Generation of Stable, Light-Driven Co-cultures of Cyanobacteria with Heterotrophic Microbes

Singh, Amit Kumar; Ducat, Daniel C.

doi:10.1007/978-1-0716-1791-5_16

Cited by 4 publications

(5 citation statements)

References 31 publications

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“…Two of the most used are barium and calcium. Barium generally produces stronger gels and has a stronger binding a nity to alginate compared than calcium and has been used in making hydrogels capsulating cyanobacteria 10,27,28 . However, barium, in its soluble form, is highly toxic 29 .…”

Section: Resultsmentioning

confidence: 99%

“…For example, arti cial consortium of sucrose secreting S. elongatus 2973 and 3-hydroxypropionic acid (3-HP) producing E. coli was constructed where relatively low light intensity was used to reduce oxidative stresses, compromising the sucrose production rate 7 . Therefore, stable and high productive consortia are di cult to optimize unless the two species form a mutualistic growth relationship 10 .…”

Section: Introductionmentioning

confidence: 99%

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Photobiological production of high-value pigments via compartmentalized co-cultures using Ca-alginate hydrogels

Zhao¹,

Sengupta

Tan³

et al. 2022

Preprint

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Synechococcus elongatus UTEX 2973 can use light and CO2 to produce sucrose, making them promising candidates to construct cocultures with heterotrophic workhorses. This envisioned process is, however, challenging to implement because of photosynthetic oxidative stress, light shading effect by heterotrophic cells, degradation of light sensitive metabolites, and high cost to separate intracellular products. Here, we demonstrated an effective ecosystem, where the sucrose producing cyanobacterium was freely grown in photo-bioreactors (PBRs), while an engineered heterotrophic workhorse (β-carotene producing Yarrowia lipolytica or indigoidine producing Pseudomonas putida) was encapsulated in calcium-alginate hydrogel beads and then placed inside the PBRs. The compartmentalization by hydrogels prevented growth interference so that the cyanobacterial culture could reach high sucrose concentrations, resulting the production of indigoidine (7.5g/L hydrogel) and β-carotene (1.3g/L hydrogel), respectively (i.e., the titers were 15 ~ 22 folds higher than that in the free cell coculture). Moreover, 13C-metabolic analysis indicated that hydrogels provided a favorable microenvironment so that the flux network of cells inside hydrogel was similar to the free culture. Finally, this novel system allowed the heterotroph- containing hydrogel beads to be easily harvested and dissolved by an EDTA solution for product and cell recovery, while the cyanobacterial culture could be continuously used for growing the next batch of immobilized workhorse heterotrophs.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Photobiological production of high-value pigments via compartmentalized co-cultures using Ca-alginate hydrogels

Zhao¹,

Sengupta

Tan³

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…Furthermore, the stability of light-driven co-cultures can be improved by using hydrogel systems to ensure the spaceefficient structure of the consortium (Gao et al, 2022). As the consortia-related methodological toolbox continues to expand (Ma et al, 2022;Singh and Ducat, 2022), the importance of the "phototroph + heterotroph" designs based on the conversion of CO 2 to organic chemicals can be expected to grow.…”

Section: Introductionmentioning

confidence: 99%

Construction of an artificial consortium of Escherichia coli and cyanobacteria for clean indirect production of volatile platform hydrocarbons from CO2

et al. 2022

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Ethylene and isoprene are essential platform chemicals necessary to produce polymers and materials. However, their current production methods based on fossil fuels are not very efficient and result in significant environmental pollution. For a successful transition more sustainable economic model, producing these key polymeric building blocks from renewable and sustainable resources such as biomass or CO2 is essential. Here, inspired by the symbiotic relationship of natural microbial communities, artificial consortia composed of E. coli strains producing volatile platform chemicals: ethylene and isoprene and two strains of cyanobacteria phototrophically synthesizing and exporting sucrose to feed these heterotrophs were developed. Disaccharide produced by transgenic cyanobacteria was used as a carbon and electron shuttle between the two community components. The E. coli cscB gene responsible for sucrose transport was inserted into two cyanobacterial strains, Thermosynechococcus elongatus PKUAC-SCTE542 and Synechococcus elongatus PCC7942, resulting in a maximal sucrose yield of 0.14 and 0.07 g/L, respectively. These organisms were co-cultured with E. coli BL21 expressing ethylene-forming enzyme or isoprene synthase and successfully synthesized volatile hydrocarbons. Productivity parameters of these co-cultures were higher than respective transgenic cultures of E. coli grown individually at similar sucrose concentrations, highlighting the positive impact of the artificial consortia on the production of these platform chemicals.

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“…Two of the most used are barium and calcium. Barium produces stronger hydrogel and has a stronger binding affinity to alginate than calcium, and it has been used in making hydrogels to encapsulate cyanobacteria 10,21,22 . However, barium in its soluble form is highly toxic 23 .…”

mentioning

confidence: 99%

“…Complementing these workhorse microbes, S. elongatus UTEX 2973 has the highest sucrose secretion capability (1.9 g/L/day) of all cyanobacterial hosts. In our setup the autotrophic strain in the co-culture provides the sugar feedstock for the heterotrophic workhorses 7,10 . However, for fast and efficient photosynthesis, UTEX 2973 requires high light intensity (> 800 μmol photons/m 2 /s) and high dissolved CO 2 (3%) conditions 28,29 .…”

mentioning

confidence: 99%

Photobiological production of high-value pigments via compartmentalized co-cultures using Ca-alginate hydrogels

Zhao

Sengupta

Tan

et al. 2022

Sci Rep

View full text Add to dashboard Cite

Engineered cyanobacterium Synechococcus elongatus can use light and CO2 to produce sucrose, making it a promising candidate for use in co-cultures with heterotrophic workhorses. However, this process is challenged by the mutual stresses generated from the multispecies microbial culture. Here we demonstrate an ecosystem where S. elongatus is freely grown in a photo-bioreactor (PBR) containing an engineered heterotrophic workhorse (either β-carotene-producing Yarrowia lipolytica or indigoidine-producing Pseudomonas putida) encapsulated in calcium-alginate hydrogel beads. The encapsulation prevents growth interference, allowing the cyanobacterial culture to produce high sucrose concentrations enabling the production of indigoidine and β-carotene in the heterotroph. Our experimental PBRs yielded an indigoidine titer of 7.5 g/L hydrogel and a β-carotene titer of 1.3 g/L hydrogel, amounts 15–22-fold higher than in a comparable co-culture without encapsulation. Moreover, 13C-metabolite analysis and protein overexpression tests indicated that the hydrogel beads provided a favorable microenvironment where the cell metabolism inside the hydrogel was comparable to that in a free culture. Finally, the heterotroph-containing hydrogels were easily harvested and dissolved by EDTA for product recovery, while the cyanobacterial culture itself could be reused for the next batch of immobilized heterotrophs. This co-cultivation and hydrogel encapsulation system is a successful demonstration of bioprocess optimization under photobioreactor conditions.

show abstract

Generation of Stable, Light-Driven Co-cultures of Cyanobacteria with Heterotrophic Microbes

Cited by 4 publications

References 31 publications

Photobiological production of high-value pigments via compartmentalized co-cultures using Ca-alginate hydrogels

Photobiological production of high-value pigments via compartmentalized co-cultures using Ca-alginate hydrogels

Construction of an artificial consortium of Escherichia coli and cyanobacteria for clean indirect production of volatile platform hydrocarbons from CO2

Photobiological production of high-value pigments via compartmentalized co-cultures using Ca-alginate hydrogels

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