Zymomonas mobilis metabolism: Novel tools and targets for its rational engineering

Kalnenieks, Uldis; Pappas, Katherine M.; Bettenbrock, Katja

doi:10.1016/bs.ampbs.2020.08.001

Cited by 9 publications

(9 citation statements)

References 181 publications

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“…In regard to native producers, volumetric productivities of 4.69 g L −1 h −1 [ 25 ] and 7.7 g L −1 h −1 [ 26 ] were achieved by G. oxydans and P. sacchari , respectively. Although higher than that obtained by Z. mobilis , this bacterium may present some advantages that can be exploited for the value-added materials production, such as the presence of an energetically uncoupled metabolism with high catabolic flux rates and the small fraction of substrate that is converted into biomass [ 50 ]. Based on the performance of ZM4 BX for xylonic acid production, this strain was chosen for further analysis.…”

Section: Resultsmentioning

confidence: 99%

Engineering Zymomonas mobilis for the Production of Xylonic Acid from Sugarcane Bagasse Hydrolysate

et al. 2021

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Sugarcane bagasse is an agricultural residue rich in xylose, which may be used as a feedstock for the production of high-value-added chemicals, such as xylonic acid, an organic acid listed as one of the top 30 value-added chemicals on a NREL report. Here, Zymomonas mobilis was engineered for the first time to produce xylonic acid from sugarcane bagasse hydrolysate. Seven coding genes for xylose dehydrogenase (XDH) were tested. The expression of XDH gene from Paraburkholderia xenovorans allowed the highest production of xylonic acid (26.17 ± 0.58 g L−1) from 50 g L−1 xylose in shake flasks, with a productivity of 1.85 ± 0.06 g L−1 h−1 and a yield of 1.04 ± 0.04 gAX/gX. Deletion of the xylose reductase gene further increased the production of xylonic acid to 56.44 ± 1.93 g L−1 from 54.27 ± 0.26 g L−1 xylose in a bioreactor. Strain performance was also evaluated in sugarcane bagasse hydrolysate as a cheap feedstock, which resulted in the production of 11.13 g L−1 xylonic acid from 10 g L−1 xylose. The results show that Z. mobilis may be regarded as a potential platform for the production of organic acids from cheap lignocellulosic biomass in the context of biorefineries.

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Section: Resultsmentioning

confidence: 99%

Engineering Zymomonas mobilis for the Production of Xylonic Acid from Sugarcane Bagasse Hydrolysate

et al. 2021

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“…Z. mobilis is facultative anaerobic and unique in using the Entner-Doudoroff pathway preferably anaerobically (Swings & De Ley, 1977), resulting in a titer, relative productivity, and yield with respect to ethanol far surpassing those of other ethanologenic microbes (Rogers et al, 1982). However, limitations in the usability and efficiency of genetic tools are still holding back the utilization of Z. mobilis in industrial applications (Kalnenieks et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

“…Z. mobilis is facultative anaerobic and unique in using the Entner–Doudoroff pathway preferably anaerobically (Swings & De Ley, 1977 ), resulting in a titer, relative productivity, and yield with respect to ethanol far surpassing those of other ethanologenic microbes (Rogers et al., 1982 ). However, limitations in the usability and efficiency of genetic tools are still holding back the utilization of Z. mobilis in industrial applications (Kalnenieks et al., 2020 ).The wild type strains are limited in their substrate and product range (Swings & De Ley, 1977 ) and in their tolerance to inhibitory substances (Franden et al., 2009 ), characteristics that should be altered through changes on the genetic level to achieve economically viable processes by metabolic engineering.…”

Section: Introductionmentioning

confidence: 99%

Construction and comparison of different vehicles for heterologous gene expression in Zymomonas mobilis

Behrendt,

Vlachonikolou,

Tietgens

et al. 2024

Microbial Biotechnology

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Zymomonas mobilis has the potential to be an optimal chassis for the production of bulk chemicals derived from pyruvate. However, a lack of available standardized and characterized genetic tools hinders both efficient engineering of Z. mobilis and progress in basic research on this organism. In this study, a series of different shuttle vectors were constructed based on the replication mechanisms of the native Z. mobilis plasmids pZMO1, pZMOB04, pZMOB05, pZMOB06, pZMO7 and p29191_2 and on the broad host range replication origin of pBBR1. These plasmids as well as genomic integration sites were characterized for efficiency of heterologous gene expression, stability without selection and compatibility. We were able to show that a wide range of expression levels could be achieved by using different plasmid replicons. The expression levels of the constructs were consistent with the relative copy numbers, as determined by quantitative PCR. In addition, most plasmids are compatible and could be combined. To avoid plasmid loss, antibiotic selection is required for all plasmids except the pZMO7‐based plasmid, which is stable also without selection pressure. Stable expression of reporter genes without the need for selection was also achieved by genomic integration. All modules were adapted to the modular cloning toolbox Zymo‐Parts, allowing easy reuse and combination of elements. This work provides an overview of heterologous gene expression in Z. mobilis and adds a rich set of standardized genetic elements to an efficient cloning system, laying the foundation for future engineering and research in this area.

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“…It can rely on the unique Entner Doudoroff (ED) pathway to anaerobically metabolize glucose to produce ethanol. Additionally, this ethanologenic bacterium has some desirable industrial characteristics, such as a high ethanol titer, high specific glucose uptake rate, high ethanol tolerance of 16% ( v / v ), and a broad pH range (3.5–7.5, especially low pH), which makes it an excellent chassis for the production of biochemicals [ 2 , 3 , 4 , 5 ]. Currently, different CRISPR–Cas systems have been established in Z. mobilis that facilitate gene editing manipulations [ 6 , 7 , 8 , 9 ].…”

Section: Introductionmentioning

confidence: 99%

Characterization and Application of the Sugar Transporter Zmo0293 from Zymomonas mobilis

Zhang

Qin

et al. 2023

IJMS

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Zymomonas mobilis is a natural ethanologen with many desirable characteristics, which makes it an ideal industrial microbial biocatalyst for the commercial production of desirable bioproducts. Sugar transporters are responsible for the import of substrate sugars and the conversion of ethanol and other products. Glucose-facilitated diffusion protein Glf is responsible for facilitating the diffusion of glucose uptake in Z. mobilis. However, another sugar transporter-encoded gene, ZMO0293, is poorly characterized. We employed gene deletion and heterologous expression mediated by the CRISPR/Cas method to investigate the role of ZMO0293. The results showed that deletion of the ZMO0293 gene slowed growth and reduced ethanol production and the activities of key enzymes involved in glucose metabolism in the presence of high concentrations of glucose. Moreover, ZMO0293 deletion caused different transcriptional changes in some genes of the Entner Doudoroff (ED) pathway in the ZM4-ΔZM0293 strain but not in ZM4 cells. The integrated expression of ZMO0293 restored the growth of the glucose uptake-defective strain Escherichia coli BL21(DE3)-ΔptsG. This study reveals the function of the ZMO0293 gene in Z. mobilis in response to high concentrations of glucose and provides a new biological part for synthetic biology.

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Zymomonas mobilis metabolism: Novel tools and targets for its rational engineering

Cited by 9 publications

References 181 publications

Engineering Zymomonas mobilis for the Production of Xylonic Acid from Sugarcane Bagasse Hydrolysate

Engineering Zymomonas mobilis for the Production of Xylonic Acid from Sugarcane Bagasse Hydrolysate

Construction and comparison of different vehicles for heterologous gene expression in Zymomonas mobilis

Characterization and Application of the Sugar Transporter Zmo0293 from Zymomonas mobilis

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