| 1. |
- Kalogeris, E, et al.
(författare)
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Performance of an intermittent agitation rotating drum type bioreactor for solid-state fermentation of wheat straw
- 2003
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Ingår i: Bioresource Technology. - 0960-8524 .- 1873-2976. ; 86:3, s. 207-213
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Tidskriftsartikel (refereegranskat)abstract
- A laboratory bioreactor, designed for solid-state fermentation of thermophilic microorganisms, was operated for production of cellulases and hemicellulases by the thermophilic fungus Thermoascus aurantiacus. The suitability of the apparatus for the effective control of important operating variables affecting growth of microbes in solid-state cultivation was determined. Application of the optimum conditions found for the moisture content of the medium, growth temperature and airflow rate produced enzyme yields of 1709 U endoglucanase, 4 U cellobiohydrolase, 79 U β-glucosidase, 5.5 U FPA, 4490 U xylanase and 45 U β-xylosidase per g of dry wheat straw. The correlation between microorganism growth and production of enzymes was efficiently described by the Le Duy kinetic model. © 2002 Elsevier Science Ltd. All rights reserved.
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| 2. |
- Topakas, E., et al.
(författare)
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Bioconversion of ferulic acid into vanillic acid by the thermophilic fungus Sporotrichum thermophile
- 2003
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Ingår i: Lebensmittel-Wissenschaft + Technologie. - 0023-6438 .- 1096-1127. ; 36:6, s. 561-565
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Tidskriftsartikel (refereegranskat)abstract
- Sporotrichum thermophile is capable of promoting the formation of vanillic acid during ferulic acid degradation. Ferulic acid metabolism by S. thermophile apparently occurred via the propenoic chain degradation and the formation of 4-hydroxy-3-methoxystyrene (4-vinylguaiacol) was observed which was presumably metabolized to vanillic acid. Guaiacol was detected in addition to the above-mentioned intermediates, usually as a result of nonoxidative decarboxylation of vanillic acid. The bioconversion of ferulic into vanillic acid was affected by the amount of ferulic acid that was treated and the carbon source on which the biomass was grown. Under optimum conditions vanillic acid production from ferulic acid by S. thermophile attained very high levels of 4798 mg/L with a molar yield of 35%
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| 3. |
- Topakas, E., et al.
(författare)
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Production of phenolics from corn cobs by coupling enzymic treatment and solid state fermentation
- 2004
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Ingår i: Engineering in Life Sciences. - : Wiley. - 1618-0240 .- 1618-2863. ; 4:3, s. 283-286
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Tidskriftsartikel (refereegranskat)abstract
- A two-stage process that combined solid-state fermentation (SSF) and subsequent enzymic treatment was used in order to release p-coumaric (p-CA) and ferulic acid (FA) from corn cobs. Sporotrichum thermophile was grown on corn cobs under SSF conditions, and the production of cinnamoyl esterases and xylanases was studied over 7 days. The time course of enzyme production showed a maximum activity of 1483 nkat/g, 0.3 nkat/g and 0.067 nkat/g for xylanase, feruloyl esterase, and p-coumaroyl esterase, respectively. The importance of the moisture level of the growth substrate was discussed. After SSF, the fermented substrate was directly exposed to autohydrolysis and the in situ produced multienzyme system was successfully used for the partial degradation of cell wall components and the liberation of p-CA and FA. A yield of 0.85 g/kg and 0.38 g/kg FA and p-CA, respectively, of dry matter of corn cobs was obtained.
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| 4. |
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| 5. |
- Topakas, E., et al.
(författare)
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Purification and characterization of a feruloyl esterase from Fusarium oxysporum catalyzing esterification of phenolic acids in ternary water–organic solvent mixtures
- 2003
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Ingår i: Journal of Biotechnology. - 0168-1656 .- 1873-4863. ; 102:1, s. 33-44
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Tidskriftsartikel (refereegranskat)abstract
- An extracellular feruloyl esterase (FAE-II) from the culture filtrates of Fusarium oxysporum F3 was purified to homogeneity by SP-Sepharose, t-butyl-HIC and Sephacryl S-200 column chromatography. The protein corresponded to molecular mass and pI values of 27 kDa and 9.9, respectively. The enzyme was optimally active at pH 7 and 45 °C. The purified esterase was fully stable at pH 7.0–9.0 and temperature up to 45 °C after 1 h incubation. Determination of kcat/Km revealed that the enzyme hydrolysed methyl sinapinate 6, 21 and 40 times more efficiently than methyl ferulate, methyl coumarate and methyl caffeate, respectively. The enzyme was active on substrates containing ferulic acid ester linked to the C-5 but inactive to the C-2 positions of arabinofuranose such as 4-nitrophenyl 5-O-trans-feruloyl-α-l-arabinofuranoside and 4-nitrophenyl 2-O-trans-feruloyl-α-l-arabinofuranoside. In the presence of Sporotrichum thermophile xylanase, there was a significant release of ferulic acid from destarched wheat bran by FAE-II, indicating a synergistic interaction between FAE-II and S. thermophile xylanase. FAE-II by itself could release only little ferulic acid from destarched wheat bran. The potential of FAE-II for the synthesis of various phenolic acid esters was tested using as a reaction system a surfactantless microemulsion formed in ternary mixture consisting of n-hexane, 1-propanol and water.
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| 6. |
- Topakas, E., et al.
(författare)
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Purification and characterization of a type B feruloyl esterase (StFAE-A) from the thermophilic fungus Sporotrichum thermophile
- 2004
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Ingår i: Applied Microbiology and Biotechnology. - : Springer Science and Business Media LLC. - 0175-7598 .- 1432-0614. ; 63:6, s. 686-690
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Tidskriftsartikel (refereegranskat)abstract
- A feruloyl esterase (StFAE-A) produced by Sporotrichum thermophile was purified to homogeneity. The purified homogeneous preparation of native StFAE-A exhibited a molecular mass of 57.0±1.5 kDa, with a mass of 33±1 kDa on SDS-PAGE. The pI of the enzyme was estimated by cation-exchange chromatofocusing to be at pH 3.1. The enzyme activity was optimal at pH 6.0 and 55–60 °C. The purified esterase was stable at the pH range 5.0–7.0. The enzyme retained 70% of activity after 7 h at 50 °C and lost 50% of its activity after 45 min at 55 °C and after 12 min at 60 °C. Determination of k cat/K m revealed that the enzyme hydrolyzed methyl p-coumarate 2.5- and 12-fold more efficiently than methyl caffeate and methyl ferulate, respectively. No activity on methyl sinapinate was detected. The enzyme was active on substrates containing ferulic acid ester linked to the C-5 and C-2 linkages of arabinofuranose and it hydrolyzed 4-nitrophenyl 5-O-trans-feruloyl-α-l-arabinofuranoside (NPh-5-Fe-Araf) 2-fold more efficiently than NPh-2-Fe-Araf. Ferulic acid (FA) was efficiently released from destarched wheat bran when the esterase was incubated together with xylanase from S. thermophile (a maximum of 34% total ferulic acid released after 1 h incubation). StFAE-A by itself could release FA, but at a level almost 47-fold lower than that obtained in the presence of xylanase. The potential of StFAE-A for the synthesis of various phenolic acid esters was tested using a ternary water-organic mixture consisting of n-hexane, 1-butanol and water as a reaction system
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| 7. |
- Vardakou, M., et al.
(författare)
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Synergy between enzymes involved in the degradation of insoluble wheat flour arabinoxylan
- 2004
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Ingår i: Innovative Food Science & Emerging Technologies. - 1466-8564 .- 1878-5522. ; 5:1, s. 107-112
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Tidskriftsartikel (refereegranskat)abstract
- Two microbial endo-β-1,4-xylanases (EXs, EC 3.2.1.8) belonging to glycanase families 10 and 11 were examined for their ability to release ferulic acid (FA) from water-unextractable arabinoxylan (WU-AX) in the presence of a feruloyl esterase (FoFAE-II) from Fusarium oxysporum. WU-AX was incubated with different levels of a Thermoascus aurantiacus family 10 (XYLI) and a Sporotrichum thermophile family 11 (XYLA) endoxylanases. At 10 g/l arabinoxylan, enzyme concentrations (KE values) needed to obtain half-maximal hydrolysis rates for FA release were 0.18 and 0.44 nM for the xylanases from T. aurantiacus and S. thermophile, respectively. Determination of Vmax/KE revealed that the family 10 enzyme performed 4.3 times more efficiently than the family 11 enzyme in liberation of FA when a feruloyl esterase is present. Molecular weights of the products formed were assessed and separation of feruloyl-oligosaccharides was achieved by anion-exchange and size-exclusion chromatography (SEC). The results showed that the degradation of the xylan backbone was influenced strongest by the action of xylanases while the presence of the esterase mainly resulted in the release of ferulic acid from the produced short chain feruloylated xylo-oligosaccharides by the action of xylanases
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