Phenotypic and genomic analysis of isopropanol and 1,3-propanediol producer Clostridium diolis DSM 15410

Sedlář, Karel; Vasylkivska, Maryna; Musilová, Jana; Branská, Barbora; Provazník, Ivo; Patáková, Petra

doi:10.1016/j.ygeno.2020.11.007

Cited by 11 publications

(14 citation statements)

References 76 publications

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“…Likewise, when grown on rhamnose, a decrease in spore formation was observed in C. beijerinckii cultures (Diallo et al 2018). Recently Sedlar et al observed that C. beijerinckii NCIMB 8052 and C. beijerinckii NRRLB 598 sporulated when glycerol was added to the medium instead of glucose (Sedlar et al 2021). Surprisingly the opposite was observed in C. beijerinckii (formerly C. diolis) DSM 15410 cultures.…”

Section: The Carbon Sourcementioning

confidence: 97%

“…Several studies (Awang et al 1992;Kolek et al 2017;Long et al 1983;Sedlar et al 2021;Woods and Jones 1987) have reported the substantial impact of medium composition on sporulation initiation and sporulation efficiency (number of spore generated and the spores that can germinate) and suggested a link between carbon source, mineral content, external pH, and the number of spores produced.…”

Section: Sporulation Triggersmentioning

confidence: 99%

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Sporulation in solventogenic and acetogenic clostridia

Diallo

Kengen

López-Contreras

2021

Appl Microbiol Biotechnol

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The Clostridium genus harbors compelling organisms for biotechnological production processes; while acetogenic clostridia can fix C1-compounds to produce acetate and ethanol, solventogenic clostridia can utilize a wide range of carbon sources to produce commercially valuable carboxylic acids, alcohols, and ketones by fermentation. Despite their potential, the conversion by these bacteria of carbohydrates or C1 compounds to alcohols is not cost-effective enough to result in economically viable processes. Engineering solventogenic clostridia by impairing sporulation is one of the investigated approaches to improve solvent productivity. Sporulation is a cell differentiation process triggered in bacteria in response to exposure to environmental stressors. The generated spores are metabolically inactive but resistant to harsh conditions (UV, chemicals, heat, oxygen). In Firmicutes, sporulation has been mainly studied in bacilli and pathogenic clostridia, and our knowledge of sporulation in solvent-producing or acetogenic clostridia is limited. Still, sporulation is an integral part of the cellular physiology of clostridia; thus, understanding the regulation of sporulation and its connection to solvent production may give clues to improve the performance of solventogenic clostridia. This review aims to provide an overview of the triggers, characteristics, and regulatory mechanism of sporulation in solventogenic clostridia. Those are further compared to the current knowledge on sporulation in the industrially relevant acetogenic clostridia. Finally, the potential applications of spores for process improvement are discussed.Key Points• The regulatory network governing sporulation initiation varies in solventogenic clostridia.• Media composition and cell density are the main triggers of sporulation.• Spores can be used to improve the fermentation process.

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Section: The Carbon Sourcementioning

confidence: 97%

Section: Sporulation Triggersmentioning

confidence: 99%

Sporulation in solventogenic and acetogenic clostridia

Diallo

Kengen

López-Contreras

2021

Appl Microbiol Biotechnol

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“…In the case of C. pasteurianum , glycerol is dehydrated via a coenzyme B 12 -dependent glycerol dehydratase (DhaB, DhaC, DhaE) to 3-hydroxypropionaldehyde (3-HPA), followed by the conversion of 3-HPA via a NADH-linked 1,3-PDO dehydrogenase (DhaT) to 1,3-PDO [ 27 ]. The glycerol reductive pathway of C. butyricum and C. beijerinckii consists of a coenzyme B 12 -independent glycerol dehydratase (DhaB1, DhaB2) for the dehydration of glycerol to 3-HPA, also followed by the conversion of 3-HPA to 1,3-PDO via a NADH-linked 1,3-PDO dehydrogenase (DhaT) [ 28 , 29 ]. However, glycerol is not only reduced to 1,3-PDO but also oxidized and used for biomass and by products e.g., acetate, butyrate, ethanol, and butanol [ 30 ].…”

Section: Introductionmentioning

confidence: 99%

Heterologous 1,3-Propanediol Production Using Different Recombinant Clostridium beijerinckii DSM 6423 Strains

et al. 2023

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1,3-propanediol (1,3-PDO) is a valuable basic chemical, especially in the polymer industry to produce polytrimethylene terephthalate. Unfortunately, the production of 1,3-PDO mainly depends on petroleum products as precursors. Furthermore, the chemical routes have significant disadvantages, such as environmental issues. An alternative is the biobased fermentation of 1,3-PDO from cheap glycerol. Clostridium beijerinckii DSM 6423 was originally reported to produce 1,3-PDO. However, this could not be confirmed, and a genome analysis revealed the loss of an essential gene. Thus, 1,3-PDO production was genetically reinstalled. Genes for 1,3-PDO production from Clostridium pasteurianum DSM 525 and Clostridium beijerinckii DSM 15410 (formerly Clostridium diolis) were introduced into C. beijerinckii DSM 6423 to enable 1,3-PDO production from glycerol. 1,3-PDO production by recombinant C. beijerinckii strains were investigated under different growth conditions. 1,3-PDO production was only observed for C. beijerinckii [pMTL83251_Ppta-ack_1,3-PDO.diolis], which harbors the genes of C. beijerinckii DSM 15410. By buffering the growth medium, production could be increased by 74%. Furthermore, the effect of four different promoters was analyzed. The use of the constitutive thlA promoter from Clostridium acetobutylicum led to a 167% increase in 1,3-PDO production compared to the initial recombinant approach.

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“…Under natural conditions, some microorganisms directly use glycerol as a substrate to efficiently synthesize 1, 3‐PDO (Casali et al ., 2012 ; Avci et al ., 2014 ; Lee et al ., 2019 ). The common strains, including Klebsiella pneumoniae (Sun et al ., 2021 ; Wang et al ., 2021a , 2021b ), Lactobacillus reuteri (Ju et al ., 2021a , 2021b ), and Clostridium butyricum (Sedlar et al ., 2021 ; Yun et al ., 2021 ) have been used to produce 1, 3‐PDO, and this study mainly focused on K . pneumoniae .…”

Section: Introductionmentioning

confidence: 99%

“…Under natural conditions, some microorganisms directly use glycerol as a substrate to efficiently synthesize 1, 3-PDO (Casali et al, 2012;Avci et al, 2014;Lee et al, 2019). The common strains, including Klebsiella pneumoniae (Sun et al, 2021;Wang et al, 2021aWang et al, ,2021b, Lactobacillus reuteri (Ju et al, 2021a(Ju et al, ,2021b, and Clostridium butyricum (Sedlar et al, 2021;Yun et al, 2021) have been used to produce 1, 3-PDO, and this study mainly focused on K. pneumoniae. In recent decades, researchers have paid much attention to obtain more advantageous strains by genetic engineering (Gonzalez et al, 2008;Przystalowska et al, 2015;Liu et al, 2018;Sun et al, 2019), and a more detailed understanding of the metabolic pathway of K. pneumoniae is expected to provide a better way to promote the transformation of glycerol into 1, 3-PDO in this system.…”

mentioning

confidence: 99%

Enhancing the capability of Klebsiella pneumoniae to produce 1, 3‐propanediol by overexpression and regulation through CRISPR‐dCas9

Wang

Zhang

Liang

et al. 2022

Microbial Biotechnology

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Klebsiella pneumoniae is a common strain of bacterial fermentation to produce 1, 3-propanediol (1, 3-PDO). In general, the production of 1, 3-PDO by wildtype K. pneumoniae is relatively low. Therefore, a new gene manipulation of K. pneumoniae was developed to improve the production of 1, 3-PDO by overexpressing in the reduction pathway and attenuating the by-products in the oxidation pathway. Firstly, dhaB and/or dhaT were overexpressed in the reduction pathway. Considering the cost of IPTG, the constitutive promoter P32 was selected to express the key gene. By comparing K.P. pET28a-P32-dhaT with the original strain, the production of 1, 3-PDO was increased by 19.7%, from 12.97 to 15.53 g l À1 (in a 250 ml shaker flask). Secondly, three lldD and budC regulatory sites were selected in the by-product pathway, respectively, using the CRISPR-dCas9 system, and the optimal regulatory sites were selected following the 1, 3-PDO production. As a result, the 1, 3-PDO production by K.P. L1-pRH2521 and K.P. B3-pRH2521 reached up to 19.16 and 18.74 g l À1 , which was increased by 47.7% and 44.5% respectively. Overexpressing dhaT and inhibiting expression of lldD and budC were combined to further enhance the ability of K. pneumoniae to produce 1, 3-PDO. The 1, 3-PDO production by K.P. L1-B3-PRH2521-P32-dhaT reached 57.85 g l À1 in a 7.5 l fermentation tank (with Na + neutralizer), which is higher than that of the original strain. This is the first time that the 1, 3-PDO production was improved in K. pneumoniae by overexpressing the key gene and attenuating by-product synthesis in the CRISPR-dCas9 system. This study reports an efficient approach to regulate the expression of genes in K. pneumoniae to increase the 1, 3-PDO production, and such a strategy may be useful to modify other strains to produce valuable chemicals.

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Phenotypic and genomic analysis of isopropanol and 1,3-propanediol producer Clostridium diolis DSM 15410

Cited by 11 publications

References 76 publications

Sporulation in solventogenic and acetogenic clostridia

Sporulation in solventogenic and acetogenic clostridia

Heterologous 1,3-Propanediol Production Using Different Recombinant Clostridium beijerinckii DSM 6423 Strains

Enhancing the capability of Klebsiella pneumoniae to produce 1, 3‐propanediol by overexpression and regulation through CRISPR‐dCas9

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