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. 2020 Oct 7;11:5035. doi: 10.1038/s41467-020-18833-7

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Construction of E. coli cells expressing lactonase, alcohol dehydrogenase (ADH2), aldehyde dehydrogenase (ALDH) and NADH oxidase (NOX). See Supplementary Fig. 3 for SDS-PAGE of whole-cell proteins of Module 3 expressed in E. coli. RSF: pRSFDuet-1; pBR322: pETDuet-1; arrow: T7 promoter. Pink filled rectangle: ADH2 gene; green filled rectangle: ALDH; blue filled rectangle: lactonase gene; purple filled rectangle: NOX gene. b Engineered E. coli cells containing enzyme module 3 (lactonase, ADH2 and ALDH, and NOX if necessary) for biotransformation of CL 4b to AA 7b. Blue column: AA. c Time course of E. coli (M3B_M3E)-catalyzed transformation of CL 4b to AA 7b at substrate concentration 100 mM. Blue line: AA; orange line: 6-HHA; green line: CL. d Time course of E. coli (M3B_M3E)-catalyzed transformation of CL 4b to AA 7b, with substrate added in three portions (200 mM in the beginning, and 200 mM and 100 mM indicated by arrows) for a total concentration of 500 mM with pH maintained by adjustment. Blue line: AA; orange line: 6-HHA; green line: CL. The data shown in b, c, and d are presented as mean value ± SD (standard deviations) of three biological replicates. Source data are provided as a Source Data file.