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Sử dụng tương tự protease Ubp1 để sản xuất hormon tăng trưởng người tái tổ hợp trong Escherichia coli
Tóm tắt
Nhiều phương pháp biểu hiện hormon tăng trưởng người (hGH) từ vi khuẩn dưới các điều kiện lên men và cảm ứng thông thường đã được mô tả. Mặc dù có những tiến bộ đáng kể trong lĩnh vực này trong vài năm qua, nhưng việc sản xuất hGH tái tổ hợp bằng cách sử dụng hệ thống biểu hiện tế bào vẫn yêu cầu tối ưu hơn nữa. Việc liên kết phân tử ubiquitin (Ub) với protein hGH đã cho phép tăng cường hiệu quả tổng hợp sinh học và cải thiện độ ổn định của protein. Ub là một protein bao gồm 76 dư lượng axit amin với khối lượng phân tử 8.6 kDa, được biểu hiện trong tất cả các eukaryote. Protein này là một phần của hệ thống sửa đổi protein phổ quát, không xảy ra ở vi khuẩn, và là một chất mang hữu ích cho các protein dị hợp thu được thông qua biểu hiện trong Escherichia coli. Quá trình tinh sạch các protein liên hợp Ub dễ dàng hơn so với protein tái tổ hợp không liên hợp, và Ub có thể được loại bỏ bằng các protease giải ubiquitin (DUBs hoặc UBPs). Trong nghiên cứu hiện tại, protease UBPD2C, một tương tự ổn định của UBP1, đã được sản xuất như một protein tái tổ hợp trong E. coli và được sử dụng để sản xuất hormon tăng trưởng người tái tổ hợp (rhGH). hGH được biểu hiện như một protein liên hợp với Ub như một thẻ. Các phát hiện của chúng tôi cho thấy protease UBPD2C rất hiệu quả trong việc loại bỏ phần Ub khỏi hGH tái tổ hợp gắn Ub. Cách tiếp cận được mô tả cho phép thu được một năng suất đáng kể của rhGH với độ tinh khiết cần thiết cho các sản phẩm dược phẩm.
Từ khóa
#hormon tăng trưởng người #hGH #Escherichia coli #ubiquitin #protein tái tổ hợp #UBPD2C #protease #tinh sạch #sinh học phân tửTài liệu tham khảo
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