酱油的罐式与池式发酵过程中关键呈味物质对比分析

从发酵罐和发酵池中选取发酵过程中的酱油进行中性蛋白酶活力、氨基酸态氮含量、游离氨基酸含量、pH值、总酸含量、有机酸含量、还原糖含量、淀粉酶活力和食盐含量的检测与分析。结果表明：池式发酵酱油的中性蛋白酶活力、氨基酸态氮含量、氨基酸总量、有机酸总量、淀粉酶活力、还原糖含量、总酸含量和pH值都高于罐式发酵酱油;食盐含量则是罐式酱油更多;其中鲜味氨基酸在发酵12 d后池式酱油的含量就高于罐式酱油的含量,尤其是池式酱油的谷氨酸在发酵45 d时已达到10.42 g/kg,而罐式酱油的谷氨酸含量仅7.91 g/kg;两种方式发酵的酱油都能检测出7种有机酸,且含量较高的两种有机酸分别是乳酸和乙酸;在发酵结束时池式酱油的还原糖含量比罐式酱油的还原糖含量高1.01 g/100 g;池式酱油的食盐含量比罐式酱油的食盐含量低0.90 g/100 g,说明在关键呈味物质上池式发酵酱油优于罐式发酵酱油。     

菌种耦合发酵米渣生酱油的品质和抗氧化活性

以高盐稀态法酿造米渣生酱油,探究菌种耦合对其品质和抗氧化活性的影响。4组不同菌种耦合发酵的米渣生酱油品质测定结果表明其各项质量指标均达到GB 18186—2000《酿造酱油》中一级酱油的标准,其中S4生酱油(米曲霉耦合黑曲霉和鲁氏酵母发酵)游离氨基酸总量达到41.81 g/L,4种米渣生酱油的总氨基酸、必需氨基酸、呈味氨基酸和抗氧化氨基酸的质量浓度均高于同时发酵的大豆生酱油S5生酱油(米曲霉发酵)。S3生酱油(米曲霉耦合鲁氏酵母发酵)和S4生酱油的抗氧化活性普遍高于S1生酱油(米曲霉发酵)和S2生酱油(米曲霉耦合黑曲霉发酵),各结果均表明S3的抗氧化活性最强,清除羟自由基的能力达到等质量浓度VC的最高175.42倍,对1,1-二苯基-2-三硝基苯肼自由基清除能力达到VC的2.16倍。抗氧化活性受发酵温度影响明显,分别在35℃和50℃发酵时出现峰值。多菌种耦合发酵对米渣生酱油品质和抗氧化活性均有明显改善,其中鲁氏酵母发挥了明显作用。     

一株戊糖片球菌在低盐固态酱油工艺中的应用

酱油发酵酱醅中筛选到一株戊糖片球菌,对该菌的生长温度、生长pH值、食盐耐受度等生长特性进行研究。采用离子交换色谱法对该菌株产有机酸的情况进行测定;将该菌株添加到低盐固态酱油后酵过程中,以不添加乳酸菌的实验组作对照,对酱油样品理化指标、经济指标进行分析,并采用顶空固相微萃取-气相色谱-质谱法(HS-SPME-GC-MS)对酱油挥发性香气成分进行对比分析。结果表明,该乳酸菌的生长条件与酱油酱醅发酵环境相适应,其发酵产酸以乳酸和乙酸为主,与酱油中有机酸主成分相一致,将该菌株用于低盐固态酱油发酵,对酱油香气和其他指标均有不同程度的促进作用。     

Chromohalobacter is a Causing Agent for the Production of Organic Acids and Putrescine during Fermentation of Ganjang,a Korean Traditional Soy Sauce

Ganjang, a Korean traditional fermented soy sauce, is prepared by soaking doenjang‐meju (fermented soybeans) in approximately 20% (w/v) solar salt solution. The metabolites and bacterial communities during ganjang fermentation were simultaneously investigated to gain a better understanding of the roles of the microbial population. The bacterial community analysis based on denaturing gradient gel electrophoresis of 16S rRNA gene sequences showed that initially, the genus Cobetia was predominant (0 to 10 d), followed by Bacillus (5 to 74 d), and eventually, Chromohalobacter became predominant until the end of the fermentation process (74 to 374 d). Metabolite analysis using ¹H‐NMR showed that carbon compounds, such as fructose, galactose, glucose, and glycerol, probably released from doenjang‐meju, increased rapidly during the early fermentation period (0 to 42 d). After removal of doenjang‐meju from the ganjang solution (42 d), the initial carbon compounds remained nearly constant without the increase of fermentation products. At this point, Bacillus species, which probably originated from doenjang‐meju, were predominant, suggesting that Bacillus is not mainly responsible for ganjang fermentation. Fermentation products including acetate, lactate, α‐aminobutyrate, γ‐aminobutyrate, and putrescine increased quickly with the rapid decrease of the initial carbon compounds, while Chromohalobacter, probably derived from the solar salts, was predominant. Multivariate redundancy analysis indicated that the Chromohalobacter population was closely correlated with the production of the organic acids and putrescine during the ganjang fermentation. These results may suggest that Chromohalobacter is a causing agent responsible for the production of organic acids and putrescine during ganjang fermentation and that the solar salts, not doenjang‐meju, is an important microbial source for ganjang fermentation.     

解淀粉芽孢杆菌SWJS22和米曲霉混合制曲在酱油发酵中的应用

以解淀粉芽孢杆菌SWJS22和米曲霉为发酵菌株,考察米曲霉单独制曲发酵、米曲酶和解淀粉芽孢杆菌曲料混合发酵以及双菌种混合制曲发酵对酱油理化指标及风味的影响。结果表明:曲料混合发酵的酱油氨基酸转化率有较大提高,色率和色深物质均低于米曲霉单独制曲发酵的酱油和双菌种混合制曲发酵的酱油,曲料混合发酵酱油和双菌种混合制曲发酵的酱油谷氨酸含量有明显提高。采用气相色谱-质谱对比分析3种不同曲料发酵酱油的风味物质,结果表明:3种酱油的风味物质种类差别不明显,重要风味成分在3种酱油中均有检出,曲料混合发酵的酱油酸类物质的含量最低。     

2 种米曲霉发酵酱油风味物质比较

为进一步比较传统菌株米曲霉沪酿3.042和前期实验诱变菌株米曲霉100-8酿造酱油的差异。通过高效液相色谱、气相色谱-质谱联用仪比较酱油中有机酸、氨基酸和风味物质成分的差异。通过发酵终期酱油有机酸的结果比较发现：米曲霉100-8发酵的酱油中苹果酸含量增加,柠檬酸和琥珀酸含量下降。通过酱油发酵在大曲阶段和后期阶段的氨基酸比较发现,使用米曲霉100-8菌株发酵的酱油大曲及其后期发酵为酱油阶段,天冬氨酸、谷氨酸和精氨酸这3 种氨基酸含量都明显升高。这些有机酸和氨基酸含量之间比例的差异性是造成酱油风味不同的原因之一。对风味物质成分的分析发现：与对照相比,米曲霉100-8发酵的酱油中醇、醛、酸和吡嗪类物质都有所增加,其中风味物质增多最明显的是吡嗪类。通过米曲霉100-8酿造的酱油与传统米曲霉沪酿3.042比较,其有机酸、氨基酸和风味物质都有明显的不同。     

Evaluation of a Malaysian soy sauce <Emphasis Type="Italic">koji</Emphasis> strain <Emphasis Type="Italic">Aspergillus oryzae</Emphasis> NSK for γ-aminobutyric acid (GABA) production using different native sugars

In this study, a selected γ-aminobutyric acid (GABA)-rich Malaysian strain Aspergillus oryzae NSK was collected from soy sauce koji. The strain was used to explore the effect of using renewable native sugar syrup, sugarcane, nipa, and molasses as fermentable substrates for developing a novel functional GABA soy sauce. We evaluated the strain using the chosen native sugars for 7 days using shake flask fermentation at 30 °C. The results showed optimum GABA concentration was achieved using cane molasses as the fermentable substrate (354.08 mg/L), followed by sugarcane syrup (320.7 mg/L) and nipa syrup (232.07 mg/L). Cane molasses was subsequently utilized as a substrate to determine the most suitable concentration for A. oryzae NSK to enhance GABA production and was determined as 50% g/L of glucose standard cane molasses. Our findings indicate that cane molasses can be used as a GABA-rich ingredient to develop a new starter culture for A. oryzae NSK soy sauce production.     

US consumers’ acceptability of soy sauce and <Emphasis Type="Italic">bulgogi</Emphasis>

This study investigated acceptability and consumer segmentation of soy sauce and famous Korean dish, bulgogi. A total of 123 participants evaluated intensity of 18 attributes and sensory liking of 4 commercial soy sauce samples and bulgogi samples made by aforementioned soy sauces. The overall results showed that appearance liking was the only significant different attribute among soy sauce samples and there were no significant differences among bulgogi samples. Furthermore, there was little correlation between overall liking of soy sauce and bulgogi samples. However, different segments of consumer groups were found by cluster analysis. Four and five subgroups of consumers were identified on evaluation of soy sauce samples and bulgogi samples, respectively. These results demonstrated that consumers’ different preference pattern. In conclusion, this study specified characteristics of commercial soy sauce perceived by consumers and consumers’ acceptability toward soy sauces and bulgogi made using soy sauces.     

光照对酱油发酵影响初步研究

研究了不同光质对发酵酱油的影响,结果显示不同光质发酵酱油在其色泽(A530)、氨氮含量、固形物含量、还原糖含量、总酸含量、红指、黄指等指标之间的变化率没有明显差异(p>0.05),经感官评价,其色泽、香气、滋味方面也均没有显著差异(p>0.05)。对无光控温发酵酱油与传统晒露发酵酱油进行比较,中性和酸性蛋白酶活力在无光控温发酵酱油中均下降得较快,但氨氮含量却增加较快。它们的总游离氨基酸含量分别为5.246g/100mL和4.480g/100mL;通过GC-MS对酱油的挥发性成分进行分析,无光控温发酵酱油中的挥发性成分较丰富,而且其大部分醇类、酸类、酯类、醛类、酚类和其他挥发性物质的含量均比传统晒露发酵酱油高。     

双菌种酿制酱油工艺研究

通过对采用3.042米曲霉和AS 3.350黑曲霉混合发酵酿制酱油工艺的研究,结果表明:80%沪酿米曲霉和20%AS 3.350黑曲霉混合发酵可使成品酱油中的谷氨酸含量提高30%,原料全氮利用率及氨基酸生成率也有所提高,而且色香味也均比单一米曲霉制曲发酵的酱油好.     

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。     

Simultaneous enhancement of free isoflavone content and antioxidant potential of soybean by fermentation with Aspergillus oryzae

In order to determine the fungal strain and fermentation period which produce the highest antioxidant activity of fermented soy product, soybean was fermented with Aspergillus oryzae NL 5 isolated from Korean traditional meju produced in Sunchang Yeastopia Land in Korea, and was subjected to analyzing isoflavone profiles and assaying antioxidant activity. Antioxidant potential of the samples was evaluated by ferric reducing antioxidant power assay, 2,2'-azinobis (2-ethylbenzthiazoline-6-sulfonic acid) radical scavenging activity and total phenolic contents. In addition, antioxidant assay using fluorescent probes, 2',7'-dichlorofluorescin diacetate and dihydroethidium, in HepG2 cells was conducted for the samples. The results showed that both high isoflavone variety Aga 3 and medium isoflavone variety Daewon of soybean fermented with Asp oryzae NL5 for 4 to 5 d showed the highest increase in free isoflavone contents, together with the highest antioxidant activity. In conclusion, the antioxidant activity of fermented soy was proportional to the concentration of free isoflavones, and was significantly influenced by total isoflavone content of soybean variety and fermentation period. Practical Application: Isoflavone profile and antioxidant capacity of soybean were widely variable during fermentation with Asp oryzae, and thereby it is recommended that meju (or koji) preparation condition, in particular, fermentation time is carefully optimized to maximize the antioxidant capacity of soy products fermented by Asp oryzae.     

An Improved HPLC-FLD for Fast and Simple Detection of Ethyl Carbamate in Soy Sauce and Prediction of Precursors

A high-performance liquid chromatography with fluorescence detection (HPLC-FLD) method for ethyl carbamate (EC) determination in Chinese soy sauce was developed and used to predict EC precursors, and extraction, derivatization, and chromatographic conditions were optimized systematically. Under optimal conditions, the limit of detection, linear range, and recoveries were 3.91 μg/L, 12.87–274.87 μg/L, and 81.5–95.4%, respectively. The method precision was less than 8.9% (RSD) and no significant difference was found between EC determinations measured by HPLC-FLD and GC/MS. Using the proposed HPLC-FLD method, the EC contents in Chinese soy sauces ranged from not detected to 64.88 μg/L (n = 19). Except for one soy sauce brand, the EC contents in low-salt solid-state fermented (LSF) soy sauces were lower than those in high-salt liquid fermented (HLF) soy sauces and were mainly affected by alcohol content. Moreover, the free ornithine and total arginine contents were significantly correlated with EC content in the soy sauce products. The results of this work provide a foundation for further study of EC formation and inhibition in Chinese soy sauces.     

Effect of lactic acid fermentation on antioxidant properties and phenolic acid contents of oyster (<Emphasis Type="Italic">Pleurotus ostreatus</Emphasis>) and chanterelle (<Emphasis Type="Italic">Cantharellus cibarius</Emphasis>) mushrooms

Fruiting bodies of Pleurotus ostreatus (oyster) and Cantharellus cibarius (chanterelle) mushrooms underwent acid fermentation using 3 strains of lactic acid bacteria (LAB) as starter cultures. Polyphenol contents, antioxidant activities, and phenolic acid contents in fresh, blanched, and fermented mushrooms were investigated. Fruiting bodies of oyster mushrooms exhibited higher total phenolic contents than chanterelle mushrooms. Blanching caused a decrease in polyphenol contents and antioxidant activities in both mushroom types. No important differences were observed in total phenolic compound contents (measured using Folin-Ciocalteau reagent) in mushrooms using different LAB strains. Lactobacillus plantarum was the most useful microorganism for lactic acid fermentation of fruiting bodies for reduction of the pH value. The highest concentrations of single phenolic acids: gallic, homogentisic, and ferulic acids were present in mushrooms fermented using L. plantarum.     

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。     

酶法制备大豆多肽及在酱油发酵中的应用

以脱脂大豆粕为原料,采用复合酶解技术制取大豆多肽酶解液,将制备的大豆多肽应用于酱油发酵前期,并从酱油理化指标、感官评价及香气分析三方面归纳其对酱油主要品质的影响。结果表明,先加中性蛋白酶（温度55 ℃,pH值为6.5,酶用量为4 500 U/g蛋白,酶解3 h）后,再加入碱性蛋白酶（温度55 ℃,pH值为9.0,酶用量为3 000 U/g蛋白,酶解1 h）。此时水解度达22%,多肽得率达10.84%。添加大豆多肽的酿造酱油色泽棕红亮,醇香、酱香浓郁,整体滋味均优于普通酿造酱油。气质联用（GC-MS）鉴定出酱油中含挥发性风味化合物40种,其中主要香气化合物为醇类69.41%、酸类23.06%、醛酮类4.45%、杂环酯类2.47%等,大豆多肽提高了酿造酱油的综合品质,滋味和香气明显优于传统发酵酱油,可用于开发功能营养型酱油。     

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.     

大型发酵酱油酿造过程中风味物质的动态变化分析

为促进大型发酵酱油产品风味的提高与改善,研究高盐稀态酱油在机械化酿造过程中风味成分的动态变化规律,采用气相色谱-飞行时间质谱联用技术针对不同酿造阶段酱醪中风味成分的种类和相对含量进行分析。结果定性鉴别出氨基酸及其衍生物、有机酸、糖类及其衍生物、醇类、酯类、胺类和其他物质共210 种。利用热图和主成分分析法探寻不同酿造阶段的样品风味物质差异,结果显示发酵过程对贡献酱油鲜甜味的氨基酸及其衍生物和糖类及其衍生物影响较大,第1天样品对贡献修饰风味的有机酸、胺类、酯类和其他类物质影响也较为显著,其中氨基酸及其衍生物是酱油含量最多的风味化合物。     

Effect of incubation temperature on variations in bacterial communities grown in fermenting meju and the nutritional quality of soy sauce

The effect of meju fermentation temperature on bacterial diversity and soy sauce quality was evaluated. Meju was fermented at 30, 40, 50, and 60°C for 30 days and soy sauce was prepared using fermented meju. Bacteria grown in fermenting meju at 30 and 40°C were mesophiles but bacteria grown at 50 and 60°C were thermophiles. Most bacteria grown at 30 and 40°C were Bacillus and 2 were Clostridium. Bacteria grown at 50 and 60°C were Ureibacillus, Geobacillus, Anoxybacillus, and Bacillus. Free amino acids, total nitrogen, and organic acid contents in soy sauce increased, but the ammonium content decreased in proportion to the cultivation temperature. Most bacteria grown in meju fermented at 50 and 60°C and only a few grown at 30 and 40°C produced extracellular proteolytic enzymes. Few bacteria grown at 30 and 40°C, but all bacteria grown at 50 and 60°C, were probably responsible for meju fermentation.     

市售国产酱油中大豆、小麦过敏原分析

为明确大豆和小麦发酵酱油中的残留过敏原,采用Tricine-十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）凝胶电泳,免疫印迹和间接性酶联免疫吸附测定（ELISA）等方法对从当地市场购得的14种酱油进行了分析,用来检测的四种血清包括实验室准备的大豆和小麦的兔血清免疫球蛋白G（immunoglobulin,IgG）以及从医院获得的大豆、小麦过敏病人血清免疫球蛋白E（IgE）。Tricine-SDS-PAGE结果表明,所有酱油样品都没有出现大豆和小麦蛋白的条带;免疫印迹结果表明,所有酱油样品中残留的大豆过敏原是β-伴大豆球蛋白的β亚基和大豆球蛋白的碱性亚基中的一种或两种,但小麦过敏原没有条带出现;酶联免疫结果表明,大豆过敏原含量与酱油酿造工艺与酱油等级有关,低盐固态酱油中检测到的大豆过敏原含量高于高盐稀态酱油,酱油的等级越高,所含过敏原含量相对越低。在所有酱油样品中,小麦过敏原含量极低甚至未检出。