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. 2020 Feb 21;8:125. doi: 10.3389/fbioe.2020.00125

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Copyright © 2020 Lü, Ge, Cao, Guo, Liu, Gao, Xu and Ma.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Construction of B. licheniformis MW3 (ΔbudCΔgdh) for acetoin production. (A) Technology roadmap for acetoin production from glucose by B. licheniformis MW3 (ΔbudCΔgdh). ALS, α-acetolactate synthase; ALDC, α-acetolactate decarboxylase; BudC, meso-2,3-butanediol dehydrogenase; GDH, (2R,3R)-2,3-butanediol dehydrogenase; UAR, unidentified acetoin reductase (based on the results of gas chromatograph in Supplementary Figure S1, this enzyme produces meso-2,3-butanediol as the reduction product of acetoin). (B) Analysis of PCR fragments to confirm budC and gdh disruption. Lane M: molecular mass standard (DNA/HindIII); Lane 1: PCR product of budC amplified with B. licheniformis MW3 genomic DNA as the template; Lane 2: PCR product of gdh amplified with B. licheniformis MW3 genomic DNA as the template; Lane 3: PCR product of budC amplified with B. licheniformis MW3 (ΔbudCΔgdh) genomic DNA as the template; Lane 4: PCR product of gdh amplified with B. licheniformis MW3 (ΔbudCΔgdh) genomic DNA as the template. The PCRs were performed with primer pairs ΔbudC-f/ΔbudC-r and Δgdh-f/Δgdh-r, respectively.