本文關(guān)鍵詞： 氨基甲酸乙酯 醬油 尿素 精氨酸 瓜氨酸 鳥氨酸　出處：《天津科技大學》2015年碩士論文　論文類型：學位論文

【摘要】：2007年,國際癌癥研究機構(gòu)正式將氨基甲酸乙酯(EC)歸類為2A類致癌物,在很多酒精飲料和發(fā)酵食品中均有檢測出微量EC。各國已對酒精飲料中EC的含量進行了限定,醬油作為中國的傳統(tǒng)調(diào)味品,在其中也檢測到了EC但我國并未對其含量進行限定,關(guān)于醬油中EC的形成機理人們也尚未進行研究。以米曲霉為種曲分別對高鹽稀態(tài)醬油和低鹽固態(tài)醬油進行發(fā)酵,跟蹤發(fā)酵過程醬汁中EC相關(guān)前體物質(zhì)尿素、精氨酸、瓜氨酸、鳥氨酸的變化規(guī)律,再結(jié)合EC的含量變化,分析醬油中EC的形成機理。選用AOAC994.07方法對EC進行檢測,尿素檢測采用高效液相色譜-二極管陣列檢測器配以氨基柱,精氨酸、瓜氨酸、鳥氨酸選用高效液相色譜-熒光檢測器進行檢測。按是否添加魯氏酵母將實驗分為未添加組與添加組,在高鹽稀態(tài)醬汁中,隨著發(fā)酵的進行兩組EC含量均呈逐步上升趨勢,未添加組EC終含量為34.22μg/L,添加組為64.49μg/L；低鹽固態(tài)醬汁中,未添加組EC含量先上升后下降,由10.63μg/L增至43.56μg/L后下降至33.26μg/L,添加組由10.63μg/L增至50.86μg/L后下降至44.86μg/L,由此證明米曲霉與魯氏酵母均為EC生成的來源,從高鹽稀態(tài)和低鹽固態(tài)醬汁中EC含量的差別可以看出生產(chǎn)工藝對EC的生成存在顯著影響,EC含量會隨著發(fā)酵時間的增長和溫度的升高而增加。隨著發(fā)酵的進行尿素含量不斷增加,這·與EC含量變化趨勢相同,說明尿素與EC之間存在一定的相關(guān)性。依據(jù)米曲霉和魯氏酵母的氨基酸序列,預測出其精氨酸代謝途徑,精氨酸隨著發(fā)酵的進行逐步被微生物分解利用,瓜氨酸和鳥氨酸的含量不斷上升,但在未添加組中瓜氨酸含量出現(xiàn)下降現(xiàn)象,說明在米曲霉中瓜氨酸可能存在其他代謝途徑。綜上所述,醬油中米曲霉與魯氏酵母均為EC生成的來源,尿素與瓜氨酸作為EC的前體物質(zhì)相互轉(zhuǎn)化,相互影響,共同影響著醬油中氨基甲酸乙酯的形成。
[Abstract]:In 2007, the International Agency for Research on Cancer (IARC) classified ethyl carbamate (EC) as a class 2A carcinogen. Trace ECs have been detected in many alcoholic beverages and fermented foods. Many countries have limited the content of EC in alcoholic beverages. Soy sauce is the traditional condiment in China. EC was also detected, but the content of EC was not limited in China. The formation mechanism of EC in soy sauce has not been studied. Using Aspergillus oryzae as a kind of koji to ferment high salt soy sauce and low salt solid soy sauce respectively, to track the urea, the precursor of EC in soy sauce. Arginine, citrulline, ornithine, combined with the change of EC content, the formation mechanism of EC in soy sauce was analyzed. EC was detected by AOAC994.07 method. Urea was determined by high performance liquid chromatography-diode array detector with amino column, arginine, citrulline. Ornithine was detected by high performance liquid chromatography-fluorescence detector. The experiment was divided into two groups according to whether the yeast was added or not. The EC content of the two groups increased gradually with the fermentation. The final EC content of the unadded group was 34.22 渭 g / L, and that of the added group was 64.49 渭 g / L; In the low salt solid sauce, EC content increased first and then decreased, from 10.63 渭 g / L to 43.56 渭 g / L, then to 33.26 渭 g / L. The addition group increased from 10.63 渭 g / L to 50.86 渭 g / L, and then decreased to 44.86 渭 g / L, which proved that Aspergillus oryzae and yeast Rubens were both sources of EC production. From the difference of EC content between high-salt and low-salt solid sauce, it can be seen that the production process has a significant effect on the formation of EC. EC content will increase with the increase of fermentation time and temperature, and with the increasing of urea content, the change trend of EC content is the same as that of EC content. According to the amino acid sequence of Aspergillus oryzae and Saccharomyces luciferus, the arginine metabolic pathway was predicted, and arginine was gradually decomposed and utilized by microorganisms with fermentation. The contents of citrulline and ornithine increased continuously, but the citrulline content decreased in the unadded group, indicating that there may be other metabolic pathways in Aspergillus ornithae. Aspergillus oryzae and yeast Rubens in soy sauce are the source of EC formation. Urea and citrullinic acid are the precursor of EC, which affect the formation of ethyl carbamate in soy sauce.
【學位授予單位】：天津科技大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：TS264.21

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