Višestruke mogućnosti primjene novih biokatalizatora pri enzimskoj obradi otpadnih voda prehrambene industrije

The cells of filamentous fungus R. oryzae entrapped in the polyvinyl alcohol cryogelare capable of producing various extracellular hydrolytic enzymes (proteases, amylases, lipases) and are used for the treatment of complex wastewaters of food industry. Five types of media simulating the wastewater of various food enterprises were treated under batch conditions for 600 h. Fats containing mostly residues of unsaturated fatty acids, as well as casein, glucose, sucrose, starch, soybean flour and various salts were the main components of the treated wastewaters. The immobilized cells concurrently possessed lipolytic, amylolytic and proteolytic activities. The level of each enzymatic activity depended on the wastewater content. The physiological state of immobilized cells was monitored by bioluminescent method. The intracellular adenosine triphosphate (ATP) concentration determined in the granules with immobilized cells was high enough and almost constant for all the period of biocatalyst application confirming thereby the active metabolic state of the cells. The study of mechanical strength of biocatalyst granules allowed revealing the differences in the values of modulus of biocatalyst elasticity at the beginning and at the end of its use for the wastewater treatment. The decrease in chemical oxygen demand of the tested media after their processing by immobilized biocatalyst was 68–79 % for one working cycle.Stanice filamentozne gljive Rhizopus oryzae ugrađene u kriogel polivinilnog alkohola mogu proizvesti razne ekstracelularne hidrolitičke enzime (proteaze, amilaze, lipaze), pa se koriste u obradi otpadnih voda prehrambene industrije. Pet vrsta podloga, koje simuliraju otpadne vode raznih prehrambenih poduzeća, obrađeno je u uvjetima šaržnog uzgoja tijekom 600 sati. Glavni su sastojci obrađenih otpadnih voda masti (najviše ostaci nezasićenih masnih kiselina), kazein, glukoza, saharoza, škrob, sojino brašno i razne soli. Utvrđeno je da imobilizirane stanice istodobno imaju lipolitička, amilolitička i proteolitička svojstva. Fiziološko stanje imobiliziranih stanica praćeno je pomoću bioluminiscentne metode. Intracelularna koncentracija adenozin trifosfata (ATP-a) u granulama s imobiliziranim stanicama je velika i skoro konstantna cijelo vrijeme primjene biokatalizatora, što dokazuje aktivni metabolički status stanica. Ispitivanje mehaničke čvrstoće granula biokatalizatora otkrilo je razliku u modulu elastičnosti prije i nakon obrade otpadnih voda. Smanjenje kemijske potrošnje kisika ispitanih podloga u jednom radnom ciklusu, nakon njihove obrade imobiliziranim biokatalizatorom, iznosilo je 68-79 %

Višestruke mogućnosti primjene novih biokatalizatora pri enzimskoj obradi otpadnih voda prehrambene industrije

The cells of filamentous fungus R. oryzae entrapped in the polyvinyl alcohol cryogelare capable of producing various extracellular hydrolytic enzymes (proteases, amylases, lipases) and are used for the treatment of complex wastewaters of food industry. Five types of media simulating the wastewater of various food enterprises were treated under batch conditions for 600 h. Fats containing mostly residues of unsaturated fatty acids, as well as casein, glucose, sucrose, starch, soybean flour and various salts were the main components of the treated wastewaters. The immobilized cells concurrently possessed lipolytic, amylolytic and proteolytic activities. The level of each enzymatic activity depended on the wastewater content. The physiological state of immobilized cells was monitored by bioluminescent method. The intracellular adenosine triphosphate (ATP) concentration determined in the granules with immobilized cells was high enough and almost constant for all the period of biocatalyst application confirming thereby the active metabolic state of the cells. The study of mechanical strength of biocatalyst granules allowed revealing the differences in the values of modulus of biocatalyst elasticity at the beginning and at the end of its use for the wastewater treatment. The decrease in chemical oxygen demand of the tested media after their processing by immobilized biocatalyst was 68–79 % for one working cycle.Stanice filamentozne gljive Rhizopus oryzae ugrađene u kriogel polivinilnog alkohola mogu proizvesti razne ekstracelularne hidrolitičke enzime (proteaze, amilaze, lipaze), pa se koriste u obradi otpadnih voda prehrambene industrije. Pet vrsta podloga, koje simuliraju otpadne vode raznih prehrambenih poduzeća, obrađeno je u uvjetima šaržnog uzgoja tijekom 600 sati. Glavni su sastojci obrađenih otpadnih voda masti (najviše ostaci nezasićenih masnih kiselina), kazein, glukoza, saharoza, škrob, sojino brašno i razne soli. Utvrđeno je da imobilizirane stanice istodobno imaju lipolitička, amilolitička i proteolitička svojstva. Fiziološko stanje imobiliziranih stanica praćeno je pomoću bioluminiscentne metode. Intracelularna koncentracija adenozin trifosfata (ATP-a) u granulama s imobiliziranim stanicama je velika i skoro konstantna cijelo vrijeme primjene biokatalizatora, što dokazuje aktivni metabolički status stanica. Ispitivanje mehaničke čvrstoće granula biokatalizatora otkrilo je razliku u modulu elastičnosti prije i nakon obrade otpadnih voda. Smanjenje kemijske potrošnje kisika ispitanih podloga u jednom radnom ciklusu, nakon njihove obrade imobiliziranim biokatalizatorom, iznosilo je 68-79 %

Optimization of the Use of His6-OPH-Based Enzymatic Biocatalysts for the Destruction of Chlorpyrifos in Soil

Applying enzymatic biocatalysts based on hexahistidine-containing organophosphorus hydrolase (His6-OPH) is suggested for the decomposition of chlorpyrifos, which is actively used in agriculture in many countries. The application conditions were optimized and the following techniques was suggested to ensure the highest efficiency of the enzyme: first, the soil is alkalinized with hydrated calcitic lime Ca(OH)2, then the enzyme is introduced into the soil at a concentration of 1000 U/kg soil. Non-equilibrium low temperature plasma (NELTP)-modified zeolite is used for immobilization of the relatively inexpensive polyelectrolyte complexes containing the enzyme His6-OPH and a polyanionic polymer: poly-l-glutamic acid (PLE50) or poly-l-aspartic acid (PLD50). The soil’s humidity is then increased up to 60–80%, the top layer (10–30 cm) of soil is thoroughly stirred, and then exposed for 48–72 h. The suggested approach ensures 100% destruction of the pesticide within 72 h in soils containing as much as 100 mg/kg of chlorpyrifos. It was concluded that using this type of His6-OPH-based enzyme chemical can be the best approach for soils with relatively low humus concentrations, such as sandy and loam-sandy chestnut soils, as well as types of soil with increased alkalinity (pH 8.0–8.4). Such soils are often encountered in desert, desert-steppe, foothills, and subtropical regions where chlorpyrifos is actively used

Transformation of Enzymatic Hydrolysates of <i>Chlorella</i>–Fungus Mixed Biomass into Poly(hydroxyalkanoates)

The production of poly(hydroxylalkanoates) (PHA) is limited by the high cost of the feedstock since various biomass wastes look attractive as possible sources for polymer production. The originality of this present study is in the biotransformation of mixed Chlorella-based substrates into PHAs. The synthetic potential of Cupriavidus necator B8619 cells was studied during the bioconversion of algae biomass in mixtures with spent immobilized mycelium of different fungi (genus Rhizopus and Aspergillus) into PHAs. The biomass of both microalgae Chlorella and fungus cells was accumulated due to the use of the microorganisms in the processes of food wastewater treatment. The biosorption of Chlorella cells by fungal mycelium was carried out to obtain mixed biomass samples (the best ratio of “microalgae:fungi” was 2:1) to convert them by C. necator B8619 into the PHA. The influence of conditions used for the pretreatment of microalgae and mixed types of biomass on their conversion to PHA was estimated. It was found that the maximum yield of reducing sugars (39.4 ± 1.8 g/L) can be obtained from the mechanical destruction of cells by using further enzymatic hydrolysis. The effective use of the enzymatic complex was revealed for the hydrolytic disintegration of treated biomass. The rate of the conversion of mixed substrates into the biopolymer (440 ± 13 mg/L/h) appeared significantly higher compared to similar known examples of complex substrates used for C. necator cells

“Nature-like” Cryoimmobilization of Phototrophic Microorganisms: New Opportunities for Their Long-Term Storage and Sustainable Use

It was found that immobilization of cells in poly(vinyl alcohol) (PVA) cryogel can be successfully applied for concurrent cryoimmobilization, cryoconservation and long-term storage of the cells of various phototrophic microorganisms (green and red microalgae, diatoms and cyanobacteria). For the first time, it was shown for 12 different immobilized microalgal cells that they can be stored frozen for at least 18 months while retaining a high level of viability (90%), and can further be used as an inoculum upon defrosting for cell-free biomass accumulation. Application of cryoimmobilized Chlorella vulgaris cells as inocula allowed the loading of a high concentration of the microalgal cells into the media for free biomass accumulation, thus increasing the rate of the process. It was shown that as minimum of 5 cycles of reuse of the same immobilized cells as inocula for cell accumulation could be realized when various real wastewater samples were applied as media for simultaneous microalgae cultivation and water purification

Impact of Perfluorocarbons with Gas Transport Function on Growth of Phototrophic Microorganisms in a Free and Immobilized State and in Consortia with Bacteria

The effects of the presence of perfluorocarbons (PFC) with a gas transport function in media with different phototrophic microorganisms on their growth rates and the accumulation of their biomass when using free and immobilized cells as inoculums were investigated. The significant increase in the average rate of biomass accumulation as well as levels of biomass accumulation in the presence of various PFCs were established for Chlorella vulgaris cells. When 1 g/L glycerol was introduced into the growth medium with PFCs and C. vulgaris cells, the increase in the rate of biomass accumulation was 9&ndash;32%. The maximum intracellular ATP concentrations corresponded to the combination of microalgae (Chlorella vulgaris) with bacterial cells (Pseudomonas esterophilus and Rhodoccus ruber) obtained with a mass ratio of 25:1. It provided for the formation of a consortium, which was able to accumulate the maximum amount of microalgae biomass for 3 days in the medium with PFCs and organophosphorus pesticide. The obtained data allow, on the one hand, predicting the growth of microalgae under environmental conditions in media with PFC pollution and, on the other hand, developing approaches to regulation of phototrophic microorganisms&rsquo; growth in order to obtain and use their high biomass yields for various purposes

Impact of Perfluorocarbons with Gas Transport Function on Growth of Phototrophic Microorganisms in a Free and Immobilized State and in Consortia with Bacteria

The effects of the presence of perfluorocarbons (PFC) with a gas transport function in media with different phototrophic microorganisms on their growth rates and the accumulation of their biomass when using free and immobilized cells as inoculums were investigated. The significant increase in the average rate of biomass accumulation as well as levels of biomass accumulation in the presence of various PFCs were established for Chlorella vulgaris cells. When 1 g/L glycerol was introduced into the growth medium with PFCs and C. vulgaris cells, the increase in the rate of biomass accumulation was 9–32%. The maximum intracellular ATP concentrations corresponded to the combination of microalgae (Chlorella vulgaris) with bacterial cells (Pseudomonas esterophilus and Rhodoccus ruber) obtained with a mass ratio of 25:1. It provided for the formation of a consortium, which was able to accumulate the maximum amount of microalgae biomass for 3 days in the medium with PFCs and organophosphorus pesticide. The obtained data allow, on the one hand, predicting the growth of microalgae under environmental conditions in media with PFC pollution and, on the other hand, developing approaches to regulation of phototrophic microorganisms’ growth in order to obtain and use their high biomass yields for various purposes

Various Biomimetics, Including Peptides as Antifungals

Biomimetics, which are similar to natural compounds that play an important role in the metabolism, manifestation of functional activity and reproduction of various fungi, have a pronounced attraction in the current search for new effective antifungals. Actual trends in the development of this area of research indicate that unnatural amino acids can be used as such biomimetics, including those containing halogen atoms; compounds similar to nitrogenous bases embedded in the nucleic acids synthesized by fungi; peptides imitating fungal analogs; molecules similar to natural substrates of numerous fungal enzymes and quorum-sensing signaling molecules of fungi and yeast, etc. Most parts of this review are devoted to the analysis of semi-synthetic and synthetic antifungal peptides and their targets of action. This review is aimed at combining and systematizing the current scientific information accumulating in this area of research, developing various antifungals with an assessment of the effectiveness of the created biomimetics and the possibility of combining them with other antimicrobial substances to reduce cell resistance and improve antifungal effects

Nanocatalysts for Oxidative Desulfurization of Liquid Fuel: Modern Solutions and the Perspectives of Application in Hybrid Chemical-Biocatalytic Processes

In this paper, the current advantages and disadvantages of using metal-containing nanocatalysts (NCs) for deep chemical oxidative desulfurization (ODS) of liquid fuels are reviewed. A similar analysis is performed for the oxidative biodesulfurization of oil along the 4S-pathway, catalyzed by various aerobic bacterial cells of microorganisms. The preferences of using NCs for the oxidation of organic sulfur-containing compounds in various oil fractions seem obvious. The text discusses the development of new chemical and biocatalytic approaches to ODS, including the use of both heterogeneous NCs and anaerobic microbial biocatalysts that catalyze the reduction of chemically oxidized sulfur-containing compounds in the framework of methanogenesis. The addition of anaerobic biocatalytic stages to the ODS of liquid fuel based on NCs leads to the emergence of hybrid technologies that improve both the environmental characteristics and the economic efficiency of the overall process. The bioconversion of sulfur-containing extracts from fuels with accompanying hydrocarbon residues into biogas containing valuable components for the implementation of C-1 green chemistry processes, such as CH4, CO2, or H2, looks attractive for the implementation of such a hybrid process

Biocatalysts in Synthesis of Microbial Polysaccharides: Properties and Development Trends

Polysaccharides synthesized by microorganisms (bacterial cellulose, dextran, pullulan, xanthan, etc.) have a set of valuable properties, such as being antioxidants, detoxifying, structuring, being biodegradable, etc., which makes them suitable for a variety of applications. Biocatalysts are the key substances used in producing such polysaccharides; therefore, modern research is focused on the composition and properties of biocatalysts. Biocatalysts determine the possible range of renewable raw materials which can be used as substrates for such synthesis, as well as the biochemistry of the process and the rate of molecular transformations. New biocatalysts are being developed for participating in a widening range of stages of raw material processing. The functioning of biocatalysts can be optimized using the following main approaches of synthetic biology: the use of recombinant biocatalysts, the creation of artificial consortia, the combination of nano- and microbiocatalysts, and their immobilization. New biocatalysts can help expand the variety of the polysaccharides&rsquo; useful properties. This review presents recent results and achievements in this field of biocatalysis