- Email: [email protected]
Zymomonas mobilis biofilm enhances bioethanol production from lignocellulosic hydrolysate
S90
Abstracts / New Biotechnology 33S (2016) S1–S213
P7-29 Optimization of bioethanol production from Manihot glaziovii by response surface methodology Ong Hwai Chyuan University of Malaya, Malaysia Bioethanol is known as a viable alternative fuel to solve both energy and environmental crisis. In this study, Manihot Glaziovii (M. Glaziovii) is chosen as the biomass for bioethanol production, whereby the carbohydrates in M. Glaziovii are first converted into sugars via hydrolysis process. Then, the reducing sugars obtained from the hydrolysis process are fermented into ethanol using Saccharomyces cerevisiae (S. Cerevisiae) yeast. The response surface methodology based on Box-Behnken experimental design is used to observe the optimum conditions and products quality, optimizations of bioethanol production. Enzymatic hydrolysis optimization was performed with selected hydrolysis parameters, including substrate loading, enzyme concentration and hydrolysis period, to be optimized. The experiment optimum values is found to be 11.789% (w/v) of substrate loading, 65.051 U/g enzyme concentration and 142.98 min of hydrolysis period, and yielded 36.23 g/L of reducing sugar. Fermentation optimization was also performed, using optimum values of 1,681 min incubation time, 37.27◦ C reaction temperature and 241 rpm agitation speed. The latter optimization produced 15.85 g/L or 31.08% of ethanol after being fermented by Saccharomyces cerevisiae. The optimizations were conducted with response surface methodology based on Box-Behnken method, and all values resulted from the experiment were very close to those predicted by the method. Several physical and chemical properties of the produced ethanol were also tested according to ASTM D4806 standard, and the observed values were fit within the specified standard. The good quality of ethanol produced from this study indicates that M. Glaziovii holds a practical and strong potential as bioethanol feedstock. http://dx.doi.org/10.1016/j.nbt.2016.06.1031
P7-30 Effect of inoculum source on anaerobic digestion of organic fraction of municipal solid waste in Mexico Maria Virginia Sillas Moreno 1,∗ , Alejandro Montesinos Castellanos 2 , Adriana Pacheco Moscoa 3 1
Tecnologico de Monterrey, Mexico Campus Monterrey, Mexico 3 Tecnológico de Monterrey, Mexico 2
The Biochemical Methane Potential (BMP) test is used to explore and determine the feasibility of different organic materials to be used as substrate in anaerobic digestion (AD), under different conditions. A factor that has been understudied is the effect of the inoculum source. The inoculum source impacts the equilibrium in microbial population and the adaptation to substrate, hence the startup of biogas production, considered one of the most critical steps in the process. The aim of this work was to compare the effect of inoculum source in a BMP test at three different inoculums to substrate ratios (ISR = 1, 2 and 3) during the anaerobic digestion of a model mixture of organic fraction of municipal solid waste from Mexico (MM-OFMSW). The BMP tests were carried out using two inoculum sources: (1) Sludge from an anaerobic wastewater treatment plant (WWS samples) and (2) a solid sample obtained from a landfill (LS samples). All tests were carried out under mesophilic
conditions. The results showed that the maximum amount of CH4 for LS samples (ISR = 2) was 311.51 ± 11.07 mL CH4 /gVS, and 282.41 ± 18.15 mL CH4 /gVS for WWS samples (ISR = 1). Although, the LS inoculum source produced a major quantity of CH4 , it showed a lag phase of 5 days, whereas WWS produced CH4 immediately, which might indicate a quick adaptation of the consortia to MMOFMSW. This suggests that a combination of both inoculum sources could reduce the startup obtained for LS and enhance the quantity of CH4 , obtaining an improved inoculum. http://dx.doi.org/10.1016/j.nbt.2016.06.1032
P7-31 Biotechnological production of optically pure 2,3-butanediol and its derivatives through metabolic engineering Cuiqing Ma Shandong University, China 2,3-butanediol (2,3-BD) is a bulk fuel bio-chemical that can be produced via biotechnological routes. It can be used as a starting material for the synthesis of bulk chemicals such as methylethylketone, gamma-butyrolactone and 1,3-butadiene. Due to its high heating value (27,200 J/g), 2,3-BD can also be used as a liquid fuel or fuel additive. There are three isomeric forms of 2,3-BD: (2R,3R)2,3-BD, meso-2,3-BD and (2S,3S)-2,3-BD. Optically pure 2,3-BD can act as an excellent building block in asymmetric synthesis of valuable chiral chemicals. Although many native microorganisms could be used to efficiently produce 2,3-BD, owing to lack of knowledge about stereoisomer formation mechanisms, the production of optically active 2,3-BD using these strains has been difficult to achieve. Thus, we have constructed three recombinant strains for the production of optically pure (2R,3R)-2,3-BD (Metab Eng, 2015, 28:19–27), meso-2,3-BD (Metab Eng, 2014, 23:22–33) and (2S,3S)-2,3-BD (Biotechnol Biofuels, 2015, 8:143), respectively. On the other hand, two important intermediates, diacetyl and acetoin, could also be produced during the 2,3-BD production process. Both diacetyl and acetoin are important flavors and are widely used in the food industry. Although diacetyl and acetoin could be produced through chemical methods, microbial production of diacetyl and acetoin is preferred over chemical synthesis due to the safety reasons. Thus, we also constructed two recombinant strains for the efficient production of diacetyl (Scientific Reports, 2015, 5:9033) and acetoin (Green Chem, 2015, http://dx.doi.org/ 10.1039/c5gc01638j) based on the 2,3-BD producer Enterobacter cloacae subsp. dissolvens strain SDM. http://dx.doi.org/10.1016/j.nbt.2016.06.1033
P7-32 Withdrawn
http://dx.doi.org/10.1016/j.nbt.2016.06.1034
P7-33 Zymomonas mobilis biofilm enhances bioethanol production from lignocellulosic hydrolysate Tatsaporn Todhanakasem Assumption University, Thailand
Abstracts / New Biotechnology 33S (2016) S1–S213
Z. mobilis demonstrates high ethanol production though the Entner-Doudoroff pathway using glucose as a substrate. Production of ethanol from lignocellulosic hydrolysate using Z. mobilis planktonic cells was commonly impeded by the presence of toxic inhibitors in the hydrolysate. This turns to be a challenging problem for the bioconversion. Z. mobilis in the biofilm form was emerged to resolve this complexity by having the ability to tolerate to these toxic inhibitors. From our study, Z. mobilis readily form biofilm on both biotic and abiotic surfaces in different types of biofilm forming manners. Z. mobilis was able to develop biofilm on abiotic surfaces; polystyrene (PS) and polyvinyl chloride (PVC) and also on biotic surfaces; DEAE cellulose and corn silk. Z. mobilis biofilm represented two fold higher in the percentage of live cells and metabolic activity than planktonic cells when it was exposed to the lignocellulosic hydrolysate containing toxic inhibitors (furfural, 5-hydroxymethyl furfural, vanillin, syringaldehyde, 4-hydroxybenzaldehyde and acetic acid). This consequently produced over 10 times higher in ethanol yield than planktonic cells and provided the theoretical yield of ethanol up to 60–80% and 70–100% when polysterene and corn silk were used as biofilm carriers respectively. Z. mobilis biofilm could be manipulated toward the cell recycle through repeated batch process of ethanol production from lignocellulosic hydrolysate when DEAE cellulose was used as a carrier. The study illustrates the potential of enhancing ethanol production from lignocellulosic material by the utilization of Z. mobilis biofilm in term of biofilm reactor in the future. http://dx.doi.org/10.1016/j.nbt.2016.06.1035
P7-34 Screening fatty acid composition of British heterotrophic microalgae thraustochytrids for production of omega-3 oils and biodiesel Loris Fossier Marchan Heriot Watt University, United Kingdom Heterotrophic microalgae thraustochytrid strains, originally isolated from the North and Malin Seas and a Scottish salt marsh, showed potential for biotechnological applications based on their production of high levels of omega-3 long-chain (=C20 ) polyunsaturated fatty acids (PUFA), including docosahexaenoic acid (DHA, 22:6?3). The isolates were screened for their fatty acid composition and compared with Sicyoidochytrium sp. NBRC 102979, isolated from Japanese sub-tropical waters. Screening was carried out at the end of exponential phase and at the stationary phase of growth in two media. At the end of the exponential growth phase, all strains exhibited fatty acid profiles rich in saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA), while PUFA were predominant in late stationary phase cells. Overall, the relative levels of DHA (as % of total fatty acids, TFA) were to our knowledge amongst the highest recorded from any thraustochytrid screening study to date, with strain TL18 reaching up to 67% DHA in modified GYP, although DHA yields were lower compared to previous studies with other strains. Media composition affected final biomass and fatty acid yields, while PUFA (%TFA) profiles remained unaltered. For biodiesel applications, the qualitative properties of the oils from these thraustochytrids were compared to the international standard (EN 14214; ASTM D6751). This study has therefore demonstrated the potential of previously undescribed British thraustochytrids for the production of DHA, aimed at potential applications in aquaculture feeding and human food, cosmetic and other uses, while showing some potential, under the culture conditions
S91
used, in the production of microbial biodiesel. Future prospects will involve scaling up the process and optimizing culture conditions. http://dx.doi.org/10.1016/j.nbt.2016.06.1036
P7-35 The effects of cloudberry extract and -carotene on lifespan of Drosophilla melanogaster Ekaterina Lashmanova 1,∗ , Olga Kuzivanova 2 , Olga Dymova 3 , Ekaterina Proshkina 4 , Alexey Moskalev 5 1
Moscow Institute of Physics and Technology (State University), Russia 2 Dolgoprudny, Russia 3 Russia 4 Institute of Biology, Russia 5 Komi Science Center, Russia Cloudberries are an important source of health benefiting chemicals (carotenoids (Car), phenols, lipids and carbohydrates). There is data that Car can modulate insulin/IGF-I-signaling pathway, which plays a key role in the process of aging. We studied the geroprotective properties of cloudberry extract and its dominating carotenoid -car. The experiments were performed on the wild-type CantonS Drosophilla melanogaster. The extract was added into nutrient medium in final concentrations 0.12 and 0.6 mg/ml. That was equivalent to concentration of Car 1 and 5 ?. After 20 days of cloudberry extract consumption the level of -car and its products of metabolism was 0.820 ng per fly in females and 0.220 ng per fly in males. The addition of cloudberry extract extended the median (2–11%) and maximum lifespan (3–19%) in males and females. The effects were more expressed in females. The same sex-dependent effects were observed in the experiments with a pure -car (Fluka). In females the addition of -car extended the median lifespan of flies by 7% in females and by 2% in males. The resistance of D. melanogaster to various types of stress (oxidative, hyperthermia and starvation) after -car treatment were studied as longevity usually correlates with stress resistance. In females the increased stress-resistance to paraquat was observed after exposure to -car. The -car exposure also effected the expression of aging-related genes (dSir2, JNK, p53, Gadd45, Hps70, Sod1). Thus, cloudberry extract extended lifespan of D. melanogaster. These lifespan extension effects were performed at least partly by -car. http://dx.doi.org/10.1016/j.nbt.2016.06.1037
P7-36 Null mutation of NRG1 gene improves tolerance of Saccharomyces cerevisiae to fermentation inhibitors Eun-Hee Park ∗ , Chan-Yeong Choi, Yun-Ji Cho, Myoung-Dong Kim Kangwon National University, Republic of Korea Budding yeast Saccharomyces cerevisiae plays an important role in producing bioethanol from lignocellulosic biomass. Pretreatment of lignocellulosic biomass releases a number of inhibitors reducing cell growth rate and ethanol productivity, which suggests an elaborate research should be performed to increase resistance of S. cerevisiae to fermentation inhibitors. Nrg1pisa transcriptional repressor recruiting the Cyc8-Tup1 complex to stress-responsive promoter and negatively regulates a variety of
Abstracts / New Biotechnology 33S (2016) S1–S213
P7-29 Optimization of bioethanol production from Manihot glaziovii by response surface methodology Ong Hwai Chyuan University of Malaya, Malaysia Bioethanol is known as a viable alternative fuel to solve both energy and environmental crisis. In this study, Manihot Glaziovii (M. Glaziovii) is chosen as the biomass for bioethanol production, whereby the carbohydrates in M. Glaziovii are first converted into sugars via hydrolysis process. Then, the reducing sugars obtained from the hydrolysis process are fermented into ethanol using Saccharomyces cerevisiae (S. Cerevisiae) yeast. The response surface methodology based on Box-Behnken experimental design is used to observe the optimum conditions and products quality, optimizations of bioethanol production. Enzymatic hydrolysis optimization was performed with selected hydrolysis parameters, including substrate loading, enzyme concentration and hydrolysis period, to be optimized. The experiment optimum values is found to be 11.789% (w/v) of substrate loading, 65.051 U/g enzyme concentration and 142.98 min of hydrolysis period, and yielded 36.23 g/L of reducing sugar. Fermentation optimization was also performed, using optimum values of 1,681 min incubation time, 37.27◦ C reaction temperature and 241 rpm agitation speed. The latter optimization produced 15.85 g/L or 31.08% of ethanol after being fermented by Saccharomyces cerevisiae. The optimizations were conducted with response surface methodology based on Box-Behnken method, and all values resulted from the experiment were very close to those predicted by the method. Several physical and chemical properties of the produced ethanol were also tested according to ASTM D4806 standard, and the observed values were fit within the specified standard. The good quality of ethanol produced from this study indicates that M. Glaziovii holds a practical and strong potential as bioethanol feedstock. http://dx.doi.org/10.1016/j.nbt.2016.06.1031
P7-30 Effect of inoculum source on anaerobic digestion of organic fraction of municipal solid waste in Mexico Maria Virginia Sillas Moreno 1,∗ , Alejandro Montesinos Castellanos 2 , Adriana Pacheco Moscoa 3 1
Tecnologico de Monterrey, Mexico Campus Monterrey, Mexico 3 Tecnológico de Monterrey, Mexico 2
The Biochemical Methane Potential (BMP) test is used to explore and determine the feasibility of different organic materials to be used as substrate in anaerobic digestion (AD), under different conditions. A factor that has been understudied is the effect of the inoculum source. The inoculum source impacts the equilibrium in microbial population and the adaptation to substrate, hence the startup of biogas production, considered one of the most critical steps in the process. The aim of this work was to compare the effect of inoculum source in a BMP test at three different inoculums to substrate ratios (ISR = 1, 2 and 3) during the anaerobic digestion of a model mixture of organic fraction of municipal solid waste from Mexico (MM-OFMSW). The BMP tests were carried out using two inoculum sources: (1) Sludge from an anaerobic wastewater treatment plant (WWS samples) and (2) a solid sample obtained from a landfill (LS samples). All tests were carried out under mesophilic
conditions. The results showed that the maximum amount of CH4 for LS samples (ISR = 2) was 311.51 ± 11.07 mL CH4 /gVS, and 282.41 ± 18.15 mL CH4 /gVS for WWS samples (ISR = 1). Although, the LS inoculum source produced a major quantity of CH4 , it showed a lag phase of 5 days, whereas WWS produced CH4 immediately, which might indicate a quick adaptation of the consortia to MMOFMSW. This suggests that a combination of both inoculum sources could reduce the startup obtained for LS and enhance the quantity of CH4 , obtaining an improved inoculum. http://dx.doi.org/10.1016/j.nbt.2016.06.1032
P7-31 Biotechnological production of optically pure 2,3-butanediol and its derivatives through metabolic engineering Cuiqing Ma Shandong University, China 2,3-butanediol (2,3-BD) is a bulk fuel bio-chemical that can be produced via biotechnological routes. It can be used as a starting material for the synthesis of bulk chemicals such as methylethylketone, gamma-butyrolactone and 1,3-butadiene. Due to its high heating value (27,200 J/g), 2,3-BD can also be used as a liquid fuel or fuel additive. There are three isomeric forms of 2,3-BD: (2R,3R)2,3-BD, meso-2,3-BD and (2S,3S)-2,3-BD. Optically pure 2,3-BD can act as an excellent building block in asymmetric synthesis of valuable chiral chemicals. Although many native microorganisms could be used to efficiently produce 2,3-BD, owing to lack of knowledge about stereoisomer formation mechanisms, the production of optically active 2,3-BD using these strains has been difficult to achieve. Thus, we have constructed three recombinant strains for the production of optically pure (2R,3R)-2,3-BD (Metab Eng, 2015, 28:19–27), meso-2,3-BD (Metab Eng, 2014, 23:22–33) and (2S,3S)-2,3-BD (Biotechnol Biofuels, 2015, 8:143), respectively. On the other hand, two important intermediates, diacetyl and acetoin, could also be produced during the 2,3-BD production process. Both diacetyl and acetoin are important flavors and are widely used in the food industry. Although diacetyl and acetoin could be produced through chemical methods, microbial production of diacetyl and acetoin is preferred over chemical synthesis due to the safety reasons. Thus, we also constructed two recombinant strains for the efficient production of diacetyl (Scientific Reports, 2015, 5:9033) and acetoin (Green Chem, 2015, http://dx.doi.org/ 10.1039/c5gc01638j) based on the 2,3-BD producer Enterobacter cloacae subsp. dissolvens strain SDM. http://dx.doi.org/10.1016/j.nbt.2016.06.1033
P7-32 Withdrawn
http://dx.doi.org/10.1016/j.nbt.2016.06.1034
P7-33 Zymomonas mobilis biofilm enhances bioethanol production from lignocellulosic hydrolysate Tatsaporn Todhanakasem Assumption University, Thailand
Abstracts / New Biotechnology 33S (2016) S1–S213
Z. mobilis demonstrates high ethanol production though the Entner-Doudoroff pathway using glucose as a substrate. Production of ethanol from lignocellulosic hydrolysate using Z. mobilis planktonic cells was commonly impeded by the presence of toxic inhibitors in the hydrolysate. This turns to be a challenging problem for the bioconversion. Z. mobilis in the biofilm form was emerged to resolve this complexity by having the ability to tolerate to these toxic inhibitors. From our study, Z. mobilis readily form biofilm on both biotic and abiotic surfaces in different types of biofilm forming manners. Z. mobilis was able to develop biofilm on abiotic surfaces; polystyrene (PS) and polyvinyl chloride (PVC) and also on biotic surfaces; DEAE cellulose and corn silk. Z. mobilis biofilm represented two fold higher in the percentage of live cells and metabolic activity than planktonic cells when it was exposed to the lignocellulosic hydrolysate containing toxic inhibitors (furfural, 5-hydroxymethyl furfural, vanillin, syringaldehyde, 4-hydroxybenzaldehyde and acetic acid). This consequently produced over 10 times higher in ethanol yield than planktonic cells and provided the theoretical yield of ethanol up to 60–80% and 70–100% when polysterene and corn silk were used as biofilm carriers respectively. Z. mobilis biofilm could be manipulated toward the cell recycle through repeated batch process of ethanol production from lignocellulosic hydrolysate when DEAE cellulose was used as a carrier. The study illustrates the potential of enhancing ethanol production from lignocellulosic material by the utilization of Z. mobilis biofilm in term of biofilm reactor in the future. http://dx.doi.org/10.1016/j.nbt.2016.06.1035
P7-34 Screening fatty acid composition of British heterotrophic microalgae thraustochytrids for production of omega-3 oils and biodiesel Loris Fossier Marchan Heriot Watt University, United Kingdom Heterotrophic microalgae thraustochytrid strains, originally isolated from the North and Malin Seas and a Scottish salt marsh, showed potential for biotechnological applications based on their production of high levels of omega-3 long-chain (=C20 ) polyunsaturated fatty acids (PUFA), including docosahexaenoic acid (DHA, 22:6?3). The isolates were screened for their fatty acid composition and compared with Sicyoidochytrium sp. NBRC 102979, isolated from Japanese sub-tropical waters. Screening was carried out at the end of exponential phase and at the stationary phase of growth in two media. At the end of the exponential growth phase, all strains exhibited fatty acid profiles rich in saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA), while PUFA were predominant in late stationary phase cells. Overall, the relative levels of DHA (as % of total fatty acids, TFA) were to our knowledge amongst the highest recorded from any thraustochytrid screening study to date, with strain TL18 reaching up to 67% DHA in modified GYP, although DHA yields were lower compared to previous studies with other strains. Media composition affected final biomass and fatty acid yields, while PUFA (%TFA) profiles remained unaltered. For biodiesel applications, the qualitative properties of the oils from these thraustochytrids were compared to the international standard (EN 14214; ASTM D6751). This study has therefore demonstrated the potential of previously undescribed British thraustochytrids for the production of DHA, aimed at potential applications in aquaculture feeding and human food, cosmetic and other uses, while showing some potential, under the culture conditions
S91
used, in the production of microbial biodiesel. Future prospects will involve scaling up the process and optimizing culture conditions. http://dx.doi.org/10.1016/j.nbt.2016.06.1036
P7-35 The effects of cloudberry extract and -carotene on lifespan of Drosophilla melanogaster Ekaterina Lashmanova 1,∗ , Olga Kuzivanova 2 , Olga Dymova 3 , Ekaterina Proshkina 4 , Alexey Moskalev 5 1
Moscow Institute of Physics and Technology (State University), Russia 2 Dolgoprudny, Russia 3 Russia 4 Institute of Biology, Russia 5 Komi Science Center, Russia Cloudberries are an important source of health benefiting chemicals (carotenoids (Car), phenols, lipids and carbohydrates). There is data that Car can modulate insulin/IGF-I-signaling pathway, which plays a key role in the process of aging. We studied the geroprotective properties of cloudberry extract and its dominating carotenoid -car. The experiments were performed on the wild-type CantonS Drosophilla melanogaster. The extract was added into nutrient medium in final concentrations 0.12 and 0.6 mg/ml. That was equivalent to concentration of Car 1 and 5 ?. After 20 days of cloudberry extract consumption the level of -car and its products of metabolism was 0.820 ng per fly in females and 0.220 ng per fly in males. The addition of cloudberry extract extended the median (2–11%) and maximum lifespan (3–19%) in males and females. The effects were more expressed in females. The same sex-dependent effects were observed in the experiments with a pure -car (Fluka). In females the addition of -car extended the median lifespan of flies by 7% in females and by 2% in males. The resistance of D. melanogaster to various types of stress (oxidative, hyperthermia and starvation) after -car treatment were studied as longevity usually correlates with stress resistance. In females the increased stress-resistance to paraquat was observed after exposure to -car. The -car exposure also effected the expression of aging-related genes (dSir2, JNK, p53, Gadd45, Hps70, Sod1). Thus, cloudberry extract extended lifespan of D. melanogaster. These lifespan extension effects were performed at least partly by -car. http://dx.doi.org/10.1016/j.nbt.2016.06.1037
P7-36 Null mutation of NRG1 gene improves tolerance of Saccharomyces cerevisiae to fermentation inhibitors Eun-Hee Park ∗ , Chan-Yeong Choi, Yun-Ji Cho, Myoung-Dong Kim Kangwon National University, Republic of Korea Budding yeast Saccharomyces cerevisiae plays an important role in producing bioethanol from lignocellulosic biomass. Pretreatment of lignocellulosic biomass releases a number of inhibitors reducing cell growth rate and ethanol productivity, which suggests an elaborate research should be performed to increase resistance of S. cerevisiae to fermentation inhibitors. Nrg1pisa transcriptional repressor recruiting the Cyc8-Tup1 complex to stress-responsive promoter and negatively regulates a variety of