酶法合成组胺的研究
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摘要
为探究生物催化法合成组胺,通过对IPTG浓度、诱导时间和酶学性质的研究,确定组氨酸脱羧酶诱导表达条件以及最适反应条件.结果表明:不同IPTG诱导浓度对酶活影响不大;酶活随着诱导时间的增加而升高;酶反应最适温度为30℃,pH为6.0;随着产物浓度增加,酶活降低,具有产物抑制现象;5%底物时转化率达到60%,3%底物时转化率达到97.7%.可见,该组氨酸脱羧酶有工业应用开发潜力.
In order to explore the biocatalytic synthesis of histamine, IPTG concentration, induction time and enzymatic properties were studied to determine the expression and optimum conditions of HDC. The results showed that IPTG concentrations had little effect on enzyme activity; the activity of HDC increased with the increase of induction time; the optimized reaction conditions were pH 6.0 and temperature 30 ℃. Due to the inhibition effect of product on HDC activity, the conversion rate was only 60% when the substrate concentration was 5%, however, the conversion rate was 97.7% when the substrate concentration was 3%. It was indicated that the histidine decarboxylase had potential to be applied in industry.
In order to explore the biocatalytic synthesis of histamine, IPTG concentration, induction time and enzymatic properties were studied to determine the expression and optimum conditions of HDC. The results showed that IPTG concentrations had little effect on enzyme activity; the activity of HDC increased with the increase of induction time; the optimized reaction conditions were pH 6.0 and temperature 30 ℃. Due to the inhibition effect of product on HDC activity, the conversion rate was only 60% when the substrate concentration was 5%, however, the conversion rate was 97.7% when the substrate concentration was 3%. It was indicated that the histidine decarboxylase had potential to be applied in industry.
引文
[1] GUTIéRREZ-VENEGAS G,RODRíGUEZ-PéREZ C E.Toll-like receptor 3 activation promotes desensitization of histamine response in human gingival fibroblasts:Poly (I:C) induces histamine receptor desensitization in human gingival fibroblasts[J].Cellular Immunology,2012,273(2):150-157.
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[7] KANKI M,YODA T,TSUKAMOTO T,et al.Histidine decarboxylases and their role in accumulation of histamine in tuna and dried saury[J].Applied and Environmental Microbiology,2007,73(5):1467-1473.
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[9] 胡家伟,高榕,曹敏杰,等.水产食品中组胺的丹磺酰氯柱前衍生反相高效液相色谱测定方法的建立及应用[J].食品科学,2014,35(8):283-288.
[10] 谢超,王阳光,邓尚贵.水产品中组胺产生机制及影响因素研究概述[J].肉类研究,2009(4):74-78.
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[12] 于长青,王长远,满永刚,等.植物乳杆菌组氨酸脱羧酶基因的克隆及序列分析[J].中国生物制品学杂志,2010,23(11):1204-1206.
[13] 吴天鹏,刘铭球,田鸿生,等.组织胺对小鼠Lewis肺癌生长和转移抑制作用的病理学研究[J].湖北医科大学学报,1994,15(3):206-209.
[2] 林哲靖.磷酸组胺、地塞米松联合咪喹莫特治疗血管瘤临床疗效观察[J].亚太传统医药,2014,10(5):108-109.
[3] 马培奇.2008年全球首次批准新药[J].上海食品药品监管情报研究,2009(3):18-22.
[4] HASHIMOTO M,EDA Y,OSANAI Y,et al.A novel decarboxylation of α-amino acids.A facile method of decarboxylation by the use of 2-cyclohexen-1-one as a catalyst[J].Chemistry Letters,1986,15(6):893-896.
[5] PLIML J,PROTIVA M.Synthetic experiments in the histamine group.III.New methods for the reduction of 4(5)-cyanomethylimidazole to histamine[J].Collection of Czechoslovak Chemical Communications,1954,19(1):184-186.
[6] 张涛.二盐酸组胺的合成方法:201610816868.8[P].2017-02-22.
[7] KANKI M,YODA T,TSUKAMOTO T,et al.Histidine decarboxylases and their role in accumulation of histamine in tuna and dried saury[J].Applied and Environmental Microbiology,2007,73(5):1467-1473.
[8] 徐丽,刘琳,王宗奇,等.高效液相色谱-紫外检测法测定水产品中组胺含量[J].食品安全质量检测学报,2014,5(12):3790-3794.
[9] 胡家伟,高榕,曹敏杰,等.水产食品中组胺的丹磺酰氯柱前衍生反相高效液相色谱测定方法的建立及应用[J].食品科学,2014,35(8):283-288.
[10] 谢超,王阳光,邓尚贵.水产品中组胺产生机制及影响因素研究概述[J].肉类研究,2009(4):74-78.
[11] PARENTE E,MARTUSCELLI M,GARDINI F S,et al.Evolution of microbial populations and biogenic amine production in drysausages produced in Southern Italy[J].Journal of Applied Microbiology,2001,90(6):882-891.
[12] 于长青,王长远,满永刚,等.植物乳杆菌组氨酸脱羧酶基因的克隆及序列分析[J].中国生物制品学杂志,2010,23(11):1204-1206.
[13] 吴天鹏,刘铭球,田鸿生,等.组织胺对小鼠Lewis肺癌生长和转移抑制作用的病理学研究[J].湖北医科大学学报,1994,15(3):206-209.