chemotaxis by bdellovibrio bacteriovorus toward prey.a chemotaxis assay system that uses a modified boyden chamber was characterized and used for measurements of chemotaxis by bdellovibrio bacteriovorus strain uki2 toward several bacterial species. bacteria tested included both susceptible and nonsusceptible cells (escherichia coli, pseudomonas fluorescens, bacillus megaterium, and b. bacteriovorus strains uki2 and d). none was attractive to bdellovibrios when present at densities below 10(7) cells per ml. chemotaxis toward e. coli was studied mos ...1977410796
liquid nitrogen cryo-impacting: a new concept for cell disruption.high-efficiency disruption of bacteria can be accomplished in 2 or more min by the new procedure of liquid nitrogen cryo-impacting. release of the dipicolinic acid-ca2+ chelate paralleled the breakage of bacillus megaterium endospores. lactate dehydrogenase activity was much better in supernates from liquid nitrogen cryo-impacting-broken escherichia coli cells than in those from sonically treated and broken e. coli cells.1975810088
cloning and sequencing of the genes encoding glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase and triosephosphate isomerase (gap operon) from mesophilic bacillus megaterium: comparison with corresponding sequences from thermophilic bacillus stearothermophilus.the structural genes encoding glyceraldehyde-3-phosphate dehydrogenase (gapdh), 3-phosphoglycerate kinase (pgk) and the n-terminal part of triosephosphate isomerase (tim) from mesophilic bacillus megaterium dsm319 have been cloned as a gene cluster (gap operon) by complementation of an escherichia coli gap amber mutant. subsequently, the entire tpi gene, encoding tim, was isolated by colony hybridization using a homologous probe. nucleotide (nt) sequence analysis revealed an unidentified open re ...19921452037
complementation of an rnase p rna (rnpb) gene deletion in escherichia coli by homologous genes from distantly related eubacteria.we report the construction of a strain of escherichia coli in which the only functional gene for the rna moiety of rnase p (rnpb) resides on a plasmid that is temperature sensitive for replication. the chromosomal rnase p rna gene was replaced with a chloramphenicol acetyltransferase gene. the conditionally lethal phenotype of this strain was suppressed by plasmids that carry rnase p rna genes from some distantly related eubacteria, including alcaligenes eutrophus, bacillus subtilis, and chromat ...19901699929
the 75-kilodalton cytoplasmic chlamydia trachomatis l2 polypeptide is a dnak-like protein.the gene coding for the 75-kilodalton cytoplasmic chlamydia trachomatis l2 polypeptide has been cloned in escherichia coli, and the nucleotide sequence has been determined. the cloned dna fragment contained the coding region as well as the putative promoter. the deduced amino acid sequence of the 1,980-base-pair open reading frame revealed 94% homology with a 75-kilodalton protein from c. trachomatis serovar d and 57% homology with the dnak proteins of e. coli and of bacillus megaterium, while a ...19902365454
antimicrobial activity of dimethyl sulfoxide against escherichia coli, pseudomonas aeruginosa, and bacillus megaterium. 19694980332
lipid synthesis in relation to the cell cycle of bacillus megaterium km and escherichia coli.lipid synthesis during the cell duplication cycle of bacillus megaterium km and escherichia coli was studied by glycerol incorporation both in synchronized cultures and in unsynchronized exponentially growing populations subsequently fractionated according to size (and age). a large transient increase in the rate of incorporation per unit cell mass was observed around the time of cell division, probably reflecting the synthesis of the division septum.19694982416
structure of the peptidoglycan in escherichia coli b and bacillus megaterium km. stereospecific synthesis of two meso-diaminopimelic acid peptides with the tetrapeptide subunit of bacterial cell wall peptidoglycan. 19704984729
ribonucleic acid synthesis in bacteria treated with toluene.escherichia coli and bacillus megaterium rendered permeable to ribonucleoside triphosphates by toluene treatment retain the capacity to synthesize discrete ribonucleic acid species.19715000311
some features of the bacterial membrane studied with the aid of a new fractionation technique.membranes of bacillus megaterium and other bacteria were bound to crystals of cadmium lauroylsarcosinate and were resolved into nine lipoprotein fractions by elution with potassium chloride and sodium deoxycholate solutions. the fractions differed widely in protein/lipid ratio. some major protein species were probably common to all the fractions, but the phospholipid composition showed some variation. electron-microscopic examination after negative staining revealed that material in certain frac ...19715000708
identification of the altered ribosomal component responsible for resistance to micrococcin in mutants of bacillus megaterium.a mutant strain of bacillus megaterium, arising spontaneously and resistant to micrococcin , possesses ribosomes which contain an altered form of protein bm-l11 (the homologue of escherichia coli protein l11). reconstitution analysis has revealed that the alteration to protein bm-l11 is the sole cause of resistance in this strain.19846144543
[polyethylene glycol induction of bacillus megaterium protoplast transformation by plasmid dna]. 19806772419
evaluation of penicillin acylase production by two strains of bacillus megaterium.penicillin acylase is a key enzyme for the production of semisynthetic beta-lactam antibiotics. the intracellular enzyme from escherichia coli has been thoroughly studied and characterized. the extracellular enzyme from bacillus megaterium, despite its potential advantages, has received less attention in the recent scientific literature. a comparative study is presented for the production of penicillin acylase with two strains of bacillus megaterium in batch fermentation in previously optimized ...19937606255
inactivation of the major extracellular protease from bacillus megaterium dsm319 by gene efficient method for gene replacement in bacillus megaterium was developed and used to inactivate the chromosomal neutral protease gene (nprm) from strain dsm319. a temperature-dependant suicide vector was constructed to allow replacement of the normal chromosomal copy with an altered version of the nprm gene. one mutant b. megaterium ms941 was selected for further characterization. measurement of extracellular protease activity from strain ms941 indicated the existence of an additional minor ...19957766087
cloning and sequencing of the pac gene encoding the penicillin g acylase of bacillus megaterium atcc 14945.the pac gene encoding the penicillin g acylase (pga) of bacillus megaterium atcc 14945 has been cloned in escherichia coli hb101 (proa, leub) using a selective minimal medium containing phenylacetyl-l-leucine instead of l-leucine. the nucleotide sequence of this gene has been determined and contains an open reading frame of 2406 nucleotides. the deduced amino acid sequence shows significant similarity with other beta-lactam acylases. although the pga of b. megaterium is extracellular, the enzyme ...19957875576
the occurrence of duplicate lysyl-trna synthetase gene homologs in escherichia coli and other procaryotes.the lysyl-trna synthetase (lysrs) system of escherichia coli k-12 consists of two genes, lyss, which is constitutive, and lysu, which is inducible. it is of importance to know how extensively the two-gene lysrs system is distributed in procaryotes, in particular, among members of the family enterobacteriaceae. to this end, the enterics e. coli k-12 and b; e. coli reference collection (ecor) isolates ec2, ec49, ec65, and ec68; shigella flexneri; salmonella typhimurium; klebsiella pneumoniae; ente ...19957896714
complementation of an enterococcus hirae (streptococcus faecalis) mutant in the alpha subunit of the h(+)-atpase by cloned genes from the same and different species.we isolated an enterococcus hirae (formerly streptococcus faecalis) mutant, designated ms117, in which 'g' at position 301 of the alpha-subunit gene of the f1f0 type of h(+)-atpase was deleted. ms117 had low h(+)-atpase activity, was deficient in the regulatory system of cytoplasmic ph, and was unable to grow at ph 6.0. when the alpha-subunit gene of e. hirae h(+)-atpase was ligated with the shuttle vector phy300plk at the downstream region of the tet gene of the vector, it was expressed without ...19938412656
multidrug pump inhibitors uncover remarkable activity of plant antimicrobials.plant antimicrobials are not used as systemic antibiotics at present. the main reason for this is their low level of activity, especially against gram-negative bacteria. the reported mic is often in the range of 100 to 1,000 micro g/ml, orders of magnitude higher than those of common broad-spectrum antibiotics from bacteria or fungi. major plant pathogens belong to the gram-negative bacteria, which makes the low level of activity of plant antimicrobials against this group of microorganisms puzzl ...200212234835
l-amino acid oxidase from trimeresurus jerdonii snake venom: purification, characterization, platelet aggregation-inducing and antibacterial l-amino acid oxidase (lao), designated as tj-lao, was purified to homogeneity from the venom of trimeresurus jerdonii by sephadex g-100 and q sepharose hp chromatography. the molecular weight of this enzyme was 110 kd as estimated by analytical gel filtration and was 55 kd by sds-polyacrylamide gel electrophoresis, suggesting that the enzyme is composed of two subunits. the enzyme has an absorption spectrum characteristic of flavoproteins, containing 2 moles of fmn per mole of enzyme. the n-t ...200212503878
chloride dependence of growth in bacteria.chloride is an abundant anion on earth but studies analyzing a possible function of chloride in prokaryotes are scarce. to address the question, we have tested 44 different gram-negative and gram-positive bacteria for a chloride dependence or chloride stimulation of growth. none required chloride for growth at their optimal growth (salt) conditions. however, in hyperosmotic media containing high concentrations of na+, 11 bacteria (aeromonas hydrophila, bacillus megaterium, bacillus subtilis, cor ...200312900036
reduced oxidative activities in escherichia coli and bacillus megaterium in relation to other changes during inhibition of growth by streptomycin. 196113711165
reconfirmation of antimicrobial activity in the coelomic fluid of the earthworm eisenia fetida andrei by colorimetric assay.a novel tetrazolium salt, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2h-tetrazolium (mts) was used in the assessment of antimicrobial activity in earthworm in the presence of phenazine methosulphate (pms) as an electron coupling reagent. this activity was purified from the coelomic fluid of the earthworm (ecf), eisenia fetida andrei (oligochaeta, lumbricidae, annelids) using a series of column chromatography techniques and was tested against three gram-negative st ...200314660872
antimicrobial effect of water extract of sumac (rhus coriaria l.) on the growth of some food borne bacteria including pathogens.the antimicrobial effect of water extracts of sumac (rhus coriaria l.) at concentrations of 0.1%, 0.5%, 1.0%, 2.5% and 5.0% (w/v), non-neutralized and after neutralization to ph 7.2+/-0.1, was studied on the growth of 12 bacterial strains (six gram positive strains and six gram negative strains), mostly food borne including pathogens. it was found to be effective against all the test organisms with gram positive strains being more sensitive than gram negative strains. significant differences (p< ...200415527919
peptidoglycan recognition protein (pgrp)-le and pgrp-lc act synergistically in drosophila innate immunity, pattern recognition molecules recognize cell wall components of microorganisms and activate subsequent immune responses, such as the induction of antimicrobial peptides and melanization in drosophila. the diaminopimelic acid (dap)-type peptidoglycan potently activates imd-dependent induction of antibacterial peptides. peptidoglycan recognition protein (pgrp) family members act as pattern recognition molecules. pgrp-lc loss-of-function mutations affect the imd-dependent induct ...200415538387
toxicological effect of indole and its azo dye derivatives on some microorganisms under aerobic conditions.azo dyes are ubiquitous commercial chemicals that present unique environmental problems. the azo dyes in particular can undergo natural anaerobic degradation to potentially carcinogenic amines. they pose a major problem for water-treatment plants downstream. one strategy to remidate polluted surface contaminants is to make use of the degradative capacity of bacteria rather than using destructive chemical reactions. therefore, pathogenic and non-pathogenic microorganisms namely; bacillus thuringi ...200616165191
[cloning and expression of the alpha-amylase gene from a bacillus sp. ws06, and characterization of the enzyme].a bacillus sp. ws06, which produces an extracellular alpha-amylase, was isolated from the cecum in a piglet. an amyf gene from this bacillus strain was cloned and its nucleotide sequence was determined. an open reading frame composed of 1581 bases, which encodes 526 amino acid residues was found. the amyf gene shows high sequence homologies with other microbial amylase genes, such as bacillus megaterium and bacillus polymyxa (93% and 53% identity). the deduced amino acid sequence revealed that f ...200516496695
insights into polymer versus oligosaccharide synthesis: mutagenesis and mechanistic studies of a novel levansucrase from bacillus megaterium.a novel levansucrase was identified in the supernatant of a cell culture of bacillus megaterium dsm319. in order to test for the contribution of specific amino acid residues to levansucrase catalysis, the wild-type enzyme along with 16 variants based on sequence alignments and structural information were heterologously produced in escherichia coli. the purified enzymes were characterized kinetically and the product spectrum of each variant was determined. comparison of the x-ray structures of th ...200717608626
antimicrobial activity of six pomegranate (punica granatum l.) varieties and their relation to some of their pomological and phytonutrient characteristics.arils from six pomegranate (punica granatum l.) varieties grown in the mediterranean region of turkey were tested for their antimicrobial properties by the agar diffusion and minimum inhibitory concentration (mic) methods against seven bacteria: (bacillus megaterium dsm 32, pseudomonas aeruginosa dsm 9027, staphylococcus aureus cowan 1, corynebacterium xerosis uc 9165, escherichia coli dm, enterococcus faecalis a10, micrococcus luteus la 2971), and threefungi (kluvyeromyces marxianus a230, rhodo ...200919471201
mbga-dependent lactose utilization by bacillus megaterium.the beta-galactosidase-encoding mbga gene has recently been cloned from bacillus megaterium. we now report that disruption of the chromosomal mbga rendered b. megaterium unable to utilize lactose as a sole carbon source. complementation of the mbga mutant with a multicopy plasmid carrying intact mbga restored the ability to utilize lactose for cell growth. crude extracts from the wild-type b. megaterium cells grown in the presence of lactose exhibited a significant level of lactose-hydrolyzing a ...200211815853
sugar uptake and carbon catabolite repression in bacillus megaterium strains with inactivated ptshi.we have determined the role played by the phosphoenolpyruvate:sugar phosphotransferase system (pts) in carbon catabolite repression (ccr) of xylose utilization in bacillus megaterium. for that purpose we have cloned, sequenced and inactivated the genes ptsh and ptsl of b. megaterium, encoding hpr and ei of the pts, respectively. while glucose uptake of a ptshi mutant is not affected at 12.5 mm of glucose, ccr of the xyl operon is reduced in this mutant from 16-fold to 3-fold. this may be attribu ...200011075936
spice extracts as dose-modifying factors in radiation inactivation of bacteria.three spices, chili, black pepper, and turmeric, were tested for the effect of their aqueous extracts on the sensitivity of three bacteria, escherichia coli, bacillus megaterium, and bacillus pumilusspores, to gamma-radiation. it was found that the extracts of the three spices offered protection to these organisms against inactivation by gamma-radiation. these spice extracts were also tested for their protection of naked plasmid dna. radiation-induced degradation of plasmid puc18 dna was reduced ...200010775394
synthesis, antibacterial activity and quantum-chemical studies of novel 2-arylidenehydrazinyl-4-arylthiazole analogues.a new series of 2-arylidenehydrazinyl-4-arylthiazole derivatives (2a-k) was designed and synthesized through a rapid, simple, and efficient methodology in excellent isolated yield. these compounds were screened for in vitro antimicrobial activities against eight bacteria, e.g. bacillus cereus, staphylococcus aureus, bacillus subtilis, bacillus megaterium, pseudomonas aeruginosa, shigella dysenteriae, salmonella typhi, escherichia coli, and three fungi e.g. aspergillus oryzae, candida albicans, a ...201121532194
p450 monooxygenase in biotechnology. i. single-step, large-scale purification method for cytochrome p450 bm-3 by anion-exchange efficient single-step purification protocol for recombinant cytochrome p450 bm-3 from bacillus megaterium, expressed in e. coli, was developed. functional crude protein was obtained by disintegrating induced e. coli dh5 alpha and removing cell debris by centrifugation. after investigating different anion-exchange matrices, elution salts and the elution procedures involving an aktaexplorer system, adsorption of the crude extract from lysed e. coli to toyopearl deae 650m anion exchanger, follow ...199910427755
expression of periplasmic alpha-amylase of xanthomonas campestris k-11151 in escherichia coli and its action on maltose.a gene encoding the periplasmic alpha-amylase of xanthomonas campestris k-11151 was cloned into escherichia coli using puc19 as a vector. an orf of 1578 bp was deduced to be the amylase structural gene. the primary structure of the enzyme had little identity with other alpha-amylases, except with the enzyme from bacillus megaterium. the enzyme was expressed in e. coli from the lac promoter of puc19 and was found to be transported to the periplasmic space. the expressed enzyme showed the same the ...19968704990
2,3-bisphosphoglycerate-independent phosphoglycerate mutase is conserved among different phylogenic kingdoms.we have previously demonstrated that maize (zea mays) 2,3-bisphosphoglycerate-independent phosphoglycerate mutase (pgam-i) is not related to 2,3-bisphosphoglycerate-dependent phosphoglycerate mutase. with the aid of specific anti-maize pgam-i antibodies, we demonstrate here the presence of a closely related pgam-i in other plants. we also describe the isolation and sequencing of a cdna-encoding almond (prunus amygdalus) pgam-i that further demonstrates this relationship among plant pgam-i. a sea ...19957584858
a new spectrophotometric assay for citrate synthase and its use to assess the inhibitory effects of palmitoyl thioesters.we have demonstrated that citrate synthase may be assayed by a simple, discontinuous, spectrophotometric procedure based on the measurement of oxaloacetate utilization with 2,4-dinitrophenylhydrazine. the assay is applicable both to the purified enzyme and to cell extracts, and has the advantage that it can be used in the presence of high concentrations of thiols and thioesters. we have used this new assay in part of our investigations into the inhibitory effects of palmitoyl thioesters on diver ...19883137924
enzymatic properties of isozymes and variants of glucose dehydrogenase from bacillus megaterium.three glucose dehydrogenases (glcdh) from bacillus megaterium, glcdh-i, glcdh-ii and glcdh-iwg3, were purified from escherichia coli cells harboring one of the hybrid plasmids, pgdk1, pgdk2 and pgda3, respectively, pgdk1 and pgdk2 contain two isozyme genes, gdhi and gdhii, respectively, from b. megaterium iam 1030 and pgda3 contains an isozyme gene from b. megaterium iwg3; glcdh-iwg3 is a variant of glcdh-i. glcdh-i and glcdh-ii have similar ph/activity profiles and the profile for glcdh-iwg3 is ...19892513190
the effect of atebrin on bacterial membrane adenosine triphosphatases in relation to the divalent cation used as substrate and/or activator.the action of atebrin on purified adenosine triphosphatase (atpase) from micrococcus lysodeikticus was studied as well as on the membrane-bound and soluble atpases from escherichia coli and bacillus megaterium. atebrin inhibited the ca(2+)-dependent activity of all these enzymes, and the inhibition was reversed by an excess of ca(2+) ions. kinetic studies carried out with the purified enzyme from m. lysodeikticus showed that the inhibition by atebrin was strongly cooperative, suggesting the comp ...1977138384
isolation of a gram-positive poly(3-hydroxybutyrate) (phb)-degrading bacterium from compost, and cloning and characterization of a gene encoding phb depolymerase of bacillus megaterium n-18-25-9.a gram-positive poly(3-hydroxybutyrate) (phb)-degrading bacterial strain was isolated from compost. this organism, identified as bacillus megaterium n-18-25-9, produced a clearing zone on opaque nb-phb agar, indicating the presence of extracellular phb depolymerase. a phb depolymerase gene, phaz(bm), of b. megaterium n-18-25-9 was cloned and sequenced, and the recombinant gene product was purified from escherichia coli. the n-terminal half region of phaz(bm) shared significant homologies with a ...200617064368
overview of bacterial expression systems for heterologous protein production: from molecular and biochemical fundamentals to commercial systems.during the proteomics period, the growth in the use of recombinant proteins has increased greatly in the recent years. bacterial systems remain most attractive due to low cost, high productivity, and rapid use. however, the rational choice of the adequate promoter system and host for a specific protein of interest remains difficult. this review gives an overview of the most commonly used systems: as hosts, bacillus brevis, bacillus megaterium, bacillus subtilis, caulobacter crescentus, other str ...200616791589
quantitatively determined uptake of cell-penetrating peptides in non-mammalian cells with an evaluation of degradation and antimicrobial effects.cell-penetrating peptides (cpps) are carriers developed to improve mammalian cell uptake of important research tools such as antisense oligonucleotides and short interfering rnas. however, the data on cpp uptake into non-mammalian cells are limited. we have studied the uptake and antimicrobial effects of the three representative peptides penetratin (derived from a non-mammalian protein), map (artificial peptide) and pvec (derived from a mammalian protein) using fluorescence hplc in four common m ...200616500001
regio- and enantioselective alkane hydroxylation with engineered cytochromes p450 bm-3.cytochrome p450 bm-3 from bacillus megaterium was engineered using a combination of directed evolution and site-directed mutagenesis to hydroxylate linear alkanes regio- and enantioselectively using atmospheric dioxygen as an oxidant. bm-3 variant 9-10a-a328v hydroxylates octane at the 2-position to form s-2-octanol (40% ee). another variant, 1-12g, also hydroxylates alkanes larger than hexane primarily at the 2-position but forms r-2-alcohols (40-55% ee). these biocatalysts are highly active (r ...200314583039
analysis of bacillus megaterium lipolytic system and cloning of lipa, a novel subfamily i.4 bacterial lipase.the lipolytic system of bacillus megaterium 370 was investigated, showing the existence of at least two secreted lipases and a cell-bound esterase. a gene coding for an extracellular lipase was isolated and cloned in escherichia coli. the cloned enzyme displayed high activity on short to medium chain length (c(4)-c(8)) substrates, and poor activity on c(18) substrates. on the basis of amino acid sequence homology, the cloned lipase was classified into subfamily i.4 of bacterial lipases.200212480114
isolation of three antibacterial peptides from pig intestine: gastric inhibitory polypeptide (7-42), diazepam-binding inhibitor (32-86) and a novel factor, peptide 3910.two antibacterial peptides, cecropin p1 and pr-39 (39-residue proline/arginine-rich peptide), from the upper part of pig small intestine have previously been isolated and characterized. we have now continued our search for antibacterial peptides in different side fractions generated during the isolation of intestinal hormones. starting from one such fraction and monitoring activity against bacillus megaterium, we isolated three homogeneous peptides by three consecutive chromatographic steps. ami ...19938375398
fall-39, a putative human peptide antibiotic, is cysteine-free and expressed in bone marrow and, a proline/arginine-rich peptide antibiotic, has been purified from pig intestine and later shown to originate in the bone marrow. intending to isolate a clone for a human counterpart to pr-39, we synthesized a pcr probe derived from the pr-39 gene. however, when this probe was used to screen a human bone marrow cdna library, eight clones were obtained with information for another putative human peptide antibiotic, designated fall-39 after the first four residues. fall-39 is a 39-residue p ...19957529412
characterization of the protein expressed in escherichia coli by a recombinant plasmid containing the bacillus megaterium cytochrome p-450bm-3 two previous reports (narhi lo, fulco aj, j. biol. chem. 261: 7160-7169, 1986; ibid., 262: 6683-6690, 1987) we described the characterization of a catalytically self-sufficient 119,000-dalton p-450 cytochrome that was induced by barbiturates in bacillus megaterium. in the presence of nadph and o2, this polypeptide (cytochrome p-450bm-3) catalyzed the hydroxylation of long-chain fatty acids without the aid of any other protein. the gene encoding this unique monooxygenase was cloned into escher ...19883131661
catabolite repression of the xyl operon in bacillus megaterium.we characterized catabolite repression of the genes encoding xylose utilization in bacillus megaterium. a transcriptional fusion of xyla encoding xylose isomerase to the spovg-lacz indicator gene on a plasmid with a temperature-sensitive origin of replication was constructed and efficiently used for single-copy replacement cloning in the b. megaterium chromosome starting from a single transformant. in the resulting strain, beta-galactosidase expression is 150-fold inducible by xylose and 14-fold ...19921569031
requirement for ribosomal protein bm-l11 in stringent control of rna synthesis in bacillus megaterium.a spontaneously occurring thiostrepton-resistant mutant of bacillus megaterium has been shown to yield ribosomes lacking protein bm-l11, a protein immunologically related to escherichia coli ribosomal protein l11. here we have demonstrated that the mutant strain has acquired the relaxed phenotype and is unable to synthesise guanosine tetraphosphate and pentaphosphate in vivo. ribosomes from the mutant strain are unable to support the synthesis of these two compounds in vitro, but this deficiency ...1979118006
production of recombinant antibody fragments in bacillus megaterium.recombinant antibodies are essential reagents for research, diagnostics and therapy. the well established production host escherichia coli relies on the secretion into the periplasmic space for antibody synthesis. due to the outer membrane of gram-negative bacteria, only a fraction of this material reaches the medium. recently, the gram-positive bacterium bacillus megaterium was shown to efficiently secrete recombinant proteins into the growth medium. here we evaluated b. megaterium for the reco ...200717224052
recent biotechnological interventions for developing improved penicillin g acylases.penicillin g acylase (pac; ec is the key enzyme used in the industrial production of beta-lactam antibiotics. this enzyme hydrolyzes the side chain of penicillin g and related beta-lactam antibiotics releasing 6-amino penicillanic acid (6-apa), which is the building block in the manufacture of semisynthetic penicillins. pac from escherichia coli strain atcc 11105, bacillus megaterium strain atcc 14945 and mutants of these two strains is currently used in industry. genes encoding for pa ...200416233581
enhancement of recombinant enzyme activity in cpxa-deficient mutant of escherichia coli.we have previously performed a systematic screening of single-gene knockout collection of escherichia coli bw25113 (the keio collection) to investigate the effect of cellular components on the activity of whole-cell catalysts (zhou et al. appl. microbiol. biotechnol. in press (2010)). in the present paper, the cpxa-deficient mutant of e.coli bw25113 was found to be able to enhance the activity of cytochrome p450s, including cyp154a1 of streptomyces coelicolor, compactin 6β-hydroxylase of strepto ...201020547366
crystallization and preliminary x-ray analyses of catabolite control protein a, free and in complex with its dna-binding site.the catabolite control protein (ccpa) from bacillus megaterium is a member of the bacterial repressor protein family galr/laci. ccpa with an n-terminal his-tag was used for crystallization. crystals of free ccpa and of ccpa in complex with the putative operator sequence (catabolite responsive elements, cre) were obtained by vapour-diffusion techniques at 291 k using the hanging-drop method. ccpa crystals grown in the presence of polyethylene glycol 8000 belong to the hexagonal space group p6(1)2 ...200010666630
isolation of bacillus megaterium mutants that produce high levels of heterologous protein, and their use to construct a highly mosquitocidal strain.a xylose-regulated plasmid expression system for producing high levels of recombinant proteins in bacillus megaterium has recently been described [appl microbiol biotechnol 35:594, 1991]. using an antibiotic resistance protein as the expressed protein, we have been able to select mutant plasmids that produce increased levels of heterologous protein. the mutant plasmids show increased segregational stability and have lost the ability to be transformed into escherichia coli. the same selection pro ...19979216879
organomercurials removal by heterogeneous merb genes harboring bacterial strains.organomercury lyase (merb) is a key enzyme in bacterial detoxification and bioremediation of organomercurials. however, the merb gene is often considered as an ancillary component of the mer operon because there is zero to three merb genes in different mer operons identified so far. in this study, organomercurials' removal abilities of native mercury-resistant bacteria that have one or multiple merb genes were examined. each heterogeneous merb genes from these bacteria was further cloned into es ...201020541123
high-level expression of recombinant glucose dehydrogenase and its application in nadph regeneration.two glucose dehydrogenase (e.c. genes, gdh223 and gdh151, were cloned from bacillus megaterium as1.223 and as1.151, and were inserted into pqe30 to construct the expression vectors, pqe30-gdh223 and pqe30-gdh151, respectively. the transformant escherichia coli m15 with pqe30-gdh223 gave a much higher glucose dehydrogenase activity than that with the plasmid pqe30-gdh151. thus it was used to optimize the expression of glucose dehydrogenase. an proximately tenfold increase in gdh activit ...200716941118
biocatalytic synthesis of (s)-4-chloro-3-hydroxybutanoate ethyl ester using a recombinant whole-cell catalyst.a cofactor regeneration system for enzymatic biosynthesis was constructed by coexpressing a carbonyl reductase from pichia stipitis and a glucose dehydrogenase from bacillus megaterium in escherichia coli rosetta (de3) plyss. transformants containing the polycistronic plasmid pet-pii-sd2-as1-b exhibited an activity of 13.5 u/mg protein with 4-chloro-3-oxobutanoate ethyl ester (cobe) as the substrate and an activity of 14.4 u/mg protein with glucose as the substrate; nad(h) was the coenzyme in bo ...201020725723
production, secretion, and cell surface display of recombinant sporosarcina ureae s-layer fusion proteins in bacillus megaterium.monomolecular crystalline bacterial cell surface layers (s-layers) have broad application potential in nanobiotechnology due to their ability to generate functional supramolecular structures. here, we report that bacillus megaterium is an excellent host organism for the heterologous expression and efficient secretion of hemagglutinin (ha) epitope-tagged versions of the s-layer protein ssla from sporosarcina ureae atcc 13881. three chimeric proteins were constructed, comprising the precursor, c-t ...201222101038
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