Todhanakasem, Tatsaporn

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Todhanakasem, Tatsaporn
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tatsaporn.to@kmitl.ac.th

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    Item type:Publication,
    Engineering Zymomonas mobilis for improving genetic transformation and stability of multi-gene biosynthetic pathways
    (2025-12-01)
    Huang, Yuhuan
    ;
    Wang, Xiaojie
    ;
    Chen, Mao
    ;
    Wu, Yanran
    ;
    Todhanakasem, Tatsaporn
    Zymomonas mobilis holds significant promise for metabolic engineering but suffers from inefficient transformation and instability of plasmids over 8 kb. In this study, an element library containing promoters and terminators was constructed by analyzing the promoter activities of Z. mobilis ZM4. Using these regulatory elements, the β-carotene gene cluster (crtEXYIB) and individual genes (crtE, crtXYI, crtB) were regulated to construct pEZ-crt1 and pEZ-crt2 plasmid series. Screening identified top-yielding strains CRT1-29 (0.93 mg/g DCW) and CRT2-22 (1.25 mg/g DCW). Additionally, three key genes determining both the genetic transformation and stability of exogenous plasmids were identified by inactivating the genes encoding Restriction-Modification (R-M) or DNA-repair systems in Z. mobilis ZM4. The knockout of mrr and hsdM not only enhanced the transformation efficiency of a 3.95-kb methylated regular plasmid but also enabled the successful electroporation of a 9.5-kb reporter plasmid harboring a β-carotene expression cassette, which could not be transformed into Z. mobilis ZM4 and Mrr via electroporation previously. Moreover, the deletion of tatD prompted the stability of exogenous plasmid DNAs. The resultant triple-deficiency mutant MHT (Δmrr-hsdM-tatD) maintained the β-carotene expression plasmid in Z. mobilis, producing 2.09 mg/g DCW of β-carotene. Our study will prompt the application of Z. mobilis in metabolic engineering and synthetic biology.
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    Economic co-production of cellulosic ethanol and microalgal biomass through efficient fixation of fermentation carbon dioxide
    (2024-03-01)
    Liu, Linpei
    ;
    Zhou, Zheng
    ;
    Gong, Guiping
    ;
    Wu, Bo
    ;
    Todhanakasem, Tatsaporn
    An integrated process for the co-production of cellulosic ethanol and microalgal biomass by fixing CO<inf>2</inf> generated from bioethanol fermentation is proposed. Specifically, over one-fifth of the fermentative carbon was converted into high-purity CO<inf>2</inf> during ethanol production. The optimal concentration of 4 % CO<inf>2</inf> was identified for the growth and metabolism of Chlorella sp. BWY-1. A multiple short-term intermittent CO<inf>2</inf> supply system was established to efficiently fix and recycle the waste CO<inf>2</inf>. Using this system, economical co-production of cellulosic ethanol by Zymomonas mobilis and microalgal biomass in biogas slurry wastewater was achieved, resulting in the production of ethanol at a rate of 0.4 g/L/h and a fixed fermentation CO<inf>2</inf> of 3.1 g/L/d. Moreover, the amounts of algal biomass and chlorophyll a increased by over 50 % and two-fold, respectively. Through techno-economic analysis, the integrated process demonstrated its cost-effectiveness for cellulosic ethanol production. This study presents an innovative approach to a low-carbon circular bioeconomy.