|
|
|
| Characterization of the Disrupted ack Genes on Fermentation by Thermoanaerobacterium calidifontis Rx1 |
| SHEN Dong-ling1, SHANG Shu-mei2, LI Wei-na1, YAN Jin-ping1, HANGAN Ir-bis1 |
1. Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China;
2. Life Science and Technology Institute, Yangtze Normal University, Chongqing 408100, China |
|
|
|
Abstract To improve the yield of ethanol by Thermoanaerobacterium calidifontis Rx1, constructed a engineered mutant Δack. First a recombinant plasmid containing mutation cassettes of pta::ack, was rebuild, and the vector was transformed to cell to disrupt the target genes on the chromosomal via the homologous recombination. Then glucose fermentation, cellobiose fermentation, xylose fermentation, acid hydrolyzate of corncob of Δack mutant and the wild strain were performed respectively to produce ethanol and lactate. The results indicate that the acetate of Δack mutant is much lower as compared with the wild. Dray cell weight of the mutant is always lower than that of the wild under four conditions. However, the yield of ethanol or lactate is more than the wild. When Δack mutant used cellobiose to produce ethanol, the yield is 3.60g/L higher than another three substrates. At the same time, it could be exist approximative 40mmol/L acetate in the hydrolysate, so the output of lactate and ethanol of the wild are more than that with xylose fermentation.
|
|
Received: 26 February 2015
Published: 25 July 2015
|
|
|
|
|
|
[1] Kumar A, Singh L K, Ghosh S, et al. Bioconversion of lignocellulosic fraction of water-hyacinth hemicelluloses acid hydrolysate to ethanol by pickier styptic. Bioresource Technology, 2009, 100(13): 3293-3297.
[2] Lynd L R, Laser M S, Bransby D, et al. How biotech can transform biofuels. Nat Biotechnol, 2008, 26(2):169-172.
[3] Menon V, Rao M. Trends in bioconversion of lignocellulose: biofuels, platform chemicals & biorefinery concept. Progress in Energy and Combustion Science, 2012, 38(14):522-550.
[4] EI-Zawawy W K, Ibrahim M M, Abdel-Fattah Y R, et al. Acid and enzyme hydrolysis to convert pretreated lignocellulosic materials into glucose for ethanol production. Carbohydrate Polymers, 2011, 84(3):865-871.
[5] Sommer P, Georgieva T, Ahring B K. Potential for using thermophilic anaerobic bacteria for bioethanol production from hemicellulose. Biochem Soc Trans, 2004, 32(2):283-289.
[6] Olson D G, McBride J E, Shaw A J, et al. Recent progress in consolidatedbioprocessing. Curr Opin Biotechnol, 2012, 23(3):396-405.
[7] Desai S G, Guerinot M L, Lynd L R. Cloning of L-lactate dehydrogenase and elimination of lactic acid production via gene knockout in Thermoanaerobacterium saccharolyticum JW/SL-YS485. Appl Microbiol Biotechnol, 2004, 65(5): 600-605.
[8] Shaw A J, Podkaminer K K, Desai S G, et al. Metabolic engineering of a thermophilic bacterium to produce ethanol at high yield. Proc Natl Acad Sci USA, 2008, 105(37): 13769-13774.
[9] Taylor M P, Eley K L, Martin S, et al. Thermophilic ethanologenesis: future prospects for second-generation bioethanol production. Trends Biotechnol, 2009, 27(7): 398-405.
[10] Yao S, Mikkelsen M J. Identification and overexpression of a bifunctional aldehyde/alcohol dehydrogenase responsible for ethanol production in Thermoanaerobacter mathranii. J Mol Microbiol Biotechnol, 2010, 19(3): 123-133.
[11] Li S, Lai C, Cai Y, et al. High efficiency hydrogen production from glucose/xylose by the ldh-deleted Thermoanaerobacterium strain. Bioresour Technol, 2010, 101(22): 8718-8724.
[12] Park J M, Kim T Y, Lee S Y. Constraints-based genome-scale metabolic simulation for systems metabolic engineering. Biotechnol Adv, 2009, 27(6): 979-988.
[13] Patnaik R. Engineering complex phenotypes in industrial strains. Biotechnol Prog, 2008, 24(1): 38-47.
[14] Shang S M, Qian L, Zhang X, et al. Thermoanaerobacterium calidifontis sp. nov., a novel anaerobic, thermophilic, ethanol producing bacterium from hot springs in China. Arch Microbiol, 2013, 195(6):439-445.
[15] Hoseki J, Yano T, Y Koyama, et al. Directed evolution of thermostable kanamycin-resistance gene: a convenient selection marker for Thermus thermophilus. J Biochem, 1999, 126(5): 951-956.
[16] Romano I, Dipasquale L, Orlando P, et al.Thermoanaerobacterium thermostercus sp.nov. a new anaerobic thermophilic hydrogen-producing bacterium from buffalo-dung. Extremophiles, 2010,14(2): 233-240.
[17] Argyros D A, Tripathi S A, Barrett T F, et al. High ethanol titers from cellulose by using metabolically engineered thermophilic, anaerobic microbes. Applied and Environmental Microbiology, 2011,77(23):8288-8294.
[18] Yang X, Lai Z, Lai, C, et al. Efficient production of l-lactic acid by an engineered Thermoanaerobacterium aotearoense with broad substrate specificity. Biotechnol Biofuels, 2013,6(1):124.
[19] Mai V, Wiegel J. Advances in development of a genetic system for Thermoanaerobacterium spp.: expression of genes encoding hydrolytic enzymes, development of a second shuttle vector, and integration of genes into the chromosome. Applied and Environmental Microbiology, 2000,66(11): 4817-4821.
[20] Tripathi S A, Olson D G, Argyros D A, et al. Development of pyrF-based genetic system for targeted gene deletion in clostridium thermocellum and creation of a pta mutant. Applied and Environmental Microbiology, 2010,76(19): 6591-6599.
[21] Shaw A J, Podkaminer K K, Desai S G, et al. Metabolic engineering of a thermophilic bacterium to produce ethanol at high yield. Proc Natl Acad Sci USA, 2008,105(37): 13769-13774.
[22] 尚淑梅,申冬玲,李坤志,等. 高温菌发酵甘露醇高效产乙醇的代谢途径研究. 中国生物工程杂志, 2013,33(10): 73-80. Shang S M, Shen D L, Li K Z, et al. Study on metabolic pathway of efficiently producting ethanol by thermophilic bacterium using mannitol. China Biotechnology, 2013, 33(10): 73-80.
[23] He Q, Lokken P M, Chen S, et al. Characterization of the impact of acetate and lactate on ethanolic fermentation by Thermoanaerobacter ethanolicus. Bioresour Technol, 2009, 100(23): 5955-5965.
[24] Mai V, Lorenz W W, Wiegel J. Transformation of Thermoanaerobacterium sp. Strain JW/SL-YS485 with plasmid pIKM1 conferring kanamycin resistance. FEMS Microbiology Letters, 1997, 148(2):163-167.
[25] Jantama K, Zhang X, Moore J C, et al. Eliminating side products and increasing succinate yields in engineered strains of Escherichia coli. Biotechnology and Bioengineering, 2008,101(5): 881-893.
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
