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Backbone and Ile-δ1, Leu, Val methyl 1H, 15N, and 13C, chemical shift assignments for Rhizopus chinensis lipase

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Lipase r27RCL is a 296-residue, 33 kDa monomeric enzyme with high ester hydrolysis activity, which has significant applications in the baking, paper and leather industries. The lipase gene proRCL from Rhizopus microsporus var. chinensis (also Rhizopus chinensis) CCTCC M201021 was cloned as a fusion construct C-terminal to a maltose-binding protein (MBP) tag, and expressed as MBP-proRCL in an Escherichia coli BL21 trxB (DE3) expression system with uniform 2H,13C,15N-enrichment and Ile-δ1, Leu, and Val 13CH3 methyl labeling. The fusion protein was hydrolyzed by Kex2 protease at the recognition site Lys-Arg between residues −29 and −28 of the prosequence, producing the enzyme form called r27RCL. Here we report extensive backbone 1H, 15N, and 13C, as well as Ile-δ1, Leu, and Val side chain methyl, NMR resonance assignments for r27RCL.

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We thank Profs. J. Hunt and T. Szyperski for helpful discussions on the RCL lipase project. Financial support from the High-end Foreign Experts Recruitment Program (GDW20123200113), Six Talent Peaks Project in Jiangsu Province (NY-010), 333 Project in Jiangsu Province (BRA2015316), NSFC (31671799), and the 111 Project (111-2-06) are greatly appreciated. This work was also supported as a Community Outreach Project of the NIH NIGMS Protein Structure Initiative, Grant U54 GM094597.

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Correspondence to Xiao-Wei Yu, Yan Xu or Gaetano T. Montelione.

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Zhang, M., Yu, XW., Swapna, G.V.T. et al. Backbone and Ile-δ1, Leu, Val methyl 1H, 15N, and 13C, chemical shift assignments for Rhizopus chinensis lipase. Biomol NMR Assign 12, 63–68 (2018).

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