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1.
Afr. J. Clin. Exp. Microbiol ; 24(2): 1-10, 2023. figures, tables
Article in English | AIM | ID: biblio-1427772

ABSTRACT

Corynebacterium diphtheriae is responsible for both endemic and epidemic diphtheria. The predisposing factor for this disease is the failure to immunize during childhood. Humans are the only hosts of the organism and is present in the upper respiratory tract. The organism is transmitted via airborne route and can cause respiratory obstruction and heart failure because of the exotoxin it produces. There is presently a resurgence of diphtheria outbreaks in Nigeria. The Nigeria Center for Disease Control (NCDC) was notified of suspected diphtheria outbreaks in Lagos and Kano States, Nigeria, in December 2022 and has been issuing monthly reports since that time. This review of the diphtheria outbreaks following online database searches on PubMed and Google Scholar as well as the NCDC/WHO websites and grey literatures, describes the current trend of the outbreaks globally, elucidated the different strains of Corynebacterium responsible for the outbreaks, identified the recent vaccine formulation developed to tackle the outbreaks, and provide information on vaccine delivery and efficacy studies in the country and globally.


Subject(s)
Humans , Actinomycetales , Diphtheria-Tetanus-Pertussis Vaccine , Disease Outbreaks , Diphtheria , Vaccination Coverage
2.
Chinese Journal of Biotechnology ; (12): 2027-2039, 2023.
Article in Chinese | WPRIM | ID: wpr-981187

ABSTRACT

The discovery of new enzymes for poly(ethylene terephthalate) (PET) degradation has been a hot topic of research globally. Bis-(2-hydroxyethyl) terephthalate (BHET) is an intermediate compound in the degradation of PET and competes with PET for the substrate binding site of the PET-degrading enzyme, thereby inhibiting further degradation of PET. Discovery of new BHET degradation enzymes may contribute to improving the degradation efficiency of PET. In this paper, we discovered a hydrolase gene sle (ID: CP064192.1, 5085270-5086049) from Saccharothrix luteola, which can hydrolyze BHET into mono-(2-hydroxyethyl) terephthalate (MHET) and terephthalic acid (TPA). BHET hydrolase (Sle) was heterologously expressed in Escherichia coli using a recombinant plasmid, and the highest protein expression was achieved at a final concentration of 0.4 mmol/L of isopropyl-β-d-thiogalactoside (IPTG), an induction duration of 12 h and an induction temperature of 20 ℃. The recombinant Sle was purified by nickel affinity chromatography, anion exchange chromatography, and gel filtration chromatography, and its enzymatic properties were also characterized. The optimum temperature and pH of Sle were 35 ℃ and 8.0, and more than 80% of the enzyme activity could be maintained in the range of 25-35 ℃ and pH 7.0-9.0 and Co2+ could improve the enzyme activity. Sle belongs to the dienelactone hydrolase (DLH) superfamily and possesses the typical catalytic triad of the family, and the predicted catalytic sites are S129, D175, and H207. Finally, the enzyme was identified as a BHET degrading enzyme by high performance liquid chromatography (HPLC). This study provides a new enzyme resource for the efficient enzymatic degradation of PET plastics.


Subject(s)
Actinomycetales/genetics , Hydrolases/metabolism , Phthalic Acids/chemistry , Polyethylene Terephthalates/metabolism
3.
Chinese Journal of Biotechnology ; (12): 3351-3363, 2023.
Article in Chinese | WPRIM | ID: wpr-1007962

ABSTRACT

Polyhydroxyalkanoate depolymerase (PHAD) can be used for the degradation and recovery of polyhydroxyalkanoate (PHA). In order to develop a PHAD with good stability under high temperature, PHAD from Thermomonospora umbrina (TumPHAD) was heterelogously expressed in Escherichia coli BL21(DE3). At the same time, a mutant A190C/V240C with enhanced stability was obtained via rational design of disulfide bonds. Characterization of enzymatic properties showed that the mutant A190C/V240C had an optimum temperature of 60 ℃, which was 20 ℃ higher than that of the wild type. The half-life at 50 ℃ was 7 hours, at 50 ℃ which was 21 times longer than that of the wild type. The mutant A190C/V240C was used for the degradation of polyhydroxybutyrate (PHB), one of the typical PHA. At 50 ℃, the degradation rate of PHB being treated for 2 hours and 12 hours was 2.1 times and 3.8 times higher than that of the wild type, respectively. The TumPHAD mutant A190C/V240C obtained in this study shows tolerance to high temperature resistance, good thermal stability and strong PHB degradation ability, which may facilitate the degradation and recovery of PHB.


Subject(s)
Thermomonospora , Actinomycetales , Escherichia coli/genetics , Polyhydroxyalkanoates
4.
Med. UIS ; 32(3): 27-33, Sep.-Dec. 2019. tab
Article in English | LILACS | ID: biblio-1114974

ABSTRACT

Abstract Genital hair is one of the secondary sexual traits that marks the beginning of puberty; its removal has been part of human culture since ancient times. This practice may lead to modifications in vaginal microbiome with potential repercussions on skin health and balance. We conducted a narrative review with the purpose of describing normal skin microbiota, its impact under microenvironment changes and genital hair removal. Menses, pathological conditions and pubic hair removal may alter vaginal microbiota, being the latter of special relevance giving the risk of hair microtrauma, irritations and potential spread of infectious agents. MÉD.UIS.2019;32(3):27-33


Resumo O cabelo genital é um dos traços sexuais secundários que marcam o início da puberdade; sua remoção faz parte da cultura humana desde os tempos antigos. Essa prática pode levar a modificações no microbioma vaginal com possíveis repercussões na saúde e equilíbrio da pele. Realizamos uma revisão narrativa com o objetivo de descrever a microbiota normal da pele, seu impacto nas alterações do microambiente e na remoção de pelos genitais. A menstruação, as condições patológicas e a remoção de pelos pubianos podem alterar a microbiota vaginal, sendo esta última de especial relevância dando o risco de microtraumatismo capilar, irritações e potencial disseminação de agentes infecciosos. MÉD.UIS.2019;32(3): 27-33


Resumen El vello genital es uno de los rasgos sexuales secundarios que marca el comienzo de la pubertad; su eliminación ha sido parte de la cultura humana desde la antigüedad. Esta práctica puede conducir a modificaciones en el microbioma vaginal con posibles repercusiones potenciales en la salud y el equilibrio de la piel. Realizamos una revisión narrativa con el propósito de describir la microbiota cutánea normal, su impacto bajo los cambios del microambiente y la depilación genital. La menstruación, las condiciones patológicas y la depilación púbica pueden alterar la microbiota vaginal, siendo esta última de especial relevancia dado el riesgo de microtraumatismos, irritaciones y posible propagación de agentes infecciosos. MÉD.UIS.2019;32(3): 27-33


Subject(s)
Humans , Female , Microbiota , Hair Removal , Rupture , Skin , Staphylococcus , Actinomycetales , Humans , Health , Risk , Puberty , Dermatology , Genitalia, Female , Hair , Infections , Menstruation , Noxae
5.
Infectio ; 23(4): 399-401, Dec. 2019. graf
Article in Spanish | LILACS, COLNAL | ID: biblio-1019867

ABSTRACT

Los miembros del género Kocuria corresponden a cocos Gram positivos ubicuos, generalmente inocuos y que hacen parte de la flora saprófita de un porcentaje importante de la población; ocasionalmente han sido descritos como los agentes responsables de patologías infecciosas, principalmente dentro del contexto de pacientes que concomitantemente cursan con enfermedades crónicas y estados de inmunosupresión. Son escasos los casos reportados como causa de endocarditis en pacientes inmunocompetentes a nivel global por especies de este género. Se expone el caso de una mujer inmunocompetente de 44 años, sin antecedentes de importancia, en quien solo el diagnóstico microbiológico permitió confirmar la presencia de Kocuria kristinae como agente causal de su endocarditis infecciosa; la literatura señala la dificultad existente al momento de diferenciar la endocarditis producida por Staphylococcus spp. versus Kocuria kristinae por su evolución clínica similar, indicando la importancia de la microbiología al momento de identificar y diagnosticar apropiadamente.


Members of the genus Kocuria correspond to ubiquitous, generally harmless, Gram-positive cocci that are part of the saprophytic flora of a significant percentage of the population; occasionally they have been described as the agents responsible for infectious pathologies, mainly in the context of patients who concomitantly have chronic diseases and are under an immunosuppression state. There are few cases reported as a cause of endocarditis in immunocompetent patients globally by species of this genus. We present the case of a 44-year-old immunocompetent woman, with no relevant history, in whom only the microbiological diagnosis confirmed the presence of Kocuria kristinae as the causative agent of her infectious endocarditis; Literature points out the difficulty existing when differentiating endocarditis produced by Staphylococcus spp. versus Kocuria kristinae because of their similar clinical evolution, indicating the importance of microbiology when identifying and diagnosing accurately.


Subject(s)
Humans , Male , Adult , Gram-Positive Cocci , Endocarditis, Bacterial , Immunocompetence , Actinomycetales , Actinobacteria , Endocarditis , Infections , Micrococcaceae
6.
Electron. j. biotechnol ; 40: 52-57, July. 2019. graf, tab
Article in English | LILACS | ID: biblio-1053462

ABSTRACT

Background: Plastic waste is a serious problem because it is difficult to degrade, thereby leading to global environment problems. Poly(lactic acid) (PLA) is a biodegradable aliphatic polyester derived from renewable resources, and it can be degraded by various enzymes produced by microorganisms. This study focused on the scale-up and evaluated the bioprocess of PLA degradation by a crude microbial enzyme produced by Actinomadura keratinilytica strain T16-1 in a 5 L stirred tank bioreactor. Results: PLA degradation after 72 h in a 5 L bioreactor by using the enzyme of the strain T16-1 under controlled pH conditions resulted in lactic acid titers (mg/L) of 16,651 mg/L and a conversion efficiency of 89% at a controlled pH of 8.0. However, the PLA degradation process inadvertently produced lactic acid as a potential inhibitor, as shown in our experiments at various concentrations of lactic acid. Therefore, the dialysis method was performed to reduce the concentration of lactic acid. The experiment with a dialysis bag achieved PLA degradation by weight loss of 99.93%, whereas the one without dialysis achieved a degradation of less than approximately 14.75%. Therefore, the dialysis method was applied to degrade a commercial PLA material (tray) with a conversion efficiency of 32%, which was 6-fold more than that without dialysis. Conclusions: This is the first report demonstrating the scale-up of PLA degradation in a 5 L bioreactor and evaluating a potential method for enhancing PLA degradation efficiency.


Subject(s)
Polyesters/metabolism , Actinomycetales/enzymology , Enzymes/metabolism , Polymers/metabolism , Biodegradation, Environmental , Lactic Acid/analysis , Bioreactors , Hydrogen-Ion Concentration
7.
Electron. j. biotechnol ; 30: 71-76, nov. 2017. graf, ilus, tab
Article in English | LILACS | ID: biblio-1021543

ABSTRACT

Background: Poly(DL-lactic acid), or PDLLA, is a biodegradable polymer that can be hydrolyzed by various types of enzymes. The protease produced by Actinomadura keratinilytica strain T16-1 was previously reported to have PDLLA depolymerase activity. However, few studies have reported on PDLLA-degrading enzyme production by bacteria. Therefore, the aims of this study were to determine a suitable immobilization material for PDLLA-degrading enzyme production and optimize PDLLA-degrading enzyme production by using immobilized A. keratinilytica strain T16-1 under various fermentation process conditions in a stirrer fermenter. Results: Among the tested immobilization materials, a scrub pad was the best immobilizer, giving an enzyme activity of 30.03 U/mL in a shake-flask scale. The maximum enzyme activity was obtained at aeration 0.25 vvm, agitation 170 rpm, 45°C, and 48 h of cultivation time. Under these conditions, a PDLLA-degrading enzyme production of 766.33 U/mL with 15.97 U/mL·h productivity was observed using batch fermentation in a 5-L stirrer fermenter. Increased enzyme activity and productivity were observed in repeated-batch (942.67 U/mL and 19.64 U/mL·h) and continuous fermentation (796.43 U/mL and 16.58 U/mL·h) at a dilution rate of 0.013/h. Scaled-up production of the enzyme in a 10-L stirrer bioreactor using the optimized conditions showed a maximum enzyme activity of 578.67 U/mL and a productivity of 12.06 U/mL·h. Conclusions: This research successfully scaled-up the enzyme production to 5 and 10 L in a stirrer fermenter and is helpful for many applications of poly(lactic acid).


Subject(s)
Polyesters/metabolism , Actinomycetales/enzymology , Enzymes/biosynthesis , Biodegradation, Environmental , Bioreactors , Enzymes/metabolism , Enzymes, Immobilized , Fermentation
8.
Braz. j. microbiol ; 48(3): 393-394, July-Sept. 2017.
Article in English | LILACS | ID: biblio-889136

ABSTRACT

Abstract Dietzia sp. 111N12-1, isolated from the seawater of South China Sea, shows strong petroleum hydrocarbons degradation activity. Here, we report the draft sequence of approximately 3.7-Mbp genome of this strain. To the best of our knowledge, this is the first genome sequence of Dietzia strain isolated from the sea. The genome sequence may provide fundamental molecular information on elucidating the metabolic pathway of hydrocarbons degradation in this strain.


Subject(s)
Seawater/microbiology , Actinomycetales/isolation & purification , Actinomycetales/genetics , Genome, Bacterial , Hydrocarbons/metabolism , Phylogeny , Biodegradation, Environmental , Actinomycetales/classification , Actinomycetales/metabolism , Petroleum/metabolism , Base Sequence , China
9.
Annals of Clinical Microbiology ; : 13-16, 2017.
Article in English | WPRIM | ID: wpr-193197

ABSTRACT

The genus Gordonia is one of the mycolic acid-containing aerobic actinomycetes. This genus has 38 named species that are widespread in the natural environment; however, Gordonia species rarely cause human infections. A 76-year-old woman presented with cough and sputum for over 1 year and was suspected of having nontuberculous mycobacterial (NTM) lung disease. An NTM isolate from the sputum was initially identified as Mycobacterium lentiflavum or Mycobacterium genavense by genotypic identification targeting internal transcribed spacer (ITS). However, the isolate was finally confirmed as Gordonia otitidis by sequencing of 16S rRNA, gyrB and secA1 genes. In patients with suspected NTM lung disease, the etiologic agent might be an organism other than NTM such as G. otitidis but still be identified as NTM without sequencing of 16S rRNA or other genes. Especially in case that a possible NTM isolate is identified as M. lentiflavum or M. genavense by the genotypic method targeting ITS, additional genotypic tests such as sequencing of 16S rRNA and other genes would be necessary for more reliable identification.


Subject(s)
Aged , Female , Humans , Actinobacteria , Actinomycetales , Cough , Lung Diseases , Lung , Methods , Mycobacterium , Nontuberculous Mycobacteria , Respiratory Tract Infections , Sputum
10.
Electron. j. biotechnol ; 19(6): 56-62, Nov. 2016. ilus
Article in English | LILACS | ID: biblio-840314

ABSTRACT

Background: Endoglucanase, one of three type cellulases, can randomly cleave internal p-1,4-linkages in cellulose polymers. Thus, it could be applied in agricultural and industrial processes. Results: A novel endoglucanase gene (JqCel5A) was cloned from Jonesia quinghaiensis and functionally expressed in Escherichia coli Rosetta (DE3). It contained 1722 bp and encoded a 573-residue polypeptide consisting of a catalytic domain of glycoside hydrolase family 5 (GH5) and a type 2 carbohydrate-binding module (CBM2), together with a predicted molecular mass of 61.79 kD. The purified JqCel5A displayed maximum activity at 55°C and pH 7.0, with 21.7 U/mg, 26.19 U/mg and 4.81 U/mg towards the substrate carboxymethyl cellulose, barley glucan and filter paper, respectively. Interestingly, JqCel5A exhibited high pH stability over a broad pH range of pH (3-11), and had good tolerance to a wide variety of deleterious chemicals including heavy metals and detergent. The catalytic mechanism of JqCel5A was also investigated by site mutagenesis and homology-modeling in this study. Conclusions: It was believed that these properties might make JqCel5A to be potentially used in the suitable industrial catalytic condition, which has a broad pH fluctuation and/or chemical disturbance.


Subject(s)
Actinomycetales/enzymology , Cellulases/chemistry , Cellulases/isolation & purification , Cellulases/genetics , Hydrogen-Ion Concentration , Mutagenicity Tests , Temperature
11.
Chinese Journal of Biotechnology ; (12): 599-609, 2016.
Article in Chinese | WPRIM | ID: wpr-337438

ABSTRACT

We isolated and identified the symbiotic and adnascent microorganisms from an unidentified sponge collected from 10-meter-deep seawater of the Paracel Islands in China. A total of 16 strains were obtained and identified. Through bacteriostatic activity assay, one of the strains, Dermacoccus sp. X4, was found to effectively inhibit the growth of Staphylococcus aureus. Subsequently, its secondary metabolites were purified by silica gel partition, octadecylsilane (ODS) reverse phase, Sephadex™LH-20 size exclusion, and C18 reverse phase chromatography. Using liquid chromatography, mass spectrometry, and nuclear magnetic resonance, three of the purified compounds were structurally characterized to be one 3-(4-hydroxybenzyl) hexahydropyrrolo [1,2-a]pyrazine-1,4-dione and two indole acid glycerides. This is the first report about indole acid glyceride isolated from microbial secondary metabolites, enriching marine drug candidate resources.


Subject(s)
Animals , Actinomycetales , Chemistry , China , Chromatography, Liquid , Indoles , Pharmacology , Magnetic Resonance Spectroscopy , Mass Spectrometry , Porifera , Microbiology , Seawater , Secondary Metabolism , Staphylococcus aureus
12.
Chinese Journal of Biotechnology ; (12): 845-856, 2015.
Article in Chinese | WPRIM | ID: wpr-240582

ABSTRACT

Nitrate not only remarkably stimulates the rifamycinbiosynthesis in Amycolatopsis mediterranei, but also influences the primary metabolisms, including the inhibition of fatty acids biosynthesis in the bacterial. This phenomenon has been designated as "Nitrate Stimulating Effect" by the late Prof. J.S. Chiaosince its discovery in the 1970's, and has been found in many other antibiotics-producing actinomycetes subsequently. Based on the research in his laboratory, we have revealed that the nitrate stimulation effect mainly manifests in two aspects over the last two decades. First, nitrate promotes the supply of rifamycin precursors, e.g., UDP-glucose, AHBA, malonyl-CoA and methylmalonyl-CoA. Specifically, the biosynthesis of fatty acids is inhibited by nitrate consequently the acetyl-CoA is shunted into malonyl-CoA. Second, nitrate facilitates the expression of genes in the rifclulsterthat encodes rifamycin biosynthetic enzymes. Following our current understanding, the future research will focus on the signals, the signal transduction pathway and the molecular mechanisms that dictate nitrate-mediated transcriptional and post-translational regulations.


Subject(s)
Actinomycetales , Classification , Metabolism , Acyl Coenzyme A , Chemistry , Anti-Bacterial Agents , Nitrates , Chemistry , Rifamycins
13.
Indian J Exp Biol ; 2014 Nov; 52(11): 1147-1151
Article in English | IMSEAR | ID: sea-153805

ABSTRACT

The study was undertaken with the aim of exploring novel and beneficial agro activities of rare actinomycetes like Microbispora sp. V2. The antagonistic activity of Microbispora sp. V2 was evaluated as a biocontrol agents against Sclerotium rolfsii, a soil-borne fungal plant pathogen. The methodology performed for evaluation of biocontrol agent was in vitro evaluation assay which comprised of three tests viz., cellophane overlay technique, seed germination test and Thiram (fungicide) tolerance of Microbispora sp. V2. The isolate was found to inhibit the fungal pathogen Sclerotium rolfsii to 91.43% in cellophane assay. In seed germination assay, Microbispora sp. V2 treated seeds resulted in 25.75% increased germination efficiency, as compared to seeds infected by Sclerotium rolfsii. The isolate Microbispora sp. V2 could tolerate 1000 µg mL-1 of Thiram (fungicide). The in vitro assay studies proved that Microbispora sp. V2 can be used as antifungal antagonist and thus posses’ great potential as biocontrol agent against southern blight caused by Sclerotium rolfsii in Zea mays L (Baby corn) which causes large economical losses.


Subject(s)
Actinomycetales/drug effects , Actinomycetales/physiology , Basidiomycota , Biomass , Drug Resistance, Bacterial , Fermentation , Fungicides, Industrial/pharmacology , Germination , In Vitro Techniques , Pest Control, Biological/methods , Phenazines/metabolism , Plant Diseases/microbiology , Plant Diseases/prevention & control , Seeds/microbiology , Seeds/physiology , Thiram/pharmacology , Zea mays/microbiology
14.
Acta Pharmaceutica Sinica ; (12): 230-236, 2014.
Article in Chinese | WPRIM | ID: wpr-297988

ABSTRACT

The crude extracts of the fermentation broth from a marine sediment-derived actinomycete strain, Saccharothrix sp. 10-10, showed significant antibacterial activities against drug-resistant pathogens. A genome-mining PCR-based experiment targeting the genes encoding key enzymes involved in the biosynthesis of secondary metabolites indicated that the strain 10-10 showed the potential to produce tetracenomycin-like compounds. Further chemical investigation of the cultures of this strain led to the identification of two antibiotics, including a tetracenomycin (Tcm) analogs, Tcm X (1), and a tomaymycin derivative, oxotomaymycin (2). Their structures were identified by spectroscopic data analysis, including UV, 1D-NMR, 2D-NMR and MS spectra. Tcm X (1) showed moderate antibacterial activities against a number of drug-resistant pathogens, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE) pathogens, with the MIC values in the range of 32-64 microg x mL(-1). In addition, 1 also displayed significant cytotoxic activities against human cancer cell lines, including HL60 (leukemia), HepG2 (liver), and MCF-7 (breast) with the IC 50 values of 5.1, 9.7 and 18.0 micromol x L(-1), respectively. Guided by the PCR-based gene sequence analysis, Tcm X (1) and oxotomaymycin (2) were identified from the genus of Saccharothrix and their 13C NMR data were correctly assigned on the basis of 2D NMR spectroscopic data analysis for the first time.


Subject(s)
Humans , Actinomycetales , Chemistry , Genetics , Anti-Bacterial Agents , Chemistry , Pharmacology , Antineoplastic Agents , Chemistry , Pharmacology , Benzodiazepinones , Chemistry , Pharmacology , Cell Line, Tumor , Data Mining , Methods , Drug Resistance, Bacterial , Enterococcus faecalis , Fermentation , Genomics , Inhibitory Concentration 50 , Marine Biology , Methicillin-Resistant Staphylococcus aureus , Microbial Sensitivity Tests , Molecular Structure , Naphthacenes , Chemistry , Pharmacology , Phylogeny , Staphylococcus epidermidis
15.
Braz. j. microbiol ; 44(2): 639-647, 2013. ilus, graf, tab
Article in English | LILACS | ID: lil-688595

ABSTRACT

The petroleum-derived degrading Dietzia cinnamea strain P4 recently had its genome sequenced and annotated. This allowed employing the data on genes that are involved in the degradation of n-alkanes. To examine the physiological behavior of strain P4 in the presence of n-alkanes, the strain was grown under varying conditions of pH and temperature. D. cinnamea P4 was able to grow at pH 7.0-9.0 and at temperatures ranging from 35 ºC to 45 ºC. Experiments of gene expression by real-time quantitative RT-PCR throughout the complete growth cycle clearly indicated the induction of the regulatory gene alkU (TetR family) during early growth. During the logarithmic phase, a large increase in transcriptional levels of a lipid transporter gene was noted. Also, the expression of a gene that encodes the protein fused rubredoxin-alkane monooxygenase was enhanced. Both genes are probably under the influence of the AlkU regulator.


Subject(s)
Actinomycetales/genetics , Actinomycetales/metabolism , Alkanes/metabolism , Gene Expression Profiling , Genes, Bacterial , Hydrocarbons/metabolism , Metabolic Networks and Pathways/genetics , Actinomycetales/growth & development , Biotransformation , Hydrogen-Ion Concentration , Real-Time Polymerase Chain Reaction , Temperature
16.
Journal of Veterinary Science ; : 491-494, 2013.
Article in English | WPRIM | ID: wpr-43056

ABSTRACT

Methods such as real time (RT)-PCR have not been developed for the rapid detection and diagnosis of Dermatophilus (D.) congolensis infection. In the present study, a D. congolensis-specific SYBR Green RT-PCR assay was evaluated. The detection limit of the RT-PCR assay was 1 pg of DNA per PCR reaction. No cross-reaction with nucleic acids extracted from Pseudomonas aeruginosa, Mycobacterium tuberculosis, Staphylococcus aureus, or Austwickia chelonae was observed. Finally, the RT-PCR assay was used to evaluate clinical samples collected from naturally infected animals with D. congolensis. The results showed that this assay is a fast and reliable method for diagnosing dermatophilosis.


Subject(s)
Animals , Cattle , Actinomycetales/isolation & purification , Actinomycetales Infections/diagnosis , Cattle Diseases/diagnosis , Fluorescent Dyes , Horse Diseases/diagnosis , Horses , Limit of Detection , Real-Time Polymerase Chain Reaction/methods , Reproducibility of Results , Sheep , Sheep Diseases/diagnosis
17.
Chinese Journal of Biotechnology ; (12): 1590-1598, 2013.
Article in Chinese | WPRIM | ID: wpr-242434

ABSTRACT

Wuxistatin, a novel and potent statin, is converted from lovastatin by Amycolatopsis sp. CGMCC1149. In the bioconversion, lovastatin is firstly hydroxylated by a hydroxylase. To obtain the critical hydroxylase, a novel hydroxylase gene was isolated from Amycolatopsis sp. CGMCC1149 by Degenerate PCR and Self-Formed Adaptor PCR and expressed in Escherichia coli. BLAST sequence analysis revealed that the gene belonged to cytochrome P450 gene superfamily and could encode a 403-amino-acid protein with a molecular weight of 44.8 kDa. The secondary structure prediction result showed that this protein contained many typical functional regions of P450, such as oxygen binding site, ion-pair region and heme binding region. Meanwhile, a catalytic function verification system was constructed by NADH, ferredoxin and ferredoxin reductase which could catalyze lovastatin hydroxylation into the target product. These would be helpful for further studies in large-scale production of wuxistatin.


Subject(s)
Actinomycetales , Genetics , Amino Acid Sequence , Butyrates , Metabolism , Cloning, Molecular , Cytochrome P-450 Enzyme System , Genetics , Metabolism , Hydroxylation , Industrial Microbiology , Lovastatin , Metabolism , Molecular Sequence Data
18.
Tuberculosis and Respiratory Diseases ; : 82-87, 2012.
Article in Korean | WPRIM | ID: wpr-101770

ABSTRACT

In 2005, a group of mycolic acid-containing bacteria was characterized as belonging to a novel genus, Segniliparus with species Segniliparus rugosus and S. rotundus. We report a case of the S. rugosus isolated from a 54-year-old woman with radiologic features mimicking that of non-tuberculous mycobacteriosis (NTM). When the patient first visited our hospital, an acid-fast bacteria (AFB) smear tested positive and Mycobacterium tuberculosis polymerase chain reaction (TB PCR) was negative in the bronchoalveolar lavage sample. After 2 months, the growing colonies were reported as NTM, but could not be identified because they had died. One year after the initial visit, induced sputum samples showed the same results, positive AFB smear and negative TB PCR. At this point, the growing colonies were identified as S. rugosus. Therefore, we should consider Segniliparus genus as a differential diagnosis for AFB in respiratory specimens in addition to the genus Mycobacterium.


Subject(s)
Female , Humans , Middle Aged , Actinomycetales , Bacteria , Bronchoalveolar Lavage , Diagnosis, Differential , Mycobacterium , Mycobacterium tuberculosis , Polymerase Chain Reaction , Sputum
19.
Chinese Journal of Biotechnology ; (12): 813-822, 2012.
Article in Chinese | WPRIM | ID: wpr-342439

ABSTRACT

In order to characterize a thermostable urate oxidase (Uox) from Microbacterium sp. strain ZZJ4-1, we cloned its gene (uox). The open reading frame of uox contained 894 base pairs and encoded a protein with 297 amino acids. Alignment of gene sequences indicated there was no obvious identity with the most reported uox and that 72% identity was found with uox from Arthrobacter globiformis. We inserted the gene into the plasmid pET-15b to construct an expression vector pET-15b-uox and got it induced expression in Escherichia coli BL21 (DE3). After the purification of the recombinant Uox by the HisBind column, we studied some properties of it. It was composed of subunits with a molecular mass of about 35 kDa. The optimal temperature and pH was 30 degrees C and pH 7.5. It was stable below 65 degrees C and from pH 8.5 to 11.0. The Km value was 0.22 mmol/L with the uric acid as the substrate. Ag+, Zn2+, CU2+ and SDS could totally inhibit its activity while Tween 20, Tween 80 and Triton X-100 had a slight promotion effect. The thermal stability of this enzyme was the most excellent among the reported recombinant Uox. Based on this property, it would be very useful in the application.


Subject(s)
Actinomycetales , Genetics , Amino Acid Sequence , Cloning, Molecular , Enzyme Stability , Escherichia coli , Genetics , Metabolism , Genetic Vectors , Genetics , Molecular Sequence Data , Recombinant Proteins , Genetics , Urate Oxidase , Genetics , Metabolism
20.
Chinese Journal of Biotechnology ; (12): 1057-1064, 2011.
Article in Chinese | WPRIM | ID: wpr-324503

ABSTRACT

Fermentation and induction conditions for recombinant Escherichia coli expressing Thermobifida fusca cutinase were optimized in flasks and 3L fermenter. Surface modification of poly (ethylene terephthalate) fibers with cutinase was also discussed. The results showed that, cutinase yield reached 128 U/mL by adding 2 g/L inducer lactose and 0.5% glycine. In the fed-batch culture in a 3 L fermenter, the maximum biomass cutinase activity was up to 506 U/mL, which is the highest bacterial cutinase activity reported by far. Recombinant cutinase was used to modify polyester fibers and terephthalic acid substance was detected by using UV analysis. The dyeing and wetting properties of cutinase treated fibers were higher than untreated fibers. Combined utilization of cutinase and Triton X-100 can significantly improve the hydrophilicity of polyester. This is the first report of surface modification on polyester fibers by bacterial cutinase.


Subject(s)
Actinomycetales , Genetics , Carboxylic Ester Hydrolases , Chemistry , Genetics , Escherichia coli , Genetics , Metabolism , Fermentation , Polyethylene Glycols , Chemistry , Polyethylene Terephthalates , Recombinant Proteins , Genetics , Surface Properties
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