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透明颤菌血红蛋白基因在刺糖多孢菌中整合表达促进多杀菌素的生物合成

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Keywords: 刺糖多孢菌,多杀菌素,透明颤菌血红蛋白,整合型载体,同源重组

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Abstract:

为解决刺糖多孢菌(Saccharopolysporaspinosa)高密度深层培养过程中的供氧不足问题,促进多杀菌素的生物合成,采用重叠PCR技术将透明颤菌(Vitreoscillastercoraria)血红蛋白基因(vgb)可读框(ORF)置于红霉素抗性基因启动子(PermE)之下,并将其克隆到整合型载体pSET152上,构建成vgb表达载体pSET152EVHB;通过接合转移方式将其导入刺糖多孢菌SP06081中,利用载体上ΦC31整合性位点通过位点特异性重组将vgb定点整合到SP06081菌株染色体上,获得一株遗传性能稳定的重组菌株S078-1101;重组工程菌的PCR与Southernblotting检测显示vgb已整合到染色体上,一氧化碳结合差光谱分析表明在S078-1101工程菌中表达了有活性的透明颤菌血红蛋白(VHb);摇瓶发酵结果显示,在正常溶氧与中度限氧状态下,vgb的表达均可显著促进多杀菌素的生物合成(P<0.01).这说明vgb在刺糖多孢菌中的整合表达改善了菌体对溶氧的吸收性能,是一种有效的定向遗传改良手段.

References

[1]  1 Toshi M, Mande S, Dikshit K. Hemoglobin biosynthesis in Vitreoscilla stercoraria DW: cloning, expression, and characterization of a new homolog of a bacterial globin gene. Appl Environ Microbiol, 1998, 64: 2220-2228
[2]  2 Orii Y, Webster D A. Photodissociation of oxygenated cytochrome o(s) (Vitreoscilla) and kinetic studies of reassociation. J Biol Chem,1986, 261: 3544-3547
[3]  3 Ramandeep H K W, Raje M, Kim K J, et al. Vitreoscilla hemoglobin. Intracellular localization and binding to membranes. J Biol Chem,2001, 276: 24781-24789??
[4]  4 Kim K J, Chi P Y, Hwang K W, et al. Study of cytochrome bo function in Vitreoscilla using a cyo-knockout mutant. J Bio Chem, 2000, 128:49-55
[5]  5 Kvist M, Ryabova E S, Nordlander E, et al. An investigation of the peroxidase activity of Vitreoscilla hemoglobin. J Biol Inorg Chem, 2007,12: 324-334??
[6]  6 Kim Y J, Sa S O, Chang Y K, et al. Overexpression of Shinorhizobium meliloti hemoprotein in Streptomyces lividans to enhance secondary metabolite production. J Microbiol Biotechnol, 2007, 17: 2066-2070
[7]  7 Anand A, Duk B, Singh S, et al. Redox-mediated interactions of VHb (Vitreoscilla haemoglobin) with OxyR: novel regulation of VHb biosynthesis under oxidative stress. Biochem J, 2010, 426: 271-280??
[8]  8 Hofmann G, Diano A, Nielsen J. Recombinant bacterial hemoglobin alters metabolism of Aspergillus niger. Metab Eng, 2009, 11: 8-12??
[9]  9 Isarankura-Na-Ayudhya C, Panpumthong P, Tangkosakul T, et al. Shedding light on the role of Vitreoscilla hemoglobin on cellular catabolic regulation by proteomic analysis. Int J Biol Sci, 2008, 4: 71-80
[10]  10 文莹, 宋渊, 李季伦. 透明颤菌血红蛋白在肉桂地链霉菌中的表达对其细胞生长及抗生素合成的影响. 生物工程学报, 2001, 17:24-28
[11]  11 Zhang L, Li Y, Wang Z, et al. Recent developments and future prospects of Vitreoscilla hemoglobin application in metabolic engineering. Biotechnol Adv, 2007, 25: 123-136??
[12]  12 Li M, Wu J, Lin J, et al. Expression of Vitreoscilla hemoglobin enhances cell growth and dihydroxyacetone production in Gluconobacter oxydans. Curr Microbiol, 2010, 61: 370-375??
[13]  13 Kurt A G, Aytan E, Ozer U, et al. Production of L-DOPA and dopamine in recombinant bacteria bearing the Vitreoscilla hemoglobin gene. Biotechnol J, 2009, 4: 1077-1088??
[14]  14 Chen H, Chu J, Zhang S, et al. Intracellular expression of Vitreoscilla hemoglobin improves s-adenosylmethionine production in a recombinant Pichia pastoris. Appl Microbiol Biotechnol, 2007, 74: 1205-1212??
[15]  15 Chien L J, Lee C K. Enhanced hyaluronic acid production in Bacillus subtilis by coexpressing bacterial hemoglobin. Biotechnol Prog, 2007,23: 1017-1022
[16]  16 Xiong X C, Xing J M, Li X, et al. Enhancement of biodesulfurization in two-liquid systems by heterogeneous expression of Vitreoscilla hemoglobin. Appl Environ Microbiol, 2007, 73: 2394-2397??
[17]  17 于慧敏, 马现锋, 罗晖, 等. D-氨基酸氧化酶与麦芽糖结合蛋白和透明颤菌血红蛋白的融合表达. 生物工程学报, 2008, 24:1004-1009
[18]  18 Suthar D H, Chattoo B B. Expression of Vitreoscilla hemoglobin enhances growth and levels of α-amylase in Schwanniomyces occidentalis. Appl Microbiol Biotechnol, 2006, 72: 94-102??
[19]  19 杨洪涛, 刘瑞华, 何建勇, 等. 透明颤菌血红蛋白基因的克隆及其在林可链霉菌中的表达. 沈阳药科大学学报, 2009, 26: 657-662
[20]  20 Feng L, Chen S W, Sun M, et al. Expression of Vitreoscilla hemoglobin in Bacillus thuringiensis improve the cell density and insecticidal crystal proteins yield. Appl Microbiol Biotechnol, 2007, 74: 390-397??
[21]  21 Su Y S, Li X, Liu Q Z, et al. Improved poly-γ-glutamic acid production by chromosomal integration of the Vitreoscilla hemoglobin gene (vgb) in Bacillus subtilis. Bioresource Technol, 2010, 101: 4733-4736??
[22]  22 Horng Y T, Chang K C, Chien C C, et al. Enhanced polyhydroxybutyrate (PHB) production via the coexpressed phaCAB and vgb genes controlled by arabinose PBAD promoter in Escherichia coli. Lett Appl Microbiol. 2010, 50: 158-167
[23]  23 Mertz F P, Yao R C. Actinomadura fibrosa sp. nov. isolated from soil. Int J Syst Bacteriol, 1990, 40: 28-33??
[24]  24 Millar N S, Denholm I. Nicotinic acetylcholine receptors: targets for commercially important insecticides. Invert Neurosci, 2007, 7: 53-66??
[25]  25 Cleveland C B, Bormett G A, Saunders D G, et al. Environmental fate of spinosad. 1. Dissipation and degradation in aqueous systems. J Agric Food Chem, 2002, 50: 3244-3256
[26]  27 Jin Z, Wu J, Zhang Y, et al. Improvement of spinosad producing Saccharopolyspora spinosa by rational screening. J Zhejiang Univ SCIENCE A, 2006, 7: 366-370??
[27]  28 罗玉双, 丁学知, 夏立秋, 等. 多杀菌素生产菌原生质体的制备条件及其再生菌株生理特性. 生物工程学报, 2009, 25: 360-367
[28]  29 Wang C, Zhang X, Chen Z, et al. Strain construction for enhanced production of spinosad via intergeneric protoplast fusion. Can J Microbiol, 2009, 55: 1070-1075??
[29]  30 Tang Y, Xia L, Ding X, et al. Molecular cloning and overexpression of partial spinosyn biosynthetic gene cluster in Saccharopolyspora spinosa enhances spinosad production. FEMS Microbiol Lett, 2011, 325: 22-29??
[30]  31 萨姆布鲁克J, 弗里奇E F, 曼尼阿蒂斯T, 著. 金冬雁, 黎孟枫, 译. 分子克隆实验指南. 第2 版. 北京: 科学出版社, 1992. 43-370??
[31]  26 Mota-Sanchez D, Hollingworth R M, Grafius E J, et al. Resistance and cross-resistance to neonicotinoid insecticides and spinosad in the Colorado potato beetle, Leptinotarsa decemlineata (Say)(Coleoptera: Chrysomelidae). Pest Manag Sci, 2006, 62: 30-37??
[32]  32 Matsushima P, Broughton M C, Turner J R, et al. Conjugal transfer of cosmid DNA from Escherichia coli to Saccharopolyspora spinosa: effects of chromosomal insertions on macrolide A83543 production. Gene, 1994, 146: 39-45??
[33]  33 Kieser T, Bibb M J, Buttner M J, et al. Practical Streptomyces Genetics. Norwich: The John Innes Foundation, Colney, 2000. 161-211
[34]  34 Bierman M, Logan R, O’Brien K, et al. Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene, 1992, 116: 43-49??
[35]  35 冯亮. 透明颤菌血红蛋白基因在苏云金芽胞杆菌中的表达及对其发酵特性的影响. 博士学位论文. 武汉: 华中农业大学, 2006
[36]  36 袁洪水, 李术娜, 朱宝成, 等. 产黄青霉发酵过程中菌体量测定的研究. 河北农业大学学报, 2007, 30: 83-86
[37]  37 Luo Y, Ding X, Xia L, et al. Comparative proteomic analysis of Saccharopolyspora spinosa SP06081 and PR2 strains reveals the differentially expressed proteins correlated with the increase of spinosad yield. Proteome Sci, 2011, 9: 40??
[38]  38 Nikodinovic J, Priestley N D. A second generation snp-derived Escherichia coli-Streptomyces shuttle expression vector that is generally transferable by conjugation. Plasmid , 2006, 56: 223-227??
[39]  39 Brünker P, Minas W, Kallio P T, et al. Genetic engineering of an industrial strain of Saccharopolyspora erythraea for stable expression of the Vitreoscilla haemoglobin gene (vhb). Microbiology ,1998, 144: 2441-2448??
[40]  40 Yang J, Webster D A, Stark B C. ArcA works with Fnr as a positive regulator of Vitreoscilla (bacterial) hemoglobin gene expression in Escherichia coli. Microbiol Res, 2005, 160: 405-415??
[41]  41 Tsai P S, Kallio P T, Bailey J E. Fnr, a global transcriptional regulator of Escherichia coli, activates the Vitreoscilla hemoglobin (VHb) promoter and intracellular VHb expression increases cytochrome d promoter activity. Biotechnol Prog. 1995, 11: 288-293
[42]  42 Priscila G, Fernández F J, Absalón A E, et al. Expression of the bacterial hemoglobin gene from Vitreoscilla stercoraria increases rifamycin B production in Amycolatopsis mediterranei. J Biosic Bioeng, 2008, 106: 493-497??

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