@misc{meng_an_li_schwaneberg_ji_davari_fang_wang_qin_nie_et al._2020, 
  title={Tunnel Engineering for Modulating The Substrate Preference in Decarbonylase P450BsβHI}, 
  DOI={10.21203/rs.3.rs-132813/v1}, 
  abstractNote={<jats:title>Abstract</jats:title>
        <jats:p>An active site normally locates inside of enzymes, substrates should go through the tunnel to access the active site. Tunnel engineering is a powerful strategy for refining the catalytic properties of enzymes. Here, P450<jats:sub>Bsβ</jats:sub>HI (Q85H/ V170I) derived from hydroxylase P450<jats:sub>Bsβ</jats:sub> from <jats:italic>Bacillus subtilis</jats:italic> was chosen as study model, which is reported as a potential decarbonylase. 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 decarbonylase activity of P450<jats:sub>Bsβ</jats:sub>HI. 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.</jats:p>}, 
  publisher={Research Square}, 
  author={Meng, Shuaiqi and An, Ruipeng and Li, Zhongyu and Schwaneberg, Ulrich and Ji, Yu and Davari, Mehdi D. and Fang, Wang and Wang, Meng and Qin, Meng and Nie, Kaili and et al.}, 
  year={2020}, 
  month={Dec}
  }