scholarly journals Identification of biological characteristics and preliminary research on genetic transformation of the Aspergillus niger TL8 strain isolated in Vietnam

2017 ◽  
Vol 33 (2S) ◽  
Author(s):  
Do Thi Binh Xuan Loc ◽  
Tran Van Tuan
Keyword(s):  
Aspergillus Niger ◽  
Extracellular Enzymes ◽  
Carbon Sources ◽  
Morphological Characteristics ◽  
Transformation Method ◽  
Plant Materials ◽  
Rdna Its ◽  
Black Mold ◽  
Gfp Reporter Gene

Aspergillus niger is a mold commonly used in industrial production of many enzymes and organic acids. Because this fungus can produce different extracellular enzymes to degrade plant materials, it also causes the damages for some agricultural products at postharvest stages. In this study, we isolated a black mold strain named TL8 from a decayed dragon fruit. Based on morphological characteristics and the rDNA ITS (internal transcribed spacer) sequence, the TL8 strain was identified as A. niger. The A. niger TL8 strain is able to use different carbon sources for the growth and decay the peel of dragon fruits in vitro. In order to establish the basis for future studies on the mechanism of plant material decomposition of the fungus, we have successfully transferred and expressed the GFP reporter gene in this A. niger strain using the Agrobacterium tumefaciens-mediated transformation method and the hygromycin resistance marker.

scholarly journals ABC Transporters Required for Hexose Uptake byClostridium phytofermentans

2019 ◽  
Vol 201 (15) ◽  
Author(s):  
Tristan Cerisy ◽  
Alba Iglesias ◽  
William Rostain ◽  
Magali Boutard ◽  
Christine Pelle ◽  
...  
Keyword(s):  
Abc Transporters ◽  
Extracellular Enzymes ◽  
Plant Biomass ◽  
Carbon Sources ◽  
Anaerobic Bacteria ◽  
Model Organism ◽  
Industrial Transformation ◽  
Content Type ◽  
Plant Matter

ABSTRACTThe mechanisms by which bacteria uptake solutes across the cell membrane broadly impact their cellular energetics. Here, we use functional genomic, genetic, and biophysical approaches to reveal howClostridium(Lachnoclostridium)phytofermentans, a model bacterium that ferments lignocellulosic biomass, uptakes plant hexoses using highly specific, nonredundant ATP-binding cassette (ABC) transporters. We analyze the transcription patterns of its 173 annotated sugar transporter genes to find those upregulated on specific carbon sources. Inactivation of these genes reveals that individual ABC transporters are required for uptake of hexoses and hexo-oligosaccharides and that distinct ABC transporters are used for oligosaccharides versus their constituent monomers. The thermodynamics of sugar binding shows that substrate specificity of these transporters is encoded by the extracellular solute-binding subunit. As sugars are not phosphorylated during ABC transport, we identify intracellular hexokinases based onin vitroactivities. These mechanisms used byClostridiato uptake plant hexoses are key to understanding soil and intestinal microbiomes and to engineer strains for industrial transformation of lignocellulose.IMPORTANCEPlant-fermentingClostridiaare anaerobic bacteria that recycle plant matter in soil and promote human health by fermenting dietary fiber in the intestine.Clostridiadegrade plant biomass using extracellular enzymes and then uptake the liberated sugars for fermentation. The main sugars in plant biomass are hexoses, and here, we identify how hexoses are taken in to the cell by the model organismClostridium phytofermentans. We show that this bacterium uptakes hexoses using a set of highly specific, nonredundant ABC transporters. Once in the cell, the hexoses are phosphorylated by intracellular hexokinases. This study provides insight into the functioning of abundant members of soil and intestinal microbiomes and identifies gene targets to engineer strains for industrial lignocellulosic fermentation.

scholarly journals Identification of biological characteristics and prelimilary research on genetic transformation of the filamentous fungus Penicillium chrysogenum originated from Vietnam

2017 ◽  
Vol 33 (2S) ◽  
Author(s):  
Vu Xuan Tao ◽  
Tran Van Tuan
Keyword(s):  
Filamentous Fungus ◽  
Internal Transcribed Spacer ◽  
Penicillium Chrysogenum ◽  
Antibiotic Production ◽  
Morphological Characteristics ◽  
Transformation Method ◽  
Culture Collection ◽  
Resistance Marker ◽  
Rdna Its ◽  
National University

Penicillium chrysogenum is a well-known filamentous fungus for production of penicillin and some valuable secondary metabolites. In this study, we indicated that two strains VTCC-F1170 and VTCC-F1172 identified as P. chrysogenum, which are preserved at Vietnam Type Culture Collection (VTCC) of Vietnam National University Hanoi, exhibited the ability of antibiotic production to inhibit the tested bacterium Staphylococcus aureus on the agar plates by diffusion assays. Additional analyses of the morphological characteristics and the rDNA ITS (internal transcribed spacer) sequence confirmed that the identification of both strains as P. chrysogenum was completely accurate. These important evidences guaranteed that the fungal strains are reliable for the researches on genetic engineering of P. chrysogenum. Experimental assays of antibiotic susceptibility showed that the growth of both strains VTCC-F1170 and VTCC-F1172 was completely inhibited by nourseothricin at 50 μg/ml and phleomycin at 150 μg/ml. Using the Agrobacterium tumefaciens-mediated transformation method, we have succesfully transferred the nourseothricin resistance marker into the genome of the VTCC-F1170 strain.  

scholarly journals Pectin as Carbon Source for Monilinia laxa Exoproteome and Expression Profiles of Related Genes

2020 ◽  
Vol 33 (9) ◽  
pp. 1116-1128 ◽  
Author(s):  
Silvia Rodríguez-Pires ◽  
Paloma Melgarejo ◽  
Antonieta De Cal ◽  
Eduardo A. Espeso
Keyword(s):  
Extracellular Enzymes ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Carbon Sources ◽  
Brown Rot ◽  
Secretory Activity ◽  
Late Response ◽  
Monilinia Laxa ◽  
Brown Rot Fungi

Pectin, as part of the fruit cell wall, can be degraded by brown rot fungi by coordinating the production, secretion, and action of extracellular enzymes. In this study, pectin utilization by the necrotroph Monilinia laxa 8L was studied by in vitro and in silico approaches. A total of 403 genes encoding carbohydrate-active enzymes (CAZymes) were identified, including 38 coding a predicted pectin-degrading activity. Analyzing the differences between M. laxa 8L exoproteomes in media containing glucose and pectin as sole carbon sources, we identified 107 pectin-specific proteins, among them, 64.48% harbor a classical secretory activity, including 42 CAZymes and six pectin-degrading proteins. Analyzing the gene-expression patterns of some pectinase families revealed their possible sequential action in pectin disassembly. We found, in vitro, an early pectin-dependent induction of MlRGAE1, MlPG1, and three members of the rhamnosidase family (MlαRHA2, MlαRHA3, and MlαRHA6) and late response of MlPG2 and MlPNL3. M. laxa 8L has the ability to use both pectin and byproducts as carbon sources, based on a functional pectinolytic machinery encoded in its genome, subjected to pectin-dependent regulation and appropriate secretion mechanisms of these pectinolytic enzymes. Differences in the secretion and transcription profile of M. laxa 8L provided insights into the different mechanisms that contribute to brown rot development.

scholarly journals The predicted secretome of Lactobacillus plantarum WCFS1 sheds light on interactions with its environment

Microbiology ◽  
2006 ◽  
Vol 152 (11) ◽  
pp. 3175-3183 ◽  
Author(s):  
Jos Boekhorst ◽  
Michiel Wels ◽  
Michiel Kleerebezem ◽  
Roland J. Siezen
Keyword(s):  
Lactobacillus Plantarum ◽  
Extracellular Enzymes ◽  
Carbon Sources ◽  
Experimental Studies ◽  
Extracellular Proteins ◽  
Primary Role ◽  
Comprehensive Overview ◽  
Plant Materials ◽  
Phylogenetic Profiling ◽  
Insight Into

The predicted extracellular proteins of the bacterium Lactobacillus plantarum were analysed to gain insight into the mechanisms underlying interactions of this bacterium with its environment. Extracellular proteins play important roles in processes ranging from probiotic effects in the gastrointestinal tract to degradation of complex extracellular carbon sources such as those found in plant materials, and they have a primary role in the adaptation of a bacterium to changing environmental conditions. The functional annotation of extracellular proteins was improved using a wide variety of bioinformatics methods, including domain analysis and phylogenetic profiling. At least 12 proteins are predicted to be directly involved in adherence to host components such as collagen and mucin, and about 30 extracellular enzymes, mainly hydrolases and transglycosylases, might play a role in the degradation of substrates by L. plantarum to sustain its growth in different environmental niches. A comprehensive overview of all predicted extracellular proteins, their domains composition and their predicted function is provided through a database at http://www.cmbi.ru.nl/secretome, which could serve as a basis for targeted experimental studies into the function of extracellular proteins.

Fungus ball paranasal sinuses: pattern of histopathology and culture characteristics

2019 ◽  
Vol 49 (20) ◽  
Author(s):  
Nugroho Suharsono
Keyword(s):  
Aspergillus Niger ◽  
Paranasal Sinus ◽  
Paranasal Sinuses ◽  
Histopathological Examination ◽  
Morphological Characteristics ◽  
Sinus Surgery ◽  
Nasal Discharge ◽  
Fungus Ball ◽  
Methenamine Silver ◽  
Fungus Balls

Background: Fungal infection of the nose and paranasal sinuses is an uncommon condition which is now being increasingly recognized. The clinical presentation is not specific with various symptoms such as nasal obstruction, purulent nasal discharge, facial pain, and chronic cough. Only unilaterality may alert the clinician. Purpose: To find the morphological characteristics of the fungus in patients with paranasal sinus fungus ball. Methods: A retrospective study of 13 paranasal sinus fungus balls cases which underwent endoscopic sinus surgery at Department of Otorhinolaryngology Head and Neck Surgery St. Vincentius A Paulo Hospital Surabaya from March, 2012 until December, 2013. Age, sex, histopathology and fungal cultur were analysed. Histopathologic sections of all the patients were stained with hematoxylin and eosin (H&E), and Gomori methenamine silver (GMS). The specimens were then cultured on Sabouraud dextrose agar plates and incubated at 30°C for 1 month. At the end of the incubation period, the samples were evaluated microscopically to detect fungi and identify their species. Results: The age reported of the 13 patients, was ranging from 36 to 63 years old. There was a significant female predominance, 10 female patients (76.92%) and 3 male patients (23.08%). Histopathological examination showed that most causative agents were Aspergillus species 92.31% (12/13). Culture test was positive for 69.23% (9/13). Aspergillus niger (61.54%, 8/13) is the most frequent fungus reported to cause fungus balls. Conclusion: Pattern of histopathologic on HE and GMS is very helpful and sensitive to identify fungi. The most common isolated mould in our study was Aspergillus niger.Keywords: fungus ball, histopathology and culture, Aspergillus nigerABSTRAK Latar Belakang: Infeksi jamur di hidung dan sinus paranasal merupakan kondisi yang jarang terjadi, namun kini lebih sering ditemukan. Gejala klinisnya tidak spesifik dapat berupa obstruksi hidung, sekret dari hidung, nyeri wajah, dan batuk kronis. Bila terjadi unilateral, patut diwaspadai oleh para klinisi. Tujuan: Untuk mengetahui karakteristik morfologi fungus yang didapati pada pasien sinusitis jamur yang kami teliti. Metode: Dilakukan penelitian retrospektif pada 13 pasien sinusitis jamur yang menjalani bedah sinus endoskopi di Departemen Otorinolaringologi-Kepala Leher Rumah Sakit St. Vincentius A Paulo Surabaya dari bulan Maret 2012 sampai dengan Desember 2013. Dilakukan analisis usia, jenis kelamin, histopatologi dan kultur jamur. Pewarnaan preparat histopatologi menggunakan Hematoxylin dan eosin (H&E) dan Gomori Methenamine Silver (GMS). Kemudian spesimen diletakkan pada piring agar Sabouraud dextrose, dan dilakukan inkubasi pada suhu 30°C selama satu bulan. Pada akhir masa inkubasi, sampel dievaluasi dengan mikroskop untuk mendeteksi jamur dan spesiesnya. Hasil: Didapati usia 13 penderita berkisar dari 36-63 tahun. Wanita lebih dominan sebanyak 10 penderita (76,92 %) dan 3 penderita laki-laki (23,08%). Hasil pemeriksaan histopatologi menunjukkan spesies Aspergillus sebagai penyebab utama (92,31%) pada 12 penderita (12/13).Tes kultur positif pada 69,23% (9/13). Jamur yang paling sering menyebabkan bola jamur pada sinus adalah Aspergillus niger (61,54%, 8/13). Kesimpulan: Pewarnaan preparat histopatologi menggunakan Hematoxylin dan eosin (H&E) dan Gomori Methenamine Silver (GMS) sangat berguna dan sensitif dalam mendeteksi adanya jamur. Jenis jamur yang paling banyak ditemukan pada penelitian kami adalah Aspergillus niger.

The experience of holding the cycles of assisted reproductive technology with defrost, a biopsy, genetic study and refreezing of embryos in patients with multiple unsuccessful implantations

2016 ◽  
pp. 166-170
Author(s):  
Y.V. Masliy ◽  
◽  
I.O. Sudoma ◽  
P.S. Mazur ◽  
D.A. Mykytenko ◽  
...  
Keyword(s):  
Chromosomal Rearrangements ◽  
Genetic Diagnosis ◽  
Morphological Characteristics ◽  
Implantation Rate ◽  
Ovarian Response ◽  
Reproductive Technologies ◽  
Comparative Genome Hybridization ◽  
Comparative Genome ◽  
Vitro Fertilization

The objective: to study the possibility of using frozen blastocysts for biopsy and genetic testing and performance measurement transfer euploeded 5–7-day-old embryos after thawing, biopsies, refreezing and thawing in patients with unsuccessful implantation. Patients and methods. The object of the study was the group of patients with repeated failure of implantation (4) in programs of auxiliary reproductive technologies (ART), subject to transfer to the uterus in total (i.e. in all the programs) for at least 6 good quality embryos based on morphological characteristics). All women had sufficient ovarian reserve. The patient was treated for infertility within the ART programs of the clinic of reproductive medicine "Nadiya" in the period from 2006 to 2016. The sample included couples who were not carriers of chromosomal rearrangements, without anomalies of the uterus (congenital and acquired: a doubling of the uterus, one-horned uterus, intrauterine membrane, synechia, submucous myoma of the uterus). All women had a positive ovarian response to controlled stimulation with gonadotropins (at least 7 oocytes) and a sufficient number of cryopreserved embryos. The first group (G1) included 64 women who trophectodermal a biopsy was performed on fresh blastocysts (in a loop controlled ovarian hyperstimulation). The second group (G2) were included 31 women who underwent thawing previously cryopreserved blastocysts trophectodermal re-biopsy and vitrification of blastocysts. Results. It was found that the performance of transfers euploid embryos that were vitrified, bioptrone and revitriphted, a little lower than those that were bioptrone fresh and vitrified only once. At the same time computationa genetic diagnosis previously vitrified blastocysts using comparative genome hybridization in patients with recurrent failed implantation allows to obtain a reasonable pregnancy rate (58%), implantation rate (33.3 %) and the birth of living children (45.1 %). Conclusion. Reprising biopropane embryos does not cause significant destructive impact and allows you to achieve pregnancy and birth of the alive child. Key words: in vitro fertilization, reusable unsuccessful implantation, a method of comparative genome hybridization, refreezing.

Effect of carbon source on Phenobarbital metabolism by Rhizopus stolonifer

2010 ◽  
Vol 3 (2) ◽  
pp. 1022-1027
Author(s):  
Kavitha K ◽  
Asha S ◽  
Hima Bindu T.V.L ◽  
Vidyavathi M
Keyword(s):  
Carbon Source ◽  
High Performance ◽  
Carbon Sources ◽  
In Vitro Models ◽  
Rhizopus Stolonifer ◽  
Safety And Efficacy ◽  
Alternative Systems ◽  
Toxicological Studies ◽  
Microbial Model

The safety and efficacy of a drug is based on its metabolism or metabolite formed. The metabolism of drugs can be studied by different in vitro models, among which microbial model became popular. In the present study, eight microbes were screened for their ability to metabolize phenobarbital in a manner comparable to humans with a model to develop alternative systems to study human drug metabolism. Among the different microbes screened, a filamentous fungi Rhizopus stolonifer metabolized phenobarbital to its metabolite which is used for further pharmacological and toxicological studies. The transformation of phenobarbital was identified by high- performance liquid chromatography (HPLC). Interestingly, Rhizopus stolonifer sample showed an extra metabolite peak at 3.11min. compared to its controls. The influence of different carbon sources in media used for growth of fungus, on metabolite production was studied, to find its effect in production of metabolite as the carbon source may influence the growth of the cell.

Formulation of Modified Release Atorvastatin Nanoparticles by Emulsion Interfacial Reaction Method

2015 ◽  
Vol 8 (3) ◽  
pp. 2937-2940
Author(s):  
Sudhakar Sekar ◽  
Shee Sim May
Keyword(s):  
Interfacial Reaction ◽  
Therapeutic Potential ◽  
Chitosan Nanoparticles ◽  
In Vitro Release ◽  
Morphological Characteristics ◽  
Preparation Technique ◽  
Modified Release ◽  
Reaction Method ◽  
Vitro Release

The aim of the study is to formulate a modified release chitosan nanoparticles for the oral delivery of atorvastatin and to study the in vitro release of atorvastatin from chitosan nanoparticles. Atorvastatin-loaded chitosan nanoparticles were prepared with different concentration of cross-linking agent (glutaraldehyde) by emulsion interfacial reaction method. The formed nanoparticles were characterized in terms of size and morphological characteristics by scanning electron microscopy (SEM) and transmission electron microscope (TEM). Spherical and regular nanoparticles with the size range of 100-250nm were formed. Atorvastatin encapsulation efficiency of nanoparticles was found to be highest in ANP3, followed by ANP2 and ANP1. The in vitro release of atorvastatin was studied by membrane diffusion technique. The resulted cumulative percentage of drug released for ANP1, ANP2 and ANP3 were 60.08%, 34.81% and 20.39% respectively. Through this study, the nanoparticles preparation technique has shown to be a promising approach for enhancing the dissolution of hydrophobic drugs like atorvastatin calcium. The application of this novel delivery system offers good therapeutic potential in the management of hypercholesterolemia and dyslipidemia.

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