Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/5020
標題: Rhodopseudomonas palustris WP3-5細胞生長、產氫與PHB累積之關係及共培養試驗
Relationship between cell growth, hydrogen production and poly-β-hydroxybutyrate (PHB) accumulation by Rhodopseudomonas palustris WP3-5 and co-culture test
作者: 陳盈孜
Chen, Ying-Tzu
關鍵字: 生物產氫
biological hydrogen production
紫色不含硫光合菌
Rhodopseudomonas palustris WP3-5
poly-β-hydroxybutyrate
溶解性微生物產物
共培養
連續流產氫
purple non-sulfur photosynthetic bacteria
Rhodopseudomonas palustris WP3-5
poly-β-hydroxybutyrate
soluble microbial product
co-culture
continuous photosynthetic hydrogen production
出版社: 環境工程學系所
引用: 經濟部能源產業技術白皮書,2010。 王炳南。2005。厭氧醱酵產氫與光合產氫之反應槽串聯可行性評估。碩士論文。國立中興大學。台中。 林瑤玓。2002。紫色不含硫光合作用細菌於連續流產氫之研究。碩士論文。國立中興大學。台中。 林鈺傑。2008。紫色不含硫光合菌結合不同生物系統產生氫氣之研究。碩士論文。國立中興大學。台中。 洪國展。2004。光合產氫之程序組合及應用。碩士論文。國立中興大學。台中。 涂良君。1999。產氫光合作用細菌之分離與篩選。碩士論文。國立中興大學。台中。 郁揆民。2003。紫色不含硫光合作用細菌產氫限制因子之研究。碩士論文。國立中興大學。台中。 陳怡傑。2009。以厭氧流體化床進行廚餘過篩液及狼尾草之氫醱酵程序研究。碩士論文。國立成功大學。台南。 蕭景庭。2000。產氫光合作用細菌之生理特性研究。碩士論文。國立中興大學。台中。 Akkerman, I., Janssen, M., Rocha, J., Wijffels. 2002. Photobiological hydrogen production: photochemical efficiency and bioreactor design. International Journal of Hydrogen Energy. 27(11-12): 1195-1208. Akkerman, I., Janssen, M. G. J., Rocha, J. M. S., Reith, J. H., Wijffels, R. H. 2003. Photobiological hydrogen production: photochemical efficiency and bioreactor design. In Bio-methane and Bio-hydrogen. Reith, J.H., Wijffels, R.H., Barten, H. eds. Dutch Biological Hydrogen Foundation. Petten. The Netherlands. Chaper 6. 124-145. Akopiants, K., Florova, G., Li, C., Reynolds, K. A. 2006. Multiple pathways for acetate assimilation in Streptomyces cinnamonensis. Journal of industrial microbiology and biotechnology. 33(2): 141-150. Anthony, C. 2011. How half a century of research was required to understand bacterial growth on C1 and C2 compounds; the story of the serine cycle and the ethylmalonyl-CoA pathway. Science Progress. 94(2): 109-137. Alber, B. E., Spanheimer, R., Ebenau-Jehle, C., Fuchs, G. 2006. Study of an alternate glyoxylate cycle for acetate assimilation by Rhodobacter sphaeroides. Molecular Microbiology. 61(2): 297-309. Argun, H., Fargi, F., Kapdan, I. K. 2008. Light fermentation of dark fermentation effluent for bio-hydrogen production by different Rhodobacter species at different initial volatile fatty acid(VFA) concentrations. International Journal of Hydrogen Energy. 33(24): 7405-7412. Basak, N., Das, D. 2007. The prospect of purple non-sulfur (PNS) photosynthetic bacteria for hydrogen production: the present state of the art. World Journal of Microbiology and Biotechnology. 23(1):31-42. Berg, I. A., Ivanovsky, R. N. 2009. Enzymes of the Citramalate Cycle in Rhodospirillum rubrum. Microbiology. 78(1): 16-24. Carlozzi, P., Sacchi, A. 2001. Biomass production and studies on Rhodopseudomonas palustris grown in an outdoor, temperature controlled, underwater tubular photobioreactor. Journal of Biotechnology. 88(3): 239-249. Çetin, D., Gündüz, U., Eroğlu, I., Yücel, M., Türker, L. 2006. Poly-β-hydroxybutyrate accumulation and releasing by hydrogen producing bacteria, Rhodobacter sphaeroides O.U.001. A transmission electron microscopic study. African Journal of Biotechnology. 5(22): 2069-2072. Choi, J. I., Lee, S. Y., Han, K. 1998. Cloning of the Alcaligenes latus polyhydroxyalkanoate biosynthesis genes and use of these genes for enhanced production of poly(3-hydroxybutyrate) in Escherichia coli. Applied and Environmental Microbiology. 64(12): 4897-4903. Das, D., Veziroğlu, T. N. 2001. Hydrogen production by biology processes: a survey of literature. International Journal of Hydrogen Energy. 26: 13-28. Dixon, R. Kahn, D. 2004. Genetic regulation of biological nitrogen fixation. Nature reviews microbiology. 2(8): 621-631. Eley, J. H., Konbloch, K., Han, T. W. 1979. Effect of growth condition on enzymes of the citric acid cycle and the glyoxylate cycle in the photosynthetic bacterium Rhodopseudomonas palustris. Antonie van Leeuwenhoek. 45(4): 532-529. Erb, T. J., Berg, I. A., Brecht, V., Müller, M., Fuchs, G., Alber, B. E. 2007. Synthesis of C5-dicarboxylic acids from C2-units involving crotonyl-CoA carboxylase/reductase: The ethylmalonyl-CoA pathway. Proceedings of the National Academy of Sciences of the United States of America. 104(25): 10631-10636. Filatova, L. V., Berg, I. A., Krasil’nikova, E. N., Tsygankov, A. A., Laurinavichene, T. V., Ivanovsky, R. N. 2005. A Study of the Mechanism of Acetate Assimilation in Purple Nonsulfur Bacteria Lacking the Glyoxylate Shunt: Acetate Assimilation in Rhodobacter sphaeroides. Microbiology. 74(3): 265-269. Fascetti, E., D''addario, E., Todini, O., Robertiello, A. 1998. Photosynthetic hydrogen evolution with volatile organic acids derived from the fermentation of source selected municipal solid waste. International Journal of Hydrogen Energy. 23(9):753-760. Franchi, E., Tosi, C., Scolla, G., Penna, G. D., Rodriguez, F., Pedroni, P. M. 2004. Metabolically Engineered Rhodobacter sphaeroides RV strains for Improved Biohydrogen Photoproduction Combined with Disposal of Food Wastes. Marine Biotechnology. 6(6): 552-565. Fuller, R. C. 2004. Polyesters and Photosynthetic Bacteria. In Anoxygenic Photosynthetic Bacteria. Blankenship, R. E., Madigan, M. T., Bauer, C. E. eds. Kluwer academic publishers. The Netherlands. Chaper 60. 1245-1256. Gasslmaier, B., Krell, C. M., Seebach, S., Holler, E. 2000. Synthetic substrates and inhibitors of β-poly(L-malate)-hydrolase (polymalatase). European Journal of Biochemistry. 267(16): 5101-5105. Greenberg, A. E., Trussel, R. R., Cleseeri, L. S. 1985. Standard Methods for the Examination of Waste and Wastewater. American Public Health Association. Washington, DC. 16th ed. Gogotov, I. N. 1986. Hydrogenases of phototrophicmic roorganisms. Biochimie. 68:181-7. Golomysova, A. N., Ivanov, P. S. 2011. Investigation of the anaerobic metabolism of Rhodobacter capsulatus by means of a flux model. Biophysics. 56(1): 74-85. Hallenbeck, P. C. 1983. Nitrogenase reduction by electron carriers: Influence of redox potential on activity and ATP/2e- ratio. Archives of Biochemistry and Biophysics. 200(2): 657-660. Haywood, G. W., Anderson, A.J., Chu, L., Dawes, E. A. 1988. The role of NADH- and NADHP-linked acetoacetyl-CoA reductases in the poly-3-hydroxybutyrate synthesizing organism Alcaligenes eutrophus. FEMS Microbiology Letters. 52(3): 259-264. Henderson, R. A., Jones, C. W. 1997. Poly-3-hydroxybutyrate production by washed cells of Alcaligenes eutrophus; purification, characterisation and potential regulatory role of citrate synthase. Archives of Microbiology. 168(6): 486-492. Hillmer, P., Gest, H. 1977. H2 metabolism in the photosynthetic bacterium Rhodopseudomonas capsulate - H2 production by growing culture. Journal of Bacteriology. 129(2): 724-731. Hustede, E., Steinbüchel, A., Schlegel, H. G. 1993. Relationship between the photoproduction of hydrogen and the accumulation of PHB in non-sulphur purple bacteria. Applied Microbiology and Biotechnology. 39(1): 87-93. Jendrossek, D., Handrick, R. 2002. Microbial degradation of polyhydroxyalkanoates. Annual Review of Microbiology. 56: 403-432. Jouanneau, Y., Wong, B., Vignais, P. M. 1985. Stimulation by light of nitrogenase synthesis in cells of Rhodopseudomonas capsulata growing in N-limited continuous cultures. Biochimica et Biophysica Acta (BBA) – Bioenergetics. 808(1): 149-155. Jung, Y. M., Park, J. S., Lee, Y. H. 2000. Metabolic engineering of Alcaligenes eutrophus through the transformation of cloned phbCAB genes for the investigation of the regulatory mechanism of polyhydroxyalkanoate biosynthesis. Enzyme and Microbial Technology. 26(2-4): 201-208. Kapdan, I. K., Kargi, F. 2006. Biohydrogen production from waste materials. Enzyme and Microbial Technology. 38: 569-582. Kars, G., Gündüz, U., Yücel, M., Rakhely, G., Kovacs, K. L., Eroğlu, I. 2009. Evaluation of hydrogen production by Rhodobacter sphaeroides O.U.001 and its hupSL deficient mutant using acetate and malate as carbon sources. International Journal of Hydrogen Energy. 34(5): 2184-2190. Kessler , B., Witholt , B. 2001. Factors involved in the regulatory network of polyhydroxyalkanoate metabolism. Journal of Biotechnology. 86(2): 97-104. Khatipov, E., Miyake, M., Miyake, M., Asada, Y. 1998. Accumulation of poly-β-hydroxybutyrate by Rhodobacter sphaeroides on various carbon and nitrogen substrates. FEMS Microbiology Letters. 162(1): 39-45. Kim, M. S., Baek, J. S., Lee, J. K. 2006. Comparison of H2 accumulation by Rhodobacter sphaeroides KD131 and its uptake hydrogenase and PHB synthase deficient mutant. International Journal of Hydrogen Energy. 31(1): 121-127. Kim, M. S., Kim, D. H., Son, H. N., Ten, L. N., Lee, J. K. 2011. Enhancing photo-fermentative hydrogen production by Rhodobacter sphaeroides KD131 and its PHB synthase deleted-mutant from acetate and butyrate. International Journal of Hydrogen Energy. Article in Press, Corrected Proof. Koku, H., Eróglu, I., Gunduz, U., Yucel, M., Türker, L. 2002. Aspect of the metabolism of hydrogen production by Rhodobacter sphaeroides. International Journal of Hydrogen Energy. 27(11-12): 1315-1329. Kondratieva, E. N. 1976. Phototrophic microorganisms as source of hydrogenase formation. In Microbial Energy Conversion. Schlegel, H. G., Barnea, J. eds. Erich Goltze KG, Göttingen. 205-216. Kranz, R. G., Gabbert, K. K., Locke, T. A., Madigan, M. T. 1997. Polyhydroxyalkanoate production in Rhodobacter capsulatus: Genes, mutants, expression, and physiology. Applied and Environmental Microbiology. 63(8): 3003-3009. Lageveen, R. G., Huisman, G. W., Preusting, H., Ketelaar, P., Eggink, G., Witholt, B. 1988. Formation of polyesters by Pseudomonas oleovorans: Effect of substrates on formation and composition of poly-(R)-3-hydroxyalkanoates and poly-(R)-3-hydroxyalkenoates. Applied and Environmental Microbiology. 54(12): 2924-2932. Lay, J. J., Lee, Y. J., Noike, T. 1999. Feasibility of biological hydrogen production from organic fraction of municipal solid waste. Water Research. 33(11): 2579-2586. Lee, I. Y., Kim, M. K., Chang, H. N., Park, Y. H. 1995. Regulation of poly-β-hydroxybutyrate biosynthesis by nicotinamide nucleotide in Alcaligenes eutrophus. FEMS Microbiology Letters. 131(1): 35-39. Lemoigne, M. 1926. Products of dehydration and of polymerization of 3-hydroxybutyric acid. Bulletin de la Société de chimie biologique. 8: 770-782. Li, C. L., Fang, H. H. P. 2007. Fermentative hydrogen production from wastewater and solid wastes by mixed cultures. Critical Reviews in Environmental Science and Technology. 37(1): 1-39. Li, R. Y., Fang, H. H. P. 2008. Hydrogen production characteristics of photoheterotrophic Rubrivivax gelatinosus L31. International Journal of Hydrogen Energy. 33(3): 974-980. Luengo, J. M., García, B., Sandoval, A., Naharro, G., Olivera, E. R. 2003. Bioplastics from microorganisms. Current Opinion in Microbiology. 6(3): 251-260. Macler, B. A., Pelroy, R. A., Bassham, J. A. 1978. Hydrogen formation in nearly stoichiometric amounts by a Rhodopseudomonas sphaeroides mutant. Journal of Bacteriology. 138(2):446-452. Madigan, M. T., Martinko, J. M., Parker, J. 2006. Brock biology of Microorganisms. 11th edition. Prentice Hall/ Pearson Education, Inc., Upper Saddle River, NJ. Madison, L. L., Huisman, G. W. 1999. Metabolic engineering of poly(3-hydroxyalkanoates): From DNA to Plastic. Microbiology and Molecular Biology Reviews. 63(1): 21-53. Mckinlay, J. B., Harwood, C. S. 2010. Carbon dioxide fixation as a central redox cofactor recycling mechanism in bacteria. Proceedings of the National Academy of Sciences of the United States of America.. 107(26): 11669-11675. Melnicki, M. R., Eroglu, E., Melis, A. 2009. Changes in hydrogen production and polymer accumulation upon sulfur-deprivation in purple photosynthetic bacteria. International Journal of Hydrogen Energy. 34(15): 6157-6170. Meyer, J., Kelley, B. C., Vignais, P. M. 1978. Effect of light on nitrogenase function and synthesis in Rhodopseudomonas capsulata. The Journal of Bacteriology. 136(1):201-8. Miyake, J. 1990. Application of photosynthetic systems for energy conversion. In Hydrogen energy progress VII. Veziroğlu, T. N., Takashashi, P. K. eds. Proceedings 8th WHEC, Hawaii. New York: Pergamon Press. 755-764. Miyake, J. 1998. The science of biohydrogen: an energetic view. In BioHydrogen. Zaborsky, O. R. eds. Plenum Press. New York. 7-18. Nair, L. S., Laurencin, C. T. 2007. Biodegradable polymers as biomaterials. Progress in Polymer Science. 32(8-9): 762-798. Nielsen, A. M., Rampsch, B. J., Sojka, G. A. 1979. Regulation of isocitrate lyase in a mutant of Rhodopesudomonas capsulata adapted to growth on acetate. Archives of Microbiology. 120(1): 43-46. Ohta, Y., Frank, Mitsui, A. 1981. Hydrogen production by marine photosynthetic bacteria: Effect of environment factors and substrate specificity on growth of a hydrogen-producing marine photosynthetic bacterium, Chromatium sp. Miami PBS 1071. Journal of Hydrogen Energy. 6(5):451-460. Okubo, Y., Hiraishi, A. 2007. Population Dynamics and Acetate Utilization Kinetics of Two Different Species of Phototrophic Purple Nonsulfur Bacteria in a Continuous Co-culture System. Microbes and Environments. 22(1): 82-87. Ostle, A. G., Holt, J. G. 1982. Nile blue A as a fluorescent stain for poly-β-hydroxybutyrate. Applied and Environmental Microbiology. 44(1): 238-241. Ozmihci, S., Kargi, F. 2010. Comparison of different mixed cultures for bio-hydrogen production from ground wheat starch by combined dark and light fermentation. Journal of Industrial Microbiology and Biotechnology. 37(4): 341-347. Park, J. S., Lee, Y. H. 1996. Metabolic characteristics of isocitrate dehydrogenase leaky mutant of Alcaligenes eutrophus and its utilization for poly-β-hydroxybutyrate production. Journal of Fermentation and Bioengineering. 81(3): 197-205. Pfennig, N. 1978. Rhodocyclus purpureus gen. nov. and sp. Nov., a ring-shaped vitamin B12-requiring member of the family Rhodospirillaceae. International Journal of Systematic Bacteriology. 28: 283-288. Pieper-Fürst, U., Madkour, M. H., Mayer, F., Steinbüchel, A. 1995. Identification of the region of a 14-kilodalton protein of Rhodococcus ruber that is responsible for the binding of this phasin to polyhydroxyalkanoic acid granules. The Journal of Bacteriology. 177(9): 2513-2523. Prieto, M. A., de Eugenio, L. I., Galán, B., Luengo, J. M., Witholt, B. 2007. Synthesis and degradation of polyhydroxyalkanoates. In Pseudomonas. vol. 5“Pseudomonas: a Model System in Biology.” Ramos, J. L., Filloux, A. eds. Springer. The Netherlands. Chaper 14. 397-428. Rehm, B. H. A. 2010. Bacterial polymers: biosynthesis, modifications and applications. Nature Reviews Microbiology. 8: 578-592. Sasikala, K., Ramana, C. V., Rao, P. R., Subrahmanyam, M. 1990. Effect of gas phase on the photoproduction of hydrogen and substrate conversion efficiency on the photosynthetic bacterium Rhodobacter sphaeroides O.U. 001. International Journal of Hydrogen Energy. 154(11): 795-797. Sasikala, K., Ramana, C. V., Rao, P. R. 1991. Environmental regulation for optimal biomass yield and photoproduction of hydrogen by Rhodobacter sphaeroides O.U. 001. International Journal of Hydrogen Energy. 16(9): 597-601. Sasikala, C. H., Ramana, C. H. V., Raghuveer, P. 1995. Regulation of simultaneous hydrogen photoproduction during growth by pH and glutamate in Rhodobacter sphaeroides O.U.001. International Journal of Hydrogen Energy. 20(2): 123-126. Satoh, H., Ramey, W. D., Koch, F. A., Oldham, W. K., Mino, T., Matsuo, T. 1996. Anaerobic substrate uptake by the enhanced biological phosphorus removal activated sludge treating real sewage. Water Science and Technology. 34(1-2): 9-16. Schirmer, A., Jendrossek, D., Schlegel, H. G. 1993. Degradation of poly(3-hydroxyoctanoic acid) [P(3HO)] by bacteria: purification and properties of a P(3HO) depolymerase from Pseudomonas fluorescens GK13. Applied and Environmental Microbiology. 59(4): 1220-1227. Steinbüchel, A., Hein, S. 2001, Biochemical and molecular basis of microbial synthesis of polyhydroxyalkanoates in microorganisms. Advances in Biochemical Engineering/Biotechnology. 71: 81-123. Steinbüchel, A., Lütke-Eversloh, T. 2003. Metabolic engineering and pathway construction for biotechnological production of relevant polyhydroxyalkanoates in microorganisms. Biochemical Engineering Journal. 16(2): 81-96. Stevens, P., Vertoghen, C., Vos, P. D., Lay, J. D. 1984. The effect of temperature and light intensity on hydrogen gas production by different Rhodopseudomonas capsulate strains. Bacteriology. 6: 277-282. Sudesh, K., Abe, H., Doi, Y. 2000. Synthesis, structure and properties of polyhydroxyalkanoates: biological polyesters. Progress in Polymer Science. 25(10): 1503-1555. Tao, Y., Chen, Y., Wu, Y., He, Y., Zhou, Z. 2007. High hydrogen yield from a two-step process of dark-and photo-fermentation of sucrose. International Journal of Hydrogen Energy. 32(2): 200-206. Uyar, B., Eroglu, I., Yucel, M., Gunduz, U., Turker, L. 2007. Effect of light intensity,wavelength and illumination protocol on hydrogen production in photobioreactors. International Journal of Hydrogen Energy. 32: 4670-4677. Verlinden, R. A. J., Hill, D. J., Kenward, M. A., Williams, C. D., Radecka, I. 2007. Bacterial synthesis of biodegradable polyhydroxyalkanoates. Journal of Applied Microbiology. 102(6): 1437-1449. Vincenzini, M., Materassi, R., Tredici, M. R., Florenzano, G. 1982. Hydrogen production by immobilized cells - I. Light dependent dissimilation of organic substances by Rhodopseudomonas palustris. International Journal of Hydrogen Energy. 7(3): 231-236. Vincenzini, M., Marchini, A., Ena, A., Philippis, R. D. 1997. H2 and poly-β-hydroxybutyrate, two alternative chemicals from purple non sulfur bacteria. Biotechnology Letters. 19(8): 759-762. Yakunin, A. F., Tsygankov, A. A., Troshina O.Y., Gogotov, I. N. 1991. Growth and nitrogenase activity of continuous cultures of the purple bacteria Rhodobacter sphaeroides depending on the presence of Mo, V and W in the medium. Mikrobiologiâ. 60(6): 41-46. Yang, C. F., Lee, C. M. 2010. Enhancement of photohydrogen production using phbC deficient mutant Rhodopseudomonas palustris strain M23. Bioresource Technology. 102(9): 5418-5424. Yilmaz, L. S., Kontur, W. S., Sanders, A. P., Sohmen, U., Donohue, T. J., Noguera, D. R. 2010. Electron partitioning during light- and nutrient-powered hydrogen production by Rhodobacter sphaeroides. BioEnergy Research. 3(1): 55-66. Yiğit, D. Ö., Gündüz, U., Türker, L., Yücel, M., Eroğlu, İ. 1999. Identification of by-products in hydrogen producing bacteria; Rhodobacter sphaeroides O.U. 001 grown in the waste water of a sugar refinery. Journal of Biotechnology. 70(1-3): 125-131. Zorin, N. A. 1986. Redox properties and active center of phototrophic bacteria hydrogenases. Biochimie. 68(1): 97-101. Zürrer, H., Bachhofen, R. 1982. Aspects of growth and hydrogen production of the photosynthetic bacterium Rhodospirillum rubrum in continuous culture. Biomass. 2(3): 165-174.
摘要: 「氫氣」是一種乾淨的能源,其熱值含量高(122 kJ/g),且燃燒後無二氧化碳排放之問題,而以紫色不含硫光合菌於光異營條件下進行生物產氫為具潛力的方式之一。近年來,許多研究嘗試提升紫色不含硫光合菌之產氫效率,poly-β-hydroxybutyrate (PHB)為胞內聚合物,其合成可能會與產氫競爭能量及電子,且產氫多半是發生在微生物進入生長靜止期時,因此,紫色不含硫光合菌於產氫、細胞生長及PHB累積之能量分配是相當重要的。此外,暗醱酵出流水含有數種有機酸,適合做為紫色不含硫光合菌產氫之基質,如何有效地以生物方式進行處理及能源回收也為重要的課題。 本篇研究便希望藉由Rhodopseudomonas palustris WP3-5之PHB synthase-deficient mutant Rps. palustris M23以批次試驗去瞭解不同條件下能量分布的情形。另一方面,也嘗試分離、純化紫色不含硫光合菌,測試其利用各種有機酸產氫之情形,評估與Rps. palustris WP3-5共培養能否提升產氫及廢水處理之效率,並以光合反應槽培養達連續產氫。 實驗結果發現,Rps. palustris WP3-5於生長過程中伴隨著PHB之累積,PHB含量累積至最大值後,有被微生物再利用之情形,可供給Rps. palustris WP3-5額外的碳源及能源。當氮源耗盡時,細胞便會停止生長,此時細胞會將多餘之能量進行產氫。碳源濃度較低時,由於耗盡較快,Rps. palustris WP3-5無多餘之碳源合成PHB,因此,與突變株Rps. palustris M23產氫情形無明顯差異。當碳源濃度提高時,Rps. palustris WP3-5產氫量較多,基質轉換率也較高,突變株Rps. palustris M23產氫情形無明顯提升,其他不同條件之實驗也可觀察到相同結果,但以蘋果酸為碳源時,Rps. palustris WP3-5利用速率慢,產氫量較低,且不累積PHB。 不同紫色不含硫光合菌對同種類碳源之代謝途徑可能有差異,導致PHB累積量皆不相同。Rps. palustris WP3-5進入生長靜止期時會將大部份能量用於合成氫氣,而其PHB累積量不高,佔微生物獲得能量5%以下,可能為導致氫氣合成與PHB累積競爭還原能關係不明顯之原因。實驗也發現,SMP合成可能也會競爭還原能,PHB synthase-deficient mutant重新分配能量時不會完全分配至氫氣合成。 本實驗從環境中分離純化出數株紫色不含硫光合菌,經產氣測試及產氫試驗,各菌株對於各種有機酸利用情形無明顯差異,共培養未改善產氫量,可能有競爭能量之情形。而Rps. palustris WP3-5以新設計之光合反應槽進行連續流操作,以適合紫色不含硫光合菌產氫之鎢絲燈為光源,且水力停留時間由2天提高至3天,以初期產氣率較佳,達317.6 mL gas/L culture-day,試程中MLSS皆在220-360 mg/L,改變操作策略無法改善細胞生長量,因操作反應槽需考量光穿透率及因高細胞濃度造成之光遮蔽問題,此反應槽連續流產氫結果不如預期,可能不適合紫色不含硫光合菌進行連續流產氫。
Hydrogen is a clean energy that generates only water upon burning and it releases high amount of energy (122 kJ/g) by combustion. Photo-biological hydrogen production by purple non-sulfur photosynthetic bacteria is one of promising methods. Purple non-sulfur photosynthetic bacteria can produce hydrogen by nitrogenase under photo-heterotrophic condition. In recent years, many researchers make efforts to improve its efficiency of biological hydrogen production. Poly-β-hydroxybutyrate (PHB) is accumulated as carbon and energy storage material under unbalanced growth. The synthesis of PHB may compete with hydrogen production for energy and reducing power. In addition, maximum hydrogen production rate usually takes place during early stationary phase. Therefore, energy distribution between cell growth, hydrogen production and PHB accumulation is an important issue. Besides, dark fermentation effluent containing various volatile organic acids is suitable for purple non-sulfur photosynthetic bacteria to produce hydrogen. To treat wastewater and recycle clean energy efficiently by biological method are also essential. In this research, Rhodopseudomonas palustris WP3-5 and its PHB synthase-deficient mutant Rps. palustris M23 were used in batch experiments to explore energy distribution between cell growth, hydrogen production and PHB accumulation under different culture conditions. This study also tried to isolate purple non-sulfur photosynthetic bacteria from environment, and tested their ability to produce hydrogen using different volatile organic acids. Then, hydrogen production and treatment efficiency in co-culture system with Rps. palustris WP3-5 were evaluated and continuous photosynthetic hydrogen production was performed. From the results, Rps. palustris WP3-5 accumulated PHB in growth phase, and PHB could be utilized as another carbon and energy source when its content reached maximum value. Microorganism ceased growing when nitrogen source was exhausted, and used excess energy to produce more hydrogen in stationary phase. Rps. palustris WP3-5 could not synthesize PHB under low concentration of carbon source because substrate was degraded rapidly, and there were no difference of hydrogen production between wild-type strain and mutant Rps. palustris M23. When concentration of carbon source was three times higher, cumulated hydrogen volume and substrate conversion efficiency of Rps. palustris WP3-5 were better than Rps. palustris M23. This result coincided with other experiments in different culture conditions. But when using malate as carbon source, Rps. palustris WP3-5 had lower substrate degrading rate and cumulated hydrogen volume, and PHB was not accumulated. Different species of purple non-sulfur photosynthetic bacteria had different substrate assimilation pathway when using same carbon source and accumulated PHB content might be different. Rps. palustris WP3-5 used most energy to produce hydrogen in stationary phase and PHB content was below 10% cell dry weight, accounting for less than 5% of the substrate electrons utilized. This portion of energy might partially redistribute to synthesize soluble microbial product (SMP). Therefore, the competition relationship between hydrogen production and PHB accumulation was insignificant. In second part, several purple non-sulfur photosynthetic bacteria were isolated, and there were no obvious difference in utilizing volatile organic acids to produce hydrogen between these strains. Hydrogen production was not improved and competition for energy might exist in co-culture system. Additionally, new designed photo-bioreactor was used for Rps. palustris WP3-5 to produce hydrogen continuously when using tungsten filament lamp as light source. Operating hydraulic retention time from 2 day to 3 day, gas production rate was highest in initial period, up to 317.6 mL gas/L culture-day. MLSS was always between 220 and 360 mg/L, and MLSS wasn't enhanced by changing operational strategies. Thinking of light transmission efficiency and problem of light shielding effect caused by high biomass concentration, experimental results was not as expected, and this photo-bioreactor might not favorable for continuous hydrogen production by purple non-sulfur photosynthetic bacteria.
URI: http://hdl.handle.net/11455/5020
其他識別: U0005-2007201118365000
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-2007201118365000
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