Catabolic Division of Labor Enhances Production of D-Lactate and Succinate From Glucose-Xylose Mixtures in Engineered Escherichia coli Co-culture Systems
Although biological upgrading of lignocellulosic sugars represents a promising and sustainable route to bioplastics, diverse and variable feedstock compositions (e.g., glucose from the cellulose fraction and xylose from the hemicellulose fraction) present several complex challenges. Specifically, su...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-05-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fbioe.2020.00329/full |
| _version_ | 1818385528998854656 |
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| author | Andrew D. Flores Hyun G. Choi Rodrigo Martinez Moses Onyeabor E. Zeynep Ayla Amanda Godar Michael Machas David R. Nielsen Xuan Wang |
| author_facet | Andrew D. Flores Hyun G. Choi Rodrigo Martinez Moses Onyeabor E. Zeynep Ayla Amanda Godar Michael Machas David R. Nielsen Xuan Wang |
| author_sort | Andrew D. Flores |
| collection | DOAJ |
| description | Although biological upgrading of lignocellulosic sugars represents a promising and sustainable route to bioplastics, diverse and variable feedstock compositions (e.g., glucose from the cellulose fraction and xylose from the hemicellulose fraction) present several complex challenges. Specifically, sugar mixtures are often incompletely metabolized due to carbon catabolite repression while composition variability further complicates the optimization of co-utilization rates. Benefiting from several unique features including division of labor, increased metabolic diversity, and modularity, synthetic microbial communities represent a promising platform with the potential to address persistent bioconversion challenges. In this work, two unique and catabolically orthogonal Escherichia coli co-cultures systems were developed and used to enhance the production of D-lactate and succinate (two bioplastic monomers) from glucose–xylose mixtures (100 g L–1 total sugars, 2:1 by mass). In both cases, glucose specialist strains were engineered by deleting xylR (encoding the xylose-specific transcriptional activator, XylR) to disable xylose catabolism, whereas xylose specialist strains were engineered by deleting several key components involved with glucose transport and phosphorylation systems (i.e., ptsI, ptsG, galP, glk) while also increasing xylose utilization by introducing specific xylR mutations. Optimization of initial population ratios between complementary sugar specialists proved a key design variable for each pair of strains. In both cases, ∼91% utilization of total sugars was achieved in mineral salt media by simple batch fermentation. High product titer (88 g L–1 D-lactate, 84 g L–1 succinate) and maximum productivity (2.5 g L–1 h–1 D-lactate, 1.3 g L–1 h–1 succinate) and product yield (0.97 g g-total sugar–1 for D-lactate, 0.95 g g-total sugar–1 for succinate) were also achieved. |
| first_indexed | 2024-12-14T03:39:36Z |
| format | Article |
| id | doaj.art-03cdc8bbc00a4643a4a7c3322715246e |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-12-14T03:39:36Z |
| publishDate | 2020-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-03cdc8bbc00a4643a4a7c3322715246e2022-12-21T23:18:30ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852020-05-01810.3389/fbioe.2020.00329530658Catabolic Division of Labor Enhances Production of D-Lactate and Succinate From Glucose-Xylose Mixtures in Engineered Escherichia coli Co-culture SystemsAndrew D. Flores0Hyun G. Choi1Rodrigo Martinez2Moses Onyeabor3E. Zeynep Ayla4Amanda Godar5Michael Machas6David R. Nielsen7Xuan Wang8Chemical Engineering, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, AZ, United StatesSchool of Life Sciences, Arizona State University, Tempe, AZ, United StatesSchool of Life Sciences, Arizona State University, Tempe, AZ, United StatesSchool of Life Sciences, Arizona State University, Tempe, AZ, United StatesChemical Engineering, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, AZ, United StatesSchool of Life Sciences, Arizona State University, Tempe, AZ, United StatesChemical Engineering, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, AZ, United StatesChemical Engineering, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, AZ, United StatesSchool of Life Sciences, Arizona State University, Tempe, AZ, United StatesAlthough biological upgrading of lignocellulosic sugars represents a promising and sustainable route to bioplastics, diverse and variable feedstock compositions (e.g., glucose from the cellulose fraction and xylose from the hemicellulose fraction) present several complex challenges. Specifically, sugar mixtures are often incompletely metabolized due to carbon catabolite repression while composition variability further complicates the optimization of co-utilization rates. Benefiting from several unique features including division of labor, increased metabolic diversity, and modularity, synthetic microbial communities represent a promising platform with the potential to address persistent bioconversion challenges. In this work, two unique and catabolically orthogonal Escherichia coli co-cultures systems were developed and used to enhance the production of D-lactate and succinate (two bioplastic monomers) from glucose–xylose mixtures (100 g L–1 total sugars, 2:1 by mass). In both cases, glucose specialist strains were engineered by deleting xylR (encoding the xylose-specific transcriptional activator, XylR) to disable xylose catabolism, whereas xylose specialist strains were engineered by deleting several key components involved with glucose transport and phosphorylation systems (i.e., ptsI, ptsG, galP, glk) while also increasing xylose utilization by introducing specific xylR mutations. Optimization of initial population ratios between complementary sugar specialists proved a key design variable for each pair of strains. In both cases, ∼91% utilization of total sugars was achieved in mineral salt media by simple batch fermentation. High product titer (88 g L–1 D-lactate, 84 g L–1 succinate) and maximum productivity (2.5 g L–1 h–1 D-lactate, 1.3 g L–1 h–1 succinate) and product yield (0.97 g g-total sugar–1 for D-lactate, 0.95 g g-total sugar–1 for succinate) were also achieved.https://www.frontiersin.org/article/10.3389/fbioe.2020.00329/fulldivision of laborco-culturebiomass conversionlactatesuccinate |
| spellingShingle | Andrew D. Flores Hyun G. Choi Rodrigo Martinez Moses Onyeabor E. Zeynep Ayla Amanda Godar Michael Machas David R. Nielsen Xuan Wang Catabolic Division of Labor Enhances Production of D-Lactate and Succinate From Glucose-Xylose Mixtures in Engineered Escherichia coli Co-culture Systems Frontiers in Bioengineering and Biotechnology division of labor co-culture biomass conversion lactate succinate |
| title | Catabolic Division of Labor Enhances Production of D-Lactate and Succinate From Glucose-Xylose Mixtures in Engineered Escherichia coli Co-culture Systems |
| title_full | Catabolic Division of Labor Enhances Production of D-Lactate and Succinate From Glucose-Xylose Mixtures in Engineered Escherichia coli Co-culture Systems |
| title_fullStr | Catabolic Division of Labor Enhances Production of D-Lactate and Succinate From Glucose-Xylose Mixtures in Engineered Escherichia coli Co-culture Systems |
| title_full_unstemmed | Catabolic Division of Labor Enhances Production of D-Lactate and Succinate From Glucose-Xylose Mixtures in Engineered Escherichia coli Co-culture Systems |
| title_short | Catabolic Division of Labor Enhances Production of D-Lactate and Succinate From Glucose-Xylose Mixtures in Engineered Escherichia coli Co-culture Systems |
| title_sort | catabolic division of labor enhances production of d lactate and succinate from glucose xylose mixtures in engineered escherichia coli co culture systems |
| topic | division of labor co-culture biomass conversion lactate succinate |
| url | https://www.frontiersin.org/article/10.3389/fbioe.2020.00329/full |
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