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用于分析暴露于多氯联苯混合物Aroclor 1254的多氯联苯降解蓝藻鱼腥藻PD-1的蛋白质组学策略。

Proteomic strategy for the analysis of the polychlorobiphenyl-degrading cyanobacterium Anabaena PD-1 exposed to Aroclor 1254.

作者信息

Zhang Hangjun, Jiang Xiaojun, Xiao Wenfeng, Lu Liping

机构信息

College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou, Zhejiang, China.

出版信息

PLoS One. 2014 Mar 11;9(3):e91162. doi: 10.1371/journal.pone.0091162. eCollection 2014.

Abstract

The cyanobacterium Anabaena PD-1, which was originally isolated from polychlorobiphenyl (PCB)-contaminated paddy soils, has capabilities for dechlorinatin and for degrading the commercial PCB mixture Aroclor 1254. In this study, 25 upregulated proteins were identified using 2D electrophoresis (2-DE) coupled with matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). These proteins were involved in (i) PCB degradation (i.e., 3-chlorobenzoate-3,4-dioxygenase); (ii) transport processes [e.g., ATP-binding cassette (ABC) transporter substrate-binding protein, amino acid ABC transporter substrate-binding protein, peptide ABC transporter substrate-binding protein, putrescine-binding protein, periplasmic solute-binding protein, branched-chain amino acid uptake periplasmic solute-binding protein, periplasmic phosphate-binding protein, phosphonate ABC transporter substrate-binding protein, and xylose ABC transporter substrate-binding protein]; (iii) energetic metabolism (e.g., methanol/ethanol family pyrroloquinoline quinone (PQQ)-dependent dehydrogenase, malate-CoA ligase subunit beta, enolase, ATP synthase β subunit, FOF1 ATP synthase subunit beta, ATP synthase α subunit, and IMP cyclohydrolase); (iv) electron transport (cytochrome b6f complex Fe-S protein); (v) general stress response (e.g., molecular chaperone DnaK, elongation factor G, and translation elongation factor thermostable); (vi) carbon metabolism (methanol dehydrogenase and malate-CoA ligase subunit beta); and (vii) nitrogen reductase (nitrous oxide reductase). The results of real-time polymerase chain reaction showed that the genes encoding for dioxygenase, ABC transporters, transmembrane proteins, electron transporter, and energetic metabolism proteins were significantly upregulated during PCB degradation. These genes upregulated by 1.26- to 8.98-fold. These findings reveal the resistance and adaptation of cyanobacterium to the presence of PCBs, shedding light on the complexity of PCB catabolism by Anabaena PD-1.

摘要

蓝藻鱼腥藻PD-1最初是从受多氯联苯(PCB)污染的稻田土壤中分离出来的,它具有脱氯和降解商业PCB混合物Aroclor 1254的能力。在本研究中,使用二维电泳(2-DE)结合基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)鉴定了25种上调蛋白。这些蛋白质参与了以下过程:(i)PCB降解(即3-氯苯甲酸-3,4-双加氧酶);(ii)转运过程[例如,ATP结合盒(ABC)转运体底物结合蛋白、氨基酸ABC转运体底物结合蛋白、肽ABC转运体底物结合蛋白、腐胺结合蛋白、周质溶质结合蛋白、支链氨基酸摄取周质溶质结合蛋白、周质磷酸盐结合蛋白、膦酸盐ABC转运体底物结合蛋白和木糖ABC转运体底物结合蛋白];(iii)能量代谢(例如,甲醇/乙醇家族吡咯喹啉醌(PQQ)依赖性脱氢酶、苹果酸-CoA连接酶亚基β、烯醇化酶、ATP合酶β亚基、FOF1 ATP合酶亚基β、ATP合酶α亚基和肌苷酸环化水解酶);(iv)电子传递(细胞色素b6f复合物铁硫蛋白);(v)一般应激反应(例如,分子伴侣DnaK、延伸因子G和翻译延伸因子耐热蛋白);(vi)碳代谢(甲醇脱氢酶和苹果酸-CoA连接酶亚基β);以及(vii)氮还原酶(一氧化二氮还原酶)。实时聚合酶链反应结果表明,在PCB降解过程中,编码双加氧酶、ABC转运体、跨膜蛋白、电子转运蛋白和能量代谢蛋白的基因显著上调。这些基因上调了1.26至8.98倍。这些发现揭示了蓝藻对PCB存在的抗性和适应性,为鱼腥藻PD-1对PCB分解代谢的复杂性提供了线索。

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