苹果醋发酵过程中L-苯丙氨酸促进乙酸-2-苯乙酯的合成
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摘要
为了初步研究食品添加剂L-苯丙氨酸对苹果醋发酵过程中的特征性香气成分乙酸-2-苯乙酯的合成途径及关键酶活的影响,采用HS-SPME-GC-MS和ELISA,检测各合成相关基质生成量和关键酶活性的变化。结果表明,不同添加量的L-苯丙氨酸有明显促进乙酸-2-苯乙酯及其相关合成基质生成量积累的作用,并且对ADH、AAT的酶活性也有一定的促进作用,而对Esterase酶活性有抑制作用。其中当L-苯丙氨酸添加量为8 g/L时,乙酸-2-苯乙酯生成量显著提高了43.76%～86.59%。因此,本试验初步将8 g/L作为L-苯丙氨酸的最佳添加量,初步推测L-苯丙氨酸通过醇酰基转移酶途径促进乙酸-2-苯乙酯的合成及其相关基质和关键酶活的生成量,赋予苹果醋产品更多独特的香气和风味,为生产工艺提供一定的参考依据。
In order to study the effects of L-phenylalanine(a food additive) on the synthetic pathway and key enzyme activity related to acetic acid-2-phenyl ethyl ester in apple vinegar fermentation,HS-SPME-GC-MS and ELISA were employed to detect the changes of the matrix concentration and key enzyme activities in the synthesis of acetic acid-2-phenyl ethyl ester.It was shown that different amounts of L-phenylalanine can significantly promote the production of acetic acid-2-phenyl ethyl ester and the synthesis of the related matrix,and L-phenylalanine can increase the enzymatic activities of ADH and AAT,however,it suppressed the activity of Esterase enzyme.When 8 g/L of L-phenylalanine was added,the concentration of acetic acid-2-phenyl ethyl ester significantly increased from 43.76% to 86.59%;Therefore,the optimum additive amount of L-phenylalanine was 8 g/L,speculating that L-phenylalanine induced the synthesis of acetic acid-2-phenyl ethyl ester and its related matrix,and increased key enzyme activities through alcohol acyltransferase pathway,which increased the unique aroma and flavor of apple vinegar products and provided a reference for the production process.
In order to study the effects of L-phenylalanine(a food additive) on the synthetic pathway and key enzyme activity related to acetic acid-2-phenyl ethyl ester in apple vinegar fermentation,HS-SPME-GC-MS and ELISA were employed to detect the changes of the matrix concentration and key enzyme activities in the synthesis of acetic acid-2-phenyl ethyl ester.It was shown that different amounts of L-phenylalanine can significantly promote the production of acetic acid-2-phenyl ethyl ester and the synthesis of the related matrix,and L-phenylalanine can increase the enzymatic activities of ADH and AAT,however,it suppressed the activity of Esterase enzyme.When 8 g/L of L-phenylalanine was added,the concentration of acetic acid-2-phenyl ethyl ester significantly increased from 43.76% to 86.59%;Therefore,the optimum additive amount of L-phenylalanine was 8 g/L,speculating that L-phenylalanine induced the synthesis of acetic acid-2-phenyl ethyl ester and its related matrix,and increased key enzyme activities through alcohol acyltransferase pathway,which increased the unique aroma and flavor of apple vinegar products and provided a reference for the production process.
引文
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[4]黄筱萍,黄国昌,金丹凤,等.酿酒酵母静息细胞转化合成2-苯乙醇[J].食品与生物技术学报,2017,36(12):1316-1323HUANG Xiao-ping,HUANG Guo-chang,JIN Dan-feng,et al.Study on bioconversion of 2-phenylethanol by Saccharomyces cerevisiae resting cell[D].Journal of Food Science and Biotechnology,2017,36(12):1316-1323
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[7]张琦,沈才洪,孙啸涛,等.浓香型白酒蒸馏过程中挥发性风味物质的变化规律[J].现代食品科技,2018,34(6):244-254ZHANG Qi,SHEN Cai-hong,SUN Xiao-tao,et al.Variation regularity of volatile substances in Luzhou flavor liquor during distillation process[J].Modern Food Science and Technology,2018,34(6):244-254
[8]李维,崔丹瑶,王建辉,等.HS-SPME-GC-MS分析高产酯低产高级醇酿酒酵母发酵酒的风味物质[J].现代食品科技,2018,34(3):248-254LI Wei,CUI Dan-yao,WANG Jian-hui,et al.HS-SPME-GC-MS analysis of flavor compounds in liquor fermentation by Saccharomyces cerevisiae stains with high ester yield and low higher-alcohol yield[J].Modern Food Science and Technology,2018,34(3):248-254
[9]吴艳艳,王辉,刑凯,等.发酵过程中添加酵母助剂对葡萄酒感官品质的影响[J].中国食品学报,2018,18(5):280-287WU Yan-yan,WANG Hui,XING Kai,et al.The influence of fermentation promoter on the sensory quality of wine during fermentation[J].Journal of Chinese Institute of Food Science and Technology,2018,18(5):280-287
[10]宋建强,程仕伟,姜文广,等.蛇龙珠葡萄园葡萄果实品质的空间变异分析[J].西北农林科技大学学报(自然科学版),2019,47(2):1-7SONG Jian-qiang,CHENG Shi-wei,JIANG Wen-guang,et al.Spatial variability in grape quality of cabernet gernischt within vineyard[J].Journal of Northwest Agricultural and Forestry University(Natural Science Edition),2019,47(2):1-7
[11]余兆硕,徐瀚麟,彭彰文,等.马铃薯醋酿造工艺优化及其香气成分分析[J].食品与机械,2017,33(4):171-219YU Zhao-shuo,XU Han-lin,PENG Zhang-wen,et al.Optimization of fermentation process for potato wine and analysis of flavor substances[J].Food and Machinery,2017,33(4):171-219
[12]陈梦圆,刘学彬,汪平,等.产酯香功能菌对酱香型酒醅的影响[J].食品科学,2018,39(10):199-205CHEN Meng-yuan,LIU Xue-bin,WANG Ping,et al.Effects of an ester aroma-producing bacterial strain on maotai-flavor fermented grains[J].Food Science,2018,39(10):199-205
[13]李树萍,邱诗棋,吴宛芹,等.气相色谱-嗅闻-质谱联用分析红枣白兰地风味成分[J].食品科学,2017,38(4):187-191LI Shu-ping,QIU Shi-qi,WU Wang-qin,et al.Analysis of flavor components in jujube brandy by gas chromato-graphyolfactometry-mass spectrometry[J].Food Science,2017,38(4):187-191
[14]焦娇,李凯,李树萍,等.HS-SPME-GC-O-MS分析红枣发酵酒中的挥发性成分[J].食品科学,2017,38(4):197-203JIAO Jiao,LI Kai,LI Shu-ping,et al.Analysis of volatile compounds in fermented Chinese jujube wine by headspace solid phase micro-extraction and gas chromatographyolfactometry-mass spectrometry[J].Food Science,2017,38(4):197-203
[15]范丽.苹果酒酵母中合成酯类化合物关键酶的研究[D].江苏:江南大学,2005FAN Li.The key enzymes from apple wine yeasts for esters synthesis[D].Jiangsu:Jiangnan University,2005
[16]Ji H L,Chao Z,Ju Y K,et al.Evaluation on anticancer effect against HL-60 cells and toxicity in vitro and in vivo of the phenethyl acetate isolated from a marine bacterium Streptomyces griseus[J].Fish Aquat Science,2015,18(1):35-44
[17]Yoshioka K,Hashimoto N.Ester formation by alcohol acetyltransferase from brewers yeast[J].Agricultural and Biological Chemistry,1981,45(10):2183-2190
[18]宋娟,张霁红,康三江,等.乙酸-2-苯乙酯的合成途径与调节研究[J].西南大学学报(自然科学版),2018,40(8):25-31SONG Juan,ZHANG Ji-hong,KANG San-jiang,et al.Astudy on the synthetic pathways and regulation of acetic acid-2-phenyl ethyl ester[J].Journal of Southwest University(Natural Science Edition),2018,40(8):25-31
[19]牛明福,李亚恒,陈金帅,等.马克斯克鲁维酵母生物转化2-苯乙醇工艺优化及耐高温特性分析[J].食品与发酵工业,2018,44(2):15-20NIU Ming-fu,LI Ya-heng,CHEN Jin-shuai,et al.Optimization and characterization of 2-phenylethanol bioconversion by thermotolerant yeast Kluyveromyces marxianus[J].Food and Fermentation Industries,2018,44(2):15-20
[20]周倩,蒋和体.干型番木瓜果酒酿造工艺研究[J].西南师范大学学报(自然科学版),2016,41(5):122-128ZHOU Qian,JIANG He-ti.On brewing technology of Carica papaya dry wine[J].Journal of Southwest China Nornal University(Natural Science Edition),2016,41(5):122-128
[21]李小慧.金刺梨发酵酒工艺研究[D].重庆:西南大学,2017LI Xiao-hui.Research on the technology of Rosa kweichonensis Var.Sterilis fermented wine[D].Chongqing:Southwest University,2017
[22]GB 5009.225-2016,酒中乙醇浓度的测定[S]GB 5009.225-2016,Determination of ethanol concentration in alcohol liquor[S]
[23]吴霏霏.枇杷果实香气检测及AAT1基因克隆表达与酶活性分析研究[D].江苏:扬州大学,2015WU Fei-fei.Aroma analysis and AAT1 gene cloning and expression and enzyme activity detection of loquat(Eriobotrya japonica(Thunb.)Lindl.)fruit[D].Jiangsu:Yangzhou University,2015
[24]魏永义,王富刚,尹军杰.苹果醋饮料感官品质的模糊综合评判研究[J].中国调味品,2015,40(3):26-27WEI Yong-yi,WANG Fu-gang,YIN Jun-jie.Study on sensory quality of apple vinegar beverage by fuzzy comprehensive evaluation[J].China Condiment,2015,40(3):26-27
[25]Etschmann M M W,Sell D,Schrader J.Production of2-phenylethanol and 2-phenylethylacetate from L-phenylalanine by coupling whole-cell biocatalysis with organophilic pervaporation[J].Biotechnology and Bioengineering,2005,92(5):624-634
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