New antioxidants from the culture broth of Coprinopsis echinospora

Background: Malaria is one of the life threatening diseases which is caused by Plasmodium sp. of protozoa and uses Anopheles mosquitos as vector. Plasmodium vivax and Plasmodium falciparum are common form of malaria parasite. Artemisinin is reported for its antimalarial activities and Artemether which is a methyl ether derivative of Artemisinin, has been found effective against P. falciparum. Methods: In the present study, bioconversion of Artemisinin into Artemether was carried out experimentally and the statistical tools like experimental factorial design and Response Surface Methodology were used to find optimal conditions (concentration of Artemisinin, age of inoculum, temperature & pH) using Cunninghamella echinulata var. elegans. Experimental conditions for maximum product recovery from culture broth were also optimized using various polar and non-polar solvents for extraction. Artemether purity was analyzed by reverse-phase HPLC. Experimental data was fitted in a quadratic model and effect of various parameters was analyzed. Results: It was found that bioconversion of Artemisinin into Artemether is growth associated process. It was observed that molasses used as carbon source supported production of Artemether to 3.4g/L. The biomass and oxygen are key element affecting of bioconversion of Artemisinin into Artemether such as higher dissolved oxygen reduced the Artemether bioconversion. The highest bioconversion of Artemisinin into Artemether was obtained at temperature 25.5oC, 5g/L concentration of Artemisinin, at age of inoculum of 44.5 h and at pH 6.0. Model suggested the highest bioconversion of Artemisinin into Artemether was 54% at shake flask level which was near about experimental finding. An optimal condition for bioconversion was also analyzed and 64% bioconversion was obtained in 5L bioreactor. Conclusion: The outcomes of the study provided optimum conditions for bioconversion of Artemisinin into Artemether.

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Controlling Unconventional Secretion for Production of Heterologous Proteins in Ustilago maydis through Transcriptional Regulation and Chemical Inhibition of the Kinase Don3

Journal of Fungi ◽

10.3390/jof7030179 ◽

2021 ◽

Vol 7 (3) ◽

pp. 179

Author(s):

Kai P. Hussnaetter ◽

Magnus Philipp ◽

Kira Müntjes ◽

Michael Feldbrügge ◽

Kerstin Schipper

Keyword(s):

Protein Production ◽

Heterologous Protein ◽

Ustilago Maydis ◽

Downstream Processing ◽

Culture Broth ◽

Yeast Cells ◽

Heterologous Protein Expression ◽

Heterologous Proteins ◽

Regulatory Systems ◽

Unconventional Secretion

Heterologous protein production is a highly demanded biotechnological process. Secretion of the product to the culture broth is advantageous because it drastically reduces downstream processing costs. We exploit unconventional secretion for heterologous protein expression in the fungal model microorganism Ustilago maydis. Proteins of interest are fused to carrier chitinase Cts1 for export via the fragmentation zone of dividing yeast cells in a lock-type mechanism. The kinase Don3 is essential for functional assembly of the fragmentation zone and hence, for release of Cts1-fusion proteins. Here, we are first to develop regulatory systems for unconventional protein secretion using Don3 as a gatekeeper to control when export occurs. This enables uncoupling the accumulation of biomass and protein synthesis of a product of choice from its export. Regulation was successfully established at two different levels using transcriptional and post-translational induction strategies. As a proof-of-principle, we applied autoinduction based on transcriptional don3 regulation for the production and secretion of functional anti-Gfp nanobodies. The presented developments comprise tailored solutions for differentially prized products and thus constitute another important step towards a competitive protein production platform.

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Two New Phomaligols from the Marine-Derived Fungus Aspergillus flocculosus and Their Anti-Neuroinflammatory Activity in BV-2 Microglial Cells

Marine Drugs ◽

10.3390/md19020065 ◽

2021 ◽

Vol 19 (2) ◽

pp. 65

Author(s):

Byeoung-Kyu Choi ◽

Duk-Yeon Cho ◽

Dong-Kug Choi ◽

Phan Thi Hoai Trinh ◽

Hee Jae Shin

Keyword(s):

Nitric Oxide ◽

Mass Spectrometry ◽

Optical Rotation ◽

Chemical Oxidation ◽

Culture Broth ◽

Membered Ring ◽

Microglial Cells ◽

No Production ◽

Nmr Data ◽

Ic50 Value

Two new phomaligols, deketo-phomaligol A (1) and phomaligol E (2), together with six known compounds (3–8) were isolated from the culture broth of the marine-derived fungus Aspergillus flocculosus. Compound 1 was first isolated as a phomaligol derivative possessing a five-membered ring. The structures and absolute configurations of the new phomaligols were determined by detailed analyses of mass spectrometry (MS), nuclear magnetic resonance (NMR) data, optical rotation values and electronic circular dichroism (ECD). In addition, the absolute configurations of the known compounds 3 and 4 were confirmed by chemical oxidation and comparison of optical rotation values. Isolated compounds at a concentration of 100 μM were screened for inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)-induced BV-2 microglial cells. Among the compounds, 4 showed moderate anti-neuroinflammatory effects with an IC50 value of 56.6 μM by suppressing the production of pro-inflammatory mediators in activated microglial cells without cytotoxicity.

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Application of Bio-Friendly Formulations of Chitinase-Producing Streptomyces cellulosae Actino 48 for Controlling Peanut Soil-Borne Diseases Caused by Sclerotium rolfsii

Journal of Fungi ◽

10.3390/jof7030167 ◽

2021 ◽

Vol 7 (3) ◽

pp. 167

Author(s):

Gaber Abo-Zaid ◽

Ahmed Abdelkhalek ◽

Saleh Matar ◽

Mai Darwish ◽

Muhammad Abdel-Gayed

Keyword(s):

Open Field ◽

Biocontrol Agent ◽

Sclerotium Rolfsii ◽

Dry Weight ◽

Culture Broth ◽

Field Conditions ◽

Damping Off ◽

Mycelium Growth ◽

Particle Size Analyzer ◽

Cell Pellet

Of ten actinobacterial isolates, Streptomyces cellulosae Actino 48 exhibited the strongest suppression of Sclerotium rolfsii mycelium growth and the highest chitinase enzyme production (49.2 U L−1 min−1). The interaction between Actino 48 and S. rolfsii was studied by scanning electron microscope (SEM), which revealed many abnormalities, malformations, and injuries of the hypha, with large loss of S. rolfsii mycelia density and mass. Three talc-based formulations with culture broth, cell-free supernatant, and cell pellet suspension of chitinase-producing Actino 48 were characterized using SEM, Fourier transform infrared spectroscopy (FTIR), and a particle size analyzer. All formulations were evaluated as biocontrol agents for reducing damping-off, root rot, and pods rot diseases of peanut caused by S. rolfsii under greenhouse and open-field conditions. The talc-based culture broth formulation was the most effective soil treatment, which decreased the percentage of peanut diseases under greenhouse and open-field conditions during two successive seasons. The culture broth formulation showed the highest increase in the dry weight of peanut shoots, root systems, and yielded pods. The transcriptional levels of three defense-related genes (PR-1, PR-3, and POD) were elevated in the culture broth formulation treatment compared with other formulations. Subsequently, the bio-friendly talc-based culture broth formulation of chitinase-producing Actino 48 could potentially be used as a biocontrol agent for controlling peanut soil-borne diseases caused by S. rolfsii.

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Evaluation of the Performance of Direct Susceptibility Test by VITEK-2 from Positively Flagged Blood Culture Broth for Gram-Negative Bacilli

Journal of Laboratory Physicians ◽

10.1055/s-0041-1732489 ◽

2021 ◽

Author(s):

Kavipriya D. ◽

Suman Susan Prakash ◽

Sarumathi Dhandapani ◽

Deepashree Rajshekar ◽

Apurba Sankar Sastry

Keyword(s):

Blood Culture ◽

Susceptibility Testing ◽

Blood Stream Infection ◽

Tertiary Care ◽

Reference Method ◽

Culture Broth ◽

Timely Initiation ◽

Gram Negative ◽

Mortality And Morbidity ◽

Vitek 2

Abstract Background Timely initiation of antimicrobial therapy in patients with blood stream infection is absolutely necessary to reduce mortality and morbidity. Most clinical microbiology laboratories use conventional methods for identification and antimicrobial susceptibility testing (AST) that involve biochemical methods for identification followed by AST by disk diffusion. The aim of the current study is to assess the various errors associated with direct susceptibility testing done from blood culture broth using automated AST system-Vitek-2 compact compared with the reference method of AST done from bacterial colonies. Materials and Methods The study was conducted in a tertiary care public sector 2,200-bedded hospital in South India for a period of 6 months. The study involved positively flagged blood culture bottles that yielded single morphotype of Gram-negative organism by Gram stain. A total of 120 bacterial isolates were collected that consisted of consecutively obtained first 60 isolates of Enterobacteriaceae family (30 Escherichia coli and 30 Klebsiella pneumoniae) and consecutively obtained first 60 nonfermenters (30 Pseudomonas aeruginosa and 30 Acinetobacter baumannii). Vitek-2 AST was done from these 120 blood culture broth, following the protocol by Biomerieux, and results were obtained. Then, Vitek-2 was done from colonies (reference method) using appropriate panel for Enterobacteriaceae and nonfermenters, and results were obtained. Both the results were compared. Results Nonfermenters showed a better categorical agreement of 97.6%, as compared to Enterobacteriaceae, which showed 97%. Among Enterobacteriaceae, both E. coli and K. pneumoniae showed categorical agreement of 97% each. Conclusion The procedure of AST directly from blood culture broth represents a simple and effective technique that can reduce the turnaround time by 24 hours, which in turn benefits the clinician in appropriate utilization of antimicrobials for better patient care.

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Two antibacterial and PPARα/γ-agonistic unsaturated keto fatty acids from a coral-associated actinomycete of the genus Micrococcus

Beilstein Journal of Organic Chemistry ◽

10.3762/bjoc.16.29 ◽

2020 ◽

Vol 16 ◽

pp. 297-304 ◽

Cited By ~ 1

Author(s):

Amit Raj Sharma ◽

Enjuro Harunari ◽

Naoya Oku ◽

Nobuyasu Matsuura ◽

Agus Trianto ◽

...

Keyword(s):

Fatty Acids ◽

Spectroscopic Analysis ◽

Culture Broth ◽

Peroxisome Proliferator ◽

Simulation Studies ◽

Fish Pathogen ◽

Chemical Structures ◽

Agonistic Activity ◽

Isoform Specificity ◽

Peroxisome Proliferator Activated Receptors

A pair of geometrically isomeric unsaturated keto fatty acids, (6E,8Z)- and (6E,8E)-5-oxo-6,8-tetradecadienoic acids (1 and 2), were isolated from the culture broth of an actinomycete of the genus Micrococcus, which was associated with a stony coral, Catalaphyllia sp. Their chemical structures were elucidated by spectroscopic analysis including NMR and MS, with special assistance of spin system simulation studies for the assignment of an E geometry at C8 in 2. As metabolites of microbes, compounds 1 and 2 are unprecedented in terms of bearing a 2,4-dienone system. Both 1 and 2 showed antibacterial activity against the plant pathogen Rhizobium radiobacter and the fish pathogen Tenacibaculum maritimum, with a contrasting preference that 1 is more effective to the former strain while 2 is so to the latter. In addition, compounds 1 and 2 displayed agonistic activity against peroxisome proliferator-activated receptors (PPARs) with an isoform specificity towards PPARα and PPARγ.

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Wenzhouxiangella marina gen. nov, sp. nov, a marine bacterium from the culture broth of Picochlorum sp. 122, and proposal of Wenzhouxiangellaceae fam. nov. in the order Chromatiales

Antonie van Leeuwenhoek ◽

10.1007/s10482-015-0458-7 ◽

2015 ◽

Vol 107 (6) ◽

pp. 1625-1632 ◽

Cited By ~ 21

Author(s):

Guanghua Wang ◽

Mingxing Tang ◽

Tao Li ◽

Shikun Dai ◽

Huanlian Wu ◽

...

Keyword(s):

Marine Bacterium ◽

Culture Broth

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Characterization of the Antibiotic Compound No. 70 Produced byStreptomycessp. IMV-70

The Scientific World JOURNAL ◽

10.1100/2012/594231 ◽

2012 ◽

Vol 2012 ◽

pp. 1-8 ◽

Cited By ~ 3

Author(s):

Lyudmila P. Trenozhnikova ◽

Almagul K. Khasenova ◽

Assya S. Balgimbaeva ◽

Galina B. Fedorova ◽

Genrikh S. Katrukha ◽

...

Keyword(s):

Optimal Growth ◽

Growth Conditions ◽

Culture Broth ◽

Producer Strain ◽

New Class ◽

New Antibiotics ◽

Actinomycete Strain ◽

Main Component

We describe the actinomycete strain IMV-70 isolated from the soils of Kazakhstan, which produces potent antibiotics with high levels of antibacterial activity. After the research of its morphological, chemotaxonomic, and cultural characteristics, the strain with potential to be developed further as a novel class of antibiotics with chemotherapeutics potential was identified asStreptomycessp. IMV-70. In the process of fermentation, the strainStreptomycesspp. IMV-70 produces the antibiotic no. 70, which was isolated from the culture broth by extraction with organic solvents. Antibiotic compound no. 70 was purified and separated into individual components by HPLC, TLC, and column chromatography methods. The main component of the compound is the antibiotic 70-A, which was found to be identical to the peptolide etamycin A. Two other antibiotics 70-B and 70-C have never been described and therefore are new antibiotics. The physical-chemical and biological characteristics of these preparations were described and further researched. Determination of the optimal growth conditions to cultivate actinomycete-producer strain IMV-70 and development of methods to isolate, purify, and accumulate preparations of the new antibiotic no. 70 enable us to research further the potential of this new class of antibiotics.

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