鲁氏不动杆菌Acinetobacter lwoffii UL产类胡萝卜素的纯化与鉴定及其抗氧化活性检测
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摘要
以鲁氏不动杆菌(Acinetobacter lwoffii) UL发酵提取得到的类胡萝卜素粗提液为试材,用薄层层析(thin layer chromatography,TLC)和硅胶柱层析纯化后得到组分1和组分2;通过溶解性试验、HPLC、傅里叶红外光谱(fourier transform infrared spectrometer,FTIR)、核磁共振氢谱(nuclear magnetic resonance,1H-NMR)和液相质谱联用仪(UPLC-Q-TOF-MS-MS)对组分2进行鉴定,结果表明该色素为脂溶性色素,且含有多个甲基和亚甲基、多烯长链、苯环以及羰基等类胡萝卜素特征结构,初步推断其含有虾红素(astacen),以及较少的虾青素(astaxanthin)。采用ABTS、FRAP、DPPH三种方法评价类胡萝卜素粗提液、纯化后的组分1和组分2的抗氧化活性,结果表明类胡萝卜素粗提液具有抗氧化性,纯化后的类胡萝卜素抗氧化活性显著提高。
The crude carotenoid extracts from Acinetobacter lwoffii UL fermentation were purified by thin layer chromatography(TLC) and silica gel column chromatography to obtain component 1 and component 2. Solubility test,high performance liquid chromatography(HPLC),fourier transform infrared spectrometer(FTIR),1 hydrogennuclear magnetic resonance(1 H-NMR),and UPLC-Q-TOF-MS-MS were used to identify component 2. The results showed that pigments in component 2 were fat soluble and had carotenoid characteristic structure,such as methyl,methylene,polyene long chain,benzene ring,and carbonyl group. It was preliminarily deduced that component 2 contained astacen(C40 H48 O4,molecular weight 592) and few astaxanthin(C40 H52 O4,molecular weight 596).ABTS,FRAP,and DPPH were used to evaluate the antioxidant activities of carotenoid crude extracts,purified component 1 and component 2. The results showed that carotenoid crude extracts had antioxidant activities,and the antioxidant activities of purified carotenoids increased significantly.
The crude carotenoid extracts from Acinetobacter lwoffii UL fermentation were purified by thin layer chromatography(TLC) and silica gel column chromatography to obtain component 1 and component 2. Solubility test,high performance liquid chromatography(HPLC),fourier transform infrared spectrometer(FTIR),1 hydrogennuclear magnetic resonance(1 H-NMR),and UPLC-Q-TOF-MS-MS were used to identify component 2. The results showed that pigments in component 2 were fat soluble and had carotenoid characteristic structure,such as methyl,methylene,polyene long chain,benzene ring,and carbonyl group. It was preliminarily deduced that component 2 contained astacen(C40 H48 O4,molecular weight 592) and few astaxanthin(C40 H52 O4,molecular weight 596).ABTS,FRAP,and DPPH were used to evaluate the antioxidant activities of carotenoid crude extracts,purified component 1 and component 2. The results showed that carotenoid crude extracts had antioxidant activities,and the antioxidant activities of purified carotenoids increased significantly.
引文
[1] SAINIR K,NILE S H,PARK S W. Carotenoids from fruits and vegetables:Chemistry, analysis, occurrence,bioavailability and biological activities[J]. Food Research International,2015,76(3):735-750.
[2]尹金凤,王志轩,吴欣森,等.三孢布拉氏霉发酵产β-胡萝卜素的研究进展[J].食品科学,2014(13):316-325.
[3]李伟昆.不同红酵母发酵产类胡萝卜素的研究[D].大连:大连工业大学,2015.
[4] SAINI R K,KEUMY. Progress in microbial carotenoids production[J]. Indian Journal of Microbiology,2017,57(1):129-130.
[5]盛冉,孙志高,黄巧娟,等.β-隐黄素的生理活性及其机理研究进展[J].食品与机械,2016(12):218-223.
[6] IVANOVEA V,GERASIMOVAE L,BRAININA K Z. Potentiometric study of antioxidant activity:development and prospects[J]. Critical Reviews in Analytical Chemistry,2015,45(4):311-322.
[7] PRIORR L,WUX L,SCHAICH K. Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements[J]. Journal of Agricultural and Food Chemistry,2005,53(10):4 290-4 302.
[8]张献忠,黄海智,钟烈洲,等.植物提取物体外抗氧化活性评价方法研究进展[J].中国粮油学报,2012(11):122-128.
[9]谯天敏,张静,赵芳,等.铜绿假单胞菌抗菌色素的结构鉴定及稳定性检测[J].南京林业大学学报(自然科学版),2015(5):14-20.
[10]李春燕,牟希,张媛,等. 1株产灵菌红素家族红色色素细菌的分离鉴定及其色素性质研究[J].食品与发酵工业,2017,43(1):1-6.
[11]张文娟.柑橘幼果功能成分提取分析及抗氧化活性研究[D].杭州:浙江大学,2015.
[12]赵甜甜.反相薄层板的制备及性能评价[D].天津:天津大学,2009.
[13]惠伯棣.类胡萝卜素化学及生物化学[M].北京:中国轻工业出版社,2005.
[14]黄水英.雨生红球藻的培养及其虾青素的提取、稳定性和应用研究[D].厦门:厦门大学,2008.
[15]耿兆艳,孙翰,管斌,等.雨生红球藻粉虾青素的提取及稳定性研究[J].中国食品学报,2017,17(7):86-95.
[16] FRASSANTIO R,CANTONATI M,FLAIM G,et al. A new method for the identification and the structural characterisation of carotenoid esters in freshwater microorganisms by liquid chromatography/electrospray ionisation tandem mass spectrometry[J]. Rapid Communications in Mass Spectrometry,2008,22(22):3 531-3 539.
[17]程红艳,陈军辉,赵恒强,等.反相高效液相色谱-电喷雾质谱法测定浒苔中的叶黄素[J].食品科学,2010,31(18):206-211.
[18]张明.几种体外抗氧化检测方法的评价研究[D].西安:陕西师范大学,2010.
[2]尹金凤,王志轩,吴欣森,等.三孢布拉氏霉发酵产β-胡萝卜素的研究进展[J].食品科学,2014(13):316-325.
[3]李伟昆.不同红酵母发酵产类胡萝卜素的研究[D].大连:大连工业大学,2015.
[4] SAINI R K,KEUMY. Progress in microbial carotenoids production[J]. Indian Journal of Microbiology,2017,57(1):129-130.
[5]盛冉,孙志高,黄巧娟,等.β-隐黄素的生理活性及其机理研究进展[J].食品与机械,2016(12):218-223.
[6] IVANOVEA V,GERASIMOVAE L,BRAININA K Z. Potentiometric study of antioxidant activity:development and prospects[J]. Critical Reviews in Analytical Chemistry,2015,45(4):311-322.
[7] PRIORR L,WUX L,SCHAICH K. Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements[J]. Journal of Agricultural and Food Chemistry,2005,53(10):4 290-4 302.
[8]张献忠,黄海智,钟烈洲,等.植物提取物体外抗氧化活性评价方法研究进展[J].中国粮油学报,2012(11):122-128.
[9]谯天敏,张静,赵芳,等.铜绿假单胞菌抗菌色素的结构鉴定及稳定性检测[J].南京林业大学学报(自然科学版),2015(5):14-20.
[10]李春燕,牟希,张媛,等. 1株产灵菌红素家族红色色素细菌的分离鉴定及其色素性质研究[J].食品与发酵工业,2017,43(1):1-6.
[11]张文娟.柑橘幼果功能成分提取分析及抗氧化活性研究[D].杭州:浙江大学,2015.
[12]赵甜甜.反相薄层板的制备及性能评价[D].天津:天津大学,2009.
[13]惠伯棣.类胡萝卜素化学及生物化学[M].北京:中国轻工业出版社,2005.
[14]黄水英.雨生红球藻的培养及其虾青素的提取、稳定性和应用研究[D].厦门:厦门大学,2008.
[15]耿兆艳,孙翰,管斌,等.雨生红球藻粉虾青素的提取及稳定性研究[J].中国食品学报,2017,17(7):86-95.
[16] FRASSANTIO R,CANTONATI M,FLAIM G,et al. A new method for the identification and the structural characterisation of carotenoid esters in freshwater microorganisms by liquid chromatography/electrospray ionisation tandem mass spectrometry[J]. Rapid Communications in Mass Spectrometry,2008,22(22):3 531-3 539.
[17]程红艳,陈军辉,赵恒强,等.反相高效液相色谱-电喷雾质谱法测定浒苔中的叶黄素[J].食品科学,2010,31(18):206-211.
[18]张明.几种体外抗氧化检测方法的评价研究[D].西安:陕西师范大学,2010.