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Recovery of red colorants from fermented broth of Penicillium purpurogenum DPUA 1275 using Colloidal Gas Aphrons generated by different surfactants
New Biotechnology · Volume 29S · September 2012
Poster 5.0.184
Poster 5.0.185
Recovery of red colorants from fermented broth of Penicillium purpurogenum DPUA 1275 using Colloidal Gas Aphrons generated by different surfactants
Revealing the hexokinase step of glycolysis in lactic acid producer fungus Rhizopus oryzae
1
Valéria Carvalho Santos-Ebinuma , Maria Francisca Teixeira2 , Adalberto Pessoa Jr.1 , Paula Jauregi3,∗
Simas
1
Department of Biochemical and Pharmaceutical Technology, University of São Paulo, Avenida Prof. Lineu Prestes 580, Bl. 16, 05508-900 São Paulo, SP, Brazil 2 Culture Collection DPUA/UFAM, Universidade Federal do Amazonas, Av. Gal. Rodrigo Octávio Jordão Ramos, 3000, 69.077-000 Manaus, AM, Brazil 3 Department of Food and Nutritional Sciences, The University of Reading, Whiteknights, Reading RG6 6AP, UK Synthetic and natural colorants are used extensively in the food, cosmetic and pharmaceutical industries. However, natural colorants have recently gained popularity over synthetic coloring agents due to safety concerns about the latter. Natural colorants can be produced by microorganisms using cultivation technology and extracted from fermented broth. The aim of this study was to investigate the recovery of red colorants from fermented broth of Penicillium purpurogenum by Colloidal Gas Aphrons (CGA) flotation; CGA are surfactant-stabilized microbubbles generated by intense stirring of a surfactant solution. CGA were generated by the following surfactants: (i) cationic, hexadecyl trimethyl ammonium bromide (CTAB); (ii) anionic, Sodium dodecyl sulphate (SDS) and (iii) non-ionic, polyoxyethylenesorbitan monolaurate (Tween 20). The effect of CGA to fermented broth volumetric ratios (VCGA /VFB ) on recovery was investigated. The VCGA /VFB ranged between 3 and 18. Higher colorants recovery was obtained with CTAB and Tween 20 than with SDS. The best results were obtained with CTAB at pH = 9 and at VCGA /VFB = 12: Recovery, Re = 70%; partition coefficient, K = 5.39; selectivity in relation to proteins and sugar SePROT , 3.75 and SeSUGAR , 7.20, respectively. These results indicate that with CTAB the recovery is mainly driven by electrostatic interactions whilst with Tween 20 the separation is driven mainly by hydrophobic interactions. Depending on the red colorants application, the use of Tween 20 is more favourable due to the toxicity of CTAB. In conclusion, the results above show that CGA is a promising separation method which can be used as the first step of purification for the recovery of red colorants from fermentation broth. Keywords: Natural colorants; Recovery; Colloidal Gas Aphrons; Penicillium purpurogenum http://dx.doi.org/10.1016/j.nbt.2012.08.624
S222
www.elsevier.com/locate/nbt
Eda Alagöz1,∗ , S¸eyda Ac¸ar2 , Meral Yücel3 , Haluk Hamamci4 1
METU, Department of Biotechnology, Turkey VU of Amsterdam, Department of Physics and Astronomy, Netherlands 3 METU, Department of Biological Sciences, Turkey 4 METU, Department of Food Engineering, Turkey 2
Rhizopus oryzae is a filamentous fungus which can grow on some renewable carbon sources and this makes it a good candidate for production of lactic acid and ethanol. However, in R. oryzae the production yields of these products are lower compared to alternative organisms. The utilization of the carbon source through glycolysis might end up in lactic acid or ethanol production. This makes the glycolytic pathway a focus of attention for our search for a way to increase the speed and efficiency of this pathway thus increasing the yield of desired end product. Hexokinase catalyse the first and irreversible step in glycolysis and implicated as one of the major control points in this pathway. In this study we are trying to investigate the regulation of glycolytic pathway of R. oryzae, in the light of studies done for Saccharomyces cerevisiae. The probable hexokinase genes were found from the annotated genome database of R. oryzae by doing a blast search using the known protein sequences of closely related S. cerevisiae hexokinases and glucokinase. Five of our ten probable hexokinase genes were obtained by using PCR. Sequencing results of three of them proved that these genes have no introns meaning they were expressed in R. oryzae. In the complementation study done in hexokinaless mutant of S. cerevisiae for one of these genes the growth of the transformant was observed on glucose or fructose containing media. Same experiments will be carried out for the rest of these genes. Keywords: Hexokinase; Rhizopus oryzae http://dx.doi.org/10.1016/j.nbt.2012.08.625 Poster 5.0.186 Availability of decolorization of dyestuff by Saccharomyces cerevisiae immobilized on cone of Pinus nigra ¸ abuk Merve Girgin∗ , Ahmet C Department of Biology, Faculty of Arts and Science, Mugla University, Mugla, Turkey Department of Biology, Faculty of Arts and Science, Eskisehir Osmangazi University, 26480 Eskisehir, Turkey In this study, the function of dyestuff affect has been investigated in Saccharomyces cerevisiae immobilized on Pinus nigra cone biomass to Yellow 85, Blue 13, Orange 13, Black 8, Red 3:1, Blue 49, Blue 72, Orange 12, Violet 1. According to screening studies, Yellow 85, Blue 13 and Orange 13 chemical dyestuffs are particularly selected for further studies. To implore of these then dyestuffs of decolorization the optimum conditions of pH, initial dyestuff concentration, shaking speed, temperature, duration of dying, the materials and amount of the biomasses defined. Apart from there analyzes of FT-IR, toxicity, potential of zeta have been
Poster 5.0.184
Poster 5.0.185
Recovery of red colorants from fermented broth of Penicillium purpurogenum DPUA 1275 using Colloidal Gas Aphrons generated by different surfactants
Revealing the hexokinase step of glycolysis in lactic acid producer fungus Rhizopus oryzae
1
Valéria Carvalho Santos-Ebinuma , Maria Francisca Teixeira2 , Adalberto Pessoa Jr.1 , Paula Jauregi3,∗
Simas
1
Department of Biochemical and Pharmaceutical Technology, University of São Paulo, Avenida Prof. Lineu Prestes 580, Bl. 16, 05508-900 São Paulo, SP, Brazil 2 Culture Collection DPUA/UFAM, Universidade Federal do Amazonas, Av. Gal. Rodrigo Octávio Jordão Ramos, 3000, 69.077-000 Manaus, AM, Brazil 3 Department of Food and Nutritional Sciences, The University of Reading, Whiteknights, Reading RG6 6AP, UK Synthetic and natural colorants are used extensively in the food, cosmetic and pharmaceutical industries. However, natural colorants have recently gained popularity over synthetic coloring agents due to safety concerns about the latter. Natural colorants can be produced by microorganisms using cultivation technology and extracted from fermented broth. The aim of this study was to investigate the recovery of red colorants from fermented broth of Penicillium purpurogenum by Colloidal Gas Aphrons (CGA) flotation; CGA are surfactant-stabilized microbubbles generated by intense stirring of a surfactant solution. CGA were generated by the following surfactants: (i) cationic, hexadecyl trimethyl ammonium bromide (CTAB); (ii) anionic, Sodium dodecyl sulphate (SDS) and (iii) non-ionic, polyoxyethylenesorbitan monolaurate (Tween 20). The effect of CGA to fermented broth volumetric ratios (VCGA /VFB ) on recovery was investigated. The VCGA /VFB ranged between 3 and 18. Higher colorants recovery was obtained with CTAB and Tween 20 than with SDS. The best results were obtained with CTAB at pH = 9 and at VCGA /VFB = 12: Recovery, Re = 70%; partition coefficient, K = 5.39; selectivity in relation to proteins and sugar SePROT , 3.75 and SeSUGAR , 7.20, respectively. These results indicate that with CTAB the recovery is mainly driven by electrostatic interactions whilst with Tween 20 the separation is driven mainly by hydrophobic interactions. Depending on the red colorants application, the use of Tween 20 is more favourable due to the toxicity of CTAB. In conclusion, the results above show that CGA is a promising separation method which can be used as the first step of purification for the recovery of red colorants from fermentation broth. Keywords: Natural colorants; Recovery; Colloidal Gas Aphrons; Penicillium purpurogenum http://dx.doi.org/10.1016/j.nbt.2012.08.624
S222
www.elsevier.com/locate/nbt
Eda Alagöz1,∗ , S¸eyda Ac¸ar2 , Meral Yücel3 , Haluk Hamamci4 1
METU, Department of Biotechnology, Turkey VU of Amsterdam, Department of Physics and Astronomy, Netherlands 3 METU, Department of Biological Sciences, Turkey 4 METU, Department of Food Engineering, Turkey 2
Rhizopus oryzae is a filamentous fungus which can grow on some renewable carbon sources and this makes it a good candidate for production of lactic acid and ethanol. However, in R. oryzae the production yields of these products are lower compared to alternative organisms. The utilization of the carbon source through glycolysis might end up in lactic acid or ethanol production. This makes the glycolytic pathway a focus of attention for our search for a way to increase the speed and efficiency of this pathway thus increasing the yield of desired end product. Hexokinase catalyse the first and irreversible step in glycolysis and implicated as one of the major control points in this pathway. In this study we are trying to investigate the regulation of glycolytic pathway of R. oryzae, in the light of studies done for Saccharomyces cerevisiae. The probable hexokinase genes were found from the annotated genome database of R. oryzae by doing a blast search using the known protein sequences of closely related S. cerevisiae hexokinases and glucokinase. Five of our ten probable hexokinase genes were obtained by using PCR. Sequencing results of three of them proved that these genes have no introns meaning they were expressed in R. oryzae. In the complementation study done in hexokinaless mutant of S. cerevisiae for one of these genes the growth of the transformant was observed on glucose or fructose containing media. Same experiments will be carried out for the rest of these genes. Keywords: Hexokinase; Rhizopus oryzae http://dx.doi.org/10.1016/j.nbt.2012.08.625 Poster 5.0.186 Availability of decolorization of dyestuff by Saccharomyces cerevisiae immobilized on cone of Pinus nigra ¸ abuk Merve Girgin∗ , Ahmet C Department of Biology, Faculty of Arts and Science, Mugla University, Mugla, Turkey Department of Biology, Faculty of Arts and Science, Eskisehir Osmangazi University, 26480 Eskisehir, Turkey In this study, the function of dyestuff affect has been investigated in Saccharomyces cerevisiae immobilized on Pinus nigra cone biomass to Yellow 85, Blue 13, Orange 13, Black 8, Red 3:1, Blue 49, Blue 72, Orange 12, Violet 1. According to screening studies, Yellow 85, Blue 13 and Orange 13 chemical dyestuffs are particularly selected for further studies. To implore of these then dyestuffs of decolorization the optimum conditions of pH, initial dyestuff concentration, shaking speed, temperature, duration of dying, the materials and amount of the biomasses defined. Apart from there analyzes of FT-IR, toxicity, potential of zeta have been