Genome-wide identification of tolerance mechanisms toward p-coumaric acid in Pseudomonas putida

被引:57
作者
Calero, Patricia [1 ]
Jensen, Sheila I. [1 ]
Bojanovic, Klara [1 ]
Lennen, Rebecca M. [1 ]
Koza, Anna [1 ]
Nielsen, Alex T. [1 ]
机构
[1] Tech Univ Denmark, Novo Nordisk Fdn Ctr Biosustainabil, Lyngby, Denmark
关键词
p-coumaric acid; Pseudomonas putida; Tn-seq; tolerance; GRAM-NEGATIVE BACTERIA; BROAD-HOST-RANGE; ESCHERICHIA-COLI; ETHANOL TOLERANCE; STRESS-RESPONSE; ESSENTIAL GENES; EFFLUX SYSTEM; INSERTION; KT2440; RESISTANCE;
D O I
10.1002/bit.26495
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The soil bacterium Pseudomonas putida KT2440 has gained increasing biotechnological interest due to its ability to tolerate different types of stress. Here, the tolerance of P. putida KT2440 toward eleven toxic chemical compounds was investigated. P. putida was found to be significantly more tolerant toward three of the eleven compounds when compared to Escherichia coli. Increased tolerance was for example found toward p-coumaric acid, an interesting precursor for polymerization with a significant industrial relevance. The tolerance mechanism was therefore investigated using the genome-wide approach, Tn-seq. Libraries containing a large number of miniTn5-Km transposon insertion mutants were grown in the presence and absence of p-coumaric acid, and the enrichment or depletion of mutants was quantified by high-throughput sequencing. Several genes, including the ABC transporter Ttg2ABC and the cytochrome c maturation system (ccm), were identified to play an important role in the tolerance toward p-coumaric acid of this bacterium. Most of the identified genes were involved in membrane stability, suggesting that tolerance toward p-coumaric acid is related to transport and membrane integrity.
引用
收藏
页码:762 / 774
页数:13
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