Albert Krastanov

Microbial degradation of phenol and phenolic derivatives

Phenol and its derivatives are one of the largest groups of environmental pollutants due to their presence in many industrial effluents and broad application as antibacterial and antifungal agents. A number of microbial species possess enzyme systems that are applicable for the decomposition of various aliphatic and aromatic toxic compounds. Intensive efforts to screen species with high‐degradation activity are needed to study their capabilities of degrading phenol and phenolic derivatives. Most of the current research has been directed at the isolation and study of microbial species of potential ecological significance. In this review, some of the best achievements in degrading phenolic compounds by bacteria and yeasts are presented, which draws attention to the high efficiency of strains of Pseudomonas, Candida tropicalis, Trichosporon cutaneum, etc. The unique ability of fungi to maintain their degradation potential under conditions unfavorable for other microorganisms is outstanding. Mathematical models of the microbial biodegradation dynamics of single and mixed aromatic compounds, which direct to the benefit of the processes studied in optimization of modern environmental biotechnology are also presented.

Composition and Antioxidant Activities of the Essential Oil of Cinnamon (Cinnamomum zeylanicum Blume) Leaves from Sri Lanka

Abstract The composition of the volatiles from leaves of Cinnamomum zeylanicum Blume from Sri Lanka was studied by GC-FID and GC-MS. The basic component of the oil was found to be eugenol (74.9%), followed by β-caryophyllene (4.1%), benzyl benzoate (3.0%), linalool (2.5%), eugenyl acetate (2.1%) and cinnamyl acetate (1.8%). The essential leaf oil from cinnamon demonstrated scavenger activity against the DPPH radical at concentrations which are lower than the concentrations of eugenol, butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA). This essential cinnamon oil showed also a significant inhibitory effect on hydroxyl radicals and acted as an iron chelator. Cinnamon leaf oil efficiently inhibited the formation of conjugated dienes and the generation of secondary products from lipid peroxidation at a concentration equivalent to that of the standard BHT.

Chemical Composition and Antioxidant Properties of Juniper Berry (Juniperus communis L.) Essential Oil. Action of the Essential Oil on the Antioxidant Protection of Saccharomyces cerevisiae Model Organism

The essential oil of juniper berries (Juniperus communis L., Cupressaceae) is traditionally used for medicinal and flavoring purposes. As elucidated by gas chromatography/flame ionization detector (GC/FID) and gas chromatography/mass spectrometry (GC/MS methods), the juniper berry oil from Bulgaria is largely comprised of monoterpene hydrocarbons such as α-pinene (51.4%), myrcene (8.3%), sabinene (5.8%), limonene (5.1%) and β-pinene (5.0%). The antioxidant capacity of the essential oil was evaluated in vitro by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging, 2,2-azino-bis-3-ethylbenzothiazoline-6 sulfonic acid (ABTS) radical cation scavenging, hydroxyl radical (ОН•) scavenging and chelating capacity, superoxide radical (•O2−) scavenging and xanthine oxidase inhibitory effects, hydrogen peroxide scavenging. The antioxidant activity of the oil attributable to electron transfer made juniper berry essential oil a strong antioxidant, whereas the antioxidant activity attributable to hydrogen atom transfer was lower. Lipid peroxidation inhibition by the essential oil in both stages, i.e., hydroperoxide formation and malondialdehyde formation, was less efficient than the inhibition by butylated hydroxytoluene (BHT). In vivo studies confirmed these effects of the oil which created the possibility of blocking the oxidation processes in yeast cells by increasing activity of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx).

Phenol and cresol mixture degradation by the yeast Trichosporon cutaneum

Most industrial wastes contain different organic mixtures, making important the investigation on the microbial destruction of composite substrates. The capability of microbes to remove harmful chemicals from polluted environments strongly depends on the presence of other carbon and energy substrates. The effect of mixtures of phenol- and methyl-substituted phenols (o-, m-, p-cresol) on the growth behaviour and degradation capacity of Trichosporon cutaneum strain was investigated. The cell-free supernatants were analysed by HPLC. It was established that the presence of o-, m- and p- cresol has not prevented complete phenol assimilation but had significant delaying effect on the phenol degradation dynamics. The mutual influence of phenol and p-cresol was investigated. We developed the kinetic model on the basis of Haldane kinetics, which used model parameters from single-substrate experiments to predict the outcome of the two-substrate mixture experiment. The interaction coefficients indicating the degree to which phenol affects the biodegradation of p-cresol and vice versa were estimated. Quantitative estimation of interaction parameters is essential to facilitate the application of single or mixed cultures to the bio-treatment of hazardous compounds.

Chitinase biotechnology: Production, purification, and application

Chitin is one of the most abundant biopolymers and is present in many organisms in different forms. Its resistance to degradation has caused many problems in industry (waste decomposition) and agriculture (as protective structures in pests); this has led to increased interest in chitin‐hydrolyzing enzymes: chitinases. Chitinases are enzymes that break down the 1→4 β‐glycoside bond of N‐acetyl d‐glucosamine in chitin to produce mono‐ and oligomers. The inducible nature of chitinases, low activity of synthesized enzymes, and inertia of the substrate are only a few of the problems that can be solved by biotechnology to meet industry demands for green, energy‐efficient, pollution‐free, and economically profitable chitin use. This review aims to present the pitfalls and successes in research and production of chitinolytic enzymes, as well as to promote the use of chitinases in everyday practice. The focus is on the biosynthesis of chitinases: inducers, type of fermentation, and media composition. Methods for purification and future perspectives are also discussed.

CARBON DIOXIDE FIXATION BY CHLORELLA MINUTISSIMA BATCH CULTURES IN A STIRRED TANK BIOREACTOR

ABSTRACT Increasing concentrations of CO2 in the atmosphere is causing severe environmental destruction to the earth. To capture the excess CO2, its microalgal sequestration is increasingly being explored. This technique could become a profitable industry that would help mitigate global warming and produce a large number of value-added products like pigments, carotenoids, carbohydrates, lipids, etc. In this regard, three microalgal strains viz., Calothrix sp., Spirulina platensis and Chlorella minutissima were studied for their growth characteristics. Based upon the kinetic parameters, possible by-products, tolerance to CO2, etc., the best strain was selected for further studies. The optimum pH, photoperiod, nitrate concentration and light intensity for this strain were experimentally determined. Also, studies were conducted with and without baffles, with varying aeration rates and with two different impellers and i.e., marine propeller and disc turbine impeller, in the presence of air alone and a mixture containing air with 15% CO2. Among the three strains studied, C. minutissima was found to be the best strain for further work. It has maximum biomass productivity at pH 6, photoperiod of 14 h light:10 h dark cycles, 5 g/l nitrate and 6000 lx light intensity. Runs with baffles yielded higher biomass. Marine propeller gave better biomass yields in both runs i.e., with and without additional CO2. Also, the optimum aeration rate was determined to be 1 lpm. The carbohydrate, lipid, protein, chlorophyll and carotenoid content of the biomass were estimated.

Tilia tomentosa pectins exhibit dual mode of action on phagocytes as β-glucuronic acid monomers are abundant in their rhamnogalacturonans I

Silver linden flowers contain different pectins (PSI-PSIII) with immunomodulating properties. PSI is a low-esterified pectic polysaccharide with predominant homogalacturonan region, followed by rhamnogalacturonan I (RGI) with arabinogalactan II and RGII (traces) domains. PSII and PSIII are unusual glucuronidated RGI polymers. PSIII is a unique high molecular weight RGI, having almost completely O-3 glucuronidated GalA units with >30% O-3 acetylation at the Rha units. Linden pectins induced reactive oxygen species (ROS) and NO generation from non-stimulated whole blood phagocytes and macrophages, resp., but suppressed OZP-(opsonized zymosan particles)-activated ROS generation, LPS-induced iNOS expression and NO production. This dual mode of action suggests their anti-inflammatory activity, which is known for silver linden extracts. PSI expressed the highest complement fixation and macrophage-stimulating activities and was active on intestinal Peyers patch cells. PSIII was active on non-stimulated neutrophils, as it induced ß2-integrin expression, revealing that acetylated and highly glucuronidated RGI exhibits immunomodulating properties via phagocytes.

Examination of the technological properties of newly isolated strains of the genus Lactobacillus and possibilities for their application in the composition of starters

The ability of four Lactobacillus strains – Lactobacillus brevis LBRZ7 (isolated from fermented cabbage), Lactobacillus plantarum LBRZ12 (isolated from fermented cabbage), Lactobacillus fermentum LBRH9 (of human origin) and Lactobacillus casei ssp. rhamnosus LBRC11 (isolated from home-made cheese) – to grow in flour/water environment and to accumulate high concentrations of viable cells was examined. Two starters for sourdough were created for lab-scale production of wheat bread: a two-strain starter and a four-strain starter. Wheat bread with improved properties – greater loaf volume, enhanced flavour and softer and brighter crumb – was obtained from the 7% four-strain starter sourdough. The addition of sourdough in the production of wheat bread affected positively the technological and organoleptic characteristics of the final bread by inhibiting the growth of wild yeasts and mold and Bacillus spores without the addition of preservatives. The inclusion of 15% of the four-strain starter sourdough in the bread-making process led to enhanced safety and longer shelf life of the baked bread.

Probiotic Strain Lactobacillus Plantarum NBIMCC 2415 with Antioxidant Activity as a Starter Culture in the Production of Dried Fermented Meat Products

ABSTRACT Lactobacillus plantarum NBIMCC 2415 resistant to low and high values of pH and pepsin, and to high concentrations of bile salts was selected. It was demonstrated that Lactobacillus plantarum NBIMCC 2415 inhibited the growth of pathogenic microorganisms such as Escherichia coli ATCC 25922, Escherichia coli ATCC 8739, Proteus vulgaris G, Salmonella sp., Salmonella abony NTCC 6017, Staphylococcus aureus ATCC 25093, Staphylococcus aureus ATCC 6538 P, Listeria monocytogenes and Listeria monocytogenes I at temperature 15–18°C in the meat products. The high antioxidant activity of this strain against peroxide radicals (ORAC results) was determined. Experimental series of raw-dried meat products were produced. Reduction of the microflora of the meat raw-dried sausages and high concentration of viable cells of L. plantarum NBIMCC 2415 were observed, which makes them functional foods.

The common lavender (Lavandula angustifolia Mill.) pectic polysaccharides modulate phagocytic leukocytes and intestinal Peyer’s patch cells

Two pectic (chPS-L1, chPS-L2) and one polyphenolic (chPP-L) fractions were obtained from lavender flowers after boiling water extraction, exhaustive removing of alcohol-soluble molecules and SEC. chPS-L1 (52.4kDa) contains mainly low-acetylated and high-methoxylated homogalacturonans (HG), and smaller rhamnogalacturonan (RG) I backbone fragments rich in 1,3,5-branched arabinan and arabinogalactan (AG) II side chains. chPS-L2 (21.8kDa) contains predominantly similarly esterified HG, followed by RGI with AGII structures and RGII. The prevalence of catechin and epicatechin in chPP-L indicates that they form weak interactions with pectins. chPS-L1 and chPS-L2 enhanced ß2-integrin expression on neutrophils, inducing ROS generation and macrophage NO production. Both the effects on ß2-integrin and high complement fixation activity of chPS-L1 were proposed for its inhibitory action against PMA- and OZP-activated ROS formation. This, together with suppression of NO generation after co-stimulation with chPS-L1 and LPS, suggested anti-inflammatory activity of studied pectins. Lavender polysaccharides expressed intestinal Peyers patch immunomodulating activity.