Integrated analysis of transcriptome and proteome for exploring the mechanism of guaiacol production by Alicyclobacillus acidoterrestris

Alicyclobacillus spp. can cause commercially pasteurized fruit juices/beverages to spoil and the spoilage is characterized by the formation of a distinct medicinal or antiseptic off-odor attributed to guaiacol. The aim of this study was to reveal the mechanism of guaiacol production in A. acidoterrestris by combining transcriptomic and proteomic approaches. RNA-sequencing and iTRAQ analyses were conducted to investigate differences in expression levels of genes and proteins in A. acidoterrestris when producing (with 500 mu M vanillic acid) and not producing (without vanillic acid) guaiacol. A total of 225 differentially expressed genes and 77 differentially expressed proteins were identified. The transcription of genes vdcBCD encoding subunits of vanillic acid decarboxylase were 626.47, 185.01 and 52.81-fold up-regulated, respectively; they were the most up-regulated genes involved in guaiacol production. Expressions of the benzoate membrane transport protein, fusaric acid resistance protein, resistance-nodulation- division transporter, some ATP-binding cassette transporters and major facilitator superfamily transporters were increased at either mRNA, protein or both levels, indicating that they participated in the uptake of vanillic acid and extrusion of guaiacol. In the metabolic process of vanillic acid to guaiacol in A. acidoterrestris, genes related to the pathway of tricarboxylic acid cycle and ribosome were up-regulated, while the expression of some genes associated with valine, leucine and isoleucine biosynthesis was decreased. These findings provide novel insight to understand the mechanism of guaiacol production in A. acidoterrestris, which will serve as an important guide for developing strategies for the control of A. acidoterrestris problems in the fruit juice industry.