Production optimization of food functional factor ergothioneine in wild-type red yeast Rhodotorula mucilaginosa DL-X01.

BACKGROUND: Ergothioneine (EGT) is a high-value food functional factor that cannot be synthesized by humans and other vertebrates, and the low yield limits its application. RESULTS: In this study, the optimal fermentation temperature, fermentation time, initial pH, inoculum age, and inoculation ratio on EGT biosynthesis of Rhodotorula mucilaginosa DL-X01 were optimized. In addition, the effects of three key precursor substances - histidine, methionine, and cysteine - on fungal EGT synthesis were verified. The optimal conditions were further obtained by response surface optimization. The EGT yield of R. mucilaginosa DL-X01 under optimal fermentation conditions reached 64.48 ± 2.30 mg L-1 at shake flask fermentation level. Finally, the yield was increased to 339.08 ± 3.31 mg L-1 (intracellular) by fed-batch fermentation in a 5 L bioreactor. CONCLUSION: To the best of our knowledge, this is the highest EGT yield ever reported in non-recombinant strains. The fermentation strategy described in this study will promote the efficient biosynthesis of EGT in red yeast and its sustainable production in the food industry. © 2024 Society of Chemical Industry.

Cost-effective production of ergothioneine using Rhodotorula mucilaginosa DL-X01 from molasses and fish bone meal enzymatic hydrolysate.

Ergothioneine (EGT) is a high-value natural antioxidant that cannot be synthesized by the human body. This study showed that Rhodotorula mucilaginosa DL-X01 can use untreated molasses and fish bone meal enzymatic hydrolysate as the substrates to synthesize EGT. By optimizing the growth conditions, the EGT yield reached 29.39 mg/L when molasses and fish bone meal (FBM) were added at 60 g/L and 400 g/L respectively. Finally, the EGT yield was increased to 216.25 mg/L by fed-batch fermentation in a 5 L bioreactor. Compared with the fermentation by yeast extract peptone dextrose medium, the feedstock cost of EGT production was reduced by 330.91 % by using molasses and FBM as substrates. These results showed that R. mucilaginosa DL-X01 can produce high-value EGT using two cheap processing by-products, molasses and FBM, which is of great significance for environmental protection and sustainable development.

The Screening and Isolation of Ethyl-Carbamate-Degrading Strains from Fermented Grains and Their Application in the Degradation of Ethyl Carbamate in Chinese Baijiu.

Ethyl carbamate (EC), a 2A carcinogen produced during the fermentation of foods and beverages, primarily occurs in distilled spirits. Currently, most studies focus on strategies for EC mitigation. In the present research, we aimed to screen strains that can degrade EC directly. Here, we report two Candida ethanolica strains (J1 and J116), isolated from fermented grains, which can reduce EC concentrations directly. These two yeasts were grown using EC as the sole carbon source, and they grew well on different carbon sources. Notably, after immobilization with chitosan, the two strains degraded EC in Chinese Baijiu by 42.27% and 27.91% in 24 h (from 253.03 ± 9.89 to 146.07 ± 1.67 and 182.42 ± 5.05 µg/L, respectively), which was better than the performance of the non-immobilized strains. Furthermore, the volatile organic compound content, investigated using gas chromatography-mass spectrometry, did not affect the main flavor substances in Chinese Baijiu. Thus, the yeasts J1 and J116 may be potentially used for the treatment and commercialization of Chinese Baijiu.

Ergothioneine: new functional factor in fermented foods.

Ergothioneine (EGT) is a high-value natural sulfur-containing amino acid and has been shown to possess extremely potent antioxidant and cytoprotective activities. At present, EGT has been widely used in food, functional food, cosmetics, medicine, and other industries, but its low yield is still an urgent problem to overcome. This review briefly introduced the biological activities and functions of EGT, and expounded its specific applications in food, functional food, cosmetic, and medical industries, introduced and compared the main production methods of EGT and respective biosynthetic pathways in different microorganisms. Furthermore, the use of genetic and metabolic engineering methods to improve EGT production was discussed. In addition, the incorporation of some food-derived EGT-producing strains into fermentation process will allow the EGT to act as a new functional factor in the fermented foods.

Mapping of Research in the Field of Forest Therapy-Related Issues: A Bibliometric Analysis for 2007-2021.

Exposure to forest environments promotes human health. The number of relevant studies in this area has increased rapidly. However, an overall review of relevant analyses from the perspectives of bibliometrics and visualization is lacking. A scientometric analysis of 2,545 publications from 2007 to 2021 via the Web of Science database was conducted to identify the knowledge structure and frontiers objectively. The publications were subsequently analyzed in terms of the distribution of journals and countries, citation bursts, major subject areas, and evolutionary stages. The findings showed that the knowledge foundation of forest therapy was multidisciplinary with most published in the fields of environmental sciences and ecology but lacking input from social disciplines. The research hotspots evolved from the early focus on individual benefits obtained from nature to increasing attention on human well-being at the social-ecological scale. More rigorous experiments with strict randomized controlled trials and blinding are needed to accommodate the trend of forest therapy toward non-pharmacological treatments. According to Shneider's four-stage theory, forest therapy research is in the third stage of the scientific research process. More future studies utilizing novel technologies and decision-making frameworks to solve practical issues are needed for introducing health into policies and promoting human well-being.

Genetic Engineering Production of Ethyl Carbamate Hydrolase and Its Application in Degrading Ethyl Carbamate in Chinese Liquor.

Ethyl carbamate (EC), classified as a Group 2A carcinogen, is most abundant in the fermented foods, such as Cachaca, Shaoxing wine, and Chinese liquor (baijiu). Although biodegradation can reduce its concentration, a high ethanol concentration and acidic environment often limit its degradation. In the present study, a novel ethyl carbamate hydrolase (ECH) with high specificity to EC was isolated from Acinetobacter calcoaceticus, and its enzymatic properties and EC degradability were investigated. ECH was immobilized to resist extreme environmental conditions, and the flavor substance changes were explored by gas chromatography-mass spectrometry (GC/MS). The specific enzymatic activity of ECH was 68.31 U/mg. Notably, ECH exhibited excellent thermal stability and tolerance to sodium chloride and high ethanol concentration (remaining at 40% activity in 60% (v/v) ethanol, 1 h). The treatment of immobilized ECH for 12 h decreased the EC concentration in liquor by 71.6 µg/L. Furthermore, the immobilized ECH exerted less effect on its activity and on the flavor substances, which could be easily filtrated during industrial production.

Functional Aerogels Composed of Regenerated Cellulose and Tungsten Oxide for UV Detection and Seawater Desalination.

Functional aerogels composed of regenerated cellulose and tungsten oxide were fabricated by implanting tungsten-oxide nanodots into regenerated cellulose fiber. This superfast photochromic property benefitted from the small size and even distribution of tungsten oxide, which was caused by the confinement effect of the regenerated cellulose fiber. The composite was characterized using XRD and TEM to illustrate the successful loading of tungsten oxide. The composite turned from pale white to bright blue under ambient solar irradiation in five seconds. The evidence of solar absorption and electron paramagnetic resonance (EPR) demonstrated the fast photochromic nature of the composite and its mechanism. Furthermore, carbon fiber filled with preferential growth tungsten-oxide nanorods was obtained by annealing the photochromic composite in a N2 atmosphere. This annealed product exhibited good absorption across the whole solar spectrum and revealed an excellent photothermal conversion performance. The water evaporation rate reached 1.75 kg m-2 h-1 under one sun illumination, which is 4.4 times higher than that of pure water. The photothermal conversion efficiency was 85%, which shows its potential application prospects in seawater desalination.

Screening of dominant strains in red sour soup from Miao nationality and the optimization of inoculating fermentation conditions.

Red sour soup is a traditional fermented product in southwest China. Currently, the existing production process mainly adopts the method of natural fermentation, with long fermentation cycles and poor stability between batches. Rapid establishment of dominant strains can accelerate the formation of lactic acid, which can inhibit the growth of miscellaneous bacteria. It is also helpful for the inhibition of nitrite accumulation, shortening of fermentation. In this study, the dominant strain H9, with lactic acid-producing ability, was isolated from the natural fermented red sour soup, and was identified as Lactobacillus buchneri, based on the 16s rRNA sequence analysis and biochemical identification. Then, the optimization of fermentation conditions was performed using L. buchneri H9 strain as external bacteria. The optimized fermentation conditions were temperature of 22°C, starch dosage of 11.24 g/L, and initial inoculation of 3.5 × 108 cfu/L. The concentration of lactic acid reached 8.029 g/L after 8 days of inoculating fermentation, which exceeded 6.221 g/L for 20 days of natural fermentation. Compared with natural fermentation, the peak of nitrite during inoculating fermentation appeared earlier and the peak height was lower. While the nitrite content in inoculating fermentation decreased to safety threshold more quickly. The volatile flavor compounds analysis showed that 41 types of volatile compounds were detected in the inoculating fermentation product, while 45 in the natural fermentation product. Over 88% compounds were overlapped, which means similar flavor between two fermentation products. These results provide a sufficient scientific basis for the industrialized production of inoculating fermentation of red sour soup.

Correction: Ambiguity in the processing of Mandarin Chinese relative clauses: One factor cannot explain it all.

[This corrects the article DOI: 10.1371/journal.pone.0178369.].

Ambiguity in the processing of Mandarin Chinese relative clauses: One factor cannot explain it all.

This study addresses the question of whether native Mandarin Chinese speakers process and comprehend subject-extracted relative clauses (SRC) more readily than object-extracted relative clauses (ORC) in Mandarin Chinese. Presently, this has been a hotly debated issue, with various studies producing contrasting results. Using two eye-tracking experiments with ambiguous and unambiguous RCs, this study shows that both ORCs and SRCs have different processing requirements depending on the locus and time course during reading. The results reveal that ORC reading was possibly facilitated by linear/temporal integration and canonicity. On the other hand, similarity-based interference made ORCs more difficult, and expectation-based processing was more prominent for unambiguous ORCs. Overall, RC processing in Mandarin should not be broken down to a single ORC (dis)advantage, but understood as multiple interdependent factors influencing whether ORCs are either more difficult or easier to parse depending on the task and context at hand.