| phylogeny of rapidly growing members of the genus mycobacterium. | the 16s rrnas from nine rapidly growing mycobacterium species were partially sequenced by using the dideoxynucleotide-terminated, primer extension method with cdna generated by reverse transcriptase. the sequences were aligned with 47 16s rrna or dna sequences that represented 30 previously described and 5 undescribed species of the genus mycobacterium, and a dendrogram was constructed by using equally weighted distance values. our results confirmed the phylogenetic separation of the rapidly and ... | 1992 | 1380284 |
| co-ordinated expression of the components of iron transport (mycobactin, exochelin and envelope proteins) in mycobacterium neoaurum. | mycobacterium neoaurum was grown with a range of iron concentrations from 0.01 to 4.0 micrograms/ml. synthesis of the extracellular siderophore, exochelin, the intracellular iron storage compound, mycobactin and the iron-repressible envelope proteins were co-ordinately expressed. all three components of the iron transport system were synthesized when low amounts of iron (0.01 to 0.2 micrograms/ml) were added to the medium and were repressed when the iron concentration was increased to 0.5 microg ... | 1989 | 2777064 |
| bacteremia caused by mycobacterium neoaurum. | an immunocompromised patient with an indwelling hickman catheter developed mycobacterium neoaurum bacteremia. this rapidly growing mycobacterium was previously isolated from soil, dust, and water but has not been described as a human pathogen. the infection responded to therapy with cefoxitin and gentamicin. it was not necessary to remove the hickman catheter. | 1988 | 3366872 |
| mycobactin and the competition for iron between mycobacterium neoaurum and m. vaccae. | two closely related species of mycobacteria, mycobacterium vaccae and m. neoaurum, were grown under conditions of iron-deficiency (0.02-0.05 microgram fe ml-1) and iron-sufficiency (2-4 micrograms fe ml-1) in a simple glycerol/asparagine medium. the strain of m. vaccae used was a nonmycobactin producer whereas m. neoaurum synthesized between 6-8% of its cell biomass as the lipid-soluble siderophore when grown under iron-limitation. the role of mycobactin for iron-acquisition was examined using b ... | 1986 | 3734752 |
| mycolic acid patterns of some rapidly-growing species of mycobacterium. | representative strains of some rapidly-growing species of mycobacterium were degraded by both acid methanolysis and alkaline hydrolysis followed by phase-transfer catalysed conversion to methyl esters. patterns of mycolic acids and other long-chain components were determined for 56 samples by two-dimensional thin-layer chromatography. representatives of mycobacterium chitae contained epoxymycolates and alpha and alpha'-mycolates, lacking oxygen functions in addition to the 3-hydroxyacid unit; a ... | 1985 | 4050199 |
| mycobacterium neoaurum infection of a hickman catheter in an immunosuppressed patient. | mycobacterium neoaurum is a rapidly growing mycobacterium that has only once before been reported as a cause of bacteremia in an immunocompromised patient. we report on a patient who developed bacteremia after an allogeneic bone marrow transplantation and from whose blood (taken via a hickman catheter) m. neoaurum was isolated on three separate occasions. there was a clinical response to therapy with ticarcillin/clavulanate and tobramycin, and the organism appeared to be susceptible to the forme ... | 1994 | 8086527 |
| exochelin-mediated iron acquisition by the leprosy bacillus, mycobacterium leprae. | exochelins, water-soluble siderophores of mycobacteria, were isolated and partially purified from culture filtrates of iron-deficiently grown cultures of mycobacterium neoaurum nctc 10439 and an armadillo-derived mycobacterium (adm 8563). two biologically active fractions mediating iron uptake were isolated from each bacterium which not only were able to transport iron into the producing organism but also into suspensions of mycobacterium leprae isolated from armadillo liver. the rate of exochel ... | 1987 | 3309144 |
| numerical analysis of relationships among rapidly growing, scotochromogenic mycobacteria. | numerical analysis of the relationships among rapidly growing, scotochromogenic mycobacteria was carried out; a dendrogram was prepared by a single linkage method and the separation between species was tested. the following were distinct species: mycobacterium alichiense; mycobacterium rhodesiae;mycobacterium chubuense; mycobacterium obuense; mycobacterium gilvum; mycobacterium duvalii; mycobacterium flavescens; and mycobacterium vaccae. mycobacterium tokaiense was reduced to a synonym of m. chu ... | 1977 | 853289 |
| determination of the structure of exochelin mn, the extracellular siderophore from mycobacterium neoaurum. | siderophores are compounds produced by bacteria to acquire iron. exochelin mn, the extracellular siderophore from mycobacterium neoaurum, is of particular interest because it has been shown to transport iron into m. leprae, which is responsible for the disease leprosy. exochelins from other species cannot mediate iron transport in m. leprae, suggesting a specific uptake mechanism involving exochelin mn. we set out to determine the structure of exochelin mn and identify the features of the molecu ... | 1995 | 9383459 |
| mycobacterium neoaurum--an unusual cause of infection of vascular catheters: case report and review. | | 1999 | 10194099 |
| identification of mycobacterium neoaurum isolated from a neutropenic patient with catheter-related bacteremia by 16s rrna sequencing. | a rapidly growing pigmented mycobacterial strain with an ambiguous biochemical profile was isolated from the blood culture taken through the hickman catheter of a 9-year-old girl with acute lymphoblastic leukemia. whole-cell fatty acid analysis showed that the best match profile was that of mycobacterium aurum, but the similarity index was only 0.217, meaning that there were no good matches between the isolate and the organisms in the database of the microbial identification system. the 16s rrna ... | 2000 | 10970421 |
| a novel method for the isolation of mycobacterial dna. | dna was isolated from mycobacteria by a simplified procedure. cells were suspended in 6 m guanidinium chloride, the suspension was cooled to -70 degrees c, then incubated at 65 degrees c for 10 min, cooled in ice, deproteinized by chloroform and dna was recovered from the supernatant. the procedure was used to obtain dna from several mycobacteria (1 x 10(9) or more cells) including mycobacterium neoaurum, m. fortuitum, m. phlei and m. smegmatis. each of the species was shown to have two ribosoma ... | 1996 | 8598279 |
| mycobacterium frederiksbergense sp. nov., a novel polycyclic aromatic hydrocarbon-degrading mycobacterium species. | a polycyclic aromatic hydrocarbon-degrading bacterium isolated from coal tar-contaminated soil in denmark was characterized by a polyphasic approach. phylogenetically and chemotaxonomically, it was related to members of the genus mycobacterium. the isolate contains chemotaxonomic markers that are diagnostic for the genus mycobacterium; i.e. the meso isomer of 2,6-diaminopimelic acid, arabinose and galactose as diagnostic whole-cell sugars, mk-9(h2) as the principal isoprenoid quinone, a mycolic ... | 2001 | 11594601 |
| growth of mycobacteria on carbon monoxide and methanol. | several mycobacterial strains, such as mycobacterium flavescens, mycobacterium gastri, mycobacterium neoaurum, mycobacterium parafortuitum, mycobacterium peregrinum, mycobacterium phlei, mycobacterium smegmatis, mycobacterium tuberculosis, and mycobacterium vaccae, were found to grow on carbon monoxide (co) as the sole source of carbon and energy. these bacteria, except for m. tuberculosis, also utilized methanol as the sole carbon and energy source. a co dehydrogenase (co-dh) assay, staining by ... | 2003 | 12486050 |
| exochelin-mediated iron uptake into mycobacterium leprae. | iron chelated to the exochelins from mycobacterium neoaurum was taken up by a suspension of m. leprae, prepared from the liver of an infected armadillo, over 15 hr. no uptake occurred when the iron was chelated with exochelins from m. bovis bcg or m. smegmatis or to a single exochelin from m. vaccae. uptake appeared to be by facilitated diffusion since it was not inhibited by either hgcl2, nan3, or 2,4-dinitrophenol. this was similar to the mode of uptake of ferriexochelin into m. neoaurum itsel ... | 1983 | 6231257 |
| iron chelation properties of an extracellular siderophore exochelin mn. | the coordination chemistry of an extracellular siderophore produced by mycobacterium neoaurum, exochelin mn (exomn), is reported along with its pk(a) values, fe(iii) and fe(ii) chelation constants, and aqueous solution speciation as determined by spectrophotometric and potentiometric titration techniques. exochelin mn is of particular interest as it can efficiently transport iron into pathogenic m. leprae, which is responsible for leprosy, in addition to its own parent cells. the fe(iii) coordin ... | 2003 | 12812507 |
| arsenic methylation by micro-organisms isolated from sheepskin bedding materials. | sudden infant death syndrome (sids) has been associated with the volatilization of arsenic, antimony or phosphorus compounds from infants' bedding material by micro-organisms, the so-called 'toxic gas hypothesis'. the volatilization of arsenic by aerobic micro-organisms isolated from new sheepskin bedding material, as well as on material used by a healthy infant and by an infant who perished of sids, was examined. three fungi were isolated from a piece of sheepskin bedding material on which an i ... | 2003 | 12856955 |
| rapidly progressive dementia due to mycobacterium neoaurum meningoencephalitis. | dementia developed in a patient with widespread neurologic manifestations; she died within 5 months. pathologic findings showed granulomatous inflammation with caseation necrosis, foreign body-type giant cells, and proliferative endarteritis with vascular occlusions. broad-range polymerase chain reaction identified mycobacterium neoaurum as the possible pathogen. central nervous system infection by m. neoaurum may result in rapidly progressive dementia. | 2004 | 15200833 |
| mycobacterium cosmeticum sp. nov., a novel rapidly growing species isolated from a cosmetic infection and from a nail salon. | four isolates of a rapidly growing mycobacterium species had a mycolic acid pattern similar to that of mycobacterium smegmatis, as determined by hplc analyses. three of the isolates were from footbath drains and a sink at a nail salon located in atlanta, ga, usa; the fourth was obtained from a granulomatous subdermal lesion of a female patient in venezuela who was undergoing mesotherapy. by random amplified polymorphic dna electrophoresis and pfge of large restriction fragments, the three isolat ... | 2004 | 15545488 |
| mycobacterium fluoranthenivorans sp. nov., a fluoranthene and aflatoxin b1 degrading bacterium from contaminated soil of a former coal gas plant. | mycobacterium strain fa4t was isolated with fluoranthene as the single carbon source from soil of a former coal gas plant, polluted with polycyclic aromatic hydrocarbons. the physiological properties, fatty acid pattern, and the 16s ribosomal rna gene sequence indicated membership to the genus mycobacterium, but were different from all type strains of mycobacterium species. based on comparative 16s rrna gene sequence analyses strain fa4t could be assigned to the mycobacterium neoaurum taxon show ... | 2004 | 15612622 |
| 14-hydroxylation of opiates: catalytic direct autoxidation of codeinone to 14-hydroxycodeinone. | codeinone (3) was efficiently and directly converted to 14-hydroxycodeinone (1) by catalytic air oxidation in aqueous solution. a number of simple manganese and copper salts were identified to be effective catalysts, including mnso4, kmno4, and cuso4. an appropriate reducing agent, such as sodium thiosulfate, is required in the reaction mixture presumably for the reduction of a detrimental peroxide intermediate. this discovery allows the more abundant codeine to be employed as the starting mater ... | 2005 | 15898757 |
| mycobacterium hodleri sp. nov., a new member of the fast-growing mycobacteria capable of degrading polycyclic aromatic hydrocarbons. | a bacterial strain isolated from a fluoranthene-polluted soil was characterized with respect to its metabolic and chemotaxonomic properties, and its phylogenetic position was determined. this bacterium exhibits all of the genus-specific properties of the genus mycobacterium and clusters phylogenetically with the group of fast-growing mycobacteria. on the basis of its unique fatty acid pattern, the distinctness of its physiological properties and the uniqueness of the primary structure of its 16s ... | 1996 | 8782676 |
| purification and characterization of an l-amino amidase from mycobacterium neoaurum atcc 25795. | an l-amino amidase from mycobacterium neoaurum atcc 25795 responsible for the enantioselective resolution of dl-alpha-methyl valine amide was purified and characterized. the purification procedure included ammonium sulfate fractionation, gel filtration, and anion-exchange chromatography, which resulted in a homogeneous preparation of the enzyme with a native molecular mass of 136 kda and a subunit molecular mass of 40 kda. the purified enzyme displayed the highest activity at 50 degrees c and at ... | 1994 | 16349147 |
| [identification of a new steroid transforming strain of mycobacteria as mycobacterium neoaurum]. | the ability to utilize sterols as a sole source of carbon was studied in 80 strains and consortia of hydrocarbon-oxidizing bacteria. one of the strains, which efficiently transformed both individual sterols and their mixtures, was identified as mycobacterium neoaurum, based on the analysis of the sequence of the 16s rrna gene. | 2003 | 12722650 |
| cutaneous mycobacterium neoaurum infection causing scarring alopecia in an immunocompetent host. | | 2007 | 17501949 |
| isolation and identification of mycobacterium neoaurum from a patient with urinary infection. | mycobacterium neoaurum is a novel species of mycobacteria, until now only isolated from catheters in immunosuppressed patients. this report describes the isolation and identification of m. neoaurum from urine obtained from a hospitalized patient. | 2001 | 11346303 |
| [preparation of androsta-1,4-diene-3,17-dione from sterols using mycobacterium neoaurum vkpm as-1656 strain]. | a product of microbiological cleavage of the sterols side chain, androsta-1,4-diene-3,17-dione, is toxic for bacteria, in particular, actinobacteria of the genera mycobacterium and arthrobacter. sterols were transformed into androsta-1,4-diene-3,17-dione by culturing the m. neoaurum vkpm an-1656 strain in a high yield, provided that a sorbent was used for elimination of contact between the bacterial cells and the product. unlike the cholesterol side chain, the more branched chains of phytosterol ... | 2007 | 17682396 |
| mycobacterium neoaurum bacteremia in a hemodialysis patient. | bacteremia due to mycobacterium neoaurum, a rapidly growing mycobacterium, is described in a diabetic woman on hemodialysis. this is the first reported case of m neoaurum bacteremia in canada. the organism initially grew on standard bact/alert sa aerobic blood cultures, and was subsequently positively identified using 16s rrna sequence analysis. the present case serves to reinforce the need for a high index of clinical suspicion of infections caused by unusual microorganisms in the context of an ... | 2003 | 18159425 |
| catheter-related bloodstream infections caused by rapidly growing nontuberculous mycobacteria: a case series including rare species. | rapidly growing nontuberculous mycobacteria (rgms) are responsible for a variety of clinical syndromes in humans including catheter-related blood stream infections (crbsis). recently, we identified a cluster of rgm-associated crbsi at our institution. we describe the epidemiologic and clinical patterns associated with these infections. we conducted a retrospective single-center review of rgm crbsi between may 2004 and june 2005. rgms isolated from blood cultures of 6 patients included mycobacter ... | 2008 | 18294801 |
| [conversion of phytosterols into androstenedione by mycobacterium neoaurum]. | optimum conditions for transformation of phytosterols by mycobacterium neoaururm, required for selective cleavage of the lateral chain into androstenedione, were shown to differ from the known conditions of animal sterol (cholesterol) transformation. complete conversion of phytosterols into androstenedione at a substrate load of no less than 20 g/l was achieved on increasing the amount of the inoculum and the concentration of glucose (by 2 and 4 times, respectively, relative to cholesterol) and ... | 2008 | 18491598 |
| mycobacterium neoaurum bloodstream infection: report of 4 cases and review of the literature. | we describe a cluster of 4 bloodstream infections with mycobacterium neoaurum and 5 additional cases from the literature. infections occurred mainly in immunocompromised hosts who had central venous catheters. fever was universal at presentation, but local signs of inflammation were rare. combination antimicrobial therapy and catheter removal resulted in clinical cure. | 2007 | 17578768 |
| [mycobacterium neoaurum bacteremia in an immunodepressed patient]. | | 2009 | 19218006 |
| exochelin production in mycobacterium neoaurum. | mycobacterium neoaurum is a soil saprophyte and obligate aerobic bacterium. this group of mycobacterium is relatively fast-growing. they form colonies on nutrient agar at 37 masculinec within 3 - 4 days. in natural soil habitats, bioavailability of iron is limited. to facilitate iron uptake, most mycobacteria produce siderophores. one example is exochelin, which is extracellular and water-soluble. in this report, the production of exochelin in m. neoaurum was induced in iron-deficiency, but repr ... | 2009 | 19333449 |
| phylogenetic analysis of mycobacterium aurum and mycobacterium neoaurum with redescription of m. aurum culture collection strains. | we examined american type culture collection (atcc) strains of mycobacterium aurum and mycobacterium neoaurum by using multilocus dna target sequencing. apart from the type strain, all 10 atcc m. aurum strains examined were classified incorrectly, with most being reclassified as belonging to the m. neoaurum-'mycobacterium lacticola' relatedness group. all four m. neoaurum strains were tightly clustered, but heterogeneity was observed within the cluster. as a result of the incorrect annotation of ... | 2009 | 19502318 |
| flow injection analysis electrospray ionization mass spectrometry for high-throughput screening of a gene library for amidase activity. | in high-throughput screening of gene and mutant libraries, high analysis speeds and short method development times are important factors. mass spectrometry (ms) is considered to be a generic analytical technique with a relatively short development time. furthermore, when applying flow injection analysis (fia) for sample introduction, the requirements for high throughput are met. in this work, the use of a single quadrupole electrospray ms instrument for assaying amidase activity in a gene librar ... | 2009 | 19615964 |
| [rapidly growing mycobacteria infection after videosurgical procedures - the glutaraldehyde hypothesis]. | between august 2006 and february 2007, in the state of rio de janeiro, brazil, a massive outbreak of rgm infections after video laparoscopy was mainly associated to the recently described mycobacterium massiliense species. all confirmed and probable cases reports described the use of high-level disinfection of medical devices by using 2% glutaraldehyde (2% ga) for 30 min before the surgical procedures. we investigated the susceptibility of the m. massiliense isolates recovered during the outbrea ... | 2009 | 20076909 |
| mycobacterium neoaurum contamination. | | 2005 | 16110582 |
| inactivation and augmentation of the primary 3-ketosteroid-{delta}1- dehydrogenase in mycobacterium neoaurum nwib-01: biotransformation of soybean phytosterols to 4-androstene- 3,17-dione or 1,4-androstadiene-3,17-dione. | 3-ketosteroid-delta(1)-dehydrogenase, ksdd(m), was identified by targeted gene disruption and augmentation from mycobacterium neoaurum nwib-01, a newly isolated strain. the difficulty of separating 4-androstene-3,17-dione (ad) from 1,4-androstadiene-3,17-dione (add) is a key bottleneck to the microbial transformation of phytosterols in industry. this problem was tackled via genetic manipulation of the ksdd-encoding gene. mutants in which ksdd(m) was inactivated or augmented proved to be good ad( ... | 2010 | 20453136 |
| nontuberculous mycobacterial blood stream and cardiac infections in patients without hiv infection. | nontuberculous mycobacteria rarely cause bacteremia in hiv-negative patients. we describe 16 cases, including the first mycobacterium neoaurum endocarditis. nine cases were line related. most patients were immunocompromised secondary to hematologic malignancy or other comorbid conditions. amikacin had the most reliable in vitro activity. combination therapy was frequently used. mortality was 25%. | 2010 | 20462726 |
| pulmonary infection with mycobacterium neoaurum identified by 16s ribosomal dna sequence. | mycobacterium neoaurum infection has rarely been found in humans, and only a limited number of cases have been reported. we describe the first case of pulmonary infection with m. neoaurum. we speculate that unrecognized aspiration and long-term corticosteroid therapy predisposed our patient to this rare mycobacterial infection. | 2007 | 17306883 |
| preparation, characterization, and biotransformation of the inclusion complex of phytosterols and hydroxypropyl-beta-cyclodextrin by mycobacterium neoaurum. | the inclusion complex of hydroxypropyl-beta-cyclodextrin (hbbetacd) and phytosterols (pss) was prepared and characterized by thermogravimetric analysis (tga) and infrared (ir) spectroscopy. biotransformation of the inclusion complex of phytosterols and hydroxypropyl-beta-cyclodextrin (pss-hbbetacd) by mycobacterium neoaurum to 1,4-androstadiene-3,17-dione and 4-androstene-3,17-dione [ad(d)] was studied. the tga and ir results indicated that the thermal stability of pss was improved in the comple ... | 2011 | 21812345 |
| [conversion of soybean sterols into 3,17-diketosteroids using actinobacteria mycobacterium neoaurum, pimelobacter simplex, and rhodococcus erythropolis]. | abstract-soybean sterols were converted into androst-4-ene-3,17-dione (ad) and 9alpha-hydroxyandrost-4-ene-3,17-dione (9-oh-ad) using three actinobacterium strains. the transformation of a microcrystallic substrate (particle size 5-15 nm) or the transformation in the presence of randomly methylated beta-cyclodextrin (mcd) were carried out by mycobacterium neoaurum with a phytosterol load of 30 g/l over 144 h with an ad content of 14.5 and 15.2 g/l, respectively. ad obtained in the presence of mc ... | 2011 | 21790029 |
| identification of phenylalkane derivatives when mycobacterium neoaurum and rhodococcus erythropolis were cultured in the presence of various phenylalkanes. | phenylalkanes are ubiquitously found in nature as pollutants originating from oil, gas oil and petrol. rising commercial demand for mineral oil fractions has led to the increased prevalence of environmental contamination, whereby these particular hydrocarbons are encountered by bacteria which have subsequently developed sophisticated metabolic routes for purposes of degradation. herein a detailed analysis of these metabolic pathways in the degradation of phenylalkanes by mycobacterium neoaurum a ... | 2011 | 21701983 |
| Cluster of non-tuberculous mycobacteraemia associated with water supply in a haemato-oncology unit. | Non-tuberculous mycobacteria (NTM) are ubiquitous environmental organisms but rarely cause infections. Clinical, microbiological and epidemiological investigations and subsequent management of a cluster of NTM bacteraemia on a haemato-oncology unit are reported. From October 2007 to July 2008, five patients being managed for haematological malignancies developed pyrexia and general malaise. Mycobacterium mucogenicum (four patients) and Mycobacterium neoaurum (one patient) were identified from th ... | 2011 | 21899922 |
| a new steroid-transforming strain of mycobacterium neoaurum and cloning of 3-ketosteroid 9alpha-hydroxylase in nwib-01. | using enrichment procedures, five strains that can utilize soybean phytosterols as the sole carbon source were isolated from steroids-contaminated soil samples. among the isolated strains, the strain nwib-01 with the highest steroid degradation ability was identified as mycobacterium neoaurum by morphological, physiological, biochemical tests and 16s rrna sequence analysis. meanwhile, the key enzyme gene, which was involved in steroid metabolism and encoding 395-amino acid 3-ketosteroid 9alpha-h ... | 2010 | 20204712 |
| mycobacterium neoaurum and mycobacterium bacteremicum sp. nov. as causes of mycobacteremia. | reference isolates of mycobacterium neoaurum, mycobacterium aurum, and the nonvalidated species "mycobacterium lacticola" were the focus of two recent molecular taxonomic studies. on the basis of this grouping, we identified 46 clinical pigmented, rapidly growing mycobacterial isolates. by 16s rrna gene sequencing, only two major taxa were identified: m. neoaurum and a previously uncharacterized "m. neoaurum-like" group. the m. neoaurum-like group exhibited only 99.7% identity to m. neoaurum by ... | 2010 | 20881180 |
| mycobacterium neoaurum infection in a patient with renal failure. | mycobacterium neoaurum, a member of the mycobacterium parafortuitum complex, has only rarely been reported as a pathogen of human infections. we report a case of catheter-related bloodstream infection (crbsi) due to m. neoaurum in a patient on hemodialysis. the isolate was identified by conventional methods as well as by 16s rrna gene analysis. the patient was successfully treated with intravenous antibiotics (meropenem and amikacin) for three weeks and the catheter was removed. m. neoaurum shou ... | 2009 | 19119036 |
| a mutant form of 3-ketosteroid-δ(1)-dehydrogenase gives altered androst-1,4-diene-3, 17-dione/androst-4-ene-3,17-dione molar ratios in steroid biotransformations by mycobacterium neoaurum st-095. | mycobacterium neoaurum st-095 and its mutant m. neoaurum jc-12, capable of transforming phytosterol to androst-1,4-diene-3,17-dione (add) and androst-4-ene-3,17-dione (ad), produce very different molar ratios of add/ad. the distinct differences were related to the enzyme activity of 3-ketosteroid-δ(1)-dehydrogenase (ksdd), which catalyzes the c1,2 dehydrogenation of ad to add specifically. in this study, by analyzing the primary structure of ksddi (from m. neoaurum st-095) and ksddii (from m. ne ... | 2016 | 26886757 |
| efficient androst-1,4-diene-3,17-dione production by co-expressing 3-ketosteroid-δ(1) -dehydrogenase and catalase in bacillus subtilis. | 3-ketosteroid-δ(1) -dehydrogenase (ksdd), a flavin adenine dinucleotide (fad)-dependent enzyme involved in sterol metabolism, specifically catalyses the conversion of androst-4-ene-3,17-dione (ad) to androst-1,4-diene-3,17-dione (add). however, the low ksdd activity and the toxic effects of hydrogen peroxide (h2 o2 ) generated during the biotransformation of ad to add with fad regeneration hinder its application on ad conversion. the aim of this work was to improve ksdd activity and eliminate th ... | 2017 | 27797429 |
| enhanced production of androst-1,4-diene-3,17-dione by mycobacterium neoaurum jc-12 using three-stage fermentation strategy. | to improve the androst-1,4-diene-3,17-dione (add) production from phytosterol by mycobacterium neoaurum jc-12, fructose was firstly found favorable as the initial carbon source to increase the biomass and eliminate the lag phase of m. neoaurum jc-12 in the phytosterol transformation process. based on this phenomenon, two-stage fermentation by using fructose as the initial carbon source and feeding glucose to maintain strain metabolism was designed. by applying this strategy, the fermentation dur ... | 2015 | 26352898 |
| molecular identification of mycobacterium species of public health importance in cattle in zimbabwe by 16s rrna gene sequencing. | mycobacterium species are naturally found in the environment as well as in domestic animals such as cattle. so far, more than 150 species of mycobacterium, some of which are pathogenic, have been identified. laboratory isolation, detection and identification of mycobacterium species are therefore critical if human and animal infections are to be controlled. the objective of this study was to identify mycobacterium species isolated in cattle in zimbabwe using 16s ribosomal rna gene amplification ... | 2015 | 26668660 |
| efficient 9α-hydroxy-4-androstene-3,17-dione production by engineered bacillus subtilis co-expressing mycobacterium neoaurum 3-ketosteroid 9α-hydroxylase and b. subtilis glucose 1-dehydrogenase with nadh regeneration. | 3-ketosteroid 9α-hydroxylase (ksh, consisting of ksha and kshb), a key enzyme in steroid metabolism, can catalyze the transformation of 4-androstene-3,17-dione (ad) to 9α-hydroxy-4-androstene-3,17-dione (9ohad) with nadh as coenzyme. in this work, ksh from mycobacterium neoaurum jc-12 was successfully cloned and overexpressed in bacillus subtilis 168. the expression and purification of ksh was analyzed by sds-page and ksh activity assay. preliminary characterization of ksh was performed using pu ... | 2016 | 27516945 |
| mycobacterium avium subsp. avium and mycobacterium neoaurum detection in an immunocompromised patient. | non-tuberculous mycobacteria are increasingly described as infectious agents in immunocompromised patients. a 17-year-old male patient suffering from secondary non-hodgkin's lymphoma and treated with chemotherapeutic agents was admitted to hospital due to pleuropneumonia. mycobacterium neoaurum was cultured repeatedly from his sputum and, mycobacterium avium subsp. avium (m. a. avium) was detected by is901 qpcr from detached fragments of his intestinal mucosa. we attempted to determine the possi ... | 2014 | 23842543 |
| bioconversion of 4-androstene-3,17-dione to androst-1,4-diene-3,17-dione by recombinant bacillus subtilis expressing ksdd gene encoding 3-ketosteroid-δ1-dehydrogenase from mycobacterium neoaurum jc-12. | the enzyme 3-ketosteroid-δ(1)-dehydrogenase (ksdd), involved in steroid metabolism, catalyzes the transformation of 4-androstene-3,17-dione (ad) to androst-1,4-diene-3,17-dione (add) specifically. its coding gene was obtained from mycobacterium neoaurum jc-12 and expressed on the plasmid pma5 in bacillus subtilis 168. the successfully expressed ksdd was analyzed by native-page. the activities of the recombinant enzyme in b. subtilis were 1.75 u/mg, which was about 5-fold that of the wild type in ... | 2013 | 23298646 |
| role identification and application of sigd in the transformation of soybean phytosterol to 9α-hydroxy-4-androstene-3,17-dione in mycobacterium neoaurum. | 9α-hydroxy-4-androstene-3,17-dione (9-ohad) is a valuable steroid pharmaceutical intermediate which can be produced by the conversion of soybean phytosterols in mycobacteria. however, the unsatisfactory productivity and conversion efficiency of engineered mycobacterial strains hinder their industrial applications. here, a sigma factor d (sigd) was investigated due to its dramatic downregulation during the conversion of phytosterols to 9-ohad. it was determined as a negative regulator in the meta ... | 2017 | 28035826 |
| bacteremia with an unusual pathogen: mycobacterium neoaurum. | mycobacterium neoaurum (m. neoaurum) is an infrequently encountered cause of infection in humans. it is a member of the rapidly growing mycobacteria family. it predominately afflicts those with a compromised immune status and a chronically indwelling vascular access. isolation of this organism is challenging yet the advent of 16s ribosomal sequencing paved the way for more sensitive detection. no treatment guidelines are available and treatment largely depends on the experience of the treating p ... | 2016 | 27807489 |
| prominent contribution of th1, th17, and tregs to the host response during m. neoaurum infection. | nontuberculous mycobacteria are ubiquitous in outside environment and animals. as for nontuberculous mycobacteria infection, there is only limited information in humans regarding infection and the subsequent immune response, especially for mycobacterium neoaurum. here, haematoxylin-eosin and ziehl-neelsen staining were used to observe pathological changes and detect acid-fast bacilli in organ samples in mouse model. flow cytometry and quantitative real-time polymerase chain reaction were perform ... | 2016 | 27706786 |
| single nucleotide polymorphism analysis for the production of valuable steroid intermediates in mycobacterium neoaurum. | to investigate single nucleotide polymorphism (snp) in the transformation process of phytosterol to valuable steroid intermediates in three steroid-producing mycobacterium neoaurum strains using deep sequencing and bioinformation analysis. | 2016 | 27571967 |
| rna-seq analysis uncovers non-coding small rna system of mycobacterium neoaurum in the metabolism of sterols to accumulate steroid intermediates. | understanding the metabolic mechanism of sterols to produce valuable steroid intermediates in mycobacterium by a noncoding small rna (srna) view is still limited. in the work, rna-seq was implemented to investigate the noncoding transcriptome of mycobacterium neoaurum (mn) in the transformation process of sterols to valuable steroid intermediates, including 9α-hydroxy-4-androstene-3,17-dione (9ohad), 1,4-androstadiene-3,17-dione (add), and 22-hydroxy-23, 24-bisnorchola-1,4-dien-3-one (1,4-bna). | 2016 | 27112590 |
| [overexpressing 3-ketosteroid-δ1-dehydrogenase for degrading phytosterols into androst-1,4-diene-3,17-dione]. | we constructed plasmid pmtac to overexpress 3-ketosteroid-δ1-dehydrogenase (ksdd) in mycobacterium neoaurum jc-12 for improving androst-1,4-diene-3,17-dione (add) production. to construct pmtac, pace promoter on pmf41 was replaced by tac promoter, and then four recombinants were constructed, which were m. neoaurum jc-12/pmf41-gfp, m. neoaurum jc-12/pmtac-gfp, m. neoaurum jc-12/pmf41-ksdd and m. neoaurum jc-12/pmtac-ksdd. fluorescence detection results show that much more green fluorescent protei ... | 2015 | 26939442 |
| an enantioselective nadp(+)-dependent alcohol dehydrogenase responsible for cooxidative production of (3s)-5-hydroxy-3-methyl-pentanoic acid. | a soil bacterium, mycobacterium sp. b-009, is able to grow on racemic 1,2-propanediol (pd). the strain was revealed to oxidize 3-methyl-1,5-pentanediol (mpd) to 5-hydroxy-3-methyl-pentanoic acid (hmpa) during growth on pd. mpd was converted into an almost equimolar amount of the s-form of hmpa (s-hmpa) at 72%ee, suggesting the presence of an enantioselective mpd dehydrogenase (mpd-dh). as expected, an nadp(+)-dependent alcohol dehydrogenase, which catalyzes the initial step of mpd oxidation, was ... | 2016 | 26923741 |
| unraveling and engineering the production of 23,24-bisnorcholenic steroids in sterol metabolism. | the catabolism of sterols in mycobacteria is highly important due to its close relevance in the pathogenesis of pathogenic strains and the biotechnological applications of nonpathogenic strains for steroid synthesis. however, some key metabolic steps remain unknown. in this study, the hsd4a gene from mycobacterium neoaurum atcc 25795 was investigated. the encoded protein, hsd4a, was characterized as a dual-function enzyme, with both 17β-hydroxysteroid dehydrogenase and β-hydroxyacyl-coa dehydrog ... | 2016 | 26898409 |
| effects of different carbon sources on growth, membrane permeability, β-sitosterol consumption, androstadienedione and androstenedione production by mycobacterium neoaurum. | effects of different carbon sources on growth, membrane permeability, β-sitosterol consumption, androstadienedione and androstenedione (ad(d)) production by mycobacterium neoaurum were investigated. the results indicated that glucose was advantageous to the growth and resulted in the adverse effects on the phytosterols consumption and ad(d) production compared to the results of propanol and isopropanol as sole carbon source. the cell wall widths of 9.76 by propanol and 8.00 nm by isopropanol wer ... | 2016 | 26298579 |
| mutation breeding of high 4-androstene-3,17-dione-producing mycobacterium neoaurum zadf-4 by atmospheric and room temperature plasma treatment. | steroid medication is used extensively in clinical applications and comprises a large and vital part of the pharmaceutical industry. however, the difficulty of separating 4-androstene-3,17-dione (ad) from 1,4-androstadiene-3,17-dione (add) restricts the application of the microbial transformation of phytosterols in the industry. a novel atmospheric and room temperature plasma (artp) treatment, which employs helium as the working gas, was used to generate mycobacterium neoaurum mutants producing ... | 2015 | 25845362 |
| effects of different carbon sources on growth, membrane permeability, β-sitosterol consumption, androstadienedione and androstenedione production by mycobacterium neoaurum. | effects of different carbon sources on growth, membrane permeability, β-sitosterol consumption, androstadienedione and androstenedione (ad(d)) production by mycobacterium neoaurum were investigated. the results indicated that glucose was advantageous to the growth and resulted in the adverse effects on the phytosterols consumption and ad(d) production compared to the results of propanol and isopropanol as sole carbon source. the cell wall widths of 9.76 by propanol and 8.00 nm by isopropanol wer ... | 2015 | 25682382 |
| genetic differences in ksdd influence on the add/ad ratio of mycobacterium neoaurum. | mycobacterium neoaurum tccc 11028 (mnr) and m. neoaurum tccc 11028 m3 (mnr m3) significantly differ in the ratio of androst-1,4-diene-3,17-dione (add) to androst-4-ene-3,17-dione (ad) produced. the large fluctuations are related to the dehydrogenation activity of 3-ketosteroid-δ(1)-dehydrogenase (ksdd). analysis of the primary structure of ksdd showed that the ser-138 of ksdd-mnr changed to leu-138 of ksdd-mnr m3 because of c413t in the ksdd gene. this phenomenon directly affected ksdd activity. ... | 2015 | 25572208 |
| draft genome sequence of mycobacterium neoaurum strain dsm 44074t. | we report the draft genome sequence of mycobacterium neoaurum strain dsm 44074(t), a nontuberculosis species responsible for opportunistic infections in immunocompromised patients. the strain described here is composed of 5,536,033 bp, with a g+c content of 66.24%, and carries 5,274 protein-coding genes and 72 rna genes. | 2014 | 25013147 |
| characterization and engineering of 3-ketosteroid-△1-dehydrogenase and 3-ketosteroid-9α-hydroxylase in mycobacterium neoaurum atcc 25795 to produce 9α-hydroxy-4-androstene-3,17-dione through the catabolism of sterols. | 3-ketosteroid-△(1)-dehydrogenase (kstd) is a key enzyme involved in the microbial catabolism of sterols. here, three homologues of kstd were characterized from mycobacterium neoaurum atcc 25795, showing distinct substrate preferences and transcriptional responses to steroids. single deletion of any mn-kstd failed to result in a stable and maximum accumulation of 9-ohad due to residual kstd activities. to develop stable 9-ohad producers, all of these mn-kstds were inactivated, which led to about ... | 2014 | 24831710 |
| pulmonary infection caused by mycobacterium neoaurum: the first case in korea. | mycobacterium neoaurum is rapidly growing mycobacteria that can cause human infections. it commonly causes bloodstream infections in immunocompromised hosts, and unlike other mycobacteria species, it rarely causes pulmonary infections. we confirmed the first pulmonary infection case in korea caused by m. neoaurum using full-length 16s rrna gene sequencing. | 2014 | 24790914 |
| accumulation of androstadiene-dione by overexpression of heterologous 3-ketosteroid δ1-dehydrogenase in mycobacterium neoaurum nwib-01. | mycobacterium neoaurum nwib-01 exhibits powerful ability to cleave the side chain of soybean phytosterols to accumulate 4-androstene-3,17-dione (ad) and 1,4-androstadiene-3,17-dione (add). the difficulty in separation of ad from add is one of the key bottlenecks to the microbial transformation of phytosterols in the industry. to enhance add quantity in products, 3-ketosteroid δ(1)-dehydrogenase genes (kstd m and kstd(a)) were obtained from m. neoaurum nwib-01 and arthrobacter simplex respectivel ... | 2014 | 24510385 |
| complete genome sequence of sterol-transforming mycobacterium neoaurum strain vkm ac-1815d. | mycobacterium neoaurum strain vkm ac-1815d produces 4-androstene-3,17-dione as a major compound from phytosterols. here, we report the complete genome sequence of the strain. the genome consists of a single circular 5,438,190-bp chromosome, with a g+c content of 66.88%, containing 5,318 putative open reading frames (orfs), 46 trnas, and 6 rrnas. arrays of cholesterol metabolism genes are randomly clustered throughout the chromosome. | 2014 | 24435872 |
| bacterial community of biofilms developed under different water supply conditions in a distribution system. | in order to understand the bacterial community characteristics of biofilms developed under different finished water supply histories in drinking water distribution systems (dwds), biofilm samples on different type of iron corrosion scales in a real dwds were collected and systematically investigated using 454 pyrosequencing of 16s rrna gene. the richness and diversity estimators showed that biofilms formed in dwds transporting finished groundwater (gw) had the lowest level of bacterial diversity ... | 2014 | 24291134 |
| catheter-related bloodstream infection due to mycobacterium neoaurum in a patient with acute leukemia. | | 2014 | 24152051 |
| mycobacterium neoaurum causing prosthetic valve endocarditis: a case report and review of the literature. | mycobacterium neoaurum is a rare cause of bacteremia, and infection usually occurs in an immunocompromised host in the setting of an indwelling catheter. prosthetic valve endocarditis due to non-tuberculous mycobacteria typically carries a dismal prognosis; we report a case of m. neoaurum prosthetic valve endocarditis with favorable response to antimicrobial therapy without surgical intervention. | 2014 | 24076109 |
| biotransformation of 4-sec-butylphenol by gram-positive bacteria of the genera mycobacterium and nocardia including modifications on the alkyl chain and the hydroxyl group. | the environmental pollutant 4-sec-butylphenol (4-sec-bp) which possesses estrogenic properties was transformed by the aerobic gram-positive bacteria mycobacterium neoaurum and nocardia cyriacigeorgica into three main products (p1-p3) which were isolated and structurally characterized in detail. two of them were products of a process resembling anaerobic metabolism of alkylphenols based on modifications of the alkyl side chain of 4-sec-bp. the first product (p1) was identified as 4-(2-hydroxy-1-m ... | 2013 | 23912120 |
| optimization of biotransformation from phytosterol to androstenedione by a mutant mycobacterium neoaurum zjuvn-08. | biotransformation of phytosterol (ps) by a newly isolated mutant mycobacterium neoaurum zjuvn-08 to produce androstenedione has been investigated in this paper. the parameters of the biotransformation process were optimized using fractional factorial design and response surface methodology. androstenedione was the sole product in the fermentation broth catalyzed by the mutant m. neoaurum zjuvn-08 strain. results showed that molar ratio of hydroxypropyl-β-cyclodextrin (hp-β-cd) to ps and substrat ... | 2013 | 23365012 |
| non-tuberculous mycobacterial pd peritonitis in australia. | peritonitis can be a severe complication of peritoneal dialysis (pd) due to associated morbidity and mortality. non-tuberculous mycobacteria (ntm) are a rare cause of pd peritonitis, with high rates of catheter removal and conversion to haemodialysis, and a reported mortality as high as 40 %. the incidence, culprit ntm species, and outcomes associated with pd peritonitis have not been described in many countries, including australia. | 2013 | 23180442 |
| identification and engineering of cholesterol oxidases involved in the initial step of sterols catabolism in mycobacterium neoaurum. | mycobacteria have been modified to transform sterols to produce valuable steroids. here, we demonstrated that the oxidation of sterols to sterones is a rate-limiting step in the catabolic pathway of sterols in mycobacterium neoaurum. two cholesterol oxidases chom1 and chom2 involved in the step were identified in m. neoaurum and the chom2 shared up to 45% identity with other cholesterol oxidases. we demonstrated that the combination of chom1 and chom2 plays a significant role in this step. accor ... | 2013 | 23164577 |
| two different primary oxidation mechanisms during biotransformation of thymol by gram-positive bacteria of the genera nocardia and mycobacterium. | thymol has antibacterial, antifungal, insecticidal, and antioxidative properties which are the basis for the wide use of this compound in the cosmetic, food, and pharmaceutical industries. although thymol is a ubiquitously occurring substance in the environment, data about its degradation and detoxification by bacteria are sparse. here, we show the existence of two different pathways for the biotransformation of thymol by nocardia cyriacigeorgica and mycobacterium neoaurum which were described f ... | 2013 | 22828982 |
| influence of hydroxypropyl-β-cyclodextrin on phytosterol biotransformation by different strains of mycobacterium neoaurum. | cyclodextrins (cds) can improve productivity in the biotransformation of steroids by increasing conversion rate, conversion ratio, or substrate concentration. however, little is known of the proportion of products formed by multi-catabolic enzymes, e.g., via sterol side chain cleavage. using three strains with different androst-1,4-diene-3,17-dione (add) to androst-4-ene-3,17-dione (ad) ratios, mycobacterium neoaurum tccc 11028 (mnr), m. neoaurum tccc 11028 m1 (mnr m1), and m. neoaurum tccc 1102 ... | 2012 | 22614451 |
| hand infection due to mycobacterium neoaurum. | | 2012 | 22457251 |
| mycobacterium sediminis sp. nov. and mycobacterium arabiense sp. nov., two rapidly growing members of the genus mycobacterium. | two novel isolates of rapidly growing, gram-stain-positive, non-chromogenic species of the genus mycobacterium, strain yim m13028(t) from a sediment sample collected from the south china sea (19° 30.261' n 111° 0.247' e) at a depth of 42 m and strain yim 121001(t) from a coastal zone sand sample collected in dubai, united arab emirates, were obtained in our laboratory. their taxonomic positions were determined by a polyphasic approach. good growth of the two strains was observed at 28 °c and ph ... | 2013 | 23728378 |
| improvement of ad biosynthesis response to enhanced oxygen transfer by oxygen vectors in mycobacterium neoaurum tccc 11979. | in steroid biotransformation, soybean oil can improve the productivity of steroids by increasing substrate solubility and strengthen the cell membrane permeability. however, little is known of its role as oxygen carrier and its mechanism of promoting the steroid biotransformation. in this work, soybean oil used as oxygen vector for the enhancement of androst-4-ene-3,17-dione (ad) production by mycobacterium neoaurum tccc 11979 (mnr) was investigated. upon the addition of 16% (v/v) soybean oil, t ... | 2017 | 28120242 |
| molecular identification of mycobacterium tuberculosis complex by region of differentiation-typing and polymerase chain reaction-restriction fragment length polymorphism method. | tuberculosis (tb) is one of the most common zoonotic infectious diseases in the world. identification of mycobacterium isolates is essential for proper treatment of tb. the aim of this study was to identify mycobacterium isolates collected from tb patients in alborz province, iran, by region of differentiation (rd)-typing. fifty samples from tuberculosis patients were cultured in pyruvate and glycerinated lowenstein-jensen medium. dna was extracted from the isolates by the van solingen method an ... | 2016 | 28043546 |
| intra- and extra-cellular proteome analyses of steroid-producer mycobacteria. | the importance of the pathogenic mycobacteria has mainly focused the omic analyses on different aspects of their clinical significance. in contrast, those industrially relevant mycobacteria have received less attention, even though the steroids market sales in 2011, in example, were estimated in $8 billion.the extra-cellular proteome, due to its relevance in the sterols processing and uptake; as well as the intra-cellular proteome, because of its role in steroids bioconversion, are the core of t ... | 2017 | 28710622 |
| genome sequencing of steroid producing bacteria using ion torrent technology and a reference genome. | the next-generation sequencing technology has enormously eased the bacterial genome sequencing and several tens of thousands of genomes have been sequenced during the last 10 years. most of the genome projects are published as draft version, however, for certain applications the complete genome sequence is required.in this chapter, we describe the strategy that allowed the complete genome sequencing of mycobacterium neoaurum nrrl b-3805, an industrial strain exploited for steroid production, usi ... | 2017 | 28710620 |
| comparative genomic analysis of mycobacterium neoaurum mn2 and mn4 substrate and product tolerance. | the microbial bioconversion of sterols can afford valuable steroid precursors, such as 4-androstene-3,17-dione (ad) and androsta-1,4-diene-3,17-dione (add). the mycobacterium neoaurum mn4 mutant strain can produce ad in high yield and can tolerate a higher concentration of the substrate phytosterol than the parent strain m. neoaurum mn2. in order to further investigate the mechanisms underlying the enhanced substrate and product tolerance, we performed a genomic analysis of the mn2 and mn4 strai ... | 2017 | 28664368 |
| site-directed mutagenesis under the direction of in silico protein docking modeling reveals the active site residues of 3-ketosteroid-δ(1)-dehydrogenase from mycobacterium neoaurum. | 3-ketosteroid-δ(1)-dehydrogenases (ksdd) from mycobacterium neoaurum could transform androst-4-ene-3,17-dione (ad) to androst-1,4-diene-3,17-dione. this reaction has a significant effect on the product of pharmaceutical steroid. the crystal structure and active site residues information of ksdd from mycobacterium is not yet available, which result in the engineering of ksdd is tedious. in this study, by the way of protein modeling and site-directed mutagenesis, we find that, y122, y125, s138, e1 ... | 2017 | 28634712 |
| improving the production of 22-hydroxy-23,24-bisnorchol-4-ene-3-one from sterols in mycobacterium neoaurum by increasing cell permeability and modifying multiple genes. | the strategy of modifying the sterol catabolism pathway in mycobacteria has been adopted to produce steroidal pharmaceutical intermediates, such as 22-hydroxy-23,24-bisnorchol-4-ene-3-one (4-hbc), which is used to synthesize various steroids in the industry. however, the productivity is not desirable due to some inherent problems, including the unsatisfactory uptake rate and the low metabolic efficiency of sterols. the compact cell envelope of mycobacteria is a main barrier for the uptake of ste ... | 2017 | 28532497 |
| scale-up of phytosterols bioconversion into androstenedione. | phytosterols, generated as a by-product of vegetable oils or wood pulp, contain the cyclopentane-perhydro-phenanthrene nucleus, and can be converted into steroid intermediates by removing the c17 side chain. this chapter shows the scale-up, from flask to fermentor, of the phytosterols bioconversion into 4-androstene-3,17-dione (androstenedione; ad) with mycobacterium neoaurum b-3805. due to the fact that phytosterols and ad are nearly insoluble in water, two-phase systems and the use of chemical ... | 2017 | 28710630 |
| genetic techniques for manipulation of the phytosterol biotransformation strain mycobacterium neoaurum nrrl b-3805. | mycobacterium neoaurum is a saprophytic, soil-dwelling bacterium. the strain nrrl b-3805 converts phytosterols to androst-4-ene-3,17-dione (androstenedione; ad), a precursor of multiple c19 steroids of importance to industry. nrrl b-3805 itself is able to convert ad to other steroid products, including testosterone (ts) and androst-1,4-diene-3,17-dione (androstadienedione; add). however to improve this strain for industrial use, genetic modification is a priority. in this chapter, we describe a ... | 2017 | 28710623 |
| obtaining of 11α-hydroxyandrost-4-ene-3,17-dione from natural sterols. | two-step one-pot microbial transformation enables obtaining of valuable steroids that are difficult to produce chemically. here we describe a method for obtaining 11α-hydroxyandrost-4-ene-3,17-dione (11α-had) from cheap and available natural sterols (phytosterols or cholesterol).11α-had is a primary adrenal steroid in mammals and also a key precursor in the syntheses of halogenated corticoids. conventional routes for its obtaining are based on chemical synthesis, or microbial hydroxylation of an ... | 2017 | 28710634 |