刘冬忍;田芳;王小飞;王景冒;戚元成;宋安东;申进文;邱立友;

• 1:河南农业大学生命科学学院农业部农业微生物酶工程重点实验室

摘要(Abstract)：

POXC是糙皮侧耳合成最多的一种漆酶。应用启动子替代技术,用构巢曲霉的甘油醛‐3‐磷酸脱氢酶基因(gpd)启动子替代POXC基因的启动子,构建了超量表达POXC糙皮侧耳转化子。转化子中POXC基因表达量比出发菌株提高了0.72–3倍。在PDA平板培养、PD摇瓶培养和棉籽壳试管培养条件下,转化子漆酶活力显著提高,比出发菌株提高了1.5倍以上。用棉籽壳栽培,转化子菇产量比出发菌株提高了16.2%,培养料中木质素含量比出发菌株减少21%。结果表明,应用高效启动子替代能够显著提高糙皮侧耳漆酶基因的表达量、漆酶活力及其木质纤维素降解能力。

关键词(KeyWords)： 糙皮侧耳;漆酶;启动子替代

Abstract:

Keywords:

基金项目(Foundation): 河南省重点科技攻关项目(082102150048);; 河南省高校科技创新团队支持计划(15IRTSTHN014)~~

作者(Authors): 刘冬忍;田芳;王小飞;王景冒;戚元成;宋安东;申进文;邱立友;

DOI: 10.13346/j.mycosystema.140301

参考文献(References)：

• Bailey JE,1991.Toward a science of metabolic engineering.Science,252(5013):1668-1675
• Baldrian P,der Wiesche CI,Gabriel J,Nerud F,Zadrazil F,2000.Influence of cadmium and mercury on activities of ligninolytic enzymes and degradation of polycyclic aromatic hydrocarbons by Pleurotus ostreatus in soil.Applied and Environmental Microbiology,66(6):2471-2478
• Castanera R,Omarini A,Santoyo F,Pérez G,Pisabarro AG,Ramírez L,2013.Non-additive transcriptional profiles underlie dikaryotic superiority in Pleurotus ostreatus laccase activity.PLo S One,8(9):e73282
• Chai R,Qiu C,Liu D,Qi Y,Gao Y,Shen J,Qiu L,2013a.β-Glucan synthase gene overexpression andβ-glucans overproduction in Pleurotus ostreatus using promoter swapping.PLo S One,8(4):e61693
• Chai R,Sun Q,Qiu L,2012.Quadruple DNA fragments fusion using touchdown-overlap extension PCR.Chinese Journal of Biochemistry and Molecular Biology,28(3):81-85(in Chinese)
• Chai R,Zhang G,Sun Q,Zhang M,Zhao S,Qiu L,2013b.Liposome-mediated mycelial transformation of filamentous fungi.Fungal Biology,117(9):577-583
• Chalamcherla VL,Muvva VL,Mangamoori LN,Ramireddy MR,2008.Use of lignocellulolytic mutants of Pleurotus ostreatus in ruminant feed formulations.Bio Resources,4(1):142-154
• Cohen R,Persky L,Hadar Y,2002.Biotechnological applications and potential of wood-degrading mushrooms of the genus Pleurotus.Applied Microbiology and Biotechnology,58(5):582-594
• Gao YQ,Gao F,Zhang SM,Wu K,2009.Screening for high yield laccase producing strains from Auricularia auricular.Chinese Agricultural Science Bulletin,25(21):301-304(in Chinese)
• Giardina P,Aurilia V,Cannio R,Marzullo L,Amoresano A,Siciliano R,Pucci P,Sannia G,1996.The gene,protein and glycan structures of laccase from Pleurotus ostreatus.European Journal of Biochemistry,235(3):508-515
• Homolka L,VolákováI,Nerud F,1995.Variability of enzymatic activities in ligninolytic fungi Pleurotus ostreatus and Lentinus tigrinus after protoplasting and UV-mutagenization.Biotechnology Techniques,9(3):157-162
• Liu CG,Gao JQ,Li CQ,2006.Study on the degradation and biotransformation of rice straw by Pleurotus ostreatus.Journal of Southwest Agricultural University(Natural Science),28(2):258-263(in Chinese)
• Martínez AT,Camarero S,Guillén F,Gutiérrez A,Mu?oz C,Varela E,Martínez MJ,Barrasa JM,Ruel K,Pelayo J,1994.Progress in biopulping of non-woody materials:chemical,enzymatic and ultrastructural aspects of wheat straw delignification with ligninolytic fungi from the genus Pleurotus.FEMS Microbiology Reviews,13(2-3):265-273
• Matcham SE,Jordan BR,Wood DA,1985.Estimation of fungal biomass in a solid substrate by three independent methods.Applied Microbiology and Biotechnology,21(1-2):108-112
• Mc Cleary WR,2009.Application of promoter swapping techniques to control expression of chromosomal genes.Applied Microbiology and Biotechnology,84(4):641-648
• Munoz C,Guillen F,Martinez AT,Martinez MJ,1997.Induction and characterization of laccase in the ligninolytic fungus Pleurotus eryngii.Current Microbiology,34(1):1-5
• Palmieri G,Giardina P,Bianco C,Fontanella B,Sannia G,2000.Copper induction of laccase isoenzymes in the ligninolytic fungus Pleurotus ostreatus.Applied and Environmental Microbiology,66(3):920-924
• Pezzella C,Lettera V,Piscitelli A,Giardina P,Sannia G,2013.Transcriptional analysis of Pleurotus ostreatus laccase genes.Applied Microbiology and Biotechnology,97(2):705-717
• Qiu LY,2008.The history and development of solid state fermentation.In:Qiu LY(ed.)The principle and application of solid-state fermentation.China Light Industry Press,Beijing.1-15(in Chinese)
• Sugiura T,Mori T,Kamei I,Hirai H,Kawagishi H,Kondo R,2012.Improvement of ligninolytic properties in the hyper lignin-degrading fungus Phanerochaete sordida YK-624 using a novel gene promoter.FEMS Microbiology Letters,331(1):81-88
• Tsukihara T,Honda Y,Watanabe T,Watanabe T,2006.Molecular breeding of white rot fungus Pleurotus ostreatus by homologous expression of its versatile peroxidase Mn P2.Applied Microbiology and Biotechnology,71(1):114-120
• Van Soest PJ,1963.Use of determents in the analysis of fibrous feeds.Journal of the Association of Official Agricultural Chemists,46(5):829-835Velázquez-Cede?o M,Anne Marie Farnet AM,Billette C,Mata G,Savoie JM,2007.Interspecific interactions with Trichoderma longibrachiatum induce Pleurotus ostreatus defence reactions based on the production of laccase isozymes.Biotechnology Letters,29(10):1583-1590
• Wang L,Li Y,Liu D,Zhang C,Qi Y,Gao Y,Shen J,Qiu L,2011.Immobilization of mycelial pellets from liquid spawn of oyster mushroom based on carrier adsorption.Hort Technology,21(1):82-86
• Wang L,Xia ML,Chen HZ,2014.Application of process engineering to remove lignocellulose fermentation inhibitors.Chinese Journal of Biotechnology,30(5):716-725(in Chinese)
• Wang LS,Zhu HL,Li CF,Chen MJ,Cai HY,2009.Screening of fungal mutant strain with high laccase yield by N+implantation and enzyme production condition optimization.Journal of Nuclear Agricultural Sciences,23(2):228-234(in Chinese)
• Yadav VG,De Mey M,Lim CG,Ajikumar PK,Stephanopoulos G,2012.The future of metabolic engineering and synthetic biology:towards a systematic practice.Metabolic Engineering,14(3):233-241
• Zhang ST,Zhao M,2002.Retrospect and prospect of research and development of edible fungi for forty years.Edible Fungi of China,21(6):3-6(in Chinese)
• Zhang YB,Jiang Q,Jiang ML,Ma LX,2004.Cloning of a laccase gene from Flammulina velutipes and study on its expression in Pichia pastoris.Acta Microbiology Sinica,44(6):775-779(in Chinese)
• 柴冉,孙强,邱立友,2012.降落-重叠PCR法四重融合构建平菇同源重组片段.中国生物化学与分子生物学报,28(3):81-85
• 高玉千,高方,张世敏,吴坤,2009.黑木耳漆酶高产菌株的筛选.中国农学通报,25(21):301-304
• 刘朝贵,高金权,李成琼,2006.糙皮侧耳(Pleurotus ostreatus)降解转化稻草秸秆研究.西南农业大学学报(自然科学版),28(2):258-263
• 邱立友,2008.固态发酵的历史和发展.邱立友主编.固态发酵工程原理及应用.北京:中国轻工业出版社.1-15
• 王岚,夏梦雷,陈洪章,2014.过程工程在木质纤维素发酵抑制物解除中的应用.生物工程学报,30(5):716-725
• 王力生,朱洪龙,李春凤,程茂基,蔡海莹,2009.N+注入选育漆酶高产菌株及其产酶优化研究.核农学报,23(2):228-234
• 张银波,姜琼,江木兰,马立新,2004.金针菇漆酶基因的克隆及其在毕赤酵母中的表达研究.微生物学报,44(6):775-779