâ1, 25.36â¯sâ1 and 27.84â¯sâ1, respectively. The t1/2 values of the rSLAC at 60â¯Â°C, 70â¯Â°C, 80â¯Â°C were 60â¯h, 32â¯h and 10â¯h, respectively. The enzyme deactivation energy (Ed) was 117.275â¯kJ/mol while ÎG, ÎH and ÎS for thermal inactivation of the rSLAC were all positive. The rSLAC decolourised more than 90% of Brilliant Blue G and Trypan Blue dye in 6â¯h without the addition of a mediator. High titres of SLAC expressed in P. pastoris enhance its potential for various industrial applications.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: International Journal of Biological Macromolecules - Volume 108, March 2018, Pages 642-649
Journal: International Journal of Biological Macromolecules - Volume 108, March 2018, Pages 642-649
نویسندگان
Deepti Yadav, Bibhuti Ranjan, Nokuthula Mchunu, Marilize Le Roes-Hill, Tukayi Kudanga,