Compounds In Soy Sauce Research Articles

Co‐culture relationship between Zygosaccharomyces rouxii and Candida versatilis and its effect on the flavour of soy sauce

SummaryZygosaccharomyces rouxii and Candida versatilis are important flavour‐producing yeasts in soy sauce fermentation. In the present study, the interaction between Z. rouxii and C. versatilis co‐cultures in YPD and raw soy sauce medium was investigated and applied to soy sauce fermentation. The concentration of Z. rouxii in the co‐culture was 0.47 times that in the mono‐culture. This observation suggested that C. versatilis inhibited the growth of Z. rouxii, likely attributed to various factors, including the production of acids and heat‐ and protease‐sensitive substances by C. versatilis. Furthermore, we examined the physicochemical properties and characteristic volatile flavour components of soy sauce by adding varying ratios of Z. rouxii and C. versatilis at different stages of fermentation. The results showed that mixed fermentation of Z. rouxii and C. versatilis enhanced the total nitrogen, amino acid nitrogen, 4‐hydroxy‐2(5)‐ethyl‐5(2)‐methyl‐3(2H)‐furanone, and volatile compounds in soy sauce compared to single‐strain fermentation. Additionally, optimal soy sauce results were obtained when yeasts were added on day 45, with an inoculation ratio of 1:1. In conclusion, during the moromi fermentation stage, the appropriate selection of the inoculation ratio and time for Z. rouxii and C. versatilis addition can effectively enhance the physicochemical properties and flavour attributes of soy sauce.

Dispersive liquid-liquid microextraction coupled with gas chromatography-mass spectrometry (GC-MS) for the determination of soy sauce aroma compounds

In the present work, a dispersive liquid-liquid microextraction (DLLME) method combined with gas chromatography-mass spectrometry (GC-MS) was successfully developed for accurate determination of aroma-active components in soy sauce. Several key experimental parameters were optimized, including the type of dispersant, vortex time and the volume ratio of extractant and dispersant. The optimal conditions were 1200 μL acetone as dispersant, 800 μL dichloromethane as extractant and 3 min of vortex mixing. The proposed method was validated, achieving good linearities with R2 ≥ 0.9941, recoveries between 82 and 129% with 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF) and 4-hydroxy-2 (or 5)-ethly-5 (or 2)-methyl-3(2H)-furanone (HEMF) within 84–103%, relative standard deviations ranging from 0.02 to 8.60% with HDMF and HEMF between 0.26 and 1.44%, and limits of detection and limits of quantification in the range of 0.08–35.95 μg/L and 0.25–119.83 μg/L, respectively. The entire sample processing with DLLME was completed in roughly 15 min with a total consumption of 2 mL of organic solvent, and 93 volatiles identified in the obtained extracts, with 87.1% (81 volatiles) showing relative standard deviations lower than 10%. Compared to conventional liquid-liquid extraction (LLE), DLLME exhibited obvious advantages of simplicity, rapidity, stability and low solvent consumption, favouring the extraction of acids and ketones. This is the first application of the DLLME technique for the analysis of aroma compounds in soy sauce and is suitable for rapid determination of key aroma compounds like HDMF and HEMF, especially for routine detection and large batch analysis of soy sauce, which can provide reliable information for soy sauce flavour analysis.

Characterization of the key aroma compounds in soy sauce by gas chromatography-mass spectrometry-olfactometry, headspace-gas chromatography-ion mobility spectrometry, odor activity value, and aroma recombination and omission analysis.

Most previous studies on volatile compounds in soy sauce were performed by gas chromatography-mass spectrometry (GC-MS). In this study, the volatile compounds of high-salt liquid-state fermentation soy sauce (HLFSS) were analyzed qualitatively and quantitatively by GC-MS and headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS). One hundred and seventy-four substances were detected using the two instruments, 87 by HS-GC-IMS and 127 by GC-MS. Aldehydes (26), ketones (28), esters (29), and alcohols (26) were the main compounds in HLFSS. In addition, ethyl pyruvate, (E)-2-pentenal and diethyl propanedioate were detected by HS-GC-IMS, which were previously not detected in HLFSS. Forty-eight aromatics including 34 key ones were identified by gas chromatography-olfactometry. Phenylacetaldehyde, methional, 2-methylbutanal, 1-octen-3-ol, ethyl acetate, 2-ethyl-4-hydroxy-5-methyl-3(2H)-furanone, 4-hydroxy-2,5-dimethyl-3(2H)-furanone and 4-ethyl guaiacol were identified as the main aroma compounds in HLFSS by aroma recombination and omission test. This study laid foundation for developing flavor assessment standards for soy sauce.

Analysis of the contribution of koji-making with Z. rouxii on volatile compounds of soy sauce

The co-inoculation fermentation of different microorganisms plays a crucial role in soy sauce fermentation in terms of enriching the composition of the volatile compounds. Therefore, the present study sought to investigate the effect of co-inoculation fermentation of Aspergillus oryzae and Zygosaccharomyces rouxii on the dynamics of volatile compounds in soy sauce using the combination of gas chromatography-mass spectrometry (GC-MS) and multivariate correlation analysis. A total of 124 volatile compounds were identified by A. oryzae and Z. rouxii inoculation alone and co-inoculation, including hydrocarbons, fatty acids, phenols, fatty esters, and benzamides. The content of phenylethyl alcohol, ethyl linoleate, and ethyl palmitate was significantly increased, and the metabolic pathways of multiple volatile compounds were affected in soy sauce inoculated with Z. rouxii. The present study results potentially contribute to the optimization of soy sauce fermentation processes.

Flavor differences of soybean and defatted soybean fermented soy sauce and its correlation with the enzyme profiles of the kojis.

Soybeans and defatted soybeans, commonly used as protein ingredients, have different flavors of their fermented soy sauce. Clarifying the differences between the two soy sauces, as well as the formation mechanism, is an important prerequisite for improving the flavor of defatted soybean soy sauce. To this goal, the aroma characteristics of two soy sauces and their volatile profiles were compared by sensory evaluation and gas chromatography-mass spectrometry, and eight enzyme activities and volatile profiles of matured koji were determined. Sensory results showed that the acids, fruity and cooked potato-like attributes were higher in whole soybean fermented soy sauce, whereas defatted soybean soy sauce exhibited higher smoky and malty attributes, closely related to the contents of aroma-active compounds in soy sauce, such as isobutyl acetate, 2/3-methylbutanal, acetic acid and 2/3-methylbutanoic acid. The content of most volatiles in the matured kojis showed a consistent trend with that of soy sauce: alcohols, acids, furan(one)s and ketones. Interestingly, acid protease and cellulase activities were 3.3 and 1.6 times higher in the whole soybean koji than in defatted soybean koji, respectively, whereas neutral protease, aminopeptidase, glucoamylase and β-glucosidase were approximately 2.0 times higher in defatted soybean koji. In summary, the flavor differences between soybean and defatted soybean fermented soy sauce were not only caused by the differences in the content of flavor precursors in the materials, but also closely related to the differences in the enzymatic profiles accumulated during the koji-making process. © 2022 Society of Chemical Industry.

Changes in the Flavor Compounds in Soy Sauce Stored in Antioxidation Containers

Changes in the Flavor Compounds in Soy Sauce Stored in Antioxidation Containers

Stir bar sorptive extraction of aroma compounds in soy sauce: Revealing the chemical diversity

Fermented soy sauce is used worldwide to enhance the flavour of many dishes. Many types of soy sauce are on the market, and their differences are mostly related to the country of origin, the production process applied and the ratio of ingredients used. Consequently, several aromas, tastes, colours, and textures are obtained. Nowadays, soy sauce can also be produced without microorganisms making the process shorter and cheaper. However, flavour may be lost. We have carried out a comprehensive metabolomics analysis of volatile compounds using stir bar sorptive extraction (SBSE)-GC–MS to relate differences in volatile content to production history and origin. The results revealed major differences between fermented and non-fermented soy sauces, and a list of volatile compounds is reported as being characteristic of each type. This study was able to relate volatiles to the production process using SBSE-GC–MS and to aroma characteristics using GC-O-MS.

Correlation analysis between the concentration of α-dicarbonyls and flavor compounds in soy sauce

Soy sauce was fermented with various sterilization temperatures. Among the Maillard reaction products, α-dicarbonyl compounds (α-DC; glyoxal, methylglyoxal, and diacetyl) and 16 aroma compounds were analyzed, and their correlations were investigated. After fermentation for 8 wk, glyoxal, methylglyoxal, and diacetyl were present in soy sauce at 0.88–2.84, 1.73–3.27, and 0.53–0.86 μg/ml, respectively. Among the 16 aroma compounds, acetic acid dominated and showed the largest change with the sterilization temperature. The quantities of α-DC and volatiles showed a strong positive correlation (maximum r = 0.912) with the sterilization temperature. To reduce the levels of α-DC in soy sauce, a 60–70 °C sterilization temperature is proposed. Partial least squares regression was done to investigate the aroma compounds affecting the formation of α-DC. Among the volatile compounds analyzed in this soy sauce model system, 1-octen-3-ol was the most likely determinant of α-DC content.

Effects of Tetragenococcus halophilus and Candida versatilis on the production of aroma-active and umami-taste compounds during soy sauce fermentation.

Soy sauce is a well-known condiment worldwide. However, the high salt content in soy sauce contributes to the high intake of sodium salt, which usually results in high blood pressure. High salt soy sauce usually has the better quality (aroma and taste) than low salt. Tetragenococcus halophilus and Candida versatilis are important starters for soy sauce fermentation. It is of urgent need to explore what the effect of these two strains on the aroma- and taste-attributes of soy sauce to achieve high quality fermentation with low salt. In this study, aroma-active and taste compounds in soy sauce were reviewed and listed. Then, soy sauce fermentation inoculated with different combinations of T. halophilus (at different inoculated time) and C. versatilis were completed. Aroma-active and taste compounds in different samples were quantified. Multivariate analysis was used to analyze these data. The aroma-active compounds which were highly related to the inoculation of T. halophilus and C. versatilis were found. Meanwhile, the addition time of T. halophilus could also be highly related to the production of aroma-active compounds. More importantly, T. halophilus was highly correlated with the production of umami-taste compounds in soy sauce, including aspartic acid, glutamic acid, alanine and N -succinyl-glutamic acid. These results will provide a better understanding of the effects of T. halophilus and C. versatilis on the formation of significant aroma-active and umami-taste constituents in soy sauce. Furthermore, it will be helpful to realize fermentative control of soy sauce with high quality at low salt. © 2020 Society of Chemical Industry.

Characterisation of sugars as the typical taste compounds in soy sauce by silane derivatisation coupled with gas chromatography–mass spectrometry and electronic tongue

SummaryA total of 214 taste compounds were identified in soy sauce by silane derivatisation coupled with gas chromatography–mass spectrometry (SD‐GC‐MS), including 74 kinds of sugars. The proportion of total sugars is highest from 37.24–77.24% among all taste compounds. In particular, rare sugars were detected and identified as the special compounds in soy sauce which come from the sugar metabolism. Rare sugars identified in soy sauce by traditional fermented technology were prominent than Japanese technology. Principal component analysis (PCA) investigation of these results showed that samples can be distinguished by sugars as the first principal component and the general evaluation index (GEI) of samples was consistent with the result of sensory evaluation. Meanwhile, sweetness had the maximum range (from −9.89 to 14.10) in all taste indexes by electronic tongue (E‐tongue) analysis.

Determination of Key Volatile Compounds Related to Long-Term Fermentation of Soy Sauce.

The changes of volatile compounds in soy sauce during long-term fermentation (12 months) were investigated using solid-phase microextraction (SPME) and stir bar sorptive extraction (SBSE). A total of 144 and 129 compounds were identified in soy sauce with long-term fermentation by SPME and SBSE, respectively. The contents of most compounds, such as acids, aldehydes, benzene and benzene derivatives, esters, lactones, pyrazines, pyrones, and pyrroles, showed a tendency to increase, whereas those of alcohols and ketones decreased according to long-term fermentation. In addition, principal component analysis and partial least squares discriminant analysis were applied to discriminate soy sauce samples according to fermentation periods and determine key volatile compounds related to long-term fermentation. The initial fermentation stages were mainly associated with some alcohols, ketones, and lactones, whereas the later stages were strongly associated with most esters, some phenols, benzene and benzene derivatives, and pyrroles. Moreover, the key volatile compounds associated with long-term fermentation in soy sauce samples were ethyl 3-methylbutanoate (ethyl isovalerate), ethyl pentanoate (ethyl valerate), 1-octen-3-yl acetate, 3-(methylthio)-1-propanol (methionol), ethyl benzoate, ethyl 2-phenylacetate, 1-(1H-pyrrol-2-yl)ethanone (2-acetylpyrrole), and 5-pentyl-2-oxolanone (γ-nonalactone). PRACTICAL APPLICATION: This study investigated changes of volatile compounds in soy sauce during long-term fermentation (12 months) using solid-phase microextraction and stir bar sorptive extraction. In addition, the key volatile compounds associated with long-term fermentation in soy sauce samples were determined. These results may help to predict the effective contributors related to long-term fermentation of soy sauce and improve the quality of soy sauce during long-term fermentation.

Effects of sonication during moromi fermentation on antioxidant activities of compounds in raw soy sauce

To elucidate the effects of sonication on the antioxidant activities of compounds in soy sauce, sonication at 68 kHz (60 W/L/10 min/8 circles) was employed during moromi fermentation. Sonication enhanced four antioxidant indices of soy sauce from 5.29% to 24.29%, which were attributed to the significantly higher levels of free amino acids (FAAs, 12.45%), total phenolics (26.71%) and 1–3 kDa peptides (12.28%) in sonicated moromi. Amongst them, FAAs contributed 65.88% and 53.45% to the enhanced metal ion chelating activity and DPPH radical scavenging activity (p < 0.05), respectively. However, Maillard reaction products and isoflavones contributed little to the enhanced antioxidant activities, because sonication slightly affected Maillard reaction products, isoflavones and β-glucosidase activities during moromi fermentation. It was concluded that cell wall of soybean was destroyed by sonication, facilitating the release of antioxidant compounds, which significantly enhanced the antioxidant activities in raw soy sauce.

Solid-phase microextraction Arrow for the volatile organic compounds in soy sauce.

A novel solid-phase microextraction Arrow was used to separate volatile organic compounds from soy sauce, and the results were verified by using gas chromatography with mass spectrometry. Solid-phase microextraction Arrow was optimized in terms of three extraction conditions: type of fiber used (polydimethylsiloxane, polyacrylate, carbon wide range/polydimethylsiloxane, and divinylbenzene/polydimethylsiloxane), extraction temperature (40, 50, and 60°C), and extraction time (10, 30, and 60min). The optimal solid-phase microextraction Arrow conditions were as follows: type of fiber=polyacrylate, extraction time=60min, and extraction temperature=50°C. Under the optimized conditions, the solid-phase microextraction Arrow was compared with conventional solid-phase microextraction to determine extraction yields. The solid-phase microextraction Arrow yielded 6-42-fold higher levels than in solid-phase microextraction for all 21 volatile organic compounds detected in soy sauce due to the larger sorption phase volume. The findings of this study can provide practical guidelines for solid-phase microextraction Arrow applications in food matrixes by providing analytical methods for volatile organic compounds.

Effect of volatile compounds on the quality of Japanese fermented soy sauce

To understand the effects of volatile compounds on the evaluation of soy sauces so their quality could be classified, samples collected over 3 consecutive years from 2015 to 2017 were evaluated then ranked. The volatile compounds of these evaluated samples were directly extracted by headspace analysis to reduce technical interference and to identify the most common compounds in soy sauce. Sixty-four volatile compounds were identified in the evaluated soy sauces, of which 35 were found in all samples. The correlation coefficients between the levels of volatile compounds detected and the sensory rankings of the evaluated soy sauces were calculated. This enabled those volatile compounds having a greater measure of influence, both negatively and positively, on the quality of the soy sauce to be identified. It was noteworthy that the quality of Japanese fermented soy sauces was negatively correlated with their levels of 4-ethyl-2methoxyphenol and pyrazines.

Enrichment of antioxidants from soy sauce using macroporous resin and identification of 4-ethylguaiacol, catechol, daidzein, and 4-ethylphenol as key small molecule antioxidants in soy sauce

The adsorption and desorption characteristics of seven macroporous resins on the antioxidants in soy sauce were investigated. SP-207 and SP-825 resins possessing good adsorption and desorption capacities were studied further. The pseudo-second-order kinetics and Langmuir isotherm models were demonstrated to be appropriate to describe the whole exothermic and physical adsorption processes of antioxidants onto resins. The 60% ethanol eluted fraction from soy sauce purified by SP-825 resin column possessed the strongest antioxidant activity. The antioxidant activities and contents of typical soy isoflavones, furanones, pyranones, and phenolic acids in soy sauce were determined. These compounds contributed to 50.02% of the total antioxidant activity of the SP-60% fraction. The key small molecule antioxidant compounds in soy sauce were identified as 4-ethylguaiacol, catechol, daidzein, and 4-ethylphenol by the antioxidants omission experiments. Additionally, the purified active fraction with high contents of antioxidants from soy sauce could be applied as bioactive ingredient in food industry.

Effects of a mixed koji culture of Aspergillus oryzae HG‐26 and Aspergillus niger HG‐35 on the levels of enzymes, antioxidants and phenolic compounds in soy sauce during the fermentation process

SummarySoy sauce contains a number of bioactive components, which have been shown to possess strong antioxidative properties. The objective of this study was to compare the enzyme activities, antioxidant activity and phenolic compounds content of soy sauces prepared from a mixed koji (SSAON, inoculated with Aspergillus oryzae HG‐26 and Aspergillus niger HG‐35), with those of soy sauces made from a koji culture containing only Aspergillus oryzae HG‐26 koji (SSAO). The total phenolic content (TPC), total flavonoid content (TFC), content of three isoflavone aglycones (daizein, glycitein and genistein) and the antioxidant activity of the SSAON were found to be higher than those of the SSAO during moromi fermentation. In addition, they showed strong positive correlations with the antioxidant potential of the soy sauce. This study has demonstrated the potential of a mixed‐culture koji, based upon A. orzyae and A. niger, for the production of soy sauce with an increased level of bioactive components.

Flavor improvement of fermented soy sauce by extrusion as soybean meal pretreatment

Flavor, an important index of soy sauce quality, includes odor and taste which mainly depend on volatile compounds and amino acids. Soy sauce made from extruded mixture of soy meal and flour (EMSF) was employed in fermentation to improve the flavor of final product. Combination of Headspace solid phase microextraction (HS-SPME) and GC-MS was employed to identify the volatile compounds, 40 volatile compounds were identified in soy sauce made from EMSF and 24 volatile compounds in soy sauce made form cooked mixture of soy meal and flour (CMSF). Pyrazines and their derivatives occupied one-third of the total content of volatile compounds. Both samples had the same amino acid number, while total content of amino acids and contents of 15 kinds of amino acids in soy sauce made from EMSF were significantly higher than CMSF. Practical applications Extrusion had the advantage in low cost and capability of continuous processing, extruded mixture of soy meal and flour were used in soy sauce fermentation. Content of amino acids was improved by extrusion, this pretreatment method also increased volatile compounds of soy sauce and produced more desirable flavor and taste soy sauce.

Reducing the Influence of the Thermally Induced Reactions on the Determination of Aroma-Active Compounds in Soy Sauce Using SDE and GC-MS/O

A method based on simultaneous distillation extraction (SDE) coupled with gas chromatography–mass spectrometry/olfactometry (GC-MS/O) was developed for the analysis of volatile profiles in soy sauce. Its optimum operating conditions were as follows: 100 mL soy sauce plus 100 mL saturated brine; 2 h extraction using 50 mL dichloromethane. Eighty-eight volatile compounds were identified. Of these, 26 aroma-active compounds were detected by GC-O including 3-(methylthio)propanal, 2-methoxy-4-vinylphenol, 2-methoxyphenol, 2-methylbutanoic acid, 2,3-butanedione, phenylacetaldehyde, 2-ethyl-6-methylpyrazine, 3-methylbutanal, 2-methylbutanal, 3-methyl-1-butanol, and 2-methyl-1-butanol (which occurred at a high odor intensity in both high-salt liquid-state fermentation soy sauce (HLFSS) and low-salt solid-state fermentation soy sauce (LSFSS). SDE analysis of the odorless soy sauce indicated 12 volatiles (mainly Strecker aldehydes and α-dicarbonyl compounds). Thus, the proportions of these compounds (1.94–54.07 %) shown in previous GC-MS results could result from thermal degradation of non-volatile compounds in soy sauce. Especially, the contribution of 3-methylbutanal, 2-methylbutanal, phenylacetaldehyde, 3-(methylthio)propanal, and 2,3-butanedione to the aroma of soy sauce could be overestimated by conventional SDE-GC-O. With a full consideration of such limitations, this study will improve the application of SDE on aroma analysis of complex food matrix.

Autolysis of Aspergillus oryzae Mycelium and Effect on Volatile Flavor Compounds of Soy Sauce.

The autolyzed mycelia of Aspergillus oryzae are rich in proteins, nucleic acids, sugar, and other biomacromolecules, and are one of the main contributors to the flavor profile of commercially important fermented goods, including soy sauce and miso. We induced autolysis of the mycelia of A. oryzae over 1 to 10 d, and found that the maximum dissolved amounts of total protein and nucleic acid ratio accounted for 28.63% and 88.93%, respectively. The organic acid content, such as citric acid, tartaric acid, succinic acid, lactic acid, and acetic acid, initially increased and then decreased as autolysis progressed, corresponding to changes in pH levels. The main characteristic flavor compounds in soy sauce, namely, ethanol, 2-phenylethanol, and 2-methoxy-4-vinylphenol, were all detected in the autolysate. Subsequently, we tested the effect of adding mycelia of A. oryzae during the fermentation process of soy sauce for 60 d, and found that addition of 1.2‰ A. oryzae mycelia provided the richest flavor. Overall, our findings suggest that compounds found in the autolysate of A. oryzae may promote the flavor compounds of soy sauce, such as alcohols, aldehydes, phenols, and esters.

Both BAT1 and ARO8 are responsible for unpleasant odor generation in halo-tolerant yeast Zygosaccharomyces rouxii.

Soy sauce yeast Zygosaccharomyces rouxii plays a central role in the production of flavor compounds in soy sauce, while the flor-forming strain spoils its quality by producing 2-methylpropanoic acid, 2-methylbutanoic acid, and 3-methylbutanoic acid, which have an unpleasant odor. To investigate the relationship between flor formation and unpleasant odor, we measured the volatile compounds that accumulated under various growth conditions. As a result, marked amounts of 2-methylpropanoic acid, 2-methylbutanoic acid, or 3-methylbutanoic acid accumulated in synthetic medium containing valine, isoleucine, or leucine, respectively, under aerobic growth conditions. These results implied that the unpleasant compounds were produced from their corresponding branched chain amino acid (BCAA) when the cell was placed under aerobic condition through flor formation. The first step in BCAA catabolism and the last step in BCAA anabolism are both catalyzed by a BCAA transaminase. A mutant lacking the BCAA transaminase gene, BAT1, resulted in valine and isoleucine auxotrophy, while a mutant lacking both BAT1 and the α-aminoadipate aminotransferase gene, ARO8, resulted in valine, isoleucine, and leucine auxotrophy. Although the bat1∆ aro8∆ double mutant formed flor similarly to the wild-type strain, the mutant exhibited less unpleasant odor generation. These results suggest that the interconversion between 4-methyl-2-oxopentanoate and leucine is catalyzed by both Bat1p and Aro8p in Z. rouxii. Taken together, these results indicate that flor formation is not seemed to be directly linked to unpleasant odor generation. These findings encourage us to breed flor-forming yeasts without an unpleasant odor.