This WangReadme.txt file was updated on 2017-05-17 by Laura Seifert ------------------- GENERAL INFORMATION ------------------- Title of Dataset: Supporting Data for “Engineering of a Highly Efficient Escherichia coli Strain for Mevalonate Fermentation through Chromosomal Integration” Author Information: Principal Investigator Contact Information Name: Kechun Zhang Institution: University of Minnesota Address: Department of Chemical Engineering and Materials Science, 421 Washington Ave SE, Minneapolis, Minnesota 55455 Email: kzhang@umn.edu Associate or Co-investigator Contact information Name: Jilong Wang Institution: University of Minnesota Address: Department of Chemical Engineering and Materials Science, 421 Washington Ave SE, Minneapolis, MN 55455 Email: wang5708@umn.edu Associate or Co-investigator Contact Information Name: Suthamat Niyompanich Institution: Srinakharinwirot University Address: Department of Biology, Faculty of Science, Bangkok, Thailand Email: suthamat@g.swu.ac.th Associate or Co-investigator Contact Information Name: Yi-Shu Tai Institution: University of Minnesota Address: Department of Chemical Engineering and Materials Science, 421 Washington Ave SE, Minneapolis, Minnesota 55455 Email: taixx035@umn.edu Associate or Co-investigator Contact Information Name: Jingyu Wang Institution: University of Minnesota Address: Department of Chemical Engineering and Materials Science, 421 Washington Ave SE, Minneapolis, Minnesota 55455 Email: wang4968@umn.edu Associate or Co-investigator Contact Information Name: Prithviraj Mahida Institution: University of Minnesota Address: Department of Chemical Engineering and Materials Science, 421 Washington Ave SE, Minneapolis, Minnesota 55455 Email: mahid002@umn.edu Associate or Co-investigator Contact Information Name: Tuo Gao Institution: University of Minnesota Address: Department of Chemical Engineering and Materials Science, 421 Washington Ave SE, Minneapolis, Minnesota 55455 Email: gaoxx381@umn.edu Date of data collection (single date, range, approximate date): 2016 Geographic location of data collection (where was data collected?): University of Minnesota Information about funding sources that supported the collection of the data: This research was supported by a grant from the National Science Foundation through the Center for Sustainable Polymers (grant CHE-1413862). -------------------------- SHARING/ACCESS INFORMATION -------------------------- 1. Licenses/restrictions placed on the data: None 2. Links to publications that cite or use the data: Wang, J., Niyompanich, S. Tai, Y.-S., Wang, J., Bai, W., Mahida, P., Gao, T., Zhang, K. Appl. Environ. Microbiol. 2016, 82, 7176-7184. DOI: 10.1128/AEM.02178-16 3. Links to other publicly accessible locations of the data: N/A 4. Links/relationships to ancillary data sets: None 5. Was data derived from another source? No 6. Recommended citation for the data: Zhang, Kechun; Wang, Jilong; Niyompanich, Suthamat; Tai, Yi-Shu; Wang, Jingyu; Mahida, Prithviraj; Gao, Tuo. (2017). Supporting Data for “Engineering of a Highly Efficient Escherichia coli Strain for Mevalonate Fermentation through Chromosomal Integration”. Retrieved from the Data Repository for the University of Minnesota, https://doi.org/10.13020/D6W89N. --------------------- DATA & FILE OVERVIEW --------------------- Purpose Statement: This document describes the high efficient production of mevalonate in engineered bacteria. Metabolic engineering is employed to integrate heterologous mevalonate pathway into Escherichia coli. Subsequent optimization strategies improve the productivity and also re-direct the distribution of carbon flux to synthesize more mevalonate. Finally, the engineered strain CMEV-7 exhibits both high maximal productivity (~1.01 g/liter/h) and high yield (86.1% of the maximum theoretical yield, 30 g/liter mevalonate from 61 g/liter glucose after 48 h in a shake flask). This work provides an example of how to tune the carbon flux for the optimal production of exogenous chemicals. General Notes: The files below includes the pathway design (Fig. 1) and the bioengineering experimental data. Figure numbers in associated paper also include raw experimental data. The concentrations of feedstock (glucose), intermediate (acetate and pyruvate), and product (mevalonate) were measured by high-performance liquid chromatography (HPLC). Files Contained: Fig 1 new pathway 5082015.cdx Short description: This figure describes the metabolic pathway design. It summarizes all the bioengineering strategies used in this work Chemdraw file. CDX files can be opened with proprietary ChemDraw software. The current version as of 20170517 is ChemDraw 12.0, distributed by CambridgeSoft. Fig 2 edit 7072015.xlsx Short description: Comparing to the plasmid-based expression of the mevalonate pathway, the chromosomal integrations of the mevalonate pathway produce more mevalonate. Excel file Fig 3 edit 7072015.xlsx Short description: To improve the productivity, the glycolytic flux which is the first module for mevalonate production, is enhanced by deletion of the atpFH genes. The productivity is increased from 0.43 to 0.92 g/liter/h. Excel file Fig 4 edit 7072015.xlsx Short description: To enhance the downstream pathway from acetyl-CoA to mevalonate, two copies of the mevalonate synthetic operon are integrated into the chromosome of E. coli. Excel file Fig 5 edit 7072015.xlsx Short description: Fed-batch fermentation by feeding the bacteria with more glucose shows that the engineered strains produce more mevalonate. Excel file Fig 6 edit 7072015.xlsx Short Description: Figure 6A shows that the best engineered strain CMEV-7 exhibits high yield (0.5 g/g glucose) in fed-batch fermentation. Figure 6B shows that further improvement of the productivity is possible, since the intermediate, pyruvate, is accumulated during the fermentation. Excel file File Relationships / Notes: Figure 1 summarizes the designed metabolic pathway and all the optimization strategies applied in this work. Figure 2 to Figure 3 show the sequential bioengineering experiments based on the optimization strategies.