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Triticale (x Triticosecale, Wittmack), an important industrial crop, with high grain yield, containing high amounts of starch, proteins and also major and minor mineral elements, is not yet sufficiently utilized. The simultaneous production of alpha-amylase and glucoamylase isoforms by Aspergillus niger on triticale grains, without any nutritive supplements, was developed, optimized and scaled up 10 fold for the first time. The specific combination of the examined effects led to the production of a novel glucoamylase isoform. Reduction of particle size, increase in oxygen availability and substrate height lead to an increase of 30 % in the production of amylases. Reduction of the relative humidity from 65 to 30 % increased glucoamylase production 2 fold and alpha-amylase production by 30%. The peak production of alpha-amylase (158 U g(-1)) and glucoamylase (170 U g(-1)) were obtained in Erlenmeyer flasks and in scaled-up trays. The obtained A. niger amylase cocktail was more efficient in raw starch hydrolysis from wheat flour, 29 % more efficient in glucose formation and 10 % more efficient in total reducing sugar formation, than the commercially available amylase cocktail SAN Super 240L, which is widely used in industry.

With the purpose of finding a suitable available inducer in combination with starvation, carbohydrate mixtures from triticale were used and compared with well-known amylase inducers in fungi. Carbohydrate mixtures from triticale induced the production of an amylase cocktail (alpha-amylase and glucoamylase) in Aspergillus niger, unlike induction with well-known inducers that induce only glucoamylase, shown by zymography and TLC analysis of the carbohydrate mixtures before and after fermentation. Glucoamylase production by A. niger was the highest in the presence of the extract obtained after auto-hydrolysis of starch from triticale (95.88 U mL(-1)). Carbohydrate mixtures from triticale induced the production of alpha-amylase in A. oryzae. More alpha-amylase isoforms were detected when using a complex carbohydrate mixture, compared to induction with maltose or starch. A 48-h induction was the most efficient using a triticale extract (101.35 U mL(-1)). Carbohydrates from triticale extracts could be used as very good cheap amylase inducers. Triticale, still not fully utilized, could be taken into consideration as an inducer in amylase production by Aspergillus sp, and in such a way, it could be used as the sole substrate in fermentation.

Phenols containing halogens, which tend to deactivate the aromatic nuclei, constitute a significant category of highly toxic and difficult-to-degrade pollutants, which arise from a wide variety of industries. The main purpose of this study was to obtain an inexpensive immobilized enzyme for the removal of phenols. Partially purified potato polyphenol oxidase (PPO) was immobilized onto different commercial and laboratory produced carriers. Three of the obtained biocatalysts, with the highest PPO activities, namely Eupergit C250L-PPO; Celite-PPO and CelluloseM-PPO, were tested in a batch reactor for the removal of phenol, 4-chlorophenol and 4-bromophenol. In the case of 2.5 mM substrates with Eupergit C250L-PPO, an around 45 % removal of 4-bromophenol was achieved, while the removals 4-chlorophenol and phenol were 35 and 20 %, respectively. The reusability of Eupergit C250L-PPO for the removal of 4-chlorophenol was tested. After eight repeated tests, the efficiency of 4-chlorophenol removal by Eupergit C250L-PPO immobilisate had decreased to 55 %., Halogenovani fenoli imaju dezaktivirano aromatično jezgro i čine značajnu kategoriju veoma toksičnih i teško razgradivih zagađivača u raznim industrijskim granama. Glavni cilj ovog rada je bio dobijanje jeftinog imobilizovanog enzima za uklanjanje fenola. Delimično prečišćena polifenol-oksidaza (PFO) iz krompira je imobilizovana na različitim komercijalnim i laboratorijski sintetizovanim nosačima. Od dobijenih biokatalizatora, tri sa najvećim aktivnostima PPO, nazvani Eupergit C250L-PFO; Celit-PFO i CelulozaM-PFO, testirani su u reaktoru za uklanjanje fenola, 4-hlorfenola i 4-bromfenola. U slučaju 2,5 mM supstrata sa Eupergit C250L-PFO, postignuto je oko 45 % razgradnje 4-bromfenola, dok su 4-hlorfenol i fenol razgrađeni 35, odnosno 20 %. Testirana je i sposobnost višestruke upotrebe Eupergit C250L-PFO imobilizata za uklanjanje 4-hlorfenola. Nakon osam ponovljenih ciklusa efikasnost Eupergit C250L-PFO imobilizata za uklanjanje 4-hlorfenola je pala na 55%.

Phenols containing halogens, which tend to deactivate the aromatic nuclei, constitute a significant category of highly toxic and difficult-to-degrade pollutants, which arise from a wide variety of industries. The main purpose of this study was to obtain an inexpensive immobilized enzyme for the removal of phenols. Partially purified potato polyphenol oxidase (PPO) was immobilized onto different commercial and laboratory produced carriers. Three of the obtained biocatalysts, with the highest PPO activities, namely Eupergit C250L-PPO; Celite-PPO and CelluloseM-PPO, were tested in a batch reactor for the removal of phenol, 4-chlorophenol and 4-bromophenol. In the case of 2.5 mM substrates with Eupergit C250L-PPO, an around 45% removal of 4-bromophenol was achieved, while the removals 4-chlorophenol and phenol were 35 and 20%, respectively. The reusability of Eupergit C250L-PPO for the removal of 4-chlorophenol was tested. After eight repeated tests, the efficiency of 4-chlorophenol removal by Eupergit C250L-PPO immobilisate had decreased to 55%., Halogenovani fenoli imaju dezaktivirano aromatično jezgro i čine značajnu kategoriju veoma toksičnih i teško razgradivih zagađivača u raznim industrijskim granama. Glavni cilj ovog rada je bio dobijanje jeftinog imobilizovanog enzima za uklanjanje fenola. Delimično prečišćena polifenol-oksidaza (PFO) iz krompira je imobilizovana na različitim komercijalnim i laboratorijski sintetizovanim nosačima. Od dobijenih biokatalizatora, tri sa najvećim aktivnostima PPO, nazvani Eupergit C250L-PFO; Celit-PFO i CelulozaM-PFO, testirani su u reaktoru za uklanjanje fenola, 4-hlorfenola i 4-bromfenola. U slučaju 2,5 mM supstrata sa Eupergit C250L-PFO, postignuto je oko 45 % razgradnje 4-bromfenola, dok su 4-hlorfenol i fenol razgrađeni 35, odnosno 20 %. Testirana je i sposobnost višestruke upotrebe Eupergit C250L-PFO imobilizata za uklanjanje 4-hlorfenola. Nakon osam ponovljenih ciklusa efikasnost Eupergit C250L-PFO imobilizata za uklanjanje 4-hlorfenola je pala na 55%.