Metabolic engineering of Clostridium tyrobutyricum for n-butanol production: effects of CoA transferase

Yu, Lean; Zhao, Jun; Xu, Mengmeng; Dong, Jie; Varghese, Saju; Yu, Mingrui; Tang, I‐Ching; Yang, Shang‐Tian

doi:10.1007/s00253-015-6566-5

Cited by 42 publications

(11 citation statements)

References 66 publications

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“…3A, Additional file Table S5A). Enough amounts of acetic acid and butyric acid accumulated, then they were re-assimilated and converted into acetyl-CoA and bytyryl-CoA, respectively, by CoA-transferase16. The corresponding ORFs of CoA-transferase (Cbei_2040, Cbei_3278, Cbei_3833 and Cbei_3834) showed different transcription.…”

Section: Resultsmentioning

confidence: 99%

Transcriptional analysis of degenerate strain Clostridium beijerinckii DG-8052 reveals a pleiotropic response to CaCO3-associated recovery of solvent production

Jiao

Zhang

Chen

et al. 2016

Sci Rep

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Degenerate Clostridium beijerinckii strain (DG-8052) can be partially recovered by supplementing CaCO3 to fermentation media. Genome resequencing of DG-8052 showed no general regulator mutated. This study focused on transcriptional analysis of DG-8052 and its response to CaCO3 treatment via microarray. The expressions of 5168 genes capturing 98.6% of C. beijerinckii NCIMB 8052 genome were examed. The results revealed that with addition of CaCO3 565 and 916 genes were significantly up-regulated, and 704 and 1044 genes significantly down-regulated at acidogenic and solventogenic phase of DG-8052, respectively. These genes are primarily responsible for glycolysis to solvent/acid production (poR, pfo), solventogensis (buk, ctf, aldh, adh, bcd) and sporulation (spo0A, sigE, sigma-70, bofA), cell motility and division (ftsA, ftsK, ftsY, ftsH, ftsE, mreB, mreC, mreD, rodA), and molecular chaperones (grpE, dnaK, dnaJ, hsp20, hsp90), etc. The functions of some altered genes in DG-8052, totalling 5.7% at acidogenisis and 8.0% at sovlentogenisis, remain unknown. The response of the degenerate strain to CaCO3 was suggested significantly pleiotropic. This study reveals the multitude of regulatory function that CaCO3 has in clostridia and provides detailed insights into degeneration mechanisms at gene regulation level. It also enables us to develop effective strategies to prevent strain degeneration in future.

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Section: Resultsmentioning

confidence: 99%

Transcriptional analysis of degenerate strain Clostridium beijerinckii DG-8052 reveals a pleiotropic response to CaCO3-associated recovery of solvent production

Jiao

Zhang

Chen

et al. 2016

Sci Rep

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“…Additionally, it has been reported that acetone can be produced through non‐enzymatic breakdown of acetoacetate, suggesting that ctfAB , converting acetoacetyl‐CoA to acetoacetate, plays an important role in acetone generation (Han et al, ; Yu et al, ). Hence, transcription levels of ctfA , ctfB1 , and ctfB2 were compared in both xylose and xylan fed cultures through RT‐qPCR.…”

Section: Resultsmentioning

confidence: 99%

Direct conversion of xylan to butanol by a wild‐type Clostridium species strain G117

Yan

Basu

et al. 2016

Biotech & Bioengineering

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Lignocellulosic biomass has great potential for use as a carbon source for the production of second-generation biofuels by solventogenic bacteria. Here we describe the production of butanol by a newly discovered wild-type Clostridium species strain G117 with xylan as the sole carbon source for fermentation. Strain G117 produced 0.86 AE 0.07 g/L butanol and 53.4 AE 0.05 mL hydrogen directly from 60 g/L xylan provided that had undergone no prior enzymatic hydrolysis. After process optimization, the amount of butanol produced from xylan was increased to 1.24 AE 0.37 g/L. In contrast to traditional acetone-butanol-ethanol (ABE) solventogenic fermentation, xylan supported fermentation in strain G117 and negligible amount of acetone was produced. The expression of genes normally associated with acetone production (adc and ctfB2) were down-regulated compared to xylose fed cultures. This lack of acetone production may greatly simplify downstream separation process. Moreover, higher amount of butanol (2.94 g/L) was produced from 16.99 g/L xylo-oligosaccharides, suggesting a major role for strain G117 in butanol production from xylan and its oligosaccharides. The unique ability of strain G117 to produce a considerable amount of butanol directly from xylan without producing undesirable fermentation byproducts opens the door to the possibility of cost-effective biofuels production in a single step.

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“…Alternatively, a routine strategy for butyrate reassimilation in C. acetobutylicum and C. beijerinckii was used under solventogenesis. The strategy of overexpressing genes involved in the CoA transferase encoded by ctfA-ctfB (CA_p0163-0164 or Cbei_3833-3834) has been successfully verified in C. acetobutylicum, Clostridium tyrobutyricum, and C. beijerinckii (6,38,39). Both the adhE1 and ctfA-ctfB genes located in C. acetobutylicum's sol operon were introduced, followed by its adjacent adc gene (CA_p0165, with native promoter) (33), since there are no known genes involved in the acetoacetate metabolism of C. cellulovorans.…”

Section: Figmentioning

confidence: 99%

Improved n -Butanol Production from Clostridium cellulovorans by Integrated Metabolic and Evolutionary Engineering

Wen

Ledesma-Amaro

Lin

et al. 2019

Appl Environ Microbiol

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Clostridium cellulovorans DSM 743B offers potential as a chassis strain for biomass refining by consolidated bioprocessing (CBP). However, its n-butanol production from lignocellulosic biomass has yet to be demonstrated. This study demonstrates the construction of a coenzyme A (CoA)-dependent acetone-butanol-ethanol (ABE) pathway in C. cellulovorans by introducing adhE1 and ctfA-ctfB-adc genes from Clostridium acetobutylicum ATCC 824, which enabled it to produce n-butanol using the abundant and low-cost agricultural waste of alkali-extracted, deshelled corn cobs (AECC) as the sole carbon source. Then, a novel adaptive laboratory evolution (ALE) approach was adapted to strengthen the n-butanol tolerance of C. cellulovorans to fully utilize its n-butanol output potential. To further improve n-butanol production, both metabolic engineering and evolutionary engineering were combined, using the evolved strain as a host for metabolic engineering. The n-butanol production from AECC of the engineered C. cellulovorans was increased 138-fold, from less than 0.025 g/liter to 3.47 g/liter. This method represents a milestone toward n-butanol production by CBP, using a single recombinant clostridium strain. The engineered strain offers a promising CBP-enabling microbial chassis for n-butanol fermentation from lignocellulose. IMPORTANCE Due to a lack of genetic tools, Clostridium cellulovorans DSM 743B has not been comprehensively explored as a putative strain platform for n-butanol production by consolidated bioprocessing (CBP). Based on the previous study of genetic tools, strain engineering of C. cellulovorans for the development of a CBP-enabling microbial chassis was demonstrated in this study. Metabolic engineering and evolutionary engineering were integrated to improve the n-butanol production of C. cellulovorans from the low-cost renewable agricultural waste of alkali-extracted, deshelled corn cobs (AECC). The n-butanol production from AECC was increased 138-fold, from less than 0.025 g/liter to 3.47 g/liter, which represents the highest titer of n-butanol produced using a single recombinant clostridium strain by CBP reported to date. This engineered strain serves as a promising chassis for n-butanol production from lignocellulose by CBP.

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Metabolic engineering of Clostridium tyrobutyricum for n-butanol production: effects of CoA transferase

Cited by 42 publications

References 66 publications

Transcriptional analysis of degenerate strain Clostridium beijerinckii DG-8052 reveals a pleiotropic response to CaCO3-associated recovery of solvent production

Transcriptional analysis of degenerate strain Clostridium beijerinckii DG-8052 reveals a pleiotropic response to CaCO3-associated recovery of solvent production

Direct conversion of xylan to butanol by a wild‐type Clostridium species strain G117

Improved n -Butanol Production from Clostridium cellulovorans by Integrated Metabolic and Evolutionary Engineering

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