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Một degron C-đầu mới được xác định trong các aldehyde decarbonylase vi khuẩn bằng phương pháp tiến hóa có định hướng
Tóm tắt
Các aldehyde decarbonylase (AD) chuyển đổi aldehyde acyl thành alkan, cung cấp giải pháp hứa hẹn để sản xuất alkan từ nguyên liệu tái tạo. Tuy nhiên, sự không ổn định của các AD cản trở ứng dụng tiếp theo của chúng. Do đó, nghiên cứu hiện tại nhằm điều tra cơ chế phân hủy của các AD và cải tiến chúng để có độ ổn định cao. Tại đây, chúng tôi mô tả sự phát hiện một nhãn phân hủy (degron) trong AD từ vi khuẩn xanh Prochlorococcus marinus bằng cách sử dụng hệ thống tiến hóa có định hướng dựa trên PCR dễ mắc lỗi. Phân tích bioinformatics cho thấy degron C-đầu này là phổ biến trong các AD vi khuẩn và xác định một mô hình C-đầu bảo tồn, RMSAYGLAAA, đại diện cho degron AD (ADcon). Hơn nữa, chúng tôi đã chứng minh rằng các protease phụ thuộc ATP ClpAP và Lon tham gia vào sự phân hủy của các protein gắn nhãn AD trong E. coli, từ đó hạn chế sản xuất alkan. Việc xóa bỏ hoặc chỉnh sửa mô hình degron đã tăng cường sản xuất alkan trong điều kiện in vivo. Công việc này đã tiết lộ sự hiện diện của một degron mới trong các AD vi khuẩn có trách nhiệm cho sự không ổn định của chúng. Các thí nghiệm in vivo đã chứng minh rằng việc loại bỏ hoặc chỉnh sửa degron có thể ổn định AD, từ đó tạo ra lượng alkan cao hơn.
Từ khóa
#aldehyde decarbonylases #degradation mechanism #stability engineering #renewable feedstock #directed evolution #bioinformaticsTài liệu tham khảo
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