不同木质底物诱导下桦剥管菌细胞内差异蛋白质分析
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摘要
【目的】研究桦剥管菌在不同木质底物诱导下细胞内蛋白的差异表达情况,探讨桦剥管菌降解木材机理。【方法】以白桦、云杉木屑为诱导底物,利用蛋白质组双向电泳技术(2-DE)结合质谱技术(MALDI-TOF/TOF MS),检测桦剥管菌在不同树种木屑诱导下与对照组(未加木屑)相比菌体胞内蛋白质变化情况,对差异蛋白进行生物信息学分析。【结果】在白桦和云杉木屑诱导下,桦剥管菌细胞内蛋白表达发生了明显变化,处理组(添加木屑)共检测到32个细胞内差异表达蛋白点,经质谱检测及数据库检索,成功鉴定17个差异蛋白点,这些蛋白具有转移酶、水解酶、蛋白结合、细胞信号转导等分子功能,涉及分解代谢、有机物质代谢、氮化合物代谢、初级代谢、细胞生物合成和刺激响应过程。【结论】桦剥管菌降解木材是多种蛋白质共同作用的结果,通过调节多种代谢过程中的相关蛋白表达来发挥作用。
【Objective】In this paper,we investigated the differential expression of intracellular proteins induced by different woody substrates,and explored the degradation mechanism of wood by Piptoporus betulinus.【Method】Wood sawdust of birch and spruce were used as inducing substrates,the protein two-dimensional electrophoresis technique( 2-DE) and mass spectrometry( MALDI-TOF/TOF MS) was used to detect the changes in Piptoporus betulinus intracellular protein induced by wood sawdust compared with the control group( without sawdust),and analyze the differences in bioinformatics.【Result】The Piptoporus betulinus intracellular protein expression changed significantly induced by birch and spruce sawdust. A total of 32 intracellular differential expression protein sites were detected in the treatment group( adding sawdust),17 differential protein sites were successfully identified by mass spectrometry and database retrieval.These proteins had transferase,hydrolase,protein binding,and cell signal transduction molecular functions,and were involved in the stimulation of organic compounds,metabolism and metabolic processes.【Conclusions】The degradation of wood by Piptoporus betulinus is the result of the interaction of various proteins,regulating the expression of related proteins in various metabolic processes.
【Objective】In this paper,we investigated the differential expression of intracellular proteins induced by different woody substrates,and explored the degradation mechanism of wood by Piptoporus betulinus.【Method】Wood sawdust of birch and spruce were used as inducing substrates,the protein two-dimensional electrophoresis technique( 2-DE) and mass spectrometry( MALDI-TOF/TOF MS) was used to detect the changes in Piptoporus betulinus intracellular protein induced by wood sawdust compared with the control group( without sawdust),and analyze the differences in bioinformatics.【Result】The Piptoporus betulinus intracellular protein expression changed significantly induced by birch and spruce sawdust. A total of 32 intracellular differential expression protein sites were detected in the treatment group( adding sawdust),17 differential protein sites were successfully identified by mass spectrometry and database retrieval.These proteins had transferase,hydrolase,protein binding,and cell signal transduction molecular functions,and were involved in the stimulation of organic compounds,metabolism and metabolic processes.【Conclusions】The degradation of wood by Piptoporus betulinus is the result of the interaction of various proteins,regulating the expression of related proteins in various metabolic processes.
引文
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[21]SNODGRASS P J. Ornithine transcarbamylase:basic science and clinical considerations[M]. London:Springer Science&Business Media,2003.
[2]刘欣,赵敏,王秋玉. 5种木材腐朽菌的生物学特性及对白桦木材腐朽能力的分析[J].东北林业大学学报,2008,36(3):41-44. DOI:10.13759/j.cnki.dlxb.2008.03.019.LIU X,ZHAO M,WANG Q Y. Biological characters of five species of wood rot fungi and decay capacity to Betula platyphylla[J]. Journal of Northeast Forestry University,2008,36(3):41-44.
[3]王丽华,田雪梅,毕旺华,等.桦剥管孔菌液体发酵环境条件的研究[J].食用菌学报,2011,18(4):37-39. DOI:10.16488/j.cnki.1005-9873.2011.04.006WANG L H,TIAN X M,BI W H,et al. Optimization of parameters affecting extracellular polysaccharide production in submerged cultures of Piptoporus betulinus[J]. Acta Edulis Fungi,2011,18(4):37-39.
[4]CYRANKA M,GRAZ M,KACZOR J,et al. Investigation of antiproliferative effect of ether and ethanol extracts of Birch polypore medicinal mushroom,Piptoporus betulinus(Bull.:Fr.)P. karst.(Higher Basidiomycetes)in vitro grown mycelium[J]. International Journal of Medicinal Mushrooms,2011,13(6):525-533.
[5]易晓雷,苗雄鹰.丝切蛋白-1和内皮细胞分化因子受体-1的研究进展[J].肿瘤药学,2012,2(4):242-248.YI X L,MIAO X Y. Recent progress in the research of CFL and EDG-1[J]. Anti-tumor Pharmacy,2012,2(4):242-248.
[6]田朝光,马延和.真菌降解木质纤维素的功能基因组学研究进展[J].生物工程学报,2010,26(10):1333-1339.TIAN C G,MA Y H. Progress in lignocellulose deconstruction by fungi[J]. Chin J Biotech,2010,26(10):1333-1339.
[7]HUANG K J,LIN S H,LIN M R,et al. Xanthone derivatives could be potential antibiotics:virtual screening for the inhibitors of enzyme I of bacterial phosphoenolpyruvate-dependent phosphotransferase system[J]. The Journal of Antibiotics,2013,66(8):453-458.
[8]SANTINI D,VINCENZI B,MASSACESI C,et al. S-adenosylmethionine(AdoMet)supplementation for treatment of chemotherapy-induced liver injury[J]. Anticancer research,2002,23(6D):5173-5179.
[9]GAJADEERA C S,ZHANG X,WEI Y,et al. Structure of inorganic pyrophosphatase from Staphylococcus aureus reveals conformational flexibility of the active site[J]. Journal of structural biology,2015.
[10]王真,欧齐星,王双山,等.流产布鲁氏菌ATP/GTP结合蛋白基因缺失株保护力及安全性研究[J].中国农业大学学报,2013,18(3):138-143.WANG Z,OU Q X,Wang S S,et al. Studies on protective efficacy and safety of Brucella abortus ATP/GTP binding protein gene deleted mutant[J]. Journal of China Agricultural University,2013,18(3):138-143.
[11]WOLFRAM F,KITOVA E N,ROBINSON H,et al. Catalytic mechanism and mode of action of the periplasmic alginate epimerase Alg G[J]. Journal of Biological Chemistry,2014,289(9):6006-6019.
[12]STEEG P S,PALMIERI D,OUATAS T,et al. Histidine kinases and histidine phosphorylated proteins in mammalian cell biology,signal transduction and cancer[J]. Cancer Letters,2003,190(1):1-12.
[13]MATSUSHIKA A,GOSHIMA T,FUJII T,et al. Characterization of non-oxidative transaldolase and transketolase enzymes in the pentose phosphate pathway with regard to xylose utilization by recombinant Saccharomyces cerevisiae[J]. Enzyme and Microbial Technology,2012,51(1):16-25.
[14]KRK Y'M,VINOVJ,NOVOTNE,et al. Salicylanilide derivatives block Mycobacterium tuberculosis through inhibition of isocitrate lyase and methionine aminopeptidase[J]. Tuberculosis,2012,92(5):434-439.
[15]HUANG J,HU H,XIE Y,et al. Effects of TUBB3,TS and ERCC1 mRNA expressions on chemoresponse and clinical outcome of advanced gastric cancer by multiplex branched-DNA liquid chip technology[J]. Translational Gastrointestinal Cancer,2013,3(1):21-28.
[16] WILLIAMS A J,WERNER-FRACZEK J,CHANG F,et al.Regulated phosphorylation of 40S ribosomal protein S6 in root tips of maize[J]. Plant Physiology,2003,132(4):2086-2097.
[17]SPAHN C M T,KIEFT J S,GRASSUCCI R A,et al. Hepatitis C virus IRES RNA-induced changes in the conformation of the 40s ribosomal subunit[J]. Science,2001,291(5510):1959-1962.
[18]汪新颖,王波,侯松涛,等.苹果酸脱氢酶的结构及功能[J].生物学杂志,2009,26(4):69-72.WANG X Y,WANG B,HOU S T,et al. Structure and function of malate dehydrogenases[J]. Journal of Biology,2009,26(4):69-72.
[19] GRALLERT A,CONNOLLY Y,SMITH D L,et al. The S.pombe cytokinesis NDR kinase Sid2 activates Fin1 NIMA kinase to control mitotic commitment through Pom1/Wee1[J]. Nature Cell Biology,2012,14(7):738-745.
[20]KURZAI O,SCHMITT C,CLAUS H,et al. Carbohydrate composition of meningococcal lipopolysaccharide modulates the interaction of Neisseria meningitidis with human dendritic cells[J].Cellular Microbiology,2005,7(9):1319-1334.
[21]SNODGRASS P J. Ornithine transcarbamylase:basic science and clinical considerations[M]. London:Springer Science&Business Media,2003.