用于调节细胞色素P450HI中底物偏好性的通道工程
Tunnel engineering for modulating the substrate preference in cytochrome P450HI.
作者信息
Meng Shuaiqi, An Ruipeng, Li Zhongyu, Schwaneberg Ulrich, Ji Yu, Davari Mehdi D, Wang Fang, Wang Meng, Qin Meng, Nie Kaili, Liu Luo
机构信息
Beijing Bioprocess Key Laboratory, Beijing University of Chemical Technology, Beijing, 100029, People's Republic of China.
Institute of Biotechnology, RWTH Aachen University, Worringerweg 3, 52074, Aachen, Germany.
出版信息
Bioresour Bioprocess. 2021 Apr 3;8(1):26. doi: 10.1186/s40643-021-00379-1.
An active site is normally located inside enzymes, hence substrates should go through a tunnel to access the active site. Tunnel engineering is a powerful strategy for refining the catalytic properties of enzymes. Here, P450HI (Q85H/V170I) derived from hydroxylase P450 from Bacillus subtilis was chosen as the study model, which is reported as a potential decarboxylase. However, this enzyme showed low decarboxylase activity towards long-chain fatty acids. Here, a tunnel engineering campaign was performed for modulating the substrate preference and improving the decarboxylation activity of P450HI. The finally obtained BsβHI-F79A variant had a 15.2-fold improved conversion for palmitic acid; BsβHI-F173V variant had a 3.9-fold improved conversion for pentadecanoic acid. The study demonstrates how the substrate preference can be modulated by tunnel engineering strategy.
活性位点通常位于酶内部，因此底物需通过一条通道才能到达活性位点。通道工程是优化酶催化特性的一种有效策略。在此，选用源自枯草芽孢杆菌羟化酶P450的P450HI（Q85H/V170I）作为研究模型，据报道它是一种潜在的脱羧酶。然而，该酶对长链脂肪酸显示出较低的脱羧酶活性。在此，开展了一项通道工程研究，以调节底物偏好并提高P450HI的脱羧活性。最终获得的BsβHI-F79A变体对棕榈酸的转化率提高了15.2倍；BsβHI-F173V变体对十五烷酸的转化率提高了3.9倍。该研究展示了如何通过通道工程策略来调节底物偏好。
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