Bioactive fractions from Streptococcus macedonicus MBF 10-2 produced in an optimized plant-based peptone medium fermentation

Background: Postbiotic fractions of several lactic acid bacteria have potential as microbial therapeutics for skin health and may also appeal to consumers who wish to avoid animal-based products. We aim to establish the optimum plant-peptone fermentation of Streptococcus macedonicus MBF10-2, which possess Bacteriocin Like-Inhibitory Substance activity in our previous study, to produce bacterial bioactive fractions. We evaluate their potential antibacterial and antioxidant actions, and as well assess the preliminary safety for human skin application. Methods: Fermentation was carried out by using plant peptone modified MRS, i.e., soy peptone and Vegitone, a non-animal-carbon sources that substitute proteose peptone in MRS medium. Fractions of MBF10-2 lysate and cell-free supernatant were collected and processed as follows, i.e. cell disruption, fraction separation and fractions freeze-drying. Fractions were confirm for antibacterial properties by the agar well diffusion method and assess for antioxidant activity using DPPH, while safety assessment was carried-out by skin patch assay. Result: Maximum growth of MBF10-2 achieved by fermentation in soy peptone- and in Vegitone-modified media was 9.00 and 7.99 g total cell mass, respectively. The antibacterial property of fractions was most effective against Micrococcus luteus T18. The lysate fraction exhibited a mild antioxidant potency (IC50 840 µg/mL), and all bioactive fractions were proven safe and non-allergenic for human skins. Conclusion: Strep. macedonicus MBF10-2 postbiotics bioactive fractions were indicated as being safe for topical application. This is the first report on the production of a safe Strep. macedonicus bioactive postbiotic possessing mild antibacterial and mild-to-weak antioxidant. Keywords: antibacterial; antioxidant; lysate; soy peptone; Streptococcus macedonicus MBF 10-2

Towards a Better Understanding of the Removal of Carbamazepine by Ankistrodesmus braunii: Investigation of Some Key Parameters

Nowadays, water pollution by pharmaceuticals is a major issue that needs an urgent solution, as these compounds, even when found at trace or ultra-trace levels, could have harmful effects on organisms. Carbamazepine (CBZ) is a pharmaceutical product that is detected as a micropollutant in many water resources. Different treatment methods were lately employed for the removal of CBZ, which are often cheap but inefficient or efficient but expensive. Yet, there are limited available studies on the elimination of this molecule by algae despite their well-known highly adaptive abilities. In this study, the biological treatment of CBZ was carried out using the green microalgae, Ankistrodesmus braunii (A. braunii), which has been reported to be particularly resistant to CBZ toxicity in the literature. The respective effects of the culture medium, the initial inoculum, and CBZ concentrations were studied on CBZ removal. Lastly, the mechanism of CBZ elimination by A. braunii was investigated. The presented data clearly demonstrates that the presence of this molecule did not completely repress A. braunii growth or the ability of these algae to remove CBZ; after 60 days of incubation, the highest percentage of CBZ elimination achieved was 87.6%. Elimination was more successful in Bold’s basal medium than in proteose peptone medium. Finally, the removal mechanism was also investigated to provide a better understanding of the transformation mechanism of this molecule. It was shown that the main removal mechanism was the bioaccumulation of CBZ by A. braunii cells, but the biotransformation of the initial CBZ into metabolites was also observed.

ETHANOL SUPPLEMENTATION AS A NEW APPROACH TO REGULATE GROWTH AND HYDROGEN PRODUCTION OF $ \textit{ESCHERICHIA COLI} $ UPON GLYCEROL FERMENTATION

Molecular hydrogen (H2) and ethanol are the main by-products of glycerol fermentation by Escherichia coli. In this study, the growth of E. coli BW25113 was investigated with the addition of small amounts (0.05 to 2 %) of ethanol alone and in a combination with glycerol The bacterial growth, the kinetic of the redox potential, and the H2 production in peptone medium, pH 7.5, were investigated upon various amounts of ethanol supplementation. In the presence of any amount of ethanol, but upon the absence of other sources of carbon, no H2 production was observed. Whereas ethanol (0.3 to 1 %) with a combination of glycerol stimulated both bacterial growth and H2 production, pH 7.5. A correlation was observed between the redox potential and stimulated by ethanol bacterial growth. The obtained results can be applied to regulate fermentation processes in biotechnology.

Phenolic substances as regulators of the intensity of lipid peroxidation processes of the strains Pleurotus eryngii, Fistulina hepatica and Agrocybe cylindracea

The work is devoted to the calculation, comparison of indicators and the development of a method for regulating the intensity of lipid peroxidation processes (LPP) of strains of basidiomycetes. The purpose of the investigation is to study the effect of phenolic type chemicals and hydrogen peroxide on the lipid peroxidation of certain strains of basidiomycetes under laboratory cultivation. Cultivation of strains of basidiomycetes was carried out by periodic surface method on glucose-peptone medium (GPM) in flasks. The influence of sodium lignosulfonate, tannin, gallic acid and hydrogen peroxide at 0.1% concentration at 24 and 48 hours of exposure on the intensity of lipid peroxidation processes of the strains Pleurotus eryngii P-er, Fistulina hepatica Fh-08 and Agrocybe cylindracea 960, fungi of the phylum Basidiomycota, orderAgaricales has been investigated. It was established that the used phenolic-type chemical compounds that are part of the lignocellulose complex of wood or are the products of its decomposition to a certain extent affect the lipid peroxidation processes of mycelial cell lipids in the studied cultures of basidium fungi. The individual reaction of LPP of cultures to the applied substance and the time of its exposure are determined. The highest degree of LPP induction was recorded upon addition of tannin – by 161%, after 48 hours of exposure in the mycelium of strain Fh-08; sodium lignosulfonate – by 192%, after 48 hours in the mycelium of strain P-er; gallic acid – by 182%, after 24 hours, in the mycelium of strain P-er; hydrogen peroxide – by 257%, after 24 hours, in the CR of strain 960. The biotechnological significance of this is the possibility of regulation (induction or repression) of LPP of producer strains. From a biomedical point of view, the possibility of changing the activity of lipid peroxidation processes of mycelium and culture fluid makes it possible to obtain and use more effective functional products of fungal origin.

Nutritional characteristics and physiological studies on the growth and development of Phlebiopsis gigantea

<p>Physiological studies pertaining to the effect of physical factors, carbon and nitrogen requirement of <em>Phlebiopsis gigantea</em> were conducted to know its behaviour <em>in vitro</em>. These studies have revealed interesting results regarding its growth and reproduction behaviour. The best medium for the optimum growth is Glucose-peptone medium. The optimum temperature (28°C) and pH (5.0) is required for the optimum mycelial production of <em>Phlebiopsis gigantea</em> after 12 days of incubation. The best carbon source for the growth of the fungus is D(+)Glucose least seen in Sucrose medium. The best inorganic nitrogen sources for the growth of the fungus is Sodium nitrate. The least mycelial growth of fungus is observed in ammonium acetate and ammonium nitrate. The best organic nitrogen compound for the mycelial growth of fungus is DL-threonine.</p>

Condition of the prooxidant-antioxidant system of some strains of Basidiomycetes

The article deals with the calculation and comparison indications of the condition of the prooxidant-antioxidant system (PAS) of strains of Basidiomycetes under periodic surface cultivation on a glucose-peptone medium. The research material consisted of the mycelium and culture filtrate (CF) from 57 strains, 52 of them belonging to 7 species of the order Agaricales and 5 belonging to 5 species of the order Polyporales. The intensity of the processes of lipid peroxidation was determined by a modified spectrophotometric method for contents of active products to thiobarbituric acid. Total antioxidant activity (АОА) of the mycological material was evaluated by intensity of inhibition from accumulated products of lipid peroxide oxidation (LPO) in a model reaction of oxidation by Twin-80 oxygen of the air. From the data obtained, indicators of prooxidant activity (POA), indicators of reserve of substrate peroxidation (SPO) and the balance coefficient of the prooxidant-antioxidant system (CbPАS) were calculated. It was established that strains of Basidiomycetes are characterized by significant predominance of prooxidant activity characteristic of PAS in the culture filtrate in comparison with the mycelium indicator. The highest values of POA in the Culture Filtrate were observed on the 12-th day of cultivation for the strain Р-089 genus Pleurotus and strain Gl-2 genus Ganoderma, and for the mycelium on the 9-th day of cultivation for the strains Р-сіtr, Р-089, Р-er and Р-082 of the genus Pleurotus. There is a direct dependence between the indicators of POA in the CF and mycelium for each strain, this dependence and level of indication do not reflect their systematic placement. We distinguished a more significant prevalence of indicators of reserve of substrates peroxidation of mycelium for most strains, than for such indicators with CF The highest value of reserve SPO of mycelium was recorded for strains Р-447, Р-998, Р-039, Р-94, Р-2175, Р-сіtr, Р-er, D-140 of the genus Pleurotus and strains F-107, F-610 of the genus Flammulina. For the majority of the strains there was a clear prevalence of the values of the balance coefficients for PAS of mycelium over those of CF. Maximum values of the balance coefficient PAS of mycelium on the 9-th day were recorded for strains Т-10 (F. fomentarius) and D-140, Р-035, Р-01, Р-105 (P. ostreatus) and on the 12-th day for strains Sc-10 (S. commune), Р-kl, Р-14 (P. ostreatus), Р-er (P. eryngii) і 960 (A. cylindracea). The abovementioned strains are distinguished by the with high level of AOA in the mycelium on certain days of cultivation and can be recommended as producers of antioxidants of fungal origin.

Respon Beberapa Genotipe Kacang Tanah terhadap Penyakit Layu Bakteri (Ralstonia solanacearum) di Rumah Kaca

<p>The objective of the study was to evaluate resistance response of groundnut genotypes against bacterial wilt (Ralstonia solanacearum) in the green house of Biochemistry Department, ICABIOGRAD, Bogor. Ralstonia solanacearum isolate was obtained from groundnut plant showing wilt symptom, collected from Karadenan village (Cibinong). Preparation of bacterial RS inoculum was done using SP (sucrose peptone) medium. Groundnut plants were inoculated by RS isolate using inoculum capacity of 107 cfu/ml. The experiment was arranged in randomized complete block design (RCBD) consisted of seven treatments with four replications, whilst Tupai and Kelinci genotypes were used as resistant and susceptible control check, respectively. The result indicated that groundnut genotypes i.e. ICGV 88262, Local Sindangbarang, PI 203395, ICG 10067, and ICG 3400, showed resistant response against bacterial wilt causing wilt damage of &lt;20%. Bacterial wilt disease could affect reductions of some groundnut phenotypic character. The plant height of resistant groundnut showed relatively tall than that of Kelinci. In addition, it showed the lowest healthy pod yield among the genotypes tested.</p><p> </p><p><strong>Abstrak</strong></p><p>Penelitian bertujuan untuk mempelajari tingkat ketahanan genotipe kacang tanah terhadap penyakit layu bakteri (Ralstonia solanacearum) di rumah kaca Kelompok Peneliti Biokimia BB-Biogen. Isolat R. solanacearum diperoleh dari tanaman kacang tanah yang menunjukkan gejala layu dari Desa Karadenan (Cibinong). Penyiapan dan penyediaan inokulum bakteri R. solanacearum menggunakan media SP (sukrose pepton). Tanaman kacang tanah diinokulasi dengan isolat R. solanacearum dengan kerapatan koloni 107 cfu/ml. Penelitian disusun menggunakan rancangan acak kelompok dengan tujuh perlakuan dan empat ulangan. Untuk pembanding tahan dan rentan masing-masing digunakan genotipe Tupai dan Kelinci. Hasil penelitian menunjukkan bahwa genotipe ICGV 88262, lokal Sindangbarang, PI 203395, ICG 10067, dan ICG 3400 tahan terhadap penyakit layu bakteri dengan nilai kerusakan &lt;20%. Penyakit layu bakteri berpengaruh terhadap penurunan beberapa karakter fenotipik kacang tanah, di mana tanaman genotipe kacang tanah tahan PLB relatif lebih tinggi dibandingkan dengan genotipe Kelinci. Polong bernas Kelinci paling sedikit di antara semua genotipe uji.</p>

ВПЛИВ МІКРОЕЛЕМЕНТІВ НА НАКОПИЧЕННЯ КАРОТИНОЇДІВ ДЕЯКИМИ ШТАМАМИ БАЗИДІОМІЦЕТІВ

<p>The effect of microelements on growth and accumulation of carotenoids highly productive strains of basidiomycetes at surface cultivation on glucose-peptone medium was investigated. The objects of research are 3 wood destroying strain. There are <em>Laetiporus sulphureus</em> (Bull.) Murrill Ls-08, <em>Fomes fomentarius</em> (L.) Fr. Ff-1201 from the order <em>Polyporales</em> and <em>Fistulina hepatica</em> (Schaeff.) Sibth Fh-18 from the order <em>Agaricales</em>. Research materials are strains mycelium and culture filtrate (CF). Absolutely dry biomass (ADB) mycelium was determined by the gravimetric method, the content of carotenoids was determined by spectrophotometric method in acetone extracts of the Vetshteyn formula. Established individual influence of microelements on the accumulation of biomass and carotenoids of basidiomycetes strains. The possibility of the regulation of these processes by introducing into the glucose-peptone medium of various Fe, Cu, Zn, Ni and Mn sulphate. So, the best to increase the intensity of the growth processes and the accumulation of carotenoids strain of<em> L. sulphureus</em> Ls-08 is an experimental environment which includes Zn sulfate in a concentration of 8 mmol/L. To induce the accumulation of ADB and carotenoids in the mycelium and CF of strain <em>F fomentarius</em> Ff-1201 making in is expedient Mn sulfate in a concentration of 1.6 mmol/L. To improve carotenogenesis of <em>F. hepatica</em> Fh-18 strain expedient entry in GPM Mn sulphate at concentration of 8 mmol/L. These allow to optimize the concentration of microelements in nutrient medium for the cultivation of carotenoids high-producing strains of Basidiomycetes.</p><p align="left"> </p>

ЗАГАЛЬНА АНТИОКСИДАНТНА АКТИВНІСТЬ ДЕЯКИХ ШТАМІВ БАЗИДІАЛЬНИХ ГРИБІВ В ДИНАМІЦІ РОСТУ

<p>The work is devoted to the study of total antioxidant activity (AOA) in the growth dynamics of basidiomycetes strains in their periodic surface cultivation on glucose-peptone medium. Subjects of research are mycelium and culture filtrate (CF) from 57 strains, 5 of which are belong to 5 types of Polyporales order, and 52 of which are belong to the 7 types of Agaricales order. In order to study the dynamics of growth used method for determining the weight of absolutely dry biomass accumulation (ADB). Total AOA of mycological material was evaluated by inhibition of lipid peroxidation products accumulation intensity in the model oxidation reaction of Tween-80 by air oxygen. It was found that the most productive in terms of the accumulation of ADB are strains <em>F. velutipes</em> F-610 and <em>P. eryngii</em> P-er. Lowest values of ADB accumulation recorded for strains <em>P. ostreatus</em> P-14 and P-192 and <em>P. citrinopileatus </em>P sіtr. Were selected the most productive strains of Basidiomycetes for the level of total AOA in mycelium and CF. There are strains <em>P. eryngii</em> P-er, <em>P. citrinopileatus</em> P sіtr, <em>P. ostreatus</em> P-035, <em>F. hepatica</em> Fh-08, <em>A. cylindracea</em> 960, <em>P. ostreatus </em>P-081, P-082, P-087, <em>P. citrinopileatus</em> P sіtr. Has not been established the dependence between the growth and the antioxidant activity of the 9- and 12-day fungal cultures. Selected producers of natural antioxidants may be used as biological agents in biotechnology.</p><p> </p>

The lipid peroxidation intensity of fungi strains from the orders Agaricales and Polyporales

This article is devoted to investigation of the dynamics of growth and level of spontaneous and induced lipid peroxidation intensity of Basidiomycetes strains grown by surface cultivation on a glucose-peptone medium. The materials of the research are mycelium and culture filtrates (CF) of 57 strains (5 belong to 5 species from the order Polyporales s.l., and 52 belong to 7 species of the order Agaricales s.l.). To study the dynamics of growth we used a weighing method for determining the accumulation of absolutely dry biomass. Intensity of lipid peroxidation was determined by a modified spectrophotometric method for content of active to thiobarbituric acid products. It was found that the most productive in absolutely dry biomass accumulation were the strains Flammulina velutipes (Curt.: Fr.) Sing. F-610 and Pleurotus eryngii (DC.: Fr.) Quél. P-er. The level of spontaneous and induced LPO intensity in mycelia of all strains was higher than this figure in the culture filtrate and increased with the duration of cultivation. Dependencies between the content of lipid peroxidation products in the mycelia and CF were not established. The lowest values were recorded for biomass accumulation by the strains Pleurotus ostreatus (Jacq.: Fr.) P. Kumm. P-14, P-192 and P. citrinopileatus Singer. Р-сіtr. Groups of basidiomycete cultures with different levels of TBA-AP were identified. Spontaneous and induced intensivity of lipid peroxidation in all studied strains of mycelia was higher than the figure in the culture filtrate. The intensity of lipid peroxidation in both mycelia and culture filtrate constantly increased, which can be explained by the growing shortage of certain nutrients (primarily carbon) and increased concentration of metabolic products in the medium. The ratio of spontaneous and induced lipid peroxidation intensity is specific to each strain and is independent of its systematic position. Shifting of prooxidant-antioxidant balance to a relatively stationary level is a mark of stress reaction. LPO-products can be both inductors and primary mediators of stress as a special class of biological systems. Selected strains with high rates of growth and LPO ntensity are promising for applications in biotechnology and ecology.