Liu Huan, Sun Wen-Bing, Liang Ru-Bing, Huang Li, Hou Jing-Li, Liu Jian-Hua
School of Life Science & Biotechnology, Shanghai Jiao Tong University, 800 Dong-Chuan Road, Shanghai 200240, China.
Department of Plastic and Aesthetic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
J Proteomics. 2015 Jun 18;123:14-28. doi: 10.1016/j.jprot.2015.03.034. Epub 2015 Apr 4.
N-octadecane, the shortest solid-state alkane, was efficiently consumed by Pseudomonas aeruginosa SJTD-1. To reveal its mechanism, the iTRAQ-LC-MS/MS strategy was applied for quantification of proteins in response to alkane. As a result, 383 alkane-responsive proteins were identified and these proteins could be linked to multiple biochemical pathways. Above all, the level of alkane hydroxylase AlkB2 has been significantly higher in alkane condition. Also, the presence of a putative novel AlmA-like monooxygenase and its role on alkane hydroxylation were firstly proposed in Pseudomonas. In addition, other proteins for chemotaxic, β-oxidation, glyoxylate bypass, alkane uptake, cross membrane transport, enzymatic steps and the carbon flow may have important roles in the cellular response to alkane. Most of those differently expressed proteins were functionally mapped into pathways of alkane degradation or metabolism thereof. In this sense, findings in this study provide critical clues to reveal biodegradation of long chain n-alkanes and rationally be important for potent biocatalyst for bioremediation in future.
We use iTRAQ strategy firstly to compare the proteomes of Pseudomonas SJTD-1 degrading alkane. Changes in protein clearly provide a comprehensive overview on alkane hydroxylation of SJTD-1, including those proteins for chemotaxis, alkane uptake, cross membrane transport, enzymatic steps and the carbon flow. AlkB2 and a putative novel AlmA-like monooxygenase have been highlighted for their outstanding contribution to alkane use. We found that several chemotaxic proteins were altered in abundance in alkane-grown cells. These results may be helpful for understanding alkane use for Pseudomonas.
正十八烷是最短的固态烷烃，能被铜绿假单胞菌SJTD-1有效消耗。为揭示其机制，采用iTRAQ-LC-MS/MS策略对响应烷烃的蛋白质进行定量分析。结果，鉴定出383种烷烃响应蛋白，这些蛋白可与多种生化途径相关联。首先，在烷烃条件下，烷烃羟化酶AlkB2的水平显著升高。此外，首次在假单胞菌中提出了一种推定的新型AlmA样单加氧酶的存在及其在烷烃羟化中的作用。此外，其他参与趋化作用、β-氧化、乙醛酸旁路、烷烃摄取、跨膜运输、酶促步骤和碳流的蛋白质可能在细胞对烷烃的反应中起重要作用。大多数差异表达的蛋白质在功能上被映射到烷烃降解或其代谢途径中。从这个意义上说，本研究的结果为揭示长链正构烷烃的生物降解提供了关键线索，并且对未来用于生物修复的高效生物催化剂具有重要意义。
我们首次使用iTRAQ策略比较降解烷烃的铜绿假单胞菌SJTD-1的蛋白质组。蛋白质的变化清楚地提供了SJTD-1烷烃羟化的全面概述，包括那些参与趋化作用、烷烃摄取、跨膜运输、酶促步骤和碳流的蛋白质。AlkB2和一种推定的新型AlmA样单加氧酶因其对烷烃利用的杰出贡献而受到关注。我们发现几种趋化蛋白在烷烃生长的细胞中丰度发生了变化。这些结果可能有助于理解假单胞菌对烷烃的利用。
