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Abstract
While many data are available on genes encoding proteins for degradation of hydrocarbons in bacteria, the impact of alkane on transporter protein expression is unclear. Pseudomonas aeruginosa SJTD-1 is a strain that can consume medium- and long-chain n-alkanes. In order to study the proteins involved in n-octadecane uptake, we use iTRAQ and label free comparative proteomics analysis to identify the proteins of alkane uptake in response to n-octadecane (C18) comparing with n-hexadecane (C16) in P. aeruginosa SJTD-1. A total of 1102 and 1249 proteins were identified by iTRAQ-based and label free quantitative methodologies, respectively. By application of 1.5 (iTRAQ) or 2-fold (label free) for upregulated and 0.65 (iTRAQ) or 0.5-fold (label free) for downregulated cutoff values, 91 and 99 proteins were found to be differentially expressed comparing SJTD-1 cultivated on C18 with C16 respectively. There are six proteins with the common differential expression by iTRAQ and label free-based methods. Results of bioinformational analysis suggested the involvement of bacterial chemotaxis in responds to C18. Additionally, quantitative reverse transcriptase PCR (qRT-PCR) results confirmed C18-induced change in levels of FleQ, FliC, NirS, FadL and FadD proteins and the role of the proteins in n-octadecane uptake was further discussed in P. aeruginosa. In conclusion, results of the present study provided information about possible target-related proteins of bacterial chemotaxis, swimming performance, alkane transport to stimulus of n-ctadecane rather than n-hexadecane in P. aeruginosa SJTD-1.
Highlights
Some microorganisms are able to use hydrocarbons, are toxic to animals and plants, as carbon and energy sources [1, 2]
Results of the present study provided information about possible target-related proteins of alkane uptake and be important for understanding alkane use in P. aeruginosa SJTD-1
P. aeruginosa SJTD-1 was precultured overnight in Luria-Bertani medium (LB) at 37 ̊C with vigorous shaking followed by being harvested by centrifugation and washed three times with 1× PBS buffer
Summary
Some microorganisms are able to use hydrocarbons, are toxic to animals and plants, as carbon and energy sources [1, 2]. N-alkanes are a large class of saturated hydrocarbons with extremely low water solubility. Quantitative proteomic analysis of Pseudomonas aeruginosa SJTD-1 in alkane uptake example Alcanivorax, Thalassolituus, Pseudomonas, Rhodococcus, or Acinetobacter, have been found possess the ability to degrade short- (C4-C8), medium- (C5–C16) and long-chain (C10-C36) linear n-alkanes [3,4,5,6]. Pseudomonads have been surveyed for their efficient utilization of n-alkanes for a long time. A strain of P. aeruginosa SJTD-1 was isolated from oil-contaminated soil; its hydrocarbon utilization capability and n-alkane breakdown efficiency were investigated [7]
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