Properties of recombinant ATP-dependent fructokinase from the halotolerant methanotroph Methylomicrobium alcaliphilum 20Z

But, Sergey Y.; Rozova, Olga N.; Khmelenina, V. N.; Reshetnikov, A. S.; Trotsenko, Yu. A.

doi:10.1134/s0006297912040086

Cited by 19 publications

(15 citation statements)

References 18 publications

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“…2) [13,14,17]. The specific intra-cellular ectoine concentrations reached 20.2 ± 4.2, 27.0 ± 2.0 and 25.8 ± 1.7 mg ectoine g biomass -1 at the end of the cultivation at high salinity in TS1, 2 and 3, respectively.…”

Section: Resultsmentioning

confidence: 95%

“…This finding was attributed to the use of ectoine as an readily biodegradable nitrogen and carbon source by M. alcaliphilum 20Z [13,14]. The restoration of NaCl salinity resulted in a gradual re-assimilation of the extracellular ectoine within the first 10 h of cultivation under high salinity.…”

Section: <Fig 2>mentioning

confidence: 96%

“…Recent studies performed in both batch and continuous bioreactors have demonstrated the ability of the methanotrophic bacterium Methylomicrobium alcaliphilum 20Z to synthesize ectoine using methane as the sole carbon and energy source [13][14][15]. The promising results obtained hitherto with methanotrophs encourage further research devoted to develop biotechnologies capable of creating value from this greenhouse gas.…”

Section: Introductionmentioning

confidence: 98%

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Ectoine bio-milking in methanotrophs: A step further towards methane-based bio-refineries into high added-value products

Cantera

Lebrero

Rodríguez

et al. 2017

Chemical Engineering Journal

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Abstract:This communication showed for the first time that the methanotrophic strain Methylomicrobium alcaliphilum 20Z can efficiently synthesize and excrete (through a tailored bio-milking process) ectoine under continuous mode using methane as the sole energy and carbon source. First, three consecutive 50 h fed batch fermentations consisting of alternating high salinity (6% NaCl for 24h) and low salinity (0 % NaCl for 24h) cultivation stages were carried out in triplicate to determine the influence of sudden modifications in media salinity on ectoine synthesis and excretion. The results demonstrated that M. alcaliphilum 20Z exhibited a rapid response to osmotic shocks, which resulted in the release of the accumulated ectoine under hyposmotic shocks and the immediate uptake of the previously excreted ectoine during hyperosmotic shocks. A second experiment was carried out under continuous cultivation mode in two sequential stirred tank reactors operated at NaCl concentrations of 0 and 6 %. Cells exhibited a constant intracellular ectoine concentration of 70.4 ± 14.3 mg g biomass -1 along the entire operation period when cultivated at a NaCl concentration of 6%. The centrifugation of the cultivation broth followed by a hyposmotic shock resulted in the excretion of ~ 70 % of the total intracellular ectoine. In brief, this research shows the feasibility of the continuous bioconversion of diluted CH 4 emissions into high added-value products such as ectoine, which can turn greenhouse gas abatement into a sustainable and profitable process.

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Section: Resultsmentioning

confidence: 95%

Section: <Fig 2>mentioning

confidence: 96%

Section: Introductionmentioning

confidence: 98%

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Ectoine bio-milking in methanotrophs: A step further towards methane-based bio-refineries into high added-value products

Cantera

Lebrero

Rodríguez

et al. 2017

Chemical Engineering Journal

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“…Since sucrose synthesis requires a significant part of the carbon and energy of a growth substrate, the study of biochemical and regulatory mechanisms of its synthesis in methantrophs is impor tant for the selection or engineering of efficient ecto ine producers. [47]. In order to define more accurately the function of these genes and the role of sucrose, M. alcaliphilum mutants defective in sps and ams genes have been obtained [48].…”

mentioning

confidence: 99%

Biosynthesis of secondary metabolites in methanotrophs: Biochemical and genetic aspects (Review)

Khmelenina

Rozova

But

et al. 2015

Appl Biochem Microbiol

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“…However, in nonphotosynthetic prokaryotes, the genes and enzymes responsible for disaccharide biosynthesis have not yet been characterized. Recently, an SPS-like open reading frame (ORF) but no SPP-coding gene has been found in the genome of the obligate methylotroph Methylobacillus flagellatus KT, which uses methanol or methylamine but not methane as sole carbon and energy sources for growth (But et al, 2012). Sucrose synthesis and ability to grow at enhanced salinity were not studied in this bacterium (Govorukhina et al, 1987).…”

Section: Introductionmentioning

confidence: 99%

Bifunctional sucrose phosphate synthase/phosphatase is involved in the sucrose biosynthesis byMethylobacillus flagellatusKT

But

Khmelenina

Reshetnikov

et al. 2013

FEMS Microbiol Lett

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The aerobic obligate methylotroph Methylobacillus flagellatus KT was shown to synthesize sucrose in the presence of 0.5-2% NaCl in the growth medium. In the genome of this bacterium, an open reading frame (ORF) encoding a predicted 84-kD polypeptide homologous to the plant and cyanobacterial sucrose phosphate synthases (SPSs) was found. Using heterologous expression of the putative sps gene in Escherichia coli, followed by affinity chromatography, pure recombinant protein SPS-His6 was obtained. The enzyme catalyzed two reactions: conversion of fructose 6-phosphate and UDP-glucose into sucrose 6-phosphate and hydrolysis of sucrose 6-phosphate to sucrose. The bifunctional sucrose phosphate synthase/phosphatase (SPS/SPP) was a 340 kDa homotetrameric Mg(2+) -dependent enzyme activated by fructose 1,6-phosphate2 and ATP but inhibited by glucose 6-phosphate, fructose 1-phosphate, AMP and inorganic phosphate. The amino acid sequence of the protein had a C-terminal domain homologous to SPPs. This correlated with the absence of the spp gene in the M. flagellatus chromosome. The ORFs homologous to the M. flagellatus SPS were found in the genomes of another obligate methylotroph Methylovorus glucosetrophus as well as the lithoautotrophic bacteria Acidithiobacillus ferrooxidans, Nitrosomonas europaea and Nitrosospira multiformis whose genomes lacked the spp genes. Thus, data extending the knowledge of biochemical properties of bacterial SPSs have been obtained.

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Properties of recombinant ATP-dependent fructokinase from the halotolerant methanotroph Methylomicrobium alcaliphilum 20Z

Cited by 19 publications

References 18 publications

Ectoine bio-milking in methanotrophs: A step further towards methane-based bio-refineries into high added-value products

Ectoine bio-milking in methanotrophs: A step further towards methane-based bio-refineries into high added-value products

Biosynthesis of secondary metabolites in methanotrophs: Biochemical and genetic aspects (Review)

Bifunctional sucrose phosphate synthase/phosphatase is involved in the sucrose biosynthesis byMethylobacillus flagellatusKT

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