| cloning and sequence analysis of the meso-diaminopimelate decarboxylase gene from bacillus methanolicus mga3 and comparison to other decarboxylase genes. | the lysa gene of bacillus methanolicus mga3 was cloned by complementation of an auxotrophic escherichia coli lysa22 mutant with a genomic library of b. methanolicus mga3 chromosomal dna. subcloning localized the b. methanolicus mga3 lysa gene into a 2.3-kb smai-ssti fragment. sequence analysis of the 2.3-kb fragment indicated an open reading frame encoding a protein of 48,223 da, which was similar to the meso-diaminopimelate (dap) decarboxylase amino acid sequences of bacillus subtilis (62%) and ... | 1993 | 8215365 |
| electron microscopic analysis and structural characterization of novel nadp(h)-containing methanol: n,n'-dimethyl-4-nitrosoaniline oxidoreductases from the gram-positive methylotrophic bacteria amycolatopsis methanolica and mycobacterium gastri mb19. | the quaternary protein structure of two methanol:n,n'-dimethyl-4-nitrosoaniline (ndma) oxidoreductases purified from amycolatopsis methanolica and mycobacterium gastri mb19 was analyzed by electron microscopy and image processing. the enzymes are decameric proteins (displaying fivefold symmetry) with estimated molecular masses of 490 to 500 kda based on their subunit molecular masses of 49 to 50 kda. both methanol:ndma oxidoreductases possess a tightly but noncovalently bound nadp(h) cofactor at ... | 1993 | 8449887 |
| purification and characterization of an oxygen-labile, nad-dependent alcohol dehydrogenase from desulfovibrio gigas. | a nad-dependent, oxygen-labile alcohol dehydrogenase was purified from desulfovibrio gigas. it was decameric, with subunits of m(r) 43,000. the best substrates were ethanol (km, 0.15 mm) and 1-propanol (km, 0.28 mm). n-terminal amino acid sequence analysis showed that the enzyme belongs to the same family of alcohol dehydrogenases as zymomonas mobilis adh2 and bacillus methanolicus mdh. | 1993 | 8491707 |
| characterization of a restriction-modification system of the thermotolerant methylotroph bacillus methanolicus. | we report the isolation of a restriction endonuclease, bmeti, an isoschizomer of bcli, that recognizes the dna sequence 5' tgatca 3'. we also report that bmeti sites are modified to tgm6atca. these findings provide the basis for devising strategies to prevent bmeti restriction of any dna introduced into bacillus methanolicus. | 1996 | 8975604 |
| genetic manipulation of bacillus methanolicus, a gram-positive, thermotolerant methylotroph. | we report the fist genetic transformation system, shuttle vectors, and integrative vectors for the thermotolerant, methylotrophic bacterium bacillus methanolicus. by using a polyethylene glycol-mediated transformation procedure, we have successfully transformed b. methanolicus with both integrative and multicopy plasmids. for plasmids with a single bmeti recognition site, dam methylation of plasmid dna (in vivo or in vitro) was found to enhance transformation efficiency from 7- to 11-fold. two l ... | 1997 | 9097439 |
| properties of an nad(h)-containing methanol dehydrogenase and its activator protein from bacillus methanolicus. | oxidation of c1-c4 primary alcohols in thermotolerant bacillus methanolicus strains is catalyzed by an nad-dependent methanol dehydrogenase (mdh), composed of ten identical 43,000-mr subunits. each mdh subunit contains a tightly, but non-covalently, bound nad(h) molecule, in addition to 1 zn2+ and 1-2 mg2+ ions. the nad(h) cofactor is oxidized and reduced by formaldehyde and methanol, respectively, while it remains bound to the enzyme. incubation of mdh with methanol and exogenous nad (coenzyme) ... | 1997 | 9119008 |
| a low-molecular-mass protein from methylococcus capsulatus (bath) is responsible for the regulation of formaldehyde dehydrogenase activity in vitro. | an 8.6 kda protein, which the authors call a modifin, has been purified from methylococcus capsulatus (bath) and has been shown to alter the substrate specificity and kinetics of nad+-linked formaldehyde dehydrogenase (fdh) isolated from the same organism. purification methods for both the modifin and fdh are presented which reliably produced pure protein for further analysis. analysis of the molecular mass and n-terminal sequence of both fdh and the modifin indicate that they are unique protein ... | 1999 | 10206695 |
| molecular, biochemical, and functional characterization of a nudix hydrolase protein that stimulates the activity of a nicotinoprotein alcohol dehydrogenase. | the cytoplasmic coenzyme nad(+)-dependent alcohol (methanol) dehydrogenase (mdh) employed by bacillus methanolicus during growth on c(1)-c(4) primary alcohols is a decameric protein with 1 zn(2+)-ion and 1-2 mg(2+)-ions plus a tightly bound nad(h) cofactor per subunit (a nicotinoprotein). mg(2+)-ions are essential for binding of nad(h) cofactor in mdh protein expressed in escherichia coli. the low coenzyme nad(+)-dependent activity of mdh with c(1)-c(4) primary alcohols is strongly stimulated by ... | 2002 | 12089158 |
| identification of a magnesium-dependent nad(p)(h)-binding domain in the nicotinoprotein methanol dehydrogenase from bacillus methanolicus. | the bacillus methanolicus methanol dehydrogenase (mdh) is a decameric nicotinoprotein alcohol dehydrogenase (family iii) with one zn(2+) ion, one or two mg(2+) ions, and a tightly bound cofactor nad(h) per subunit. the mg(2+) ions are essential for binding of cofactor nad(h) in mdh. a b. methanolicus activator protein strongly stimulates the relatively low coenzyme nad(+)-dependent mdh activity, involving hydrolytic removal of the nmn(h) moiety of cofactor nad(h) (kloosterman, h., vrijbloed, j. ... | 2002 | 12351635 |
| dissimilation of [(13)c]methanol by continuous cultures of bacillus methanolicus mga3 at 50 degrees c studied by (13)c nmr and isotope-ratio mass spectrometry. | using a continuous culture of bacillus methanolicus mga3 limited by 100 mm methanol in the feed and growing at a dilution rate d=0.25 h(-1), transients in dissolved methanol were studied to determine the effects of methanol toxicity and the pathway of methanol dissimilation to co(2). steady-state cultures were disturbed by pulses of methanol resulting in a rapid change in concentration of 6.4-12.8 mm. b. methanolicus mga3 responded to a sudden increase in available methanol by a transient declin ... | 2002 | 12368456 |
| role of the bacillus methanolicus citrate synthase ii gene, city, in regulating the secretion of glutamate in l-lysine-secreting mutants. | the thermotolerant, restrictive methylotroph bacillus methanolicus mga3 (atcc 53907) can secrete 55 g of glutamate per liter (maximum yield, 0.36 g/g) at 50 degrees c with methanol as a carbon source and a source of ammonia in fed-batch bioreactors. a homoserine dehydrogenase mutant, 13a52-8a66, secreting up to 35 g of l-lysine per liter in fed-batch fermentations had minimal 2-oxoglutarate dehydrogenase activity [7.3 nmol min(-1) (mg of protein)(-1)], threefold-increased pyruvate carboxylase ac ... | 2003 | 12839772 |
| plasmid-dependent methylotrophy in thermotolerant bacillus methanolicus. | bacillus methanolicus can efficiently utilize methanol as a sole carbon source and has an optimum growth temperature of 50 degrees c. with the exception of mannitol, no sugars have been reported to support rapid growth of this organism, which is classified as a restrictive methylotroph. here we describe the dna sequence and characterization of a 19,167-bp circular plasmid, designated pbm19, isolated from b. methanolicus mga3. sequence analysis of pbm19 demonstrated the presence of the methanol d ... | 2004 | 14973041 |
| reclassification of brevibacillus brevis strains ncimb 13288 and dsm 6472 (=nrrl nrs-887) as aneurinibacillus danicus sp. nov. and brevibacillus limnophilus sp. nov. | comparison of the hypervariable region (269-279 bases in length) at the 5' end of the 16s rdna sequences of 29 bacterial strains that were identified previously as brevibacillus brevis showed that 13 strains clustered with aneurinibacillus species, eight strains clustered with bacillus species and eight strains clustered with brevibacillus species. based on dna-dna hybridization results, 27 strains, not including [brevibacillus brevis] ncimb 13288 and [brevibacillus brevis] dsm 6472, were reiden ... | 2004 | 15023954 |
| growth of bacillus methanolicus in seawater-based media. | bacillus methanolicus has been proposed as a biocatalyst for the low cost production of commodity chemicals. the organism can use methanol as sole carbon and energy source, and it grows aerobically at elevated temperatures. methanol can be made available from off-shore conversion of natural gas to methanol, through gas-to-liquid technology. growth of the organism in seawater-based medium would further reduce the costs of chemical production performed near an off-shore natural gas source. the gro ... | 2005 | 15726441 |
| bacillus alveayuensis sp. nov., a thermophilic bacterium isolated from deep-sea sediments of the ayu trough. | two thermophilic, spore-forming strains, tm1(t) and tm5, were isolated from deep-sea sediment (4000 m below sea level) of the ayu trough in the western pacific ocean. cells of the two strains were gram-positive, motile and rod-shaped. their spores were ellipsoidal, subterminal to terminal and occurred in swollen sporangia. the two strains grew at temperatures up to 65 degrees c and in the ph range 6.5-9.0. the nacl concentration for optimal growth was 3.0 % (w/v) and growth was inhibited by 5 % ... | 2005 | 15879257 |
| cloning and sequence analysis of the meso-diaminopimelate decarboxylase gene from bacillus methanolicus mga3 and comparison to other decarboxylase genes. | [this corrects the article on p. 2935 in vol. 59.]. | 1993 | 16349139 |
| upregulated transcription of plasmid and chromosomal ribulose monophosphate pathway genes is critical for methanol assimilation rate and methanol tolerance in the methylotrophic bacterium bacillus methanolicus. | the natural plasmid pbm19 carries the key mdh gene needed for the oxidation of methanol into formaldehyde by bacillus methanolicus. five more genes, glpx, fba, tkt, pfk, and rpe, with deduced roles in the cell primary metabolism, are also located on this plasmid. by using real-time pcr, we show that they are transcriptionally upregulated (6- to 40-fold) in cells utilizing methanol; a similar induction was shown for two chromosomal genes, hps and phi. these seven genes are involved in the fructos ... | 2006 | 16585766 |
| bacillus methanolicus: a candidate for industrial production of amino acids from methanol at 50 degrees c. | amino acids are among the major products in biotechnology in both volume and value, and the global market is growing. microbial fermentation is the dominant method used for industrial production, and today the most important microorganisms used are corynebacteria utilizing sugars. for low-prize bulk amino acids, the possibility of using alternative substrates such as methanol has gained considerable interest. in this mini review, we highlight the unique genetics and favorable physiological trait ... | 2007 | 17216461 |
| lysine production from methanol at 50 degrees c using bacillus methanolicus: modeling volume control, lysine concentration, and productivity using a three-phase continuous simulation. | a simulation was developed based on experimental data obtained in a 14-l reactor to predict the growth and l-lysine accumulation kinetics, and change in volume of a large-scale (250-m(3)) bacillus methanolicus methanol-based process. homoserine auxotrophs of b. methanolicus mga3 are unique methylotrophs because of the ability to secrete lysine during aerobic growth and threonine starvation at 50 degrees c. dissolved methanol (100 mm), ph, dissolved oxygen tension (0.063 atm), and threonine level ... | 1996 | 18626859 |
| overexpression of wild-type aspartokinase increases l-lysine production in the thermotolerant methylotrophic bacterium bacillus methanolicus. | aspartokinase (ak) controls the carbon flow into the aspartate pathway for the biosynthesis of the amino acids l-methionine, l-threonine, l-isoleucine, and l-lysine. we report here the cloning of four genes (asd, encoding aspartate semialdehyde dehydrogenase; dapa, encoding dihydrodipicolinate synthase; dapg, encoding aki; and yclm, encoding akiii) of the aspartate pathway in bacillus methanolicus mga3. together with the known akii gene lysc, dapg and yclm form a set of three ak genes in this or ... | 2009 | 19060158 |
| bacillus methanolicus pyruvate carboxylase and homoserine dehydrogenase i and ii and their roles for l-lysine production from methanol at 50 degrees c. | we here present the pyc gene encoding pyruvate carboxylase (pc), and the hom-1 and hom-2 genes encoding two active homoserine dehydrogenase (hd) proteins, in methylotrophic bacillus methanolicus mga3. in general, both pc and hd are regarded as key targets for improving bacterial l-lysine production; pc plays a role in precursor oxaloacetate (oaa) supply while hd controls an important branch point in the l-lysine biosynthetic pathway. the hom-1 and hom-2 genes were strongly repressed by l-threoni ... | 2010 | 20372887 |
| analysis and manipulation of aspartate pathway genes for l-lysine overproduction from methanol by bacillus methanolicus. | we investigated the regulation and roles of six aspartate pathway genes in l-lysine overproduction in bacillus methanolicus: dapg, encoding aspartokinase i (aki); lysc, encoding akii; yclm, encoding akiii; asd, encoding aspartate semialdehyde dehydrogenase; dapa, encoding dihydrodipicolinate synthase; and lysa, encoding meso-diaminopimelate decarboxylase. analysis of the wild-type strain revealed that in vivo lysc transcription was repressed 5-fold by l-lysine and induced 2-fold by dl-methionine ... | 2011 | 21724876 |
| metabolic engineering of corynebacterium glutamicum for methanol metabolism. | methanol is already an important carbon feedstock in the chemical industry, but it has found only limited application in biotechnological production processes. this can be mostly attributed to the inability of most microbial platform organisms to utilize methanol as a carbon and energy source. with the aim to turn methanol into a suitable feedstock for microbial production processes, we engineered the industrially important but nonmethylotrophic bacterium corynebacterium glutamicum toward the ut ... | 2015 | 25595770 |
| genome-based genetic tool development for bacillus methanolicus: theta- and rolling circle-replicating plasmids for inducible gene expression and application to methanol-based cadaverine production. | bacillus methanolicus is a thermophilic methylotroph able to overproduce amino acids from methanol, a substrate not used for human or animal nutrition. based on our previous rna-seq analysis a mannitol inducible promoter and a putative mannitol activator gene mtlr were identified. the mannitol inducible promoter was applied for controlled gene expression using fluorescent reporter proteins and a flow cytometry analysis, and improved by changing the -35 promoter region and by co-expression of the ... | 2016 | 27713731 |
| genome sequence of thermotolerant bacillus methanolicus: features and regulation related to methylotrophy and production of l-lysine and l-glutamate from methanol. | bacillus methanolicus can utilize methanol as its sole carbon and energy source, and the scientific interest in this thermotolerant bacterium has focused largely on exploring its potential as a biocatalyst for the conversion of methanol into l-lysine and l-glutamate. we present here the genome sequences of the important b. methanolicus model strain mga3 (atcc 53907) and the alternative wild-type strain pb1 (ncimb13113). the physiological diversity of these two strains was demonstrated by a compa ... | 2012 | 22610424 |
| characterization of fructose 1,6-bisphosphatase and sedoheptulose 1,7-bisphosphatase from the facultative ribulose monophosphate cycle methylotroph bacillus methanolicus. | the genome of the facultative ribulose monophosphate (rump) cycle methylotroph bacillus methanolicus encodes two bisphosphatases (glpx), one on the chromosome (glpx(c)) and one on plasmid pbm19 (glpx(p)), which is required for methylotrophy. both enzymes were purified from recombinant escherichia coli and were shown to be active as fructose 1,6-bisphosphatases (fbpases). the fbpase-negative corynebacterium glutamicum δfbp mutant could be phenotypically complemented with glpx(c) and glpx(p) from ... | 2013 | 24013630 |
| functional characterization of key enzymes involved in l-glutamate synthesis and degradation in the thermotolerant and methylotrophic bacterium bacillus methanolicus. | bacillus methanolicus wild-type strain mga3 secretes 59 g/liter(-1) of l-glutamate in fed-batch methanol cultivations at 50°c. we recently sequenced the mga3 genome, and we here characterize key enzymes involved in l-glutamate synthesis and degradation. one glutamate dehydrogenase (gdh) that is encoded by yweb and two glutamate synthases (gogats) that are encoded by the gltab operon and by glta2 were found, in contrast to bacillus subtilis, which has two different gdhs and only one gogat. b. met ... | 2013 | 23811508 |
| 6-phosphofructokinase and ribulose-5-phosphate 3-epimerase in methylotrophic bacillus methanolicus ribulose monophosphate cycle. | d-ribulose-5-phosphate-3-epimerase (rpe) and 6-phosphofructokinase (pfk) catalyse two reactions in the ribulose monophosphate (rump) cycle in bacillus methanolicus. the b. methanolicus wild-type strain mga3 possesses two putative rpe and pfk genes encoded on plasmid pbm19 (rpe1-mga3 and pfk1-mga3) and on the chromosome (rpe2-mga3 and pfk2-mga3). the wild-type strain pb1 also encodes putative rpe and pfk genes on plasmid pbm20 (rpe1-pb1 and pfk1-pb1*); however, it only harbours a chromosomal pfk ... | 2017 | 28213736 |
| characterization and evolution of an activator-independent methanol dehydrogenase from cupriavidus necator n-1. | methanol utilization by methylotrophic or non-methylotrophic organisms is the first step toward methanol bioconversion to higher carbon-chain chemicals. methanol oxidation using nad-dependent methanol dehydrogenase (mdh) is of particular interest because it uses nad(+) as the electron carrier. to our knowledge, only a limited number of nad-dependent mdhs have been reported. the most studied is the bacillus methanolicus mdh, which exhibits low enzyme specificity to methanol and is dependent on an ... | 2016 | 26846745 |
| dynamet: a fully automated pipeline for dynamic lc-ms data. | dynamic isotope labeling data provides crucial information about the operation of metabolic pathways and are commonly generated via liquid chromatography-mass spectrometry (lc-ms). metabolome-wide analysis is challenging as it requires grouping of metabolite features over different samples. we developed dynamet for fully automated investigations of isotope labeling experiments from lc-high-resolution ms raw data. dynamet enables untargeted extraction of metabolite labeling profiles and provides ... | 2015 | 26366644 |
| core pathways operating during methylotrophy of bacillus methanolicus mga3 and induction of a bacillithiol-dependent detoxification pathway upon formaldehyde stress. | bacillus methanolicus mga3 is a model facultative methylotroph of interest for fundamental research and biotechnological applications. previous research uncovered a number of pathways potentially involved in one-carbon substrate utilization. here, we applied dynamic (13) c labeling to elucidate which of these pathways operate during growth on methanol and to uncover potentially new ones. b. methanolicus mga3 uses the assimilatory and dissimilatory ribulose monophosphate (rump) cycles for convers ... | 2015 | 26303953 |
| methanol-based cadaverine production by genetically engineered bacillus methanolicus strains. | methanol is regarded as an attractive substrate for biotechnological production of value-added bulk products, such as amino acids and polyamines. in the present study, the methylotrophic and thermophilic bacterium bacillus methanolicus was engineered into a microbial cell factory for the production of the platform chemical 1,5-diaminopentane (cadaverine) from methanol. this was achieved by the heterologous expression of the escherichia coli genes cada and ldcc encoding two different lysine decar ... | 2015 | 25644214 |
| cloning, expression, and sequence analysis of the bacillus methanolicus c1 methanol dehydrogenase gene. | the gene (mdh) coding for methanol dehydrogenase (mdh) of thermotolerant, methylotroph bacillus methanolicus c1 has been cloned and sequenced. the deduced amino acid sequence of the mdh gene exhibited similarity to those of five other alcohol dehydrogenase (type iii) enzymes, which are distinct from the long-chain zinc-containing (type i) or short-chain zinc-lacking (type ii) enzymes. highly efficient expression of the mdh gene in escherichia coli was probably driven from its own promoter sequen ... | 1992 | 1644761 |
| bacillus methanolicus sp. nov., a new species of thermotolerant, methanol-utilizing, endospore-forming bacteria. | the generic position of 14 strains of gram-positive bacteria able to use methanol as a growth substrate was determined. all are obligately aerobic, thermotolerant organisms that are able to grow at temperatures of 35 to 60 degrees c. nine of the strains produce oval spores at a subterminal-to-central position in slightly swollen rod-shaped cells. dna-dna hybridization studies, 5s rrna sequence analysis, and physiological characteristics revealed that all 14 strains cluster as a well-defined grou ... | 1992 | 1380290 |
| engineering escherichia coli for methanol conversion. | methylotrophic bacteria utilize methanol and other reduced one-carbon compounds as their sole source of carbon and energy. for this purpose, these bacteria evolved a number of specialized enzymes and pathways. here, we used a synthetic biology approach to select and introduce a set of "methylotrophy genes" into escherichia coli based on in silico considerations and flux balance analysis to enable methanol dissimilation and assimilation. we determined that the most promising approach allowing the ... | 2015 | 25596507 |
| characterization of two transketolases encoded on the chromosome and the plasmid pbm19 of the facultative ribulose monophosphate cycle methylotroph bacillus methanolicus. | transketolase (tkt) is a key enzyme of the pentose phosphate pathway (ppp), the calvin cycle and the ribulose monophosphate (rump) cycle. bacillus methanolicus is a facultative rump pathway methylotroph. b. methanolicus mga3 harbors two genes putatively coding for tkts; one located on the chromosome (tkt(c)) and one located on the natural occurring plasmid pbm19 (tkt(p)). | 2014 | 24405865 |
| the methylotrophic bacillus methanolicus mga3 possesses two distinct fructose 1,6-bisphosphate aldolases. | the thermotolerant gram-positive methylotroph bacillus methanolicus is able to grow with methanol, glucose or mannitol as a sole carbon and energy source. fructose 1,6-bisphosphate aldolase (fba), a key enzyme of glycolysis and gluconeogenesis, is encoded in the genome of b. methanolicus by two putative fba genes, the chromosomally located fba(c) and fba(p) on the naturally occurring plasmid pbm19. their amino acid sequences share 75 % identity and suggest a classification as class ii aldolases. ... | 2013 | 23760818 |
| methylotrophic bacillus methanolicus encodes two chromosomal and one plasmid born nad+ dependent methanol dehydrogenase paralogs with different catalytic and biochemical properties. | bacillus methanolicus can utilize methanol as the sole carbon source for growth and it encodes an nad(+)-dependent methanol dehydrogenase (mdh), catalyzing the oxidation of methanol to formaldehyde. recently, the genomes of the b. methanolicus strains mga3 (atcc53907) and pb1 (ncimb13113) were sequenced and found to harbor three different putative mdh encoding genes, each belonging to the type iii fe-nad(+)-dependent alcohol dehydrogenases. in each strain, two of these genes are encoded on the c ... | 2013 | 23527128 |
| bacillus salitolerans sp. nov., a novel bacterium isolated from a salt mine in xinjiang province, china. | a novel aerobic bacterium, kc1(t), was isolated from a salt mine in kuche county, xinjiang province, china. cells were observed to be gram-positive, rod-shaped, endospore-forming and motile with flagella. strain kc1(t) was found to grow at 25-45 °c (optimum 37 °c), ph 6.5-9.0 (optimum 8.0) and nacl 0-10 % (v/v) (optimum 4 %). the major fatty acids were identified as anteiso-c15:0 and anteiso-c17:0. menaquinone-7 (mk-7) was found to be the predominant isoprenoid quinone. the cell-wall diamino aci ... | 2015 | 26076748 |
| l-lysine production by bacillus methanolicus: genome-based mutational analysis and l-lysine secretion engineering. | bacillus methanolicus is a methylotrophic bacterium with an increasing interest in academic research and for biotechnological applications. this bacterium was previously applied for methanol-based production of l-glutamate, l-lysine and the five-carbon diamine cadaverine by wild type, classical mutant and recombinant strains. the genomes of two different l-lysine secreting b. methanolicus classical mutant strains, noa2#13a52-8a66 and m168-20, were sequenced. we focused on mutational mapping in g ... | 2017 | 28163092 |
| production of carbon-13-labeled cadaverine by engineered corynebacterium glutamicum using carbon-13-labeled methanol as co-substrate. | methanol, a one-carbon compound, can be utilized by a variety of bacteria and other organisms as carbon and energy source and is regarded as a promising substrate for biotechnological production. in this study, a strain of non-methylotrophic corynebacterium glutamicum, which was able to produce the polyamide building block cadaverine as non-native product, was engineered for co-utilization of methanol. expression of the gene encoding nad+-dependent methanol dehydrogenase (mdh) from the natural m ... | 2015 | 26276544 |
| expression of heterologous non-oxidative pentose phosphate pathway from bacillus methanolicus and phosphoglucose isomerase deletion improves methanol assimilation and metabolite production by a synthetic escherichia coli methylotroph. | synthetic methylotrophy aims to develop non-native methylotrophic microorganisms to utilize methane or methanol to produce chemicals and biofuels. we report two complimentary strategies to further engineer a previously engineered methylotrophic e. coli strain for improved methanol utilization. first, we demonstrate improved methanol assimilation in the presence of small amounts of yeast extract by expressing the non-oxidative pentose phosphate pathway (ppp) from bacillus methanolicus. second, we ... | 2018 | 29203223 |
| quantitative metabolomics of the thermophilic methylotroph bacillus methanolicus. | the gram-positive bacterium bacillus methanolicus mga3 is a promising candidate for methanol-based biotechnologies. accurate determination of intracellular metabolites is crucial for engineering this bacteria into an efficient microbial cell factory. due to the diversity of chemical and cell properties, an experimental protocol validated on b. methanolicus is needed. here a systematic evaluation of different techniques for establishing a reliable basis for metabolome investigations is presented. | 2016 | 27251037 |
| growth of bacillus methanolicus in 2 m methanol at 50 °c: the effect of high methanol concentration on gene regulation of enzymes involved in formaldehyde detoxification by the ribulose monophosphate pathway. | bacillus methanolicus mga3 is a gram-positive aerobic methylotroph growing optimally at 50-53°c. methylotrophy in b. methanolicus is encoded on pbm19 and by two chromosomal copies of the methanol dehydrogenase (mdh), hexulose phosphate synthase (hps) and phosphohexuloisomerase (phi) genes. however, there are no published studies on the regulation of methylotrophy or the dominant mechanism of detoxification of intracellular formaldehyde in response to high methanol concentration. the µ max of b. ... | 2015 | 25952117 |
| transcriptome analysis of thermophilic methylotrophic bacillus methanolicus mga3 using rna-sequencing provides detailed insights into its previously uncharted transcriptional landscape. | bacillus methanolicus mga3 is a thermophilic, facultative ribulose monophosphate (rump) cycle methylotroph. together with its ability to produce high yields of amino acids, the relevance of this microorganism as a promising candidate for biotechnological applications is evident. the b. methanolicus mga3 genome consists of a 3,337,035 nucleotides (nt) circular chromosome, the 19,174 nt plasmid pbm19 and the 68,999 nt plasmid pbm69. 3,218 protein-coding regions were annotated on the chromosome, 22 ... | 2015 | 25758049 |
| genomics of methylotrophy in gram-positive methylamine-utilizing bacteria. | gram-positive methylotrophic bacteria have been known for a long period of time, some serving as model organisms for characterizing the specific details of methylotrophy pathways/enzymes within this group. however, genome-based knowledge of methylotrophy within this group has been so far limited to a single species, bacillus methanolicus (firmicutes). the paucity of whole-genome data for gram-positive methylotrophs limits our global understanding of methylotrophy within this group, including the ... | 2015 | 27682081 |
| methylotrophy in the thermophilic bacillus methanolicus, basic insights and application for commodity production from methanol. | using methanol as an alternative non-food feedstock for biotechnological production offers several advantages in line with a methanol-based bioeconomy. the gram-positive, facultative methylotrophic and thermophilic bacterium bacillus methanolicus is one of the few described microbial candidates with a potential for the conversion of methanol to value-added products. its capabilities of producing and secreting the commercially important amino acids l-glutamate and l-lysine to high concentrations ... | 2015 | 25431011 |
| complete genome sequence of bacillus methanolicus mga3, a thermotolerant amino acid producing methylotroph. | bacillus methanolicus mga3 was isolated from freshwater marsh soil and characterised as a thermotolerant and methylotrophic l-glutamate producer. the complete genome consists of a circular chromosome and the two plasmids pbm19 and pbm69. it includes genomic information about c1 metabolism and amino acid biosynthetic pathways. | 2014 | 25152427 |
| in vitro activation of nad-dependent alcohol dehydrogenases by nudix hydrolases is more widespread than assumed. | in the gram-positive methylotroph bacillus methanolicus, methanol oxidation is catalyzed by an nad-dependent methanol dehydrogenase (mdh) that belongs to the type iii alcohol dehydrogenase (adh) family. it was previously shown that the in vitro activity of b. methanolicus mdh is increased by the endogenous activator protein act, a nudix hydrolase. here we show that this feature is not unique, but more widespread among type iii adhs in combination with act or other act-like nudix hydrolases. in a ... | 2014 | 24928437 |
| proteomic analysis of the thermophilic methylotroph bacillus methanolicus mga3. | bacillus methanolicus mga3 is a facultative methylotroph of industrial relevance that is able to grow on methanol as its sole source of carbon and energy. the gram-positive bacterium possesses a soluble nad(+) -dependent methanol dehydrogenase and assimilates formaldehyde via the ribulose monophosphate (rump) cycle. we used label-free quantitative proteomics to generate reference proteome data for this bacterium and compared the proteome of b. methanolicus mga3 on two different carbon sources (m ... | 2014 | 24452867 |
| expression of recombinant green fluorescent protein in bacillus methanolicus. | microbial biocatalysts are used in a wide range of industries to produce large scale quantities of proteins, amino acids, and commodity chemicals. while the majority of these processes use glucose or other low-cost sugars as the substrate, bacillus methanolicus is one example of a biocatalyst that has shown sustained growth on methanol as a carbon source at elevated temperature (50-53°c optimum) resulting in reduced feed and utility costs. specifically, the complete chemical process enabled by t ... | 2012 | 22275315 |