Abstract
Tuber melanosporum, known as the black diamond of cuisine, is highly appreciated for its unique and characteristic aroma, which is mainly due to its volatile organic sulfur-containing compounds (VOSCs). In this work, by adding 5 g/L l-methionine to the fermentation medium, the activities of aminotransferase and α-ketoacid decarboxylase were significantly enhanced by 103 and 250 %, respectively, while the activities of alcohol dehydrogenase and demethiolase were decreased by 277 and 39 %. Then, the six VOSCs, i.e., methanethiol (MTL), dimethyl sulfide (DMS), dimethyl disulfide (DMDS), dimethyl trisulfide (DMTS), 3-(methylthio)propanal (methional), and 3-(methylthio)-1-propanol (methionol), were first detected in the submerged fermentation of T. melanosporum. These results indicated that the biosynthesis of VOSCs was triggered by aminotransferase and α-ketoacid decarboxylase. The production of methional and methionol increased with the increased concentrations of l-methionine (i.e., 5, 10, 15, and 20 g/L) before day 4 of the culture protocol, and methionol was the major product in the Ehrlich pathway. The production of MTL was significantly decreased after day 4 with a significantly increased DMDS, and DMDS was the major product of the demethiolation pathway. Compared with the demethiolation pathway with a total flux of sulfur of 11.33–24.32 μM, the Ehrlich pathway with a total flux of sulfur of 6,149–10,330 μM was considered the major pathway for the biosynthesis of VOSCs. This is the first report linking the metabolism of l-methionine to the biosynthesis of VOSCs by the Ehrlich and demethiolation pathways during the submerged fermentation of T. melanosporum.
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Acknowledgments
Financial supports from the National Natural Science Foundation of China (NSFC, Project Nos. 21176059, 21206035, and 21376066), Hubei Provincial Natural Science Foundation for Agriculture (2012DBA20001), and High-Tech Industry Development Program for Innovative Research Team in Wuhan Municipality (2013070204020049) are gratefully acknowledged. Ya-Jie Tang also thank the Chutian Scholar Program (Hubei Provincial Department of Education, China) (2006), Training Program for the Youth Leading Talents by Ministry of Science and Technology, Program for New Century Excellent Talents in University (NCET-11-0961), and Training Program for Top Talents in Hubei Province.
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Rui-Sang Liu and Huan Zhou contributed equally to this work.
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Liu, RS., Zhou, H., Li, HM. et al. Metabolism of l-methionine linked to the biosynthesis of volatile organic sulfur-containing compounds during the submerged fermentation of Tuber melanosporum . Appl Microbiol Biotechnol 97, 9981–9992 (2013). https://doi.org/10.1007/s00253-013-5224-z
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DOI: https://doi.org/10.1007/s00253-013-5224-z