Please use this identifier to cite or link to this item:
標題: 以Bacillus-Yeast共培養策略建構纖維素乙醇生產系統
Construction of cellulosic ethanol production process via Bacillus-yeast co-culturing strategy
作者: 何政育
Ho, Cheng-Yu
關鍵字: artificial cellulosome
cellulosic ethanol
出版社: 生命科學系所
引用: Abdel-Banat B M, Nonklang S, Hoshida H, Akada R. Random and targeted gene integrations through the control of non-homologous end joining in the yeast Kluyveromyces marxianus. Yeast 2010; 27: 29-39. Abouzied MM, Reddy CA. Direct fermentation of potato starch to ethanol by cocultures of Aspergillus niger and Saccharomyces cerevisiae. Appl Environ Microbiol 1986; 52:1055-59. Aminov RI, N. Golovchenko P, Ohmiya K. Expression of a celE gene from Clostridium thermocellum in Bacillus. J Ferment Bioeng 1995; 79:530-537. Arantes and Saddler. Access to cellulose limits the efficiency of enzymatic hydrolysis: the role of amorphogenesis. Biotechnol Biofuels. 2010; 23:3-4. Aslanidis C, de Jong PJ. Ligation-independent cloning of PCR products (LIC-PCR). Nucleic Acids Res 1990;18: 6069-6074. Banat IM, Nigam P, Singh D, Marchant R, McHale AP. Ethanol production at elevated temperatures and alcohol concentrations: part I—yeasts in general. World J Microbiol Biotechnol 1998; 14:809-821. Banat IM, Nigam P, Marchant R. Isolation of thermotolerant, fermentative yeasts growing at 52 °C and producing ethanol at 45°C and 50°C. World J Microbiol Biotechnol 1992; 8:259-63. Banat IM, Singh D, Marchant R. The use of a thermotolerant fermentative Kluyveromyces marxianus IMB3 yeast strain for ethanol production. Acta Biotechnol 2004; 2:215-23. Bajpai P, Margaritis A. Ethanol inhibition kinetics of Kluyveromyces marxianus grown on Jerusalem artichoke juice. Appl Environ Microbiol 1982; 44:1325-29. Bayer EA, Shimon LJ, Shoham Y, Lamed R. Cellulosomes-structure and ultrastructure. J Struct Biol. 1998; 124:221-234 Bayer EA, Lamed R, Himmel ME. The potential of cellulases and cellulosomes for cellulosic waste management. Curr Opin Biotechnol 2007; 18:237-45. Beukes N, Pletschke BI. Effect of sulfur-containing compounds on Bacillus cellulosome-associated ''CMCase'' and ''Avicelase'' activities. FEMS Microbiol Lett 2006; 264: 226-31. Berry DR, Brown C. Physiology of yeast growth. In: Berry DR, Russel I, Steward GG (eds) Yeast biotechnology. Allen and Uwin London 1987; 159-99. Belem MA, Lee BH. Production of bioingredients from Kluyveromyces marxianus grown on whey: an alternative. Crit Rev Food Sci Nutr 1998; 38: 565-98. Behera ES, Mohanty RC, Ray RC. Comparative study of bio-ethanol production from mahula (Madhuca latifolia L.) flowers by Saccharomyces cerevisiae and Zymomonas mobilis. Appl Energy 2010; 7: 2352-2355. Bitinaite J, Rubino M, Varma KH, Schildkraut I, Vaisvila R, Vaiskunaite R. USER friendly DNA engineering and cloning method by uracil excision. Nucleic Acids Res 2007; 35: 1992-2002. Bootten TJ, Joblin KN, McArdle BH, Harris PJ. Degradation of lignified secondary cell walls of lucerne (Medicago sativa L.) by rumen fungi growing in methanogenic co-culture. J Appl Microbiol 2011; 111: 1086-1096. Cha J, Matsuoka S, Chan H, Yukawa H, Inui M, Doi RH. Effect of multiple copies of cohesins on cellulase and hemicellulase activities of Clostridium cellulovorans mini-cellulosomes. J Microbiol Biotechnol 2007; 17:1782-1788. Caspi J, Irwin D, Lamed R, Li Y, Fierobe HP, Wilson DB, Bayer EA. Conversion of Thermobifida fusca free exoglucanases into cellulosomal components: comparative impact on cellulose-degrading activity. J Biotechnol Jul 2008; 135: 351-357. Chapelle FH, O''Neill K, Bradley PM, Methe BA, Ciufo SA, Knobel LL, Lovley DR. A hydrogen-based subsurface microbial community dominated by methanogens. Nature 2002; 415: 312-315. Chang JJ, Chou CH, Ho CY, Chen WE, Lay JJ, Huang CC. Syntrophic co- culture of aerobic Bacillus and anaerobic Clostridium for bio-fuels and bio-hydrogen production. Int J Hydrogen Energy 2008; 33:5137-5146. Chang ACC, Tu YH, Huang MH, Lay CH, Lin CY. Hydrogen production by the anaerobic fermentation from acid hydrolyzed rice straw hydrolysate. Int J Hydrogen Energy 2011; 21: 14280-14288. Chang JJ, Lin JJ, Ho CY, Chin WC, Huang CC. Establishment of rumen-mimic bacterial consortia: A functional union for bio-hydrogen production from cellulosic bioresource. Int J Hydrogen Energy 2010; 35: 13399-13406. Chang JJ, Ho CY, Ho FJ, Tsai TY, Ke HM, Wang Christine HT, Chen HL, Shih MC, Huang CC, Li WH. Synthetic Biological Technique for Yeast Genome Engineering: Promoter-based Gene Assembly and Simultaneous Overexpression Method. Bioresour Technol 2012. (submitted) Chen HL, Lu MY, Chen YC, Chang JJ, Wang HT, Wang TY, Ruan SK, Wang TY, Hung KY, Cho HY, Ke HM, Lin WT, Shih MC, Li WH.A highly efficient β-glucosidase from the calf lumen fungus Neocallimastix patriciarum W5. Bio-resource technology Bioresour Technol 2012. (submitted) Cho HY, Yukawa H, Inui M, Doi RH, Wong SL. Production of minicellulosomes from Clostridium cellulovorans in Bacillus subtilis WB800. Appl Environ Microbiol 2004; 70:5704-5707. Cotter JL, Chinn MS, Grunden AM. Ethanol and acetate production by Clostridium ljungdahlii and Clostridium autoethanogenum using resting cells. Bioprocess Biosyst Eng 2009; 32:369-380. Cocolin L, Bisson LF, Millsa DA. Direct profiling of the yeast dynamics in wine fermentations. FEMS Microbiol Lett 2000; 189: 81-7. Colussi PA & Taron CH. Kluyveromyces lactis LAC4 Promoter Variants That Lack Function in Bacteria but Retain Full Function in K. lactis. Appl Environ Microbiol 2005; 71:7092-8. Coughlan MP. The properties of fungal and bacterial cellulases with comment on their production and application. Biotechnology and genetic engineering reviews Newcastle-upon-Tyne: InterscienceRussell GE 1985; 3:37-109. Daianova L, Dotzauer E, Thorin E, Yan J. Evaluation of a regional bioenergy system with local production of biofuel for transportation, integrated with a CHP plant. Appl Energy 2012; 92: 739-749. Demain AL, Newcomb M, Wu JH. Cellulase, Clostridia, and ethanol. Microbiol Mol Biol Rev 2005; 69:124-54. Diep NQ, Fujimoto S, Minowa T, Sakanishi K, Nakagoshi N. Estimation of the potential of rice straw for ethanol production and the optimum facility size for different regions in Vietnam. Appl Energy 2012; Available online 21 January. Dimitrellou D, Tsaousi K, Kourkoutas Y, Panas P, Kanellaki M, Koutinas AA. Fermentation efficiency of thermally dried immobilized kefir on casein as starter culture. Process Biochem 2008; 43:1323-9. Dimitrellou D, Kourkoutas Y, Koutinas AA, Kanellaki M. Thermally-dried immobilized kefir on casein as starter culture in dried whey cheese production. Food Microbiol Dec 2009; 26:809-20. Dubois M., Gilles K.A., Hamilton J.K., Rebers P.A., Smith F., 1956. Colorimetric method for determination of sugars and related substances. Anal Chem 28, 350-356. Etschmann MM, Schrader J. An aqueous-organic two-phase bioprocess for efficient production of the natural aroma chemicals 2- phenylethanol and 2-phenylethylacetate with yeast. Appl Microbiol Biotechnol 2006; 71:440-3. Fabre CE, Blanc PJ, Goma G. Screening of yeasts producing 2-phenylethylalcohol. Biotechnol Tech 1997; 11: 523-525. Fierobe HP, Bayer EA, Tardif C, Czjzek M, Mechaly A, Belaich A, Lamed R, Shoham Y, Belaich JP. Degradation of cellulose substrates by cellulosome chimeras. Substrate targeting versus proximity of enzyme components. J Biol Chem 2002; 277: 49621-49630. Fierobe HP, Mingardon F, Mechaly A, Belaich A, Rincon MT, Pages S, Lamed R, Tardif C, Belaich JP, Bayer EA. Action of designer cellulosomes on homogeneous versus complex substrates: controlled incorporation of three distinct enzymes into a defined trifunctional scaffoldin. J Biol Chem 2005; 280:16325-16334. Fonseca GG, Heinzle E, Wittmann C, Gombert AK. The yeast Kluyveromyces marxianus and its biotechnological potential. Appl Microbiol Biotechnol 2008; 79: 339-54. Fujita, Y., Takahashi, S., Ueda, M., Tanaka, A., Okada, H., Morikawa, Y., Kawaguchi, T., Arai, M., Fukuda, H., Kondo, A., Direct and efficient production of ethanol from cellulosic material with a yeast strain displaying cellulolytic enzymes. Appl and Environ Microbiol. 2002; 68: 5136-5141. Garcez WS, Martins D, Garcez FR, Marques MR, Pereira AA, Oliveira LA, Rondon JN, Peruca AD. Effect of spores of saprophytic fungi on phytoalexin accumulation in seeds of frog-eye leaf spot and stem canker-resistant and -susceptible soybean (Glycine max L.) cultivars. J Agric Food Chem 2000; 48: 3662-5. Gibson DG, Benders GA, Axelrod KC, Zaveri J, Algire MA, Moodie M, Montague MG, Venter JC, Smith HO, Hutchison CA. One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome. Proc Natl Acad Sci U S A 2008; 105: 20404-20409. Gibson DG, Young L, Chuang RY, Venter JC, Hutchison CA, Smith HO. Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat methods 2009; 6: 343-345. Gouveia L, Oliveira AC. Microalgae as a raw material for biofuels production. J Ind Microbiol Biotechnol 2009; 36:269-274. Gold ND and Martin VJJ. Global View of the Clostridium thermocellum Cellulosome Revealed by Quantitative Proteomic Analysis. J Bacteriol 2007; 6787-6795 Hari Krishna S, Chowdary GV. Optimization of simultaneous saccharification and fermentation for the production of ethanol from lignocellulosic biomass. J Agric Food Chem 2000; 48:1971-1976. Hang YD, Woodams EE, Hang LE. Utilization of corn silage juice by Klyuveromyces marxianus. Bioresour Technol 2003; 86:305-307. Hartley JL, Temple GF, Brasch MA. DNA cloning using in vitro site-specific recombination. Genome Res 2000; 10: 1788-95. Hensing M, Vrouwenvelder H, Hellinga C, Baartmans R, Dijken H. Production of extracellular inulinase in high-cell-density fed-batch cultures of Kluyveromyces marxianus. Appl Microbiol Biotechnol 1994; 42:516-521. Hill J, Nelson E, Tilman D, Polasky S, Tiffany D. Environmental, economic, and energetic costs and benefits of biodiesel and ethanol biofuels. Proc Natl Acad Sci U.S.A. 2006; 103:11206-11210. Hiromi S. Studies of Metabolism in Tuberculous Lesions II. Further Studiesonthe N-acetyltyramine Formation by Mycobacterium Tuberculosis. Acta Tuberc Jpn 1956; 6:63-8. Hitzeman RA,Hagie FE, Levine HL, Goeddel DV, Ammerer G , Hall BD. Expression of a human gene for interferon in yeast. Nature 1981; 293:717-22. Holt RA, Warren R, Flibotte S, Missirlis PI, Smailus DE. Rebuilding microbial genomes. Bioessays 2007; 29:580-90. Ho CY, Chang JJ, Chin TY, Chen HL, Wang Christine HT, Ho FJ, Tsai TY, Wu SY, Wu YC, Li WH, Huang CC. A novel artificial cellulosomes is established for simultaneous saccharification-fermentation by dual microbe co-culturing system (American Society for Microbiology and the General Meeting Planning Committee, 2011) Ho CY, Chang JJ, Huang YR, Wu YC, Li WH, Shih MC, Huang CC. Isolation and characterization of a flavor production kefir yeast Kluyveromyces marxianus KY3: a potential strain for developing co-cultural consolidated bioprocess. Biomass & Bioenergy 2011. (submitted) Ho CY, Chang JJ, Lin JJ, Chin TY, Mathew Gincy M, Huang CC. Establishment of functional rumen bacterial consortia (FRBC) for simultaneous biohydrogen and bioethanol production from lignocellulose Int J Hydrogen Energy 2011; 6: 12168-76. Hong J, Wang Y, Kumagai H, Tamaki H. Construction of thermotolerant yeast expressing thermostable cellulase genes. J Biotechnol 2007; 130: 114-23. Hoshino E, Sasaki Y, Okazaki M, Nisizawa K, Kanda T. Mode of action of exo- and endo-type cellulases from Irpex lacteus in the hydrolysis of cellulose with different crystallinities. J Biochem 1993; 114:230-235. Itaya M, Tsuge K, Koizumi M, Fujita K. Combining two genomes in one cell: stable cloning of the Synechocystis PCC6803 genome in the Bacillus subtilis 168 genome. Proc Natl Acad Sci 2005; 102:15971-15976. Ivanova LA, Pavlova MV, Archakova LI. Therapy strategy in the treatment of patients with drug-resistant pulmonary tuberculosis. Probl Tuberk Bolezn Legk 2003; 5:14-6. Ingram LO, Aldrich HC, Borges AC, Causey TB, Martinez A, Morales F, Saleh A, Underwood SA, Yomano LP, York SW, Zaldivar J, Zhou S. Enteric bacterial catalysts for fuel ethanol production. Biotechnol Prog 1999; 15: 855-66. Johnson EA, Sakajoh M, Halliwell G, Madia A, Demain AL. Saccharification of Complex Cellulosic Substrates by the Cellulase System from Clostridium thermocellum. Appl Environ Microbiol 1982; 43:1125-1132. Katja Z, Andreja G. Mathematical modeling of ethanol production by mixed kefir grains yeast population as a function of temperature variations. Biochem Eng J 2009; (online publish). Kegel A, Martinez P, Carter SD, Astrom SU. Genome wide distribution of illegitimate recombination events in Kluyveromyces lactis. Nucleic Acids Res 2006; 34: 1633-1645. Klinke HB, Ahring BK, Schmidt AS, Thomsen AB. Characterization of degradation products from alkaline wet oxidation of wheat straw. Bioresour Technol 2002; 82:15-26. Klock HE, Koesema EJ, Knuth MW, Lesley SA. Combining the polymerase incomplete primer extension method for cloning and mutagenesis with microscreening to accelerate structural genomics efforts. Proteins. 2008; 71: 982-994. Kruus K, Wang WK, Ching J, Wu JH. Exoglucanase activities of the recombinant Clostridium thermocellum CelS, a major cellulosome component. J Bacteriol 1995;177:1641-1644. Kumar M, Goyal Y, Sarkar A, Gayen K. Comparative economic assessment of ABE fermentation based on cellulosic and non-cellulosic feedstocks. Appl Energy 2012; Available online Kurtzman CP, Robnett CJ. Phylogenetic relationships among yeasts of the Saccharomyces complex determined from multigene sequence analyses. FEMS Yeast Res 2003; 3:417-432. Lee J. Biological conversion of lignocellulosic biomass to ethanol. J Biotechnol 1997; 56:1-24. Li MZ, Elledge SJ. MAGIC, an in vivo genetic method for the rapid construction of recombinant DNA molecules. Nat Genet 2005; 37, 311-319. Li MZ, Elledge SJ. Harnessing homologous recombination in vitro to generate recombinant DNA via SLIC. Nat methods 2007; 4: 251-256. Liu Q, Li MZ, Leibham D, Cortez D, Elledge SJ. The univector plasmid-fusion system, a method for rapid construction of recombinant DNA without restriction enzymes. Current biology : CB. 1998; 8: 1300-1309. Longo MA, Sanroman MA. Production of food aroma compounds. Food Technol Biotechnol 2006; 44:335-53. Lopes TS, de Wijs IJ, Steenhauer SI, Verbakel J, Planta RJ. Factors affecting the mitotic stability of high-copy-number integration into the ribosomal DNA of Saccharomyces cerevisiae. Yeast 1996; 12: 467-477. Lynd LR, van Zyl WH, McBride JE, Laser M. Consolidated bioprocessing of cellulosic biomass: an update. Curr Opin Biotechnol 2005; 16:577-83. Marsischky G, LaBaer J. Many paths to many clones: a comparative look at high-throughput cloning methods. Genome Res 2004; 14: 2020-2028. Medeiros A, Pandey A, Christen P, Fontoura PSG, Freitas RJS, Soccol CR. Aroma compounds produced by Kluyveromyces marxianus in solid-state fermentation on packed bed column bioreactor. World J Microbiol Biotechnol 2001; 17:767-71. Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugars. Anal Chem 1959; 31:426-28. Mingardon F, Chanal A, Tardif C, Bayer EA, Fierobe HP. Exploration of new geometries in cellulosome-like chimeras. Appl Environ Microbiol 2007; 73:7138-49. Monika G, Dhamija SS, Gera R, Yadav BS. Isolation and genetic analysis of ethanol-sensitive mutants of thermotolerant Kluyveromyces marxianus. Biotechnology Techniques 2004; 11:343-6. Murashima K, Chen CL, Kosugi A, Tamaru Y, Doi RH, Wong SL. Heterologous production of Clostridium cellulovorans engB, using protease-deficient Bacillus subtilis, and preparation of active recombinant cellulosomes. J. Bacteriol. 2002; 184:76-81. Narendranath NV, Thomas KC, Ingledew WM. Effects of acetic acid and lactic acid on the growth of Saccharomyces cerevisiae in a minimal medium. J. Ind. Microbiol Biotechnol 2001; 26:171-7. Nielsen AT, Liu WT, Filipe C, Grady L Jr, Molin S, Stahl DA. Identification of a novel group of bacteria in sludge from a deteriorated biological phosphorus removal reactor. Appl Environ Microbiol 1999; 65:1251-8. Nisizawa K, Kanda T, Shikata S, Wakabayashi K. Mutarotation of hydrolysis products by different types of exo-cellulases from Trichoderma viride. J Biochem 1978; 83:1625-1630. Nishizaki T, Tsuge K, Itaya M, Doi N, Yanagawa H. Metabolic engineering of carotenoid biosynthesis in Escherichia coli by ordered gene assembly in Bacillus subtilis. Appl Environ Microbiol 2007 ; 73:1355-1361. Nofsinger GW, Bothast RJ. Ethanol production by Zymomonas mobilis and Saccharomyces uvarum on aflatoxin-contaminated and ammonia- detoxified com. Can J Microbiol 1981; 27:162-7. Nonklang S, Abdel-Banat BM, Cha-aim K, Moonjai N, Hoshida H, Limtong S, Yamada M, Akada R. High-temperature ethanol fermentation and transformation with linear DNA in the thermotolerant yeast Kluyveromyces marxianus DMKU3-1042. Appl Environ Microbiol 2008; 74: 7514-21. Pachuk CJ, Samuel M, Zurawski JA, Snyder L, Phillips P, Satishchandran C. Chain reaction cloning: a one-step method for directional ligation of multiple DNA fragments. Gene 2000; 243: 19-25. Pages S, Belaich A, Belaich JP, Morag E, Lamed R, Shoham Y, Bayer EA. Species-specificity of the cohesin-dockerin interaction between Clostridium thermocellum and Clostridium cellulolyticum: prediction of specificity determinants of the dockerin domain. Proteins. 1997; 29:517-527. Pintado M, Lopez da Silva J, Fernandes P, Malcato F, Hogg T. Microbiological and rheological studies on Portuguese kefir grains. Int J Food Sci Technol 1996; 31:15-26. Quan J, Tian J. Circular polymerase extension cloning of complex gene libraries and pathways. PloS one. 2009; 4: e6441. Rasmussen MA, Hespell RB, White BA, Bothast RJ. Inhibitory Effects of Methylcellulose on Cellulose Degradation by Ruminococcus flavefaciens. Appl Environ Microbiol1988; 54: 890-897. Robson LM, Chambliss GH. Characterization of the cellulolytic activity of a Bacillus isolate. . Appl Environ Microbiol1984; 47:1039-1046. Romanos MA, Scorer CA, Clare JJ. Foreign gene expression in yeast: a review. Yeast 1992; 8:423-88. Romero S, Merino E, Bolivar F, Gosset G, Martinez A. Metabolic engineering of Bacillus subtilis for ethanol production: lactate dehydrogenase plays a key role in fermentative metabolism. Appl Environ Microbiol 2007; 73:5190-5198. Saga K, Imou K, Yokoyama S, Minowa T, Net energy analysis of bioethanol production system from high-yield rice plant in Japan. Appl Energy 2010; 7: 2164-2168. Sabathe F, and Soucaille P. Characterization of the CipA scaffolding protein and in vivo production of a minicellulosome in Clostridium acetobutylicum. J. Bacteriol. 2003; 185:1092-1096. Saitou N and Nei M. The Neighbor-joining Method: A New Method for Reconstructing Phylogenetic Trees. Mol Biol Evol 1987; 4:406-25. Sanchez M, Iglesias FJ, Santamaria C, Dominguez A. Transformation of Kluyveromyces lactis by Electroporation. Appl Environ Microbiol. 1993 ; 59: 2087-2092. Shao Z, Zhao H. DNA assembler, an in vivo genetic method for rapid construction of biochemical pathways. Nucleic acids Res 2009; 37: e16. Schallmey M, Singh A, Ward OP. Developments in the use of Bacillus species for industrial production. Can J Microbiol 2004; 50:1-17. Schwarz WH. The cellulosome and cellulose degradation by anaerobic bacteria. Appl Microbiol Biotechnol 2001; 56:634-649. Shimizu H, Mizuguchi T, Tanaka E, Shioya S. Nisin production by a mixed-culture system consisting of Lactococcus lactis and Kluyveromyces marxianus. Appl Environ Microbiol 1999; 65:3134-3141. Shuldiner AR, Scott LA, Roth J. PCR-induced (ligase-free) subcloning: a rapid reliable method to subclone polymerase chain reaction (PCR) products. Nucleic acids Res 1990; 18: 1920. Siekstele R, Bartkeviciute D, Sasnauskas K. Cloning, targeted disruption and heterologous expression of the Kluyveromyces marxianus endopolygalacturonase gene (EPG1). Yeast 1999; 15:311-22. Sigurbjornsdottir MA, Orlygsson J. Combined hydrogen and ethanol production from sugars and lignocellulosic biomass by Thermoanaerobacterium AK54, isolated from hot spring. Appl Energy 2011; Available online 9 December, ISSN 0306-2619. Singh D, Nigam P, Banat IM, Marchant R, McHale AP. Ethanol production at elevated temperatures and alcohol concentrations: part II—use of Kluyveromyces marxianus IMB3. World J Microbiol Biotechnol 1998; 14:823-834. Suzuki M, Giovannoni SJ. Bias caused by template annealing in the amplification of 16S rRNA genes by the PCR. Appl Environ Microbiol 1996; 62:625-630. Tsuge, K. and Itaya M. Recombinational transfer of 100-kilobase genomic DNA to plasmid in Bacillus subtilis 168. J. Bacteriol. 2001; 183:5453-5458. Tsuge K, Matsui K, Itaya M. One step assembly of multiple DNA fragments with a designed order and orientation in Bacillus subtilis plasmid. Nucleic acids Res. 2003; 31: e133. Walhout AJ, Temple GF, Brasch MA, Hartley JL, Lorson MA, van den Heuvel S, Vidal M. GATEWAY recombinational cloning: application to the cloning of large numbers of open reading frames or ORFeomes. Methods Enzymol 2000; 328: 575-592. Wang EQ, Li SZ, Tao L, Geng X, Li TC. Modeling of rotating drum bioreactor for anaerobic solid-state fermentation. Appl Energy 2010; 9: 2839-2845. Wilson DB. Cellulases and biofuels. Curr Opin Biotechnol 2009; 20:295-299. Wu JHD. Clostridium thermocellum cellulosome: new mechanisticoncept for cellulose degradation. ACS Symp Ser 1993; 516:251-264. Yanase S, Hasunuma T, Yamada R, Tanaka T, Ogino C, Fukuda H, Kondo A. Direct ethanol production from cellulosic materials at high temperature using the thermotolerant yeast Kluyveromyces marxianus displaying cellulolytic enzymes. Appl Microbiol Biotechnol 2010; 88:381-388. Yamada R, Taniguchi N, Tanaka T, Ogino C, Fukuda H, Kondo A. Direct ethanol production from cellulosic materials using a diploid strain of Saccharomyces cerevisiae with optimized cellulase expression. Biotechnol Biofuels 2011; 4: 8. Yamada R, Taniguchi N, Tanaka T, Ogino C, Fukuda H, Kondo A. Cocktail delta-integration: a novel method to construct cellulolytic enzyme expression ratio-optimized yeast strains. Microb Cell Fact2010; 9: 32. Ye R, Yang LP, and Wong SL. Construction of protease-deficient Bacillus subtilis strains for expression studies: inactivation of seven extracellular proteases and the intracellular LonA protease. 1996; 160-169. Zhou J, Liu X, Jiang H, Dong M. Analysis of the microflora in Tibetan kefir grains using denaturing gradient gel electrophoresis. Food Microbiol 2009; 26:770-5.
摘要: Kefir酵母菌Kluyveromyces marxianus KY3具有很廣泛的基質利用性、很好的高溫發酵潛力及優異的異源基因表現能力。我們建構了一個以酵母菌為宿主的合成生物學技術,可以同時轉入多個基因於宿主染色體中,我們稱它為Promoter-based Gene Assembly and Simultaneous Overexpression〔PGASO〕,此方法利用特定的推動子重疊區域作為多基因同源重組的依據,PGASO已成功應用於酵母菌K. marxianus KY3,當我們同時轉入五個基因片段時,呈現相當高的轉形效率約63%。另外,K. marxianus KY3-NpaBGS帶有牛胃真菌β-glucosidase (NpaBGS)基因,這兩個基因轉殖菌種將被用在後續乙醇生產系統中。 『可設計的纖維素分解酵素複合體』是一個製作人工纖維素分解酵素複合體的重要議題,可以不同的酵素比例來面對不同的木質纖維素降解情況,參考前人針對Clostridium thermocellum ATCC27405的蛋白質體學之研究,設計了面對結晶狀纖維素(Avicel)與纖維雙糖(cellobiose)的兩種操作組以仿生式模擬C. thermocellum分解纖維質時的原始表現狀態於枯草桿菌168中表現,八個C. thermocellum的纖維素分解酵素複合體相關基因:一個支架蛋白(cipA)、一個細胞膜上連接蛋白(sdbA)、兩個纖維外切酵素(celK and celS)、兩個內切酵素(celA and cel R)、兩個聚木糖分解酵素(xynC and xynZ),利用方法Ordered Gene Assembly in B. subtilis method (OGAB)共同表現於枯草菌中。 為達成高效纖維乙醇生產製程,本研究建構了一個新的Bacillus /yeast共培養系統,兩個基轉酵母菌(KY3-NpaBGS and KR5)可以分泌纖維素分解酵素,用來與B. subtilis type I共培養,以提高纖維素糖化及發酵效率。所有Bacillus /yeast共培養系統皆可表現優於單獨CBP菌種KR5的同步纖維素糖化及乙醇轉化效率。由結果顯示,以β-glucan為碳源時,共培養組合KY3-NpaBGS+Bacillus Type I可以分別呈現6倍、34%、2.5倍的生質乙醇產量之增加。由我們的結果顯示參與菌種之特性賦予Bacillus /yeast共培養系統之纖維乙醇生產能力,且具有很好的潛力應用於同步糖化發酵的製程。
The kefir yeasts that produce ethanol and higher aromatic alcohols not only create kefir’s distinctive characteristics but could also serve as a good cell-based bioethanol production and biorefinery platform. This Kluyveromyces marxianus KY3 strain possesses a broad spectrum of substrate utilization, has a high potential for bioethanol production at elevated temperatures and suitable for expressing heterologous genes. We developed one synthetic biology techniques to transform genes into the host genome, named Promoter-based Gene Assembly and Simultaneous Overexpression 〔PGASO〕, which employed overlapping oligonucleotides for recombinatorial assembly of gene cassettes with individual unique promoters. As an example of application, PGASO was used to engineer yeast K. marxianus KY3. Our data showed high transformation efficiency and accuracy because ~63% of the transformants were found to carry the correct five-gene cassette assembly. And K. marxianus KY3-NpaBGS, which carries a β-glucosidase (NpaBGS) gene from rumen fungus. The two transgenic strains were employed to process ethanol production. “Designer cellulosomes” is a concept for making an artificial cellulosome that can be proposed as a tool for regulating the cellulosomal enzyme ratio to apply on different lignocellulose degradation. Two types of “designer operon” were expressed in Bacillus subtilis 168 via biomimetic approach according to a proteome-wide analysis of Clostridium thermocellum ATCC27405, which induced by avicel and cellobiose respectively. Eight celulosomal genes including one scaffolding protein gene (cipA), one cell-surface anchor gene (sdbA), two exoglucosidase genes (celK and celS), two endoglucanase genes (celA and cel R), and two xylanase genes (xynC and xynZ) of C. thermocellum were cloned and co-expressed on the polycistronic operons in desire order via an Ordered Gene Assembly in B. subtilis method (OGAB). A novel dual-microbe Bacillus/yeast co-culture system is developed for cellulosic bioethanol production. Two engineered yeasts, KY3-NpaBGS and KR5, which possess secretive cellulolytic enzymes, were used to co-cultivate with the B. subtilis type I strain for improving the cellulose digestion and fermentation efficiency. All Bacillus/yeast co-culture systems could achieve the cellulose saccharification and ethanol conversion simultaneously better than KR5 alone. In our result, the co-culturing of KY3-NpaBGS with Bacillus Type I appeared 6 times, 34% and 2.5 times ethanol production increasing than KY3+TypeI, KR5+ Type I and KY3-NpaBGS+host when utilized β-glucan as carbon source. Our results suggest that the dual-microbe Bacillus/yeast co-culturing system could leverage the advantages from both microbes and have a great potential for integrating into consolidated bioprocessing system.
其他識別: U0005-1506201209502600
Appears in Collections:生命科學系所



Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.