Recombinant Lipase Engineered with Amphipathic and Coiled-Coil Peptides

详细信息    查看全文

• 作者：Kyung Seok Yang ; Bong Hyun Sung ; Myung Keun Park ; Jun Hyoung Lee ; Ki Jung Lim ; Sung Chul Park ; Soo Jin Kim ; Hyung Kwoun Kim ; Jung-Hoon Sohn ; Ho Min Kim ; Sun Chang Kim
• 刊名：ACS Catalysis
• 出版年：2015
• 出版时间：September 4, 2015
• 年：2015
• 卷：5
• 期：9
• 页码：5016-5025
• 全文大小：469K
• ISSN：2155-5435

文摘

Lipases have been utilized industrially to produce biodiesel, oleochemicals, and pharmaceuticals. Many efforts such as metagenomics, directed evolution, and enzyme immobilization have been devoted to enhance the lipase activity. Here, we designed a recombinant lipase, NKC-M37-MAT, that was generated by incorporating an N-terminal amphipathic peptide (NKC) and a C-terminal coiled-coil peptide (MAT) into Photobacterium lipolyticum M37 lipase. The hydrophobic face of NKC improve the accessibility (K_m), and catalytic efficiency (K_cat/K_m) of the soluble lipase toward the hydrophobic substrate and tetrameric MAT further enhanced lipase catalytic activity (U/mg) through cooperative binding to its substrate such that the catalytic activity (U/mg) of NKC-M37-MAT was increased by a maximum of 54-fold compared with the wild-type, which decreased the biodiesel production time 5-fold from 30 to 6 h. This novel approach shows promise as a platform technology to increase lipase catalytic efficiency for industrial-scale production of biodiesel and biochemicals synthesized from hydrophobic substrates.