HSCCC结合硅胶柱层析分离法夫酵母类胡萝卜素
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摘要
法夫酵母是天然类胡萝卜素的重要来源之一,研究从法夫酵母中分离纯化并鉴定类胡萝卜素的方法,为分离鉴定法夫酵母中类胡萝卜素提供理论依据和技术支持。采用高速逆流色谱(HSCCC)结合硅胶柱层析对法夫酵母类胡萝卜进行分离纯化,进一步采用高效液相色谱(HPLC)、质谱及与标准品比对进行类胡萝卜素鉴定。结果表明,所得的类胡萝卜素分别为β-胡萝卜素、海胆酮和虾青素。采用高速逆流色谱和硅胶柱层析可高效获得类胡萝卜素,将为类胡萝卜素的进一步研究提供高效制备方法。
Phaffia rhodozyma was one of the important sources of natural carotenoids. The study provided the theoretical basis and technological support for the separation and identification of carotenoids from P. rhodozyma. High-speed countercurrent chromatography(HSCCC) and silica gel column chromatography were used to separate and purify carotenoids from P. rhodozyma. High performance liquid chromatography(HPLC), mass spectrometry(MS) and standards of carotenoids were used to identify the obtained carotenoids. Three compounds including β-carotene, echinenon and astaxanthin were successfully separated, respectively. HSCCC and silica gel column chromatography were suitable for the efficient separation of carotenoids from P. rhodozyma, and an efficient preparation method for the further study of carotenoids was provided.
Phaffia rhodozyma was one of the important sources of natural carotenoids. The study provided the theoretical basis and technological support for the separation and identification of carotenoids from P. rhodozyma. High-speed countercurrent chromatography(HSCCC) and silica gel column chromatography were used to separate and purify carotenoids from P. rhodozyma. High performance liquid chromatography(HPLC), mass spectrometry(MS) and standards of carotenoids were used to identify the obtained carotenoids. Three compounds including β-carotene, echinenon and astaxanthin were successfully separated, respectively. HSCCC and silica gel column chromatography were suitable for the efficient separation of carotenoids from P. rhodozyma, and an efficient preparation method for the further study of carotenoids was provided.
引文
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[19] LEONCINI E, EDEFONTI V, HASHIBE M, et al.Carotenoid intake and head and neck cancer:a pooled analysis in the International Head and Neck Cancer Epidemiology Consortium[J]. European Journal of Epidemiology, 2016,31(4):369-383.
[20] JIANG G L, ZHOU L Y, WANG Y T, et al. Astaxanthin from Jerusalem artichoke:Production by fed-batch fermentation using Phaffia rhodozyma and application in cosmetics[J].Process Biochemistry, 2017, 63:16-25.
[21] SCHEWE H, KREUTZER A, SCHMIDT I, et al. High concentrations of biotechnologically produced astaxanthin by lowering pH in a Phaffia rhodozyma, bioprocess[J].Biotechnology and Bioprocess Engineering, 2017, 22(3):319-326.
[22]黄莹.法夫酵母类胡萝卜素的提取纯化及生物活性研究[D].厦门:集美大学, 2017.
[23]黄莹,王凯,杜希萍,等.二甲亚砜法破壁条件对法夫酵母虾青素提取效果的影响[J].食品工业科技, 2017, 38(15):212-217.
[24] DU X P, DONG C C, WANG K, et al. Separation and purification of astaxanthin from Phaffia rhodozyma by preparative high-speed count-current chromatography[J]. Journal of Chromatography B, 2016, 1(1029/1030):191-197.
[25] VAN BREEMEN R B, DONG L L, PAJKOVIC N D.Atmospheric pressure chemical ionization tandem mass spectrometry of carotenoids[J]. International Journal of Mass Spectrometry, 2012, 312:163-172.
[26] RIVERA S M, CHRISTOU P, CANELA-GARAYOA R.Identification of carotenoids using mass spectrometry[J]. Mass Spectrometry Reviews, 2014, 33(5):353-372.
[27] ENZELL C R, FRANCIS G W, LIAAEN-JENSEN S.Mass spectrometric studies of carotenoids[J]. Pure and Applied Chemistry, 1969, 20(4):497-516.
[2]杨青杰.黄龙胆类胡萝卜素生物合成基因启动子的动能分析[D].长春:东北师范大学, 2013.
[3]靳青,毕宇霖,刘晓牧,等.类胡萝卜素代谢及功能研究进展[J].动物营养学报, 2014, 26(12):3561-3571.
[4] BARREDO J L, GARCíAESTRADA C, KOSALKOVA K,et al. Biosynthesis of astaxanthin as a main carotenoid in the heterobasidiomycetous yeast Xanthophyllomyces dendrorhous[J].Journal of Fungi, 2017, 3(44):1-17.
[5] JAMES D, JOHNSON. Do carotenoids serve as transmembrane radical channels[J]. Free Radical Biology&Medicine, 2009,47:321-323.
[6]王彬,林亮,陈敏氡,等.南瓜类胡萝卜素含量的超高效液相色谱分析[J].农学学报, 2017, 7(12):22-27.
[7]孙玉敬,乔丽萍,钟烈洲,等.类胡萝卜素生物活性的研究进展[J].中国食品学报, 2012, 12(1):160-166.
[8] CHOI J, RAHMAN M M, LEE S Y, et al. Effects of dietary inclusion of fermented soybean meal with Phaffia rhodozyma on growth, muscle pigmentation, and antioxidant activity of juvenile rainbow trout(Oncorhynchus mykiss)[J]. Turkish Journal of Fisheries and Aquatic Sciences, 2016, 16:91-101.
[9] HAVAUX M. Carotenoid oxidation products as stress signals in plants[J]. The Plant Journal, 2013, 79(4):597-606.
[10] AMBATI R R, MOI P S, RAVI S, et al. Astaxanthin:Sources, extraction, stability, biological activities and its commercial applications a review[J]. Marine Drugs, 2014,12(1):128-152.
[11] NOGUEIRA M B, PRESTES C F, BURKERT J F M.Microencapsulation by lyophilization of carotenoids produced by Phaffia rhodozyma with soy protein as the encapsulating agent[J]. Food Science and Technology, 2017, 37(9):1-4.
[12] HOU X, RIVERS J, LEóN P, et al. Synthesis and function of apocarotenoid signals in plants[J]. Trends in Plant Science,2016, 21(9):792-803.
[13] BHUVANESWARI S, YOGALAKSHMI B, SREEJA S, et al. Astaxanthin reduces hepatic endoplasmic reticum stress and nuclear factor-B-mediated inflammation in high fructose and high fat diet-fed mice[J]. Cell Stress Chaperones, 2014, 19(2):183-191.
[14] SILA A, KAMOUN Z, GHLISSI Z, et al. Ability of natural astaxanthin from shrimp by products to attenuate liver oxidative stress in diabetic rats[J]. Pharmacological Reports,2015, 67:310-316.
[15]梁新乐.法夫酵母生物合成虾青素的研究[D].杭州:浙江大学, 2001.
[16] ROBERTS R L, GREEN J, LEWIS B. Lutein and zeaxanthin in eye and skin health[J]. Clinics in Dermatology, 2009, 27(2):195-201.
[17]马乐,林晓明.叶黄素干预对长期荧屏光暴露者视功能的影响[J].营养学报, 2008, 30(5):438-442.
[18] FRASER P D, BRAMLEY P M. The biosynthesis and nutritional uses of carotenoids[J]. Progress in Lipid Research,2004, 43(3):228-265.
[19] LEONCINI E, EDEFONTI V, HASHIBE M, et al.Carotenoid intake and head and neck cancer:a pooled analysis in the International Head and Neck Cancer Epidemiology Consortium[J]. European Journal of Epidemiology, 2016,31(4):369-383.
[20] JIANG G L, ZHOU L Y, WANG Y T, et al. Astaxanthin from Jerusalem artichoke:Production by fed-batch fermentation using Phaffia rhodozyma and application in cosmetics[J].Process Biochemistry, 2017, 63:16-25.
[21] SCHEWE H, KREUTZER A, SCHMIDT I, et al. High concentrations of biotechnologically produced astaxanthin by lowering pH in a Phaffia rhodozyma, bioprocess[J].Biotechnology and Bioprocess Engineering, 2017, 22(3):319-326.
[22]黄莹.法夫酵母类胡萝卜素的提取纯化及生物活性研究[D].厦门:集美大学, 2017.
[23]黄莹,王凯,杜希萍,等.二甲亚砜法破壁条件对法夫酵母虾青素提取效果的影响[J].食品工业科技, 2017, 38(15):212-217.
[24] DU X P, DONG C C, WANG K, et al. Separation and purification of astaxanthin from Phaffia rhodozyma by preparative high-speed count-current chromatography[J]. Journal of Chromatography B, 2016, 1(1029/1030):191-197.
[25] VAN BREEMEN R B, DONG L L, PAJKOVIC N D.Atmospheric pressure chemical ionization tandem mass spectrometry of carotenoids[J]. International Journal of Mass Spectrometry, 2012, 312:163-172.
[26] RIVERA S M, CHRISTOU P, CANELA-GARAYOA R.Identification of carotenoids using mass spectrometry[J]. Mass Spectrometry Reviews, 2014, 33(5):353-372.
[27] ENZELL C R, FRANCIS G W, LIAAEN-JENSEN S.Mass spectrometric studies of carotenoids[J]. Pure and Applied Chemistry, 1969, 20(4):497-516.