常见乳酸菌降解亚硝酸盐机理探讨
|
摘要
该实验对Lactobacillus plantarum、Lactobacillus brevis、Lactobacillus rhamnosus的产酸特性、NO_2~-降解能力、16S r DNA序列及其亚硝酸盐还原酶(nitrite reductase,NiR)蛋白序列进行了研究与分析。结果表明,Lactobacillus brevis降解NO_2~-能力最强,可能是该菌有nir基因,在NO_2~-的诱导下产生NiR;该菌产酸量较低,较少的H~+和NiR降解NO_2~-生成的氨类物质,使发酵环境一直处于NiR最适作用p H值(5.0～6.0),从而使NO_2~-大量被NiR分解。Lactobacillus plantarum与Lactobacillus brevis亲缘关系较近,当环境p H值>4.5时,它主要在NiR作用下分解NO_2~-;但Lactobacillus plantarum产酸能力较强,能快速使环境p H值<4.0,从而迅速进入NO_2~-被H~+降解阶段。Lactobacillus rhamnosus与Lactobacillus brevis和Lactobacillus plantarum亲缘关系较远,没有找到nir基因,但Lactobacillus rhamnosus产酸能力较强,其降解NO_2~-机理可能主要是酸降解。
The acidogenic characteristics,NO_2~-degradation abilities,16S r DNA sequences,and amino acid sequences of nitrite reductases(NiR)of Lactobacillus plantarum,Lactobacillus brevis,and Lactobacillus rhamnosus were analyzed.It was found that L.brevis had the strongest ability to degrade nitrite as it may contain nir that could be translated into NiR under NO_2~-induction.Nevertheless,it had low acid yield.Therefore,less H~+and NiR degraded ammonia-formed substances produced by NO_2~-,causing the fermentation environment was always at the optimum p H of NiR(5.0-6.0),resulting large amount of NO_2~-could be decomposed by NiR.L.plantarum was closely related to L.brevis.When the pH value was>4.5,it mainly decomposed NO_2~-by NiR.However,L.plantarum had strong acid-producing ability and could quickly make the pH to be lower than 4.0,and NO_2~-was consequently degraded by H~+.L.rhamnosus had a remote relationship with L.brevis and L.plantarum.It had no nir,but its acid production ability was strong.Therefore,its NO_2~-degradation mechanism might mainly be acid degradation.
The acidogenic characteristics,NO_2~-degradation abilities,16S r DNA sequences,and amino acid sequences of nitrite reductases(NiR)of Lactobacillus plantarum,Lactobacillus brevis,and Lactobacillus rhamnosus were analyzed.It was found that L.brevis had the strongest ability to degrade nitrite as it may contain nir that could be translated into NiR under NO_2~-induction.Nevertheless,it had low acid yield.Therefore,less H~+and NiR degraded ammonia-formed substances produced by NO_2~-,causing the fermentation environment was always at the optimum p H of NiR(5.0-6.0),resulting large amount of NO_2~-could be decomposed by NiR.L.plantarum was closely related to L.brevis.When the pH value was>4.5,it mainly decomposed NO_2~-by NiR.However,L.plantarum had strong acid-producing ability and could quickly make the pH to be lower than 4.0,and NO_2~-was consequently degraded by H~+.L.rhamnosus had a remote relationship with L.brevis and L.plantarum.It had no nir,but its acid production ability was strong.Therefore,its NO_2~-degradation mechanism might mainly be acid degradation.
引文
[1]贺纪正,张丽梅.氨氧化微生物生态学与氮循环研究进展[J].生态学报,2009,29(1):406-415.
[2]龚骏,张晓黎.微生物在近海氮循环过程的贡献与驱动机制[J].微生物学通报,2013(1):44-58.
[3]SINDELAR J J,MILKOWSKI A L. Human safety controversies surrounding nitrate and nitrite in the diet[J]. Nitric Oxide,2012,26(4):259-266.
[4]BRYANA N S,ALEXANDERB D D,COUGHLINC J R,et al. Ingested nitrate and nitrite and stomach cancer risk:An updated review[J]. Food and Chemical Toxicology,2012,50(10):3 646-3 665.
[5]皇甫超申,史齐,李延红,等.亚硝酸盐对人体健康的利害分析[J].环境与健康杂志,2010(8):733-736.
[6]卢尧.肉制品腌制过程中亚硝酸盐变化规律及生物抗氧化剂对亚硝酸盐的抑制作用研究[D].长春:吉林大学,2016.
[7]王旭东. 16例急性亚硝酸盐中毒的临床分析[J].大家健康(学术版),2013,7(24):137.
[8]陈志诚,陈立亚,陈素哲.亚硝酸盐中毒31例临床分析[J].临床荟萃,2011,26(4):329-330.
[9]赵静,王娜,冯叙桥,等.蔬菜中硝酸盐和亚硝酸盐检测方法的研究进展[J].食品科学,2014,35(8):42-49.
[10]彭小云.亚硝酸盐在水产养殖中的危害[J].渔业致富指南,2015(12):63-64.
[11]周平.氨氮对水产养殖的危害及防治措施[J].中国水产,2013(8):63-64.
[12]MONSEES,H,KLATT L,KLOAS W,et al. Chronic exposure to nitrate significantly reduces growth and affects the health status of juvenile Nile tilapia(Oreochromis niloticus L.)in recirculating aquaculture systems[J]. Aquaculture Research,2017,48(7):3 482-3 492.
[13]谢燕丹,刘零怡,楼乔明,等.加工蔬菜中亚硝酸盐的消除技术研究进展[J].食品与发酵工业,2016,42(8):279-286.
[14]赵慧娟,程旭.亚硝酸盐降解进展研究[J].吉林农业,2016(22):84.
[15]崔珏,孙会刚,朱兆丽,等.酸菜中降解亚硝酸盐乳酸菌的筛选、鉴定及其在发酵香肠中的应用[J].食品工业科技,2016,37(18):251-255.
[16]于华,刘有晴,黄丹,等.泡菜中高产酸、高效降解亚硝酸盐的乳酸菌分离鉴定及应用[J].中国调味品,2016,41(6):34-38.
[17]张庆芳,迟乃玉,郑学仿,等.短乳杆菌(Lactobacillus brevis)去除亚硝酸盐的研究[J].微生物学通报,2004,31(2):55-60.
[18]柳念,陈佩,高冰,等.乳酸菌降解亚硝酸盐的研究进展[J].食品科学,2017(7):290-295.
[19]张庆芳,迟乃玉,郑艳,等.乳酸菌降解亚硝酸盐机理的研究[J].食品与发酵工业,2002,28(8):27-31.
[20]李星云.乳酸菌的分离筛选及在羊肉发酵香肠中的应用[D].呼和浩特:内蒙古农业大学,2010.
[21]张庆峰,吴祖芳,翁佩芳,等.浙东腌冬瓜优势乳酸菌的分离及产酶特性分析[J].现代食品科技,2016,(3):119-125.
[22]栗永乐.酸菜中乳酸菌的分离鉴定以及益生特性的研究[D].呼和浩特:内蒙古农业大学,2012.
[23]武昕媛,赵云冬,孙丽媛.东北发酵食物中乳杆菌的研究[J].食品工业,2017(5):204-208.
[24]迟雪梅,张庆芳.食品中常见乳酸菌高效降解NO2-发酵性能评价[J].食品与发酵工业,2017,43(6):78-84.
[25]周通,徐波.乳酸菌亚硝酸还原酶代谢调控研究进展[J].江西科学,2014,32(2):139-143.
[26]梁书诚,赵敏,卢磊,等.好氧反硝化菌脱氮特性研究进展[J].应用生态学报,2010,21(6):1 581-1 588.
[27]ZUMFT W G. Cell biology and molecular basis of denitrification[J]. Microbiology and Molecular biology reviews,1997,61(4):533-616.
[28]龚钢明,吕玉涛,管世敏,等.乳酸菌亚硝酸盐还原酶制备及酶学性质[J].中国酿造,2011(1):58-60.
[29]ADHIKARI U K,RAHMAN M M. Comparative analysis of amino acid composition in the active site of nirk gene encoding copper-containing nitrite reductase(Cu NiR)in bacterial spp.[J]. Computational Biology and Chemistry,2017,67:102-113.
[30]王英,周剑忠,李清,等.植物乳杆菌SD-7的分离鉴定及其亚硝酸盐降解特性[J].中国食品学报,2016,16(7):97-104.
[31]孔得杨.微生物氮转化途径综述[J].西部皮革,2017(4):42-47.
[32]何婷婷.植物乳杆菌亚硝酸盐还原酶编码基因的研究[D].上海:上海师范大学,2012.
[33]杨丽娜,迟雪梅,迟乃玉,等.常用食品乳酸菌发酵蔬菜的研究[J].食品与发酵工业,2017,43(3):134-137; 143.
[34]张庆芳,迟乃玉,孟宪军,等.蔬菜腌渍发酵乳酸菌剂的研究[J].食品与发酵工业,2004,30(1):13-16.
[35]魏日华,桂荣,塔娜.牧草中异型发酵乳酸菌的分离与鉴定[J].草业科学,2010,27(10):149-153.
[36]韩宏娇,丛敏,李欣蔚,等.自然发酵酸菜化学成分含量和微生物数量的动态变化及其相关性分析[J/OL].食品工业科技,2019,40(2):148-153.
[37]李春,孙迪,刘丽波,等.鼠李糖乳杆菌降解亚硝酸盐影响因素研究[J].中国酿造,2011,30(7):64-66.
[38]张君超,谢远红,金君华,等.产乳酸菌素菌株的筛选鉴定及其特性分析[J].食品工业科技,2016,37(9):169-174.
[39]辛玉峰,赵天颖,曲晓华.两类产NO的亚硝酸盐还原酶的分布,结构与序列比较及宏基因组分析[J].微生物学报,2017,57(4):597-608.
[40]CUTRUZZOLF. Bcaterial nitric oxide synthesis[J].Biochimica et biophysica Acta,1999,1 411(2-3):231-249.
[41]COYNE M S,ARUNAKUMARI A L A,AVERILL B A,et al. Immunological identification and distribution of dissimilatory heme cd1 and nonheme copper nitrite reductases in denitrifying bacteria[J]. Applied and Environmental Microbiology,1989,55(11):2 924-2 931.
[42]贺璟,聂乾忠,邓洁红.鼠李糖乳杆菌功能特性的研究进展[J].农产品加工(学刊),2012(3):117-120.
[2]龚骏,张晓黎.微生物在近海氮循环过程的贡献与驱动机制[J].微生物学通报,2013(1):44-58.
[3]SINDELAR J J,MILKOWSKI A L. Human safety controversies surrounding nitrate and nitrite in the diet[J]. Nitric Oxide,2012,26(4):259-266.
[4]BRYANA N S,ALEXANDERB D D,COUGHLINC J R,et al. Ingested nitrate and nitrite and stomach cancer risk:An updated review[J]. Food and Chemical Toxicology,2012,50(10):3 646-3 665.
[5]皇甫超申,史齐,李延红,等.亚硝酸盐对人体健康的利害分析[J].环境与健康杂志,2010(8):733-736.
[6]卢尧.肉制品腌制过程中亚硝酸盐变化规律及生物抗氧化剂对亚硝酸盐的抑制作用研究[D].长春:吉林大学,2016.
[7]王旭东. 16例急性亚硝酸盐中毒的临床分析[J].大家健康(学术版),2013,7(24):137.
[8]陈志诚,陈立亚,陈素哲.亚硝酸盐中毒31例临床分析[J].临床荟萃,2011,26(4):329-330.
[9]赵静,王娜,冯叙桥,等.蔬菜中硝酸盐和亚硝酸盐检测方法的研究进展[J].食品科学,2014,35(8):42-49.
[10]彭小云.亚硝酸盐在水产养殖中的危害[J].渔业致富指南,2015(12):63-64.
[11]周平.氨氮对水产养殖的危害及防治措施[J].中国水产,2013(8):63-64.
[12]MONSEES,H,KLATT L,KLOAS W,et al. Chronic exposure to nitrate significantly reduces growth and affects the health status of juvenile Nile tilapia(Oreochromis niloticus L.)in recirculating aquaculture systems[J]. Aquaculture Research,2017,48(7):3 482-3 492.
[13]谢燕丹,刘零怡,楼乔明,等.加工蔬菜中亚硝酸盐的消除技术研究进展[J].食品与发酵工业,2016,42(8):279-286.
[14]赵慧娟,程旭.亚硝酸盐降解进展研究[J].吉林农业,2016(22):84.
[15]崔珏,孙会刚,朱兆丽,等.酸菜中降解亚硝酸盐乳酸菌的筛选、鉴定及其在发酵香肠中的应用[J].食品工业科技,2016,37(18):251-255.
[16]于华,刘有晴,黄丹,等.泡菜中高产酸、高效降解亚硝酸盐的乳酸菌分离鉴定及应用[J].中国调味品,2016,41(6):34-38.
[17]张庆芳,迟乃玉,郑学仿,等.短乳杆菌(Lactobacillus brevis)去除亚硝酸盐的研究[J].微生物学通报,2004,31(2):55-60.
[18]柳念,陈佩,高冰,等.乳酸菌降解亚硝酸盐的研究进展[J].食品科学,2017(7):290-295.
[19]张庆芳,迟乃玉,郑艳,等.乳酸菌降解亚硝酸盐机理的研究[J].食品与发酵工业,2002,28(8):27-31.
[20]李星云.乳酸菌的分离筛选及在羊肉发酵香肠中的应用[D].呼和浩特:内蒙古农业大学,2010.
[21]张庆峰,吴祖芳,翁佩芳,等.浙东腌冬瓜优势乳酸菌的分离及产酶特性分析[J].现代食品科技,2016,(3):119-125.
[22]栗永乐.酸菜中乳酸菌的分离鉴定以及益生特性的研究[D].呼和浩特:内蒙古农业大学,2012.
[23]武昕媛,赵云冬,孙丽媛.东北发酵食物中乳杆菌的研究[J].食品工业,2017(5):204-208.
[24]迟雪梅,张庆芳.食品中常见乳酸菌高效降解NO2-发酵性能评价[J].食品与发酵工业,2017,43(6):78-84.
[25]周通,徐波.乳酸菌亚硝酸还原酶代谢调控研究进展[J].江西科学,2014,32(2):139-143.
[26]梁书诚,赵敏,卢磊,等.好氧反硝化菌脱氮特性研究进展[J].应用生态学报,2010,21(6):1 581-1 588.
[27]ZUMFT W G. Cell biology and molecular basis of denitrification[J]. Microbiology and Molecular biology reviews,1997,61(4):533-616.
[28]龚钢明,吕玉涛,管世敏,等.乳酸菌亚硝酸盐还原酶制备及酶学性质[J].中国酿造,2011(1):58-60.
[29]ADHIKARI U K,RAHMAN M M. Comparative analysis of amino acid composition in the active site of nirk gene encoding copper-containing nitrite reductase(Cu NiR)in bacterial spp.[J]. Computational Biology and Chemistry,2017,67:102-113.
[30]王英,周剑忠,李清,等.植物乳杆菌SD-7的分离鉴定及其亚硝酸盐降解特性[J].中国食品学报,2016,16(7):97-104.
[31]孔得杨.微生物氮转化途径综述[J].西部皮革,2017(4):42-47.
[32]何婷婷.植物乳杆菌亚硝酸盐还原酶编码基因的研究[D].上海:上海师范大学,2012.
[33]杨丽娜,迟雪梅,迟乃玉,等.常用食品乳酸菌发酵蔬菜的研究[J].食品与发酵工业,2017,43(3):134-137; 143.
[34]张庆芳,迟乃玉,孟宪军,等.蔬菜腌渍发酵乳酸菌剂的研究[J].食品与发酵工业,2004,30(1):13-16.
[35]魏日华,桂荣,塔娜.牧草中异型发酵乳酸菌的分离与鉴定[J].草业科学,2010,27(10):149-153.
[36]韩宏娇,丛敏,李欣蔚,等.自然发酵酸菜化学成分含量和微生物数量的动态变化及其相关性分析[J/OL].食品工业科技,2019,40(2):148-153.
[37]李春,孙迪,刘丽波,等.鼠李糖乳杆菌降解亚硝酸盐影响因素研究[J].中国酿造,2011,30(7):64-66.
[38]张君超,谢远红,金君华,等.产乳酸菌素菌株的筛选鉴定及其特性分析[J].食品工业科技,2016,37(9):169-174.
[39]辛玉峰,赵天颖,曲晓华.两类产NO的亚硝酸盐还原酶的分布,结构与序列比较及宏基因组分析[J].微生物学报,2017,57(4):597-608.
[40]CUTRUZZOLF. Bcaterial nitric oxide synthesis[J].Biochimica et biophysica Acta,1999,1 411(2-3):231-249.
[41]COYNE M S,ARUNAKUMARI A L A,AVERILL B A,et al. Immunological identification and distribution of dissimilatory heme cd1 and nonheme copper nitrite reductases in denitrifying bacteria[J]. Applied and Environmental Microbiology,1989,55(11):2 924-2 931.
[42]贺璟,聂乾忠,邓洁红.鼠李糖乳杆菌功能特性的研究进展[J].农产品加工(学刊),2012(3):117-120.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |