ABSTRACT
The recently described bioluminescent system from fungi has great potential for developing highly efficient tools for biomedical research. Luciferase enzyme is one of the most crucial components of this system. The luciferase from Neonothopanus nambi fungus belongs to the novel still undescribed protein family. The structure data for this protein is almost absent. A detailed study of the N. nambi luciferase properties is necessary for the improvement of analytical methods based on the fungal bioluminescent system. Here we present the positions of key amino acid residues and their effect on enzyme function described using bioinformatic and experimental approaches. These results are useful for further fungal luciferase structure determination.
Subject(s)
Agaricales/enzymology , Fungal Proteins/chemistry , Luciferases/chemistry , Agaricales/genetics , Amino Acid Sequence , Catalytic Domain , Computational Biology/methods , Fungal Proteins/genetics , Fungal Proteins/metabolism , Luciferases/genetics , Luciferases/metabolism , Luminescence , Models, Molecular , Mutagenesis , Mutation , Sequence Homology, Amino Acid , Structure-Activity RelationshipABSTRACT
AIM: To study the synthesis of phytohormones (auxins, cytokinins, abscisic acid) under cultivation of Nocardia vaccinii IMV B-7405 (surfactants producer) in media containing different carbon sources (glycerol, refined sunflower oil, as well as waste oil after frying potatoes and meat). METHODS: Phytohormones were extracted from supernatants of culture liquid (before or after surfactant separation) by ethylacetate (auxins, abscisic acid) and n-butanol (cytokinins), concentrated and purified by thin-layer chromatography, then quantitative determination was performed using a scanning Sorbfil spectrodensitometer. RESULTS: While growing in medium with refined oil IMV B-7405 strain synthesized 1.8 ± 0.09 g/l extracellular surfactant, also maximum amount of auxins (245-770 µ/l) and cytokinins (134-348 µl). Cultivation of N. vaccini LMV B-7405 on waste oil was accompanied by decreasing amount of phytohormones to 23-84 µ/l (auxins) and 16-90 µ/l (cytokinins) and increasing surfactant concentration to 2.3-2.6 g/l. The level of abscisic acid synthesis was practically not dependent on the nature of growth substrate, was substantially lower than that of auxins and cytokinins and ranged from 2 to 12 µ/l. CONCLUSIONS: Obtained data demonstrate the possibility of using oil-containing industrial waste for the simultaneous synthesis of both surfactants and phytohormones, and indicate the need for studies of the effect of producer cultivation conditions on the biological properties of the target products of microbial synthesis.
Subject(s)
Abscisic Acid/biosynthesis , Cytokinins/biosynthesis , Indoleacetic Acids/metabolism , Nocardia/metabolism , Plant Growth Regulators/metabolism , Surface-Active Agents/metabolism , 1-Butanol , Abscisic Acid/isolation & purification , Acetates , Culture Media/chemistry , Cytokinins/isolation & purification , Fermentation , Glycerol/metabolism , Indoleacetic Acids/isolation & purification , Industrial Microbiology , Industrial Oils/analysis , Industrial Waste/analysis , Plant Growth Regulators/isolation & purification , Plant Oils/metabolism , Solvents , Sunflower Oil , Surface-Active Agents/isolation & purificationABSTRACT
Attachment of the cells of some bacteria, yeasts, and micromycetes to various surfaces (catheters, dentures, plastic, polyvinyl chloride, tiles, and steel) treated with the surfactants fromAcinetobacter calcoace- ticus IMB B-7241, Rhodococcus erythropolis IMB Ac-5017, and Nocardia vaccinii IMB B-7405 was studied. Adhesion of microorganisms to all the studied surfaces depended on the surfactant concentration and purity, kind of surface, and the test culture. Treatment with the surfactants from N. vaccinii IMB B-7405 (0.005- 0.05 mg/mL), A. calcoaceticus IMB B-7241 (0.003-0.036 mg/mL), and R. erythropolis IMB Ac-5017 (0.03- 0.12 mg/mL) resulted in adhesion decreased respectively by 35-75, 60-75, and 25-90% for bacteria (Es- cherichia coli IEM-1, Bacillus subtilis BT-2, etc.), by 80-85, 55-90, and 15-60% for yeasts Candida albicans D-6, and by 40-50, 35-45, and 10-20% for micromycetes (Aspergillus niger P-3 and Fusarium culmorum T-7).
