Purification of recombinant neutral protease (NPRC10) by partitioning inaqueous two-phase systems

 Nội dung tóm tắt: A study was made of the partition and purification of recombinant neutral protease (NPRC10) from a culture supernatant of E. coli BL21 (DE3) in the polyethylene glycol (PEG)/potassium phosphate aqueous two-phase systems. Factors that influenced the partition of the enzyme in these systems, including the PEG molecular weight and concentration, potassium phosphate concentration, incubation time and temperature, and pH were investigated. The optimum condition of the primary aqueous two-phase extraction was 35% PEG 6000/30% potassium phosphate (pH 6.5) at 30oC for 25 min. The partitioning coefficient for protease (Kprotease) was 4.69 with a partitioning yield (Y) of 94.93%, a purification fold (PF) of 1.7, and protease specific activity (SAprotease T) of 694.61 unit/mg protein in top phase. The protease, which was concentrated in the top phase, was further mixed with 35% potassium phosphate in combination with 3% potassium chloride at room temperature to elute the bottom phase (salt-rich phase). Using this step, the PF of enzyme achieved a higher value of 2.38 with a recovery yield of 76.17% and SAprotease B of 983.04 unit/mg protein.

 Thể loại: Không thuộc ISI/SCOPUS, có ISSN/ISBN

 Tác giả: Nguyễn Hoàng Lộc, Ho Thi Thu Lien, Hoàng Tấn Quảng

 Đơn vị: Phòng thí nghiệm Công nghệ Gen

 Đăng tại: European Journal of Experimental Biology

 Tập, (số), trang: 3, 2, 252-257

 Số ISSN/ISBN: 2248 –9215

 Chỉ số Impact factor:

 Điểm số theo QĐ của HĐ chức danh GS Nhà nước:

 Năm công bố: 2013

 File đính kèm:

 Số lần xem: 61

 Ngày cập nhật:

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