Identification and characterization of Clostridium paraputrificum M-21, a chitinolytic, mesophilic and hydrogen-producing bacterium

Evvyernie, D; Yamazaki, Shinichi; Morimoto, Kenji; Karita, Shuichi; Kimura, Takeshi; Sakka, Kazuo; Ohmiya, Kunio

doi:10.1016/s1389-1723(00)80063-8

Cited by 83 publications

(28 citation statements)

References 15 publications

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“…It is widely distributed in nature as the integument of insects and crustaceans and as a component of fungi and algae [1]. Currently, chitin is produced from commercial crab or shrimp processing waste, which are usually discarded to sea and result in high environmental pollution in many maritime states [2,3]. Hydrolysis of chitin by mineral acids or enzymes leads to the monomeric constituent GlcNAc [4].…”

Section: Introductionmentioning

confidence: 99%

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Production of lipid from N‐acetylglucosamine by Cryptococcus curvatus

Zhao

et al. 2010

Euro J Lipid Sci & Tech

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N-Acetylglucosamine (GlcNAc), the monomeric constituent of chitin, is rarely used as a carbon source for fermentation technology. In this study, we demonstrate that the oleaginous yeast Cryptococcus curvatus ATCC 20509 can produce intracellular lipid during the cultivation process and total lipid content can reach 54% on a GlcNAc-based medium. Culture of C. curvatus under various conditions indicated that lipid accumulation also occurred at a relatively broad range of temperatures as well as relatively high initial GlcNAc concentrations. Fatty acid analysis indicated that the product was rich in palmitic acid, stearic acid, and oleic acid, closely resembling the composition of palm oil. More importantly, the lipid sample produced at 22 8C had a total saturated fatty acid content of 54.2 wt%, suggesting that it may be explored as cocoa-butter equivalent. Our data suggested that GlcNAc could be used as a feedstock for industrial biotechnology and that C. curvatus ATCC 20509 is a strain capable of accumulating high intracellular lipid using this nitrogen-rich renewable material.Practical applications: Microbial lipid is a versatile material, especially for biodiesel production. Stable and abundant renewable raw substrates remain to be explored for large-scale production of microbial lipid. The present work reports lipid production using N-acetylglucosamine (GlcNAc) by the oleaginous yeast Cryptococcus curvatus ATCC 20509 to yield up to 54% intracellular lipid content. More significantly, the lipid sample produced at 22 8C had a total saturated fatty acid content of 54.2 wt%, suggesting that it may be explored as cocoa-butter equivalent. Our technology provides the opportunity to effectively convert GlcNAc, available from one of the most abundant renewable materials chitin, into lipid. This procedure should prove valuable in terms of renewable energy production as well as environmental pollution control.

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Section: Introductionmentioning

confidence: 99%

“…As the chitin resource is abundant and seafood processing waste presents environmental problems, it is pivotal to explore GlcNAc as a feedstock for industrial biotechnology. However, to the best of our knowledge, studies on GlcNAc as a carbon source for microbial culture are scarce [2].…”

Section: Introductionmentioning

confidence: 99%

Production of lipid from N‐acetylglucosamine by Cryptococcus curvatus

Zhao

et al. 2010

Euro J Lipid Sci & Tech

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“…According to our search, there is only one report on hydrogen production from chitin or chitin-containing materials. Evvyernie et al has reported hydrogen production from a mesophilic bacterium, Clostridium paraputrificum M-21, using N-acetylglucosamine, chitin, and chitincontaining wastes (45,46). From 1 g of raw chitinous wastes, the evolved hydrogen was reported as 5.2 to 7.6 mmol.…”

Section: Discussionmentioning

confidence: 99%

Engineering of the Hyperthermophilic Archaeon Thermococcus kodakarensis for Chitin-Dependent Hydrogen Production

Aslam

Horiuchi

Simons

et al. 2017

Appl Environ Microbiol

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Thermococcus kodakarensis is a hyperthermophilic archaeon that harbors a complete set of genes for chitin degradation to fructose 6-phosphate. However, wild-type T. kodakarensis KOD1 does not display growth on chitin. In this study, we developed a T. kodakarensis strain that can grow on chitin via genetic and adaptive engineering. First, a chitinase overproduction strain (KC01) was constructed by replacing the chitinase gene promoter with a strong promoter from the cell surface glycoprotein gene, resulting in increased degradation of swollen chitin and accumulation of N-,N=-diacetylchitobiose in the medium. To enhance N-,N=-diacetylchitobiose assimilation in KC01, genes encoding diacetylchitobiose deacetylase, exo-␤-D-glucosaminidase, and glucosamine-6-phosphate deaminase were also overexpressed to obtain strain KC04. To strengthen the glycolytic flux of KC04, the gene encoding Tgr (transcriptional repressor of glycolytic genes) was disrupted to obtain strain KC04Δt. In both KC04 and KC04Δt strains, degradation of swollen chitin was further enhanced. In the culture broth of these strains, the accumulation of glucosamine was observed. KC04Δt was repeatedly inoculated in a swollen-chitin-containing medium for 13 cultures. This adaptive engineering strategy resulted in the isolation of a strain (KC04ΔtM1) that showed almost complete degradation of 0.4% (wt/vol) swollen chitin after 90 h. The strain produced high levels of acetate and ammonium in the culture medium, and, moreover, molecular hydrogen was generated. This strongly suggests that strain KC04ΔtM1 has acquired the ability to convert chitin to fructose 6-phosphate via deacetylation and deamination and further convert fructose 6-phosphate to acetate via glycolysis coupled to hydrogen generation. IMPORTANCE Chitin is a linear homopolymer of ␤-1,4-linked N-acetylglucosamine and is the second most abundant biomass next to cellulose. Compared to the wealth of research focused on the microbial degradation and conversion of cellulose, studies addressing microbial chitin utilization are still limited. In this study, using the hyperthermophilic archaeon Thermococcus kodakarensis as a host, we have constructed a strain that displays chitin-dependent hydrogen generation. The apparent hydrogen yield per unit of sugar consumed was slightly higher with swollen chitin than with starch. As gene manipulation in T. kodakarensis is relatively simple, the strain constructed in this study can also be used as a parent strain for the development and expansion of chitin-dependent biorefinery, in addition to its capacity to produce hydrogen.

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“…Our results suggest that butyrate formation deficiency improved ethanol production but not hydrogen production, indicating the importance of butyrate formation pathway forIntroduction Biological production of hydrogen by different microorganisms has been attracting interest due to concerns regarding the environmentally benign alternatives to current fossil energy sources [1,2]. Since other reducing metabolites, such as propionate, butyrate, and sometimes lactate and ethanol, are produced during anaerobic fermentation, the hydrogen yields of dark fermentation have been reported to range from 0.6 to 1.9 mol/mol glucose [3][4][5][6], which are substantially less than the theoretical yield (4 mol/mol glucose). Previous studies have attempted to improve hydrogen yield by engineering related metabolic pathways, particularly those related to the composition of liquid metabolites [7][8][9][10][11][12][13].…”

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confidence: 99%

Blocking the butyrate-formation pathway impairs hydrogen production in <italic>Clostridium perfringens</italic>

Yu¹,

Wang²,

Bi³

et al. 2013

ABBS

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Inactivating competitive pathways will improve fermentative hydrogen production by obligate anaerobes, such as those of genus Clostridium. In our previous study, the hydrogen yield of Clostridium perfringens W13 in which L-lactate dehydrogenase was inactivated increased by 44% when compared with its original strain W12. In this study, we explored whether blocking butyrate formation pathway would increase hydrogen yield. The acetyl-CoA acetyltransferase gene (atoB) encodes the first enzyme in this pathway, which ultimately forms butyrate. Clostridium perfringens W14 and W15 were constructed by inactivating atoB in W13 and W12, respectively. The hydrogen yield of W14 and W15 was 44% and 33% of those of W13 and W12, respectively. Inactivation of atoB decreased the pyruvate synthesis and its conversion to acetyl-CoA in both mutants, and increased ethanol formation in W14 and W15. Proteomic analysis revealed that the expressions of five proteins involved in butyrate formation pathway were up-regulated in W14. Our results suggest that butyrate formation deficiency improved ethanol production but not hydrogen production, indicating the importance of butyrate formation pathway for hydrogen production in C. perfringens.

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Identification and characterization of Clostridium paraputrificum M-21, a chitinolytic, mesophilic and hydrogen-producing bacterium

Cited by 83 publications

References 15 publications

Production of lipid from N‐acetylglucosamine by Cryptococcus curvatus

Production of lipid from N‐acetylglucosamine by Cryptococcus curvatus

Engineering of the Hyperthermophilic Archaeon Thermococcus kodakarensis for Chitin-Dependent Hydrogen Production

Blocking the butyrate-formation pathway impairs hydrogen production in <italic>Clostridium perfringens</italic>

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Identification and characterization of Clostridium paraputrificum M-21, a chitinolytic, mesophilic and hydrogen-producing bacterium

Cited by 83 publications

References 15 publications

Production of lipid from N‐acetylglucosamine by Cryptococcus curvatus

Production of lipid from N‐acetylglucosamine by Cryptococcus curvatus

Engineering of the Hyperthermophilic Archaeon Thermococcus kodakarensis for Chitin-Dependent Hydrogen Production

Blocking the butyrate-formation pathway impairs hydrogen production in &lt;italic&gt;Clostridium perfringens&lt;/italic&gt;

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Blocking the butyrate-formation pathway impairs hydrogen production in <italic>Clostridium perfringens</italic>