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Truffle ( Tuber spp.), also known as 'underground gold,' is popular in various cuisines because of its unique and characteristic aroma. Currently, truffle fruiting bodies are mostly obtained from nature and semi-artificial cultivation. However, the former source is scarce, and the latter is time-consuming, usually taking 4 to 12 years before harvest of the fruiting body. The truffle submerged fermentation process was first developed in Tang's lab as an alternative to its fruiting bodies. To the best of our knowledge, most reports of truffle submerged fermentation come from Tang's group. This review examines the current state of the truffle submerged fermentation process. First, the strategy to optimize the truffle submerged fermentation process is summarized; the final conditions yielded not only the highest reported truffle biomass but also the highest production of extracellular and intracellular polysaccharides. Second, the comparison of metabolites produced by truffle fermentation and fruiting bodies is presented, and the former were superior to the latter. Third, metabolites (i.e., volatile organic compounds, equivalent umami concentration, and sterol) derived from truffle fermentation could be regulated by fermentation process optimization. These findings indicated that submerged fermentation of truffles can be used for commercial production of biomass and metabolites as a promising alternative to generating its fruiting bodies in bioreactor. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The aroma active compounds of three Tuber fruiting bodies (i.e., Tuber himalayense, Tuber indicum, and Tuber sinense) were firstly systematically evaluated by instrumental gas chromatography-olfactometry combining with quantitative analysis, aroma reconstitution, and omission tests. Twelve aroma active compounds were characterized by aroma extract dilution analysis, and 3-(methylthio) propanal, 3-methylbutanal, and 1-octen-3-ol with the highest flavor dilution (FD) factor (i.e., 1,024-2,048) were suggested as key contributors to the aroma. Odor activity value (OAV) was employed to determine the relative contribution of each compound to the aroma, and the compound with the highest FD factor also had the highest OAV (i.e., 10,234-242,951). Then, the synthetic blends of odorants (aroma reconstitution) were prepared with OAV larger than 15, and their aromas were very similar to the originals. Omission tests were carried out to verify the significance of 3-(methylthio) propanal, 1-octen-3-ol, and 3-methylbutanal as key compounds in the aroma of tested Tuber fruiting bodies. [ABSTRACT FROM AUTHOR]

Abstract: A novel method using Plackett–Burman design (PBD) coupled with Draper–Lin small composite design (SCD) and desirability function (DF) was developed to optimize Tuber melanosporum fermentation medium. Firstly, sucrose, yeast extract, peptone and Mg2+ were identified to be key medium components by PBD. Secondly, in order to evaluate the relationships between responses and the identified key components, mathematical models were developed by SCD. Finally, by using DF, the responses were optimized simultaneously and the optimal concentration was located to be 73g/L sucrose, 11g/L yeast extract, 8g/L peptone, and 46mM Mg2+. Under the identified optimal conditions, the maximal biomass, the production of extracellular (EPS) and intracellular (IPS) polysaccharides was 25.10±0.12g/L, 3.88±0.23g/L and 2.87±0.32g/L, respectively, which agreed with the predicted values well. Compared with the basic medium, the biomass, the production of EPS and IPS was enhanced by 54.4%, 71.7% and 124.2%, respectively. [Copyright &y& Elsevier]

Abstract: For the first time, a fed-batch fermentation process of Tuber melanosporum was developed for the efficient production of bioactive mycelia and Tuber polysaccharides. Each 1.67g/L of peptone and 8.33g/L of yeast extract were added on day 3, 6, and 9, respectively, and sucrose was fed to maintain its concentration around 35–5g/L when its residual level decreased to 10–5g/L. Then, the maximal biomass, the production of extracellular polysaccharides (EPS) and intracellular polysaccharides (IPS) reached 53.72±2.57g DW/L, 7.09±0.62 and 4.43±0.21g/L, respectively. Compared with the batch culture conducted in the enriched medium, the biomass, the production of EPS and IPS were enhanced by 55.8%, 222.3% and 103.2%, respectively. Not only the cell density but also the production of EPS and IPS were the highest ever reported in truffle fermentation, and the biomass was also the highest as ever reported in mushroom fermentation. [Copyright &y& Elsevier]

Abstract: Truffles are hypogeous ascomycetes and ectomycorrhizal fungi with unique organoleptic properties. Effects of nitrogen source and its concentration were studied during the submerged fermentation of Chinese truffle Tuber sinense. Yeast extract was identified to be the most favorable for the cell growth, and the next was peptone. The maximal dry cell weight of 19.53±0.13g/L obtained at an initial yeast extract concentration of 25g/L was higher by 40.9% compared to that at 10g/L. Within the range of initial peptone concentration (i.e., 10, 20, 30 and 40g/L) as investigated, the maximal biomass and the production of extracellular polysaccharide (EPS) were obtained at 30g/L, while 20g/L was favorable for the production of intracellular polysaccharide (IPS). A mathematical model, which was constructed by response surface methodology combination with Box-Behnken design (BBD), was applied to study the synergic effect of glucose, yeast extract and peptone. According to the model prediction, the optimal initial concentration of glucose, yeast extract and peptone was 60, 30 and 10g/L, respectively. The validation experiment showed the experimental values agreed with the predicted values well. The highest biomass (i.e., 23.94±0.49g/L), the production of EPS (i.e., 1.59±0.06g/L) and IPS (i.e., 1.48±0.26g/L) were obtained under the predicted optimal conditions, which were higher by 22.6%, 42.0% and 16.5% compared with the best results of one-variable-at-a-time experiments, respectively. [Copyright &y& Elsevier]

The aroma attributes of sulfurous, mushroom and earthy are the most important characteristics of the aroma of Tuber melanosporum. However, these three aroma attributes are absent in the T. melanosporum fermentation system. To improve the quality of the aroma, repeated freeze-thaw treatment (RFTT) was adopted to affect the interplay of volatile organic compounds (VOCs). Using RFTT, not only was the score on the hedonic scale of the aroma increased from the 'liked slightly' to the 'liked moderately' grade, but the aroma attributes of sulfurous, mushroom and earthy could also be smelled in the T. melanosporum fermentation system for the first time. A total of 29 VOCs were identified, and 9 compounds were identified as the key discriminative volatiles affected by RFTT. Amino acid analysis revealed that methionine, valine, serine, phenylalanine, isoleucine and threonine were the key substrates associated with the biosynthesis of the 9 key discriminative VOCs. This study noted that amino acid metabolism played an important role in the regulation of the aroma of the T. melanosporum fermentation system. [ABSTRACT FROM AUTHOR]

Abstract: Besides the identified key factors of pH-shift from 3.0 to 4.5 on day 4 and dissolved oxygen tension (DOT) shift from 25 to 10% on day 6, lower shear stress was also demonstrated to be a key factor in scaling-up the fed-batch fermentation of the medicinal mushroom Ganoderma lucidum from a shake flask to a stirred-tank bioreactor. The maximal biomass (22.62g/L), intercellular polysaccharide production (4.74g/L), and ganoderic acid production (798.0mg/L) were attained at a low impeller tip speed of 1.234m/s. Based on the identified key factors of pH-shift, DOT-shift and lower shear stress, the fed-batch fermentation of G. lucidum was successfully step-wise scaled-up from 7.5- to 200-L stirred-tank bioreactor. Through the qualitative and quantitative side-by-side comparison of G. lucidum growth and metabolites accumulation, it was concluded that the comparability of the fed-batch fermentation process were well controlled and the products were maintained during the process scale-up. [ABSTRACT FROM AUTHOR]

Abstract: During the submerged fermentation of medicinal mushroom Chinese truffle Tuber sinense, there was no significant effect of metal ion on the cell growth and the production of intracellular polysaccharides, while metal ion and its concentration significantly affected the production of extracellular polysaccharides (EPS). By using the approach of “one-variable-at-a-time”, 50mM Mg2+ was identified to be the most favorable for EPS production, and the next was 10mM K+. A mathematical model, constructed by response surface methodology combination with full factorial design, was applied to study the synergic effect of Mg2+ and K+. EPS production reached its peak value of 5.86g/L under their optimal combination of 30mM Mg2+ and 5mM K+ predicted by the model, which was higher by 130.7% compared with the basal fermentation medium without metal ion. The validation experiment showed the experimental values agreed with the predicted values well. EPS production obtained in this work was the highest reported in the culture of T. sinense. [Copyright &y& Elsevier]