Soy Sauce and Its Umami Taste: A Link from the Past to Current Situation

ABSTRACT: Soy sauce taste has become a focus of umami taste research. Umami taste is a 5th basic taste, which is associated to a palatable and pleasurable taste of food. Soy sauce has been used as an umami seasoning since the ancient time in Asia. The complex fermentation process occurred to soy beans, as the raw material in the soy sauce production, gives a distinct delicious taste. The recent investigation on Japanese and Indonesian soy sauces revealed that this taste is primarily due to umami components which have molecular weights lower than 500 Da. Free amino acids are the low molecular compounds that have an important role to the taste, in the presence of sodium salt. The intense umami taste found in the soy sauces may also be a result from the interaction between umami components and other tastants. Small peptides are also present, but have very low, almost undetected umami taste intensities investigated in their fractions.     

Chemical and sensory characteristics of low molecular weight fractions obtained from three types of Japanese soy sauce (shoyu) – Koikuchi, tamari and shiro shoyu

Three typical forms of Japanese soy sauce (shoyu), koikuchi, tamari and shiro shoyu, can be differentiated, primarily due to their different compositions of soybeans and wheat used for their productions. To evaluate and compare the chemical characteristics of the low molecular weight (MW) fractions of three types of shoyu with their sensory data, gel filtration fractions of ultrafiltration products with MW less than 500 Da (F-500) were subjected to chemical and sensory analyses. The results showed that salty and umami tastes were characteristic of all F-500 fractions, however, the umami taste intensities of those of koikuchi and tamari shoyu were found to be twice as large as that of shiro shoyu. After separation by gel filtration, it was found that the tastiest fractions of the three types of shoyu were those containing sodium salt, free l-glutamic acid and most other free amino acids, especially sweet taste-eliciting amino acids, at concentrations above their thresholds. In some umami fractions of koikuchi and shiro shoyu, that predominantly contained salt and phenylalanine but had a relatively low free l-glutamic acid content, a potential synergistic effect among free l-glutamic acid, salt and phenylalanine was obvious. This first report offers new insights into soy sauce research.     

酱油中鲜味二肽的分离鉴定及其呈味特性研究

为了明确酱油中鲜味肽的结构序列,本文对酱油中鲜味肽进行分离鉴定,并系统地研究了其呈味特性。酱油经超滤膜(膜通量:5 ku、3 ku和1 ku)分离获得4个组分(F1,F2,F3和F4),通过感官评定筛选出鲜味最强的组分F4(1 ku)。该组分通过Sephadex G-15凝胶层析色谱继续分离得到8个组分(P1、P2、P3、P4、P5、P6、P7和P8),经感官评定筛选出鲜味最强的组分P2。采用超高压液相色谱串联质谱技术对P2组分进行多肽结构鉴定,通过手动De novo测序得到4条新的鲜味肽,其序列分别是Asn-Pro(230.1135 u)、Ala-His(227.1026 u)、Gly-Pro(173.0929 u)和Gly-Leu(189.1230 u)。然后采用固相合成技术合成四条肽并通过感官评定和电子舌分析,结果表明四条肽均具有明显的鲜味或鲜味增强作用。进一步研究发现,酱油的鲜味不仅来自于谷氨酸和天冬氨酸等鲜味氨基酸,小分子肽类也是构成酱油鲜味的重要成分之一。     

甜面酱的鲜味成分分析

采用国标方法分别对7种市售甜面酱样品的水分、粗脂肪、总氮和粗蛋白的含量进行了测定,利用氨基酸自动分析仪、高效液相色谱仪分别对甜面酱中的游离氨基酸及核苷酸含量进行了测定,利用凝胶色谱柱对甜面酱中小分子肽的分子量分布进行了测定,同时分析计算了甜面酱的水解度、平均肽链长度和氨基酸味道强度值,及其对呈味的贡献。研究结果表明:7种市售甜面酱的水分、粗脂肪、总氮和粗蛋白等平均含量分别为51.42%,1.84%,1.284%和7.32%;甜面酱中游离氨基酸总含量为1.55~2.78 g/100 g,含量相对较高的为脯氨酸、谷氨酸、亮氨酸、天门冬氨酸,其中谷氨酸的味道强度值(TAV值)最高(7.00~25.67),对鲜味具有重要的贡献;甜面酱中的核苷酸主要为次黄嘌呤(3.19~4.87 mg/100 g),但其TAV值均小于0.5,对甜面酱的滋味没有直接影响;甜面酱中小分子肽的含量为0.55~3.31 g/100 g,平均肽链长度为2~4,分子量主要分布于221~445 u之间,且对鲜味具有一定的贡献。     

pH值对酱油中呈味肽种类和呈味特性的影响

利用固相萃取技术结合超高效液相色谱-串联质谱法对不同pH值发酵酱油的酸性组分进行分离、鉴定和呈味特性分析.结果表明,从调控pH值发酵的酱油中鉴定出14 种呈味肽,从自然pH值发酵的酱油中鉴定出12 种呈味肽,其中8 种呈味肽在两种酱油中均鉴定出来.调控pH值发酵的酱油中含有较多良好呈味效果的小分子肽,其中ED、EE、E...     

The effect of the Corynebacterium glutamicum on the shortening of fermentation time,physicochemical and sensory properties of soy sauce

This study for the first time explores the possibility of using Corynebacterium glutamicum Ⅲ (C. glutamicum) to shorten the soy sauce fermentation period without compromising product quality. C. glutamicum was added to soy sauce that had been fermented with Aspergillus oryzae for 30 days (SS30) to initiate a further 3-day fermentation (yielding SS33). The 3-day fermentation exerted similar impacts on SS30 as the standard fermentation for further 60 days (yielding SS90): that is, increased the contents of total nitrogen, total free amino acids (FAAs), umami FAAs, acids, esters, ketones, furans and sulphur-containing compounds, whilst decreasing the contents of alcohols and phenols. This explained the similarities in taste, aroma and overall liking between SS33 and SS90. The SS90 was the darkest among the three soy sauces. Compared with SS90, SS33 was less salty and smoky, more umami and kokumi, softer and more mellow, higher overall liking, with more 0.5–1 kDa peptides and fewer peptides <0.5 kDa. Therefore, a 3-day fermentation with C. glutamicum after an initial 30 days of a normal soy sauce fermentation represents a simple and effective way to shorten the soy sauce production time from 90 to 33 days whilst increasing greatly the umami taste and mellow sensation. This approach enhances product competitiveness via lowering manufacturing cost.     

Suppression of Umami Aftertaste by Polysaccharides in Soy Sauce

Umami is one of 5 basic tastes that make foods savory and palatable. The umami aftertaste is a long‐lasting taste sensation that is important for Japanese broth (dashi) utilized for various Japanese foods. Soy sauce is usually added when making dashi‐based dishes; however, different soy sauces produce distinct effects on the umami aftertaste. In this study, we attempted to identify the substances that cause the suppression of the umami aftertaste in soy sauce by combining sensory analysis, size fractionation, chemical analysis, and enzymatic treatment. The suppressive substance was revealed to be polysaccharides with molecular weights between 44900 and 49700. The results of acid hydrolysis and enzymatic treatment suggested that the polysaccharides were cellulose. These results indicate that a type of water‐soluble cellulose derived from soybean, wheat, or microorganisms has a suppressive effect on the umami aftertaste of soy sauce. Future studies should focus on developing a strategy that regulates the amount of these polysaccharides generated during soy sauce production, to maintain or enhance the umami aftertaste.     

不同结构改性与美拉德反应前后鲜味二肽(Asn-Pro和Ala-His)呈味特性的对比分析

本文为了探讨不同结构改性(氨基酸顺序改变、增加鲜味氨基酸、串联肽)与美拉德反应对鲜味二肽(Asn-Pro和Ala-His)鲜味与增鲜效果的影响,利用电子舌分析了Asn-Pro和Ala-His及其结构改性后获得的新肽和美拉德反应产物的呈味特性,评价了Asn-Pro和Ala-His结构改性前后以及美拉德反应对味精(MSG)和酱油的增鲜效果。结果表明,Ala-His序列改性后获得的新二肽His-Ala的鲜味评分由4.09分上升到5.08分,对MSG和酱油具有明显的增鲜作用,其鲜味评分分别提高了1.40分和0.48分,而其他结构改性获得的9条新肽(Pro-Asn、Glu-Asn-Pro、Asn-Pro-Glu、Asp-Asn-Pro、Asn-Pro-Asp、Asn-Pro-Asn-Pro、Ala-His-Ala-His、Asn-Pro-Ala-His和Ala-His-Asn-Pro)相对于原二肽的鲜味均有所下降,对MSG和酱油具有明显的鲜味抑制作用。美拉德反应可显著提升Asn-Pro和Ala-His的鲜味特性,其鲜味评分分别由7.65分和4.09分上升到9.48分和6.37分,美拉德反应产物对MSG和酱油具有明显的增鲜作用,MSG的鲜味评分分别提高了3.96分和2.29分,酱油的鲜味评分分别提高了3.77分和1.49分。氨基酸序列改变、增加鲜味氨基酸以及串联肽能显著影响多肽鲜味及增鲜效果,美拉德反应是提升多肽鲜味与增鲜效果的有效途径。     

Evaluation of non-volatile taste components in commercial soy sauces

The dominating non-volatile taste compounds in commercial brewed soy sauces were determined by HPLC and evaluated on the contributions to overall taste. Aspartic acid and glutamic acid accounted for 8.77 to 147.98 mg/mL in ten commercial soy sauces samples. Lactic acid (ranging from 0.83 to 13.19 mg/mL) and pyroglutamic acid (ranging from 0 to 12.80 mg/mL) were the dominant organic acids, contributing to the acidity and ensuring a balance in taste of soy sauces. 5?-Inosine monophosphate was the most abundant nucleotide, followed by 5?-guanosine monophosphate, and they accounted for 0.30 to 3.54 mg/mL in ten soy sauces. According to the determination of non-volatile taste compounds in soy sauce samples, taste activity value (TAV) and equivalent umami concentration (EUC) of different soy sauces were calculated and compared. An exclusive cluster analysis based on TAV was proposed to classify the commercial soy sauces. The EUC value of new class A is much higher than other classes.     

Taste of Water-Soluble Extracts Obtained from Over-Fermented Tempe

Over-fermented tempe, known as tempe semangit, is popular in Indonesian culture, especially in Java, as an umami seasoning in traditional foods. The objective of this study was to characterize taste-active compounds in water soluble extracts of over-fermented tempe. Over-fermented tempe was prepared from fresh tempe in which the fermentation was prolonged (0 to 96 h). Free amino acids in the water soluble extract were analyzed and characterized for their taste activity values. Water soluble extracts contained umami and bitter tasting free amino acids at relatively high concentrations. Their umami and bitter taste activity values were higher than the taste threshold concentration. Water soluble extract from 72 h over-fermentation had a higher umami taste activity value than bitter taste activity, exhibiting the highest umami taste dilution factor. The high-performance liquid chromatography profile of water soluble extract fractions obtained using Sephadex G-25 gel filtration chromatography demonstrated that fractions having higher umami taste intensity had more hydrophilic components than hydrophobic components.     

米渣生酱油和大豆生酱油的风味表征及比较

对3种不同菌种发酵的米渣生酱油和一种大豆生酱油的风味进行比较,以阐释鲁氏酵母、植物乳杆菌参与发酵对米渣生酱油风味的影响以及米渣生酱油和大豆生酱油的风味差别。结果表明单菌种发酵的大豆生酱油及鲁氏酵母和植物乳杆菌分别参与发酵的米渣生酱油游离氨基酸含量、味道强度值、挥发性风味成分及相对含量均明显高于米曲霉单菌种发酵的米渣生酱油,鲁氏酵母参与发酵的米渣生酱油中4-乙烯基愈创木酚相对含量达9.71%;电子舌呈味分析表明4种生酱油的主成分存在明显差异,鲁氏酵母参与发酵的米渣生酱油和大豆生酱油呈味非常相似,鲜味突出而酸味相对较弱,植物乳杆菌参与发酵的米渣生酱油各种味道相对均衡,综合感官质量最好。米渣酱油和大豆酱油的整体风味有一定差别,优化菌种耦合发酵明显有利于增强米渣生酱油风味。     

黄豆酱油与黑豆酱油抗氧化活性及风味物质的比较

本文研究了相同工艺下,黄豆、黑豆两种蛋白原料制备传统高盐稀态酱油的差异。比较两种酱油基本成分及抗氧化活性的差异,并利用顶空-固相微萃取(HS-SPME)和气相色谱-质谱联用(GC-MS)分析两种酱油挥发性风味物质。结果表明,黑豆酱油总氮、氨基态氮、还原糖等显著高于黄豆酱油(p0.05),黄豆酱油鲜味氨基酸含量更高。黑豆酱油抗氧化活性高于黄豆酱油,相关性分析表明总酚、总黄酮及类黑精是酱油抗氧化活性的重要物质基础。两种酱油挥发性风味物质种类及含量差异显著(p0.05),共鉴定出98种物质,黄豆酱油、黑豆酱油总风味物质种类分别有63种、59种,总峰面积分别为297.6×10~7、213.6×10~7。综合分析表明黄豆酱油醇香突出,黑豆酱油醇、酯及醛酮类主要挥发性成分较均衡,风味协调。     

甜油和酱油中氨基酸和还原糖含量的比较研究

探讨甜油和酱油中两种关键营养成分氨基酸和还原糖的含量差异,分析原料不同、工艺相近的两种调味品特征指标存在差异的原因,并提出甜油和酱油存在的优势和缺点。结果表明,甜油中氨基酸总量和鲜味氨基酸谷氨酸低于酱油,甜油和酱油中氨基酸总量分别为15.86～23.21 mmol/L和31.69～46.04 mmol/L,其中谷氨酸含量分别为2.94～7.54 mmol/L和20.62～35.34 mmol/L;但每类氨基酸含量差距不大,比较协调;甜油中甜味氨基酸比例较大,为49.41%,酱油中的鲜味氨基酸比例较大,为71.19%;甜油中还原糖含量（14.24～27.53 g/L）高于酱油（4.80～8.26 g/L）。因此,酱油因高氨基酸和谷氨酸含量鲜味明显、口感厚重,适合烧制菜肴,而甜油口感协调、丰满,适合炒菜和凉拌菜;另外,甜油中高含量的还原糖含量能够改善菜肴质量、色泽和口感。     

热加工暗纹东方鲀肌肉中呈味肽分离鉴定及呈味特性研究

为明确养殖暗纹东方鲀煮制时关键呈味肽的结构序列,本文对热加工暗纹东方鲀肌肉中呈味肽进行分离鉴定并对其呈味特性进行研究。热加工暗纹东方鲀肌肉水提物经超滤(3 ku)、纳滤(200 u)及Sephadex G-15凝胶过滤色谱分离得到4个多肽组分(F1、F2、F3和F4),感官评价结合电子舌分析筛选出滋味特性最强组分F2。利用半制备RP-HPLC对F2组分进一步分离得到3个组分(F2-1、F2-2和F2-3),电子舌评价F2-1组分鲜味较强。对组分F2-1中呈味肽的氨基酸序列采用基质辅助激光解析电离飞行时间质谱(MALDI-TOF/TOF MS/MS)进行测定,得到了两种新的呈味五肽和六肽结构:Cys-Ala-Leu-Thr-Pro(CA,503.9253 u)及Arg-Pro-Leu-Gly-Asn-Cys(RP,659.7747 u),2条肽链中均含有的亲水性Cys残基,可能是造成肽具有浓厚感和鲜味的关键。两个肽段经固相合成后测定其呈味特性,肽段RP具有较强鲜味,且肽段RP还与氯化钠和谷氨酸钠具有滋味协同作用,可增强鲜味强度。     

Isolation and identification of flavour peptides from Puffer fish (Takifugu obscurus) muscle using an electronic tongue and MALDI-TOF/TOF MS/MS

To clarify the key flavour peptides that account for the cooked taste of puffer fish, this study was performed to examine flavour peptides extracted from the flesh of puffer fish (Takifugu obscurus). Peptides fractions (P1, P2, P3, P4 and P5) were purified from an aqueous extract of T. obscurus muscle by ultrafiltration and Sephadex G-15 gel filtration chromatography (GFC). P2 was further fractionated into P2a, P2b, and P2c by reverse phase high performance liquid chromatography (RP-HPLC). Fraction P2b elicited umami and sweet taste. The amino acid sequence of P2b subfraction was identified as Tyr-Gly-Gly-Thr-Pro-Pro-Phe-Val (836.4Da) by matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF/TOF MS/MS). Hydrophilic amino acids residues Tyr, Gly, Gly, Thr, and Phe are likely to contribute to the umami and sweet taste of this octapeptide. The results of this study suggest this peptide is one of important components of the 'mellowness' and 'tenderness' taste of the T. obscurus.     

白腐乳中呈味肽的分离与鉴定

目的:从白腐乳中寻找呈味肽,分析其氨基酸序列,人工合成肽样品,研究其滋味特性。方法:白腐乳提取液经过超滤、Sephedex G-15葡萄糖凝胶过滤层析,分离出3个呈味组分,通过感官评价筛选出呈鲜效果最优的组分,采用反相高效液相色谱对其进一步分离纯化,选择峰面积最大的组分,使用基质辅助激光解析电离-飞行时间质谱结合De Novo软件分析呈味肽的氨基酸序列,人工合成肽样品,对合成肽样品的呈味特性进行分析。结果:在白腐乳中发现并确定序列的肽链3条,其氨基酸序列分别为:Asp-Phe-Lys-Arg-Glu-Pro、Asp-Arg-Glu-Lys-Phe-AspGlu、Asp-Glu-Asp-Phe-Lys-Arg-Glu-Pro。其中肽Asp-Phe-Lys-Arg-Glu-Pro具有鲜味,肽Asp-Glu-Asp-Phe-Lys-ArgGlu-Pro兼具鲜味与酸味,肽Asp-Arg-Glu-Lys-Phe-Asp-Glu虽无明显特征滋味,但在味精溶液中则体现出较强的增鲜效果。结论:腐乳的鲜味不仅仅来自于谷氨酸等氨基酸成分,还与小分子肽类的呈鲜作用有关。     

Immunological functions of soy sauce: hypoallergenicity and antiallergic activity of soy sauce

Soy sauce is a traditional fermented seasoning of East Asian countries and is available throughout the world. In Japanese soy sauce (shoyu), soybeans and wheat are the two main raw materials, used in almost the same quantity. Proteins of the raw materials are completely degraded into peptides and amino acids by microbial proteolytic enzymes after fermentation, and no allergens of the raw materials are present in soy sauce. In contrast, polysaccharides originating from the cell wall of soybeans are resistant to enzymatic hydrolyses. These polysaccharides are present in soy sauce even after fermentation and termed shoyu polysaccharides (SPS). Soy sauce generally contains about 1% (w/v) SPS and SPS exhibit potent antiallergic activities in vitro and in vivo. Furthermore, an oral supplementation of SPS is an effective intervention for patients with allergic rhinitis in two double-blind placebo-controlled clinical studies. In conclusion, soy sauce would be a potentially promising seasoning for the treatment of allergic diseases through food because of its hypoallergenicity and antiallergic activity.     

A study of chemical characteristics of soy sauce and mixed soy sauce: chemical characteristics of soy sauce

Mixed soy sauce is one among the Korean traditional fermented soy sauces consumed as condiment for cooking in Korea, especially mixed soy sauce is made of the fresh seafood and the meat different from the soy sauce. Therefore, mixed soy sauce provides the strong and complicated umami taste, which is stronger than that of soy sauce. The objective of this study was to investigate for the chemical characteristics changes of mixed soy sauce in comparison with soy sauce during fermentation process (0, 120, 240 and 360 days) at different fermentation temperatures (4 and 20 °C), change in moisture, total nitrogen, crude fat, ash, pH, titratable acidity, total acidity, salt content and reducing sugar of mixed soy sauce in comparison with soy sauce. Especially, histidine, arginine, threonine, methionine, phenylalanine, and lysine, essential amino acid and among adenosine triphosphate (ATP)-related compounds, inosine (HxR) and hypoxantine (Hx) were prominent in mixed soy sauce at 20 °C. Otherwise, 2,4-dinitrophenylhydrazine (DNPH) derivative and 3DG content were prominent in soy sauce at 20 °C. The content of 2,4-DNPH derivative and 3DG in soy sauce is correlated with its browning intensity.     

养殖暗纹东方纯肌肉中呈味肽的分离鉴定

本文旨在从生鲜暗纹东方纯肌肉中提取呈味肽从而对暗纹东方纯肌肉中的呈味肽的结构特性进行相应研究。将生鲜养殖暗纹东方鱼屯肌肉水提物经Sephadex G15凝胶柱分离,得到3个多肽组分（分别为0-F1、0-F2和0-F3）,感官评价结合电子舌选择出具有最强烈的鲜味和kokumi感的组分0-F1。采用反相高效液相色谱（RP-HPLC）对其进行进一步分离得到2个组分0．F1．1和0．F1．2,电子舌鉴定显示O．F1．1组分有较强的鲜味。最后利用基质辅助激光解析电离飞行时间质谱（MALDI-TOF-MS）对RP-HPLC分离得到组分0-F1-1中呈味肽氨基酸序列进行测定,鉴定得到一种新的呈味七肽结构：Pro-A-A1a-B-Met＊-Cys-Arg（810．9046 Da,A和B均为氨基酸代码,Met＊表示氧化型甲硫氨酸）虽然肽段中含有大量的疏水性氨基酸,但该肽不具有苦味,而且有显著的浓厚感,其中含有的Cys残基可能是其具有kokumi感的关键。     

养殖暗纹东方鲀肌肉中呈味肽的分离鉴定

本文旨在从生鲜暗纹东方鲀肌肉中提取呈味肽从而对暗纹东方鲀肌肉中的呈味肽的结构特性进行相应研究。将生鲜养殖暗纹东方鲀肌肉水提物经Sephadex G15凝胶柱分离,得到3个多肽组分(分别为0-F1、0-F2和0-F3),感官评价结合电子舌选择出具有最强烈的鲜味和kokumi感的组分0-F1。采用反相高效液相色谱(RP-HPLC)对其进行进一步分离得到2个组分0-F1-1和0-F1-2,电子舌鉴定显示0-F1-1组分有较强的鲜味。最后利用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)对RP-HPLC分离得到组分0-F1-1中呈味肽氨基酸序列进行测定,鉴定得到一种新的呈味七肽结构:Pro-A-Ala-B-Met*-Cys-Arg(810.9046 Da,A和B均为氨基酸代码,Met*表示氧化型甲硫氨酸)虽然肽段中含有大量的疏水性氨基酸,但该肽不具有苦味,而且有显著的浓厚感,其中含有的Cys残基可能是其具有kokumi感的关键。