- Email: [email protected]
Enhanced production and recovery of microbial muconic acid
S84
Abstracts / New Biotechnology 33S (2016) S1–S213
P7-4 Enhanced production and recovery of microbial muconic acid Jiumn-Yih Wu 1,∗ , Wen-Chien Lee 2 1 2
I-Shou University, Taiwan National Chung Cheng University, Taiwan
Bio-based commodity chemicals including C3–C5 bio-materials attract global interest recently. Muconic acid (MA) with unsaturated dicarboxylic groups, an alternate precursor of adipic acid, is one kind of special chemical. We screened a MA-accumulating strain, Pseudomonas sp. identified by BCRC. After undergoing several chemical mutations and adopting an orchestrated screening strategy, the mutant has the ability of yielding ex-cellular MA and achieving 70% (g/g) of benzoate conversion. The mutated Pseudomonas sp. is able to co-metabolize benzoate (B) to MA in the presence of glucose (G), succinic acid (S) and yeast extract (Y) together as the growth substrate. The productivity of MA in the optimized RSM experiment is 3-fold of the original predesigned state. The oxygen transfer of the fermenter has a strong impact on the MA productivity during the fermentation. Above the critical kL a (800 h−1 ), the MA productivity is proportional to the system oxygen transfer rate. The MA reaches 0.525 g/L/h on condition that the kL a of the fermenter is controlled at 1200 h−1 . It was also found that BYS as feeding medium is the best fed-batch combination for getting high MA productivity. The feeding strategy with enhanced oxygen transfer was carried out in the 5L fermenter; the muconic acid reached the maximum output of 19 g/L. We apply the chemical treatment to the MA separation. Using the appropriate operating period with a competent pH adjustment in this separation process can achieve the compromise between yield and purity of recovery and avoid the unfavorable results. http://dx.doi.org/10.1016/j.nbt.2016.06.1008
P7-5 Withdrawn
http://dx.doi.org/10.1016/j.nbt.2016.06.1009
P7-6 Activation of pentose phosphate pathway in Escherichia coli for the co-production of hydrogen and ethanol from glucose Eunhee Seol ∗ , Balaji Sundara Sekar, Yeonhee Kim, Sunghoon Park Pusan National University, Republic of Korea To address low hydrogen (H2 ) production yield by dark fermentation, co-production of H2 and ethanol by Escherichia coli from glucose, was suggested in this study. Moreover, co-production of these valuable biofuels in a single process is beneficial. Glucose consumption through Embden-Meyerhof-Parnas (EMP) pathway could yield only 2 mol NAD(P)H mol−1 glucose resulting in the production of 2 mol H2 , 1 mol ethanol and 1 mol acetate. Therefore, activation of pentose-phosphate (PP) pathway was attempted which could yield 3.67 mol mol−1 NAD(P)H to improve the ethanol production yield. In order to divert the carbon flow from EMP pathway to PP pathway, downregulation or disruption of EMP pathway flux along with overexpression of key enzymes in PP pathway was attempted. pfkA coding for phosphofructokinase-I was
deleted to downregulate EMP pathway and complete disruption of EMP pathway was attempted by deleting phosphoglucose isomerase (pgi) in E. coli (E. coli BW25113 hycA hyaAB hybBC ldhA frdAB). Additionally, glucose-6-phosphate dehydrogenase (Zwf) and 6-phosphogluconate dehydrogenase (Gnd) were overexpressed in order to activate PP pathway. The pfkA strains were adapted for improving its anaerobic growth whereas Zwf has to be overexpressed in pgi strain to facilitate its anaerobic growth. Further, the expression level of Zwf and Gnd were optimized to improve the co-production yield. pgi ZG strain showed the maximum co-production yield (1.69 for H2 and 1.50 mol mol−1 ethanol) which corresponds to ∼85% energy recovery from glucose showing the possibility of operating PP pathway under anaerobic condition. http://dx.doi.org/10.1016/j.nbt.2016.06.1010
P7-7 Ethanol fermentation from sweet sorghum stem juice under high gravity condition by newly Saccharomyces cerevisiae strain Suntaree Suporn 1 , Lakkana Laopaiboon 2,3 , Preekamol Klanrit 2 , Pattana Laopaiboon 2,4,∗ 1
Graduate School, Khon Kaen University, Khon Kaen 40002, Thailand Department of Biotechnology, Faculty of Technology, Khon Kaen University, Khon Kaen 40002, Thailand 3 Fermentation Research Center for Value-Added Agricultural Products, Khon Kaen University, Khon Kaen 40002, Thailand 4 Center for Alternative Energy Research and Development, Khon Kaen University, Khon Kaen 40002, Thailand 2
E-mail address: [email protected] (P. Laopaiboon). Sweet sorghum (Sorghum bicolor{L}Moench), has recently become a promising source for bioethanol production because of its outstanding attribute, including large amounts of fermentable sugars in its stalk. In this research, ethanol production efficiency from sweet sorghum stem juice under high gravity (HG, 240 g/L of total sugar) condition using three ethanol producing yeast isolates, SCJ04 and SCJ14 isolated from sugar cane juice and SSJ01 isolated from sweet sorghum stem juice, were investigated. Saccharomyces cerevisiae NP01 was used as a reference strain. The fermentation was carried out at 30◦ C, 200 rpm and the initial cell concentration of the juice was 3.0 × 107 cells mL−1 . The results showed that SSJ01 gave the highest ethanol production efficiency. The ethanol concentration (PE ), yield (Yp/s ) and productivity (Qp ) were 90.1 g/L, 0.50 g/g and 1.88 g/L.h, respectively. When 1.54 g/L of urea (nitrogen content equivalent to that in 6 g/L of yeast extract) was added into the juice, the PE , Yp/s and Qp significantly increased to 112.5 g/L (14.3%, v/v), 0.51 and 2.34 g/L.h, with 92.9% sugar consumption. These values were slightly higher than those of the reference strain (PE , 110.3 g/L, Yp/s , 0.50 g/g and Qp , 2.30 g/L.h). Based on molecular taxonomic analysis using sequences of the D1/D2 domain of a large subunit of 26S rDNA and the ITS 1 and ITS2 regions, SSJ01 was identified as S. cerevisiae. From the obtained results, it can be concluded that S. cerevisiae SSJ01 exhibits a high potential to be used in industrial ethanol fermentation process under VHG conditions. http://dx.doi.org/10.1016/j.nbt.2016.06.1011
Abstracts / New Biotechnology 33S (2016) S1–S213
P7-4 Enhanced production and recovery of microbial muconic acid Jiumn-Yih Wu 1,∗ , Wen-Chien Lee 2 1 2
I-Shou University, Taiwan National Chung Cheng University, Taiwan
Bio-based commodity chemicals including C3–C5 bio-materials attract global interest recently. Muconic acid (MA) with unsaturated dicarboxylic groups, an alternate precursor of adipic acid, is one kind of special chemical. We screened a MA-accumulating strain, Pseudomonas sp. identified by BCRC. After undergoing several chemical mutations and adopting an orchestrated screening strategy, the mutant has the ability of yielding ex-cellular MA and achieving 70% (g/g) of benzoate conversion. The mutated Pseudomonas sp. is able to co-metabolize benzoate (B) to MA in the presence of glucose (G), succinic acid (S) and yeast extract (Y) together as the growth substrate. The productivity of MA in the optimized RSM experiment is 3-fold of the original predesigned state. The oxygen transfer of the fermenter has a strong impact on the MA productivity during the fermentation. Above the critical kL a (800 h−1 ), the MA productivity is proportional to the system oxygen transfer rate. The MA reaches 0.525 g/L/h on condition that the kL a of the fermenter is controlled at 1200 h−1 . It was also found that BYS as feeding medium is the best fed-batch combination for getting high MA productivity. The feeding strategy with enhanced oxygen transfer was carried out in the 5L fermenter; the muconic acid reached the maximum output of 19 g/L. We apply the chemical treatment to the MA separation. Using the appropriate operating period with a competent pH adjustment in this separation process can achieve the compromise between yield and purity of recovery and avoid the unfavorable results. http://dx.doi.org/10.1016/j.nbt.2016.06.1008
P7-5 Withdrawn
http://dx.doi.org/10.1016/j.nbt.2016.06.1009
P7-6 Activation of pentose phosphate pathway in Escherichia coli for the co-production of hydrogen and ethanol from glucose Eunhee Seol ∗ , Balaji Sundara Sekar, Yeonhee Kim, Sunghoon Park Pusan National University, Republic of Korea To address low hydrogen (H2 ) production yield by dark fermentation, co-production of H2 and ethanol by Escherichia coli from glucose, was suggested in this study. Moreover, co-production of these valuable biofuels in a single process is beneficial. Glucose consumption through Embden-Meyerhof-Parnas (EMP) pathway could yield only 2 mol NAD(P)H mol−1 glucose resulting in the production of 2 mol H2 , 1 mol ethanol and 1 mol acetate. Therefore, activation of pentose-phosphate (PP) pathway was attempted which could yield 3.67 mol mol−1 NAD(P)H to improve the ethanol production yield. In order to divert the carbon flow from EMP pathway to PP pathway, downregulation or disruption of EMP pathway flux along with overexpression of key enzymes in PP pathway was attempted. pfkA coding for phosphofructokinase-I was
deleted to downregulate EMP pathway and complete disruption of EMP pathway was attempted by deleting phosphoglucose isomerase (pgi) in E. coli (E. coli BW25113 hycA hyaAB hybBC ldhA frdAB). Additionally, glucose-6-phosphate dehydrogenase (Zwf) and 6-phosphogluconate dehydrogenase (Gnd) were overexpressed in order to activate PP pathway. The pfkA strains were adapted for improving its anaerobic growth whereas Zwf has to be overexpressed in pgi strain to facilitate its anaerobic growth. Further, the expression level of Zwf and Gnd were optimized to improve the co-production yield. pgi ZG strain showed the maximum co-production yield (1.69 for H2 and 1.50 mol mol−1 ethanol) which corresponds to ∼85% energy recovery from glucose showing the possibility of operating PP pathway under anaerobic condition. http://dx.doi.org/10.1016/j.nbt.2016.06.1010
P7-7 Ethanol fermentation from sweet sorghum stem juice under high gravity condition by newly Saccharomyces cerevisiae strain Suntaree Suporn 1 , Lakkana Laopaiboon 2,3 , Preekamol Klanrit 2 , Pattana Laopaiboon 2,4,∗ 1
Graduate School, Khon Kaen University, Khon Kaen 40002, Thailand Department of Biotechnology, Faculty of Technology, Khon Kaen University, Khon Kaen 40002, Thailand 3 Fermentation Research Center for Value-Added Agricultural Products, Khon Kaen University, Khon Kaen 40002, Thailand 4 Center for Alternative Energy Research and Development, Khon Kaen University, Khon Kaen 40002, Thailand 2
E-mail address: [email protected] (P. Laopaiboon). Sweet sorghum (Sorghum bicolor{L}Moench), has recently become a promising source for bioethanol production because of its outstanding attribute, including large amounts of fermentable sugars in its stalk. In this research, ethanol production efficiency from sweet sorghum stem juice under high gravity (HG, 240 g/L of total sugar) condition using three ethanol producing yeast isolates, SCJ04 and SCJ14 isolated from sugar cane juice and SSJ01 isolated from sweet sorghum stem juice, were investigated. Saccharomyces cerevisiae NP01 was used as a reference strain. The fermentation was carried out at 30◦ C, 200 rpm and the initial cell concentration of the juice was 3.0 × 107 cells mL−1 . The results showed that SSJ01 gave the highest ethanol production efficiency. The ethanol concentration (PE ), yield (Yp/s ) and productivity (Qp ) were 90.1 g/L, 0.50 g/g and 1.88 g/L.h, respectively. When 1.54 g/L of urea (nitrogen content equivalent to that in 6 g/L of yeast extract) was added into the juice, the PE , Yp/s and Qp significantly increased to 112.5 g/L (14.3%, v/v), 0.51 and 2.34 g/L.h, with 92.9% sugar consumption. These values were slightly higher than those of the reference strain (PE , 110.3 g/L, Yp/s , 0.50 g/g and Qp , 2.30 g/L.h). Based on molecular taxonomic analysis using sequences of the D1/D2 domain of a large subunit of 26S rDNA and the ITS 1 and ITS2 regions, SSJ01 was identified as S. cerevisiae. From the obtained results, it can be concluded that S. cerevisiae SSJ01 exhibits a high potential to be used in industrial ethanol fermentation process under VHG conditions. http://dx.doi.org/10.1016/j.nbt.2016.06.1011