Antihypertensive mechanism of a peptide-enriched soy sauce-like seasoning: the active constituents and its suppressive effect on renin-angiotensin-aldosterone system

We previously reported that a peptide-enriched soy sauce-like seasoning called fermented soybean seasoning (FSS) demonstrated antihypertensive effects both in spontaneously hypertensive rats (SHR) and Dahl salt-sensitive rats. Angiotensin I-converting enzyme (ACE) inhibitory substances (9 kinds of dipeptides and a nicotianamine) were identified from FSS. In the present study, we clarified the mechanisms underlying the antihypertensive effects of FSS in SHR. FSS was divided into the nicotianamine fraction and the peptide fraction. The peptide fraction was found to exert a more prevalent antihypertensive effect than the nicotianamine fraction in SHR. Among the peptides, we identified Gly-Tyr and Ser-Tyr as the 2 primary substances in FSS that contributed to the antihypertensive effect in SHR. These peptides were neither degraded by acid nor gastrointestinal proteases, and were absorbed into the circulating blood. FSS (2000 mg/kg) exerted ACE inhibitory activity in the lung of rats and provided a decrease (P= 0.0067) in the level of serum aldosterone after a single oral administration in SHR, resulting in the antihypertensive effect. The antihypertensive mechanism was found to be similar to therapeutic ACE inhibitors and other food-derived ACE inhibitory peptides, which are in wide use and are recognized as safe.     

Analysis of volatile compounds in Chinese soy sauces moromi cultured by different fermentation processes

The volatile compounds in Chinese soy sauce moromi cultured by different fermentation processes [lowsalt solid-state fermentation (LSSF), high-salt dilute-state fermentation (HSDF), and high-salt constant temperature fermentation (HSCT)] were determined by GC-MS. The LSSF moromi had the highest total concentration of volatile compounds, followed by HSCT and HSDF moromi. Volatiles such as ethyl linoleate and ethyl oleate dominated in HSCT moromi. 2,3-Butanediol and 2,6-dimethylpyrazine were present at high content in LSSF moromi. The hierarchal cluster analysis (HCA) and principal component analysis (PCA) were employed to investigate the fermentation process effect on the category of samples. Based on HCA, HSCT (a1 and a2) and HSDF (b1 and b2) clustered in 1 group, and LSSF (c1 and c2) located in another group. PCA illustrated that each kind of samples correlating with specific volatile compound groups were clearly differentiated according to their fermentation processes. The results suggest that GC-MS together with multivariate analyses could provide practical reference to recognize different Chinese soy sauces moromi.     

Changes in the chemical composition of traditional Chinese‐type soy sauce at different stages of manufacture and its relation to taste

Chemical composition of traditional Chinese‐type soy sauce (TCSS) at different stages of manufacture (moromi fermentation and pasteurisation) were analysed. Results showed that total nitrogen, formaldehyde nitrogen, total titratable acid, reducing sugar, NaCl, water soluble peptides, free amino acids (FAAs) and Maillard reaction products in TCSS changed significantly during manufacture. Most of FAAs were released during initial stage (0–15 days) of moromi fermentation, peptides (1–5 kDa) kept dominant (69.34–80.88%) among all peptides during manufacture and Maillard reaction products increased progressively during moromi fermentation and increased sharply during pasteurisation. According to hierarchical cluster analysis and sensory evaluation, there were obvious correlation between FAAs and sensory score of over‐all in TCSS during manufacture, which indicated that balanced FAAs composition might be mainly responsible for the taste formation of TCSS. Besides, pasteurisation can further improve the over‐all taste of TCSS.     

Production of tyramine in "moromi" mash during soy sauce fermentation

The concentrations of 7 non-volatile amines, tyramine (Tym), histamine (Him), phenethylamine (Phm), putrescine (Put), cadaverine (Cad), spermidine (Spd) and spermine (Spm) in the liquid part of "moromi" mash during soy sauce fermentation were studied. These amines, except for him and Cad, were detected during fermentation by the conventional production method in the laboratory. Put and Spd were detected at the beginning, and Tym, Phm and Spm appeared later; these 5 amines increased gradually during the fermentation. Put, Spd, Spm and Cad were present in the raw starting material for soy sauce; thus, Tym and Phm were produced by the fermentation. When "moromi" mash was added to liquid medium and cultivated, Tym was detected in some "moromi" mash and the other amines were not detected. Tym-producing bacterial strains were isolated from the liquid culture media of Tym-positive "moromi" mash. The Tym-producing strain was a gram-positive coccus. The conditions for production of amines by Tym-producing bacterial strains were examined. These strains grew and produced tyramine under various conditions, which may occur during soy sauce fermentation. Namely, Tym was produced at pH 5-10, at salt concentrations of less than 8%, under either aerobic or anaerobic conditions. During soy sauce fermentation, it is assumed that Tym would be produced by these strains during the early stages of soy sauce aging within a short period when the salt concentration and pH conditions are optimal for growth. Based on the bacteriological properties, the strains were identified as Enterococcus faecium. With the exception of Phm and Him, which did not exist in the starting raw material, non-volatile amines (including Put, Cad, Spd and Spm) were not produced and microorganisms producing them are not believed to be present during "moromi" fermentation.     

耐盐乳酸菌和酵母菌强化技术对高盐稀态发酵酱醪品质的影响

分别将耐盐乳酸菌Tetragenococcus halophilus CGMCC 3792、耐盐酵母菌Zygosaccharomyces rouxii CGM-CC 3791和Candida versatilis CGMCC 3790菌悬液接入酱醪中,利用顶空-固相微萃取偶联气质联用技术(HS-SPME/GC-MS)检测了发酵125 d酱醪中挥发性组分的变化。研究结果表明,T.halophilus CGMCC 3792强化的酱醪中酸类、醇类、酮类、呋喃类、吡嗪类和其他组分的相对含量降低,酯类、醛类和酚类组分的相对含量增加,其中苯乙醇、乙酸乙酯、辛酸乙酯、癸酸乙酯和4-乙基愈创木酚等的相对含量显著增加,而Z.rouxii CGMCC 3791、C.versatilis CGMCC 3790共培养强化的酱醪中醇类、酯类、醛类和其他组分的相对含量增加,酸类、酚类、呋喃类和吡嗪类组分的相对含量降低,其中苯乙醇、长链脂肪酸乙酯(月桂酸乙酯、棕榈酸乙酯、亚油酸乙酯等)的相对含量显著增加。此外,本研究中T.halophilus CGMCC 3792强化的酱醪中4-乙基愈创木酚的相对含量大幅增加,且在整个发酵过程中其含量也单调增加,对于T.halophilus CGMCC 3792是否具有分泌4-乙基愈创木酚的能力以及是否冷温工艺赋予该菌株合成4-乙基愈创木酚的能力需进一步探究。     

耐盐米曲霉CICIM F0899在酱油酿造过程中的应用

耐高盐环境的蛋白酶对酱油工业有着积极作用。该实验对1株产耐盐蛋白酶的米曲霉CICIM F0899在高盐稀态酱油发酵过程中的作用进行了研究,并同沪酿3042进行了比较。结果表明,在酱醪发酵阶段,微生物代谢产酸使酱醪pH持续下降,影响了蛋白酶的活性,导致酱油中氨基态氮和风味物质含量的变化,在180天时氨基态氮含量达到0.62 g/100 mL,比沪酿3042高11.11%。利用气相色谱-质谱联用(GC-MS)对挥发性成分进行了分析鉴定,检测出酱醪中7大类共49种挥发性化合物,其中酯类、杂环类化合物的含量明显高于沪酿3042,而醛类物质的含量相对较少。因此,米曲霉CICIM F0899在氨基态氮含量和风味组成方面都具有更加明显的优势。     

促酱醪发酵芽孢杆菌的筛选及应用

用酪素平板法从高盐稀态酱醪中筛选出56 株蛋白酶活性较好的菌株,牛津杯法复筛后制曲测定酶活性,得到淀粉酶活性最高的菌株CS1.11、蛋白酶活性最高的菌株CS1.13及纤维素酶活性最高的菌株CS1.17;16S rRNA序列测定结合形态学分析,CS1.11、CS1.13、CS1.17分别鉴定为贝莱斯芽孢杆菌（Bacillus velezensis）、甲基营养型芽孢杆菌（B. methylotrophicus）、枯草芽孢杆菌（B. subtilis）;分别将3 株实验菌与对照菌枯草芽孢杆菌CS1.03单独进行制曲后盐卤发酵,发现CS1.13发酵酱油氨态氮质量浓度最高,为5.12 g/L,CS1.11发酵酱油还原糖质量浓度最高,为27.20 g/L,与其酶活性结果一致,4 株菌对酱油总酸含量影响不大;顶空固相微萃取气相色谱-质谱联用内标法定性及定量分析发酵酱油发现,3 株实验菌与对照菌均具有产生吡嗪类物质及其前体乙偶姻和2,3-丁二醇的优势,且3 株实验菌优于对照菌;吡嗪类物质能赋予酱油良好的风味及健康因子,是芽孢杆菌酱油发酵的特征风味物质。筛选自高盐稀态酱醪的芽孢杆菌CS1.11、CS1.13、CS1.17有促酱醪发酵、丰富酱油风味、增进健康因子的潜力,具备良好的应用前景。     

提高酱醪质量生产高档本酿造酱油

本文通过改变和调整高盐稀发酵酱油工艺的发酵条件、管理方法、生产菌种以及拌曲盐水量、浓度等达到提高酱醪捏及生酱油出品率的目的,从而生产出高档本酿造酱油。     

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

Soy 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.     

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...     

糖化增香曲对高盐稀态酱油发酵过程中理化性质影响的研究

本文通过添加糖化增香曲混合制曲(KG)生产高盐稀态酱油,对比纯种米曲霉制曲(KP),研究糖化增香曲对高盐稀态酱油发酵过程中理化性质的影响.研究结果表明:添加糖化增香曲混合制曲可显著提高高盐稀态酱油的还原糖和可溶性无盐固形物含量,发酵90 d时分别较纯种制曲提高47%和7%:也可显著提高发酵初期曲料中各类物质的溶出速率;...     

ROLES OF THE MAILLARD REACTION IN BROWNING DURING MOROMI PROCESS OF THAI SOY SAUCE

Moromi samples of traditional Thai soy sauce from two conventional manufacturers A and B were used to investigate the browning and chemical changes related to the Maillard reaction (i. e. reactive amino compound, soluble protein, 5% trichloroacetic acid precipitated protein, reducing sugar, and 5-hydroxymethyl-2-furaldehyde). The samples were collected at specific intervals during the 60-day process of moromi fermentation. The browning (OD 420 nm; y) was expressed as powered regression with fermentation time-course (day; x), y=0.5191 x^0.2367, r²=0.978 and y=0.6188 x^0.1596, r²=0.821 for moromi A and B, respectively. The browning rate was high at the first 3 days and declined in the later stage, while the accumulation of 5-hydroxy-2-furaldehyde, which is the Maillard reaction intermediate, increased linearly. The size of proteins and peptides observed on sodiumdodecyl sulphate-polyacrylamide gel electrophoresis and the measurement of the ratio of 5% trichloroacetic acid precipitated protein to total soluble protein (PP/SP). This PP/SP ratio showed the linear correlation to browning rate with r²=0.921 and 0.965 for moromi A and B, respectively. The results suggested that the rate of browning of the moromi fermentation did not entirely depend upon the Maillard reaction rate. It appeared that the browning is enhanced with the proper size of proteins and peptides.     

Effect of different halophilic microbial fermentation patterns on the volatile compound profiles and sensory properties of soy sauce moromi

The effect of inoculated or co-cultured halophilic lactic bacteria (Tetragenococcus halophilus) and yeasts (Zygosaccharomyces rouxii and Candida versatilis) on the quality of soy sauce moromi was investigated. Results showed no significant change in the major physicochemical properties of soy sauce moromi such as amino nitrogen, total acid, and reducing sugar at the end of fermentation, whereas the composition of free amino acids slightly changed with different fermentation patterns. In the sample inoculated with T. halophilus, the concentration of proline and tryptophan decreased by 18.83 and 50.87 % compared with the blank sample, whereas tyrosine and histidine increased by 32.41 and 27.04 %. In the sample inoculated with Z. rouxii and C. versatilis, the concentration of bitter-tasting lysine, tyrosine, and phenylalanine increased by more than 20 %, and the concentration of tryptophan decreased by 56.71 %. The proportion of amino acids with sweet taste increased by 0.8 % through the co-culture of the three strains. Meanwhile, the total amount of volatile compounds increased by 117.66, 23.28, and 216.88 % in the T. halophilus inoculated sample, two yeast strain inoculated sample, and three strain inoculated sample, respectively. Increased odour activity values of the nine compounds that mainly contributed to the flavour of moromi ranged from 1 to 7.87 times in the three strain inoculated sample. These findings revealed that the co-culture of the three strains during moromi fermentation was beneficial to the improvement of flavour of soy sauce moromi.     

盐水浓度及pH对高盐稀态酱油酿造初期酱醪理化性质影响的研究

本文以高盐稀态酱油为研究对象,分析了不同盐水浓度及pH值对其酿造前30 d酱醪理化特性的影响.结果表明:在自然pH(7.2),盐水浓度为16%、20%、24%条件下,随着盐水浓度的增加,酱醪中总氮、氨基酸态氮、总酸的增加速率以及pH值下降速率均减小耳还原糖含量增加.酿造30 d时,16%和20%盐水浓度的酱醪总氮相比2...     

固稀发酵酱油的生产工艺浅析

先固后稀发酵酱油比高盐稀态酱油生产周期短,风味比低盐固态酱油好,是当前中小企业提高酱油风味,投资小、见效快的途径之一。固稀发酵生产工艺可以利用低盐固态发酵设备,采取低温制曲,低温发酵,生产接近广式淡色味鲜的生抽类酱油,收效较好。     

高盐稀态酱醪中功能性酵母菌的筛选、鉴定及发酵特性

从高盐稀态酱醪中筛选酵母菌株,其中14 株分离鉴定的酵母菌株可分为4 个菌种,选择代表性菌株进行生理生化鉴定并探究其生长特性,并将分离的14 株酵母菌单独发酵产香,结合气相色谱-质谱和嗅闻感官评价,探究各菌株对酱油风味的功能作用。结果表明,胶红酵母和似平滑假丝酵母对酱油风味具有一定的提升效果,可能与2-苯乙酸乙酯、3-甲硫基-1-丙醇、2,6-二甲基吡嗪等物质含量的增加相关,为潜在风味功能酵母。而粉状米勒氏酵母和近平滑假丝酵母对酱油风味具有负面影响,特别是粉状米勒氏酵母,菌落表面干燥褶皱,其繁殖代谢将大幅提高酱油中2-辛酮、2-壬酮和1,2,3-三甲氧基苯的含量,带来明显的不良风味,是酱油风味的有害酵母。本研究明确了4 种不同酵母菌对酱油风味的作用差异,对酱油发酵的风味调控具有指导意义。     

Changes in volatile aroma compounds of traditional Chinese-type soy sauce during moromi fermentation and heat treatment

Considering the important influence of longtime (150 day) moromi fermentation and heat treatment on the aroma formation of traditional Chinese-type soy sauce (TCSS), volatile compounds in samples taken from different stages of moromi fermentation and heat treatment were analyzed by solid phase microextraction coupled with gas chromatography-mass spectrometry. Results showed that a total of 76 volatile compounds were identified in all the samples, and most of the volatile compounds were common. During 150 day of moromi fermentation, relative contents of acids, alcohols, aldehydes and ketones, esters, and furan(one)s along with all the sensory attributes of acidic, alcoholic, fruity, caramel-like, smoky, and malty changed greatly. Notably, relative contents of alcohols, aldehydes and ketones along with the sensory intensities of alcoholic, caramel-like, and smoky of heated sample (80°C/60 min) decreased markedly, whereas there were slight increases in relative contents of furan(one)s, phenols, and sulfur-containing compounds of it. Long-time moromi fermentation and heat treatment have significant influence on the formation and relative contents of volatile compounds in TCSS, whereas changes in volatile compounds and their relative contents of the samples were responsible for the differences in sensory attributes.     

不同淀粉质原料对高盐稀态酱油香气品质的影响

为了研究不同淀粉质原料(面粉、麸皮)对高盐稀态酱油香气品质的影响,本文利用定量描述分析(QDA)对面粉类酱油(FSS)、麸皮类酱油(WSS)的感官特征进行分析,通过顶空-固相微萃取(HS-SPME)和液液萃取(LLE)结合气相色谱-质谱联用(GC-MS)定量对比分析了两种酱油挥发性化合物组成。FSS中酸香、土豆香、麦芽香显著性高于WSS,焦糖香、烟熏香显著性低于WSS(p0.05)。GC-MS共分离鉴定出94种挥发性化合物,其中以酸类、酮类、醛类占主要比例,HS-SPME法能萃取出较多的醛类、含硫化合物,LLE法对呋喃(酮)类、酸类化合物萃取效果较好。通过对酱油中17种关键香气活性物质定量分析以及香气活性值(OAV)的计算,有9种化合物OAV值大于20。其中3-甲基丁醛OAV值最高,HDMF、愈创木酚均有较高的OAV值,且在WSS中高于FSS,与感官分析中的焦糖香、烟熏香结果一致;FSS中2-甲基丁醛、3-甲硫基丙醛的OAV值高于WSS,与麦芽香、土豆香感官分析结果相符。     

提高酱油质量缩短发酵周期

根据酱油成分复杂，发酵需要有多种酶系参与的客观实际，提出“多菌种低温发酵”是影响酱油质量的主要工艺条件，并论证了“固态无盐低温发酵”能提高酱油质量缩短发酵周期的可行性。     

甘氨酸甜菜碱及复合纤维素酶对酱油发酵的影响

将甘氨酸甜菜碱作为渗透压保护剂,于酱醪上罐时(0d)或酱醪发酵30d后添加到高盐稀态酱油发酵的酱醪中,同时添加外源复合纤维素酶,观察其对酱醪发酵的影响。结果表明：在酱醪上罐时(0d)或酱醪发酵30d后同时添加不同质量分数的甘氨酸甜菜碱(0.20%、0.30%)及质量分数0.15%的复合纤维素酶(105EGu/100g酱醪),均能有效提高发酵酱油中蛋白质转化率,达2.26%～7.92%。添加甘氨酸甜菜碱能显著改善高盐稀态发酵酱油头油品质,平均缩短发酵时间15d左右;另外,在酱醪发酵30d后再添加甘氨酸甜菜碱和外源复合纤维素酶其效果更佳,同0d时添加相比,蛋白质转化率平均提高了3.17%。