葡萄柚中糖基转移酶对槲皮素及其类似物催化机制探讨
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摘要
研究了葡萄柚中糖基转移酶(FGT)作为催化酶,以UDP-葡萄糖、UDP-N-乙酰-D-氨基葡萄糖、UDP-甘露糖为糖基供体,槲皮素、柚皮素、柚皮苷为糖基受体,研究糖基化合成情况。通过高效液相、质谱及核磁共振氢谱对产物反应进行检测。确定了槲皮素与UDP-葡萄糖、UDP-N-乙酰-D-氨基葡萄糖可以发生糖基化反应,生成的槲皮素糖苷分子量分别为464、505。根据核磁结果鉴定,所得到的产物结构分别为槲皮素-3-O-β-D葡萄糖苷、槲皮素3-O-β-D-N-乙酰-D-氨基葡萄糖苷。以FGT酶为催化酶,对槲皮素进行了糖基化修饰。
Using Glycosyltransferase(FGT)in grapefruit as catalytic enzyme,UDP-glucose,UDP-N-acetyl-D-glucosamine,and UDP-mannose as glycosyl donor,quercetin and naringin as glycosylation receptor,glycosylation was studied. The reaction of the product was detected by high performance liquid chromatography,mass spectrometry and nuclear magnetic resonance(NMR)hydrogen spectrum.The glycosylation reaction of quercetin with UDP-glucose and UDP-N-acetyl-D-glucosamine was confirmed. The molecular weight of quercetin glycoside was 464 and 505,respectively. According to the results of nuclear magnetic resonance,the structures of the products were quercetin-3-O-beta-D glucoside and quercetin-3-O-beta-D-N-acetyl-D-glucosamine. It was verified that FGT enzyme was used to modify quercetin glycosylation.
Using Glycosyltransferase(FGT)in grapefruit as catalytic enzyme,UDP-glucose,UDP-N-acetyl-D-glucosamine,and UDP-mannose as glycosyl donor,quercetin and naringin as glycosylation receptor,glycosylation was studied. The reaction of the product was detected by high performance liquid chromatography,mass spectrometry and nuclear magnetic resonance(NMR)hydrogen spectrum.The glycosylation reaction of quercetin with UDP-glucose and UDP-N-acetyl-D-glucosamine was confirmed. The molecular weight of quercetin glycoside was 464 and 505,respectively. According to the results of nuclear magnetic resonance,the structures of the products were quercetin-3-O-beta-D glucoside and quercetin-3-O-beta-D-N-acetyl-D-glucosamine. It was verified that FGT enzyme was used to modify quercetin glycosylation.
引文
[1]Charnock SJ, Henrissat B, Davies GJ. Three-dimensional structures of UDP-sugar glycosyltransferases illuminate the biosynthesis of plant polysaccharides[J]. Plant Physiol, 2001, 125(2):527-531.
[2]Gachon CMM, Langlois-Meurinne M, Saindrenan P. Plant secondary metabolismglycosyltransferases:theemergingfunctional analysis[J]. Trends Plant Sci, 2005, 10(11):542-549.
[3]Coutinho PM, Deleury E, Davies GJ, et al. An evolving hierarchical family classification for glycosyltransferases[J]. Mol Biol, 2003,328(2):307-317.
[4]尹恒,王文霞,赵小明,等.植物糖生物学研究进展[J].植物学报, 2010, 45(5):521-529.
[5]Khorsandi L, Orazizadeh M, Niazvand F, et al. Quercetin induces apoptosis and necroptosis in MCF-7 breast cancer cells[J]. Bratisl Lek Listy, 2017, 118(2):123-128.
[6]Yu D, Ye T, Xiang Y, et al. Quercetin inhibits epithelialmesenchymal transition, decreases invasiveness and metastasis, and reverses IL-6induced epithelial-mesenchymal transition, expression of MMP by inhibiting STAT3 signaling in pancreatic cancer cells[J]. Onco Targets Ther, 2017, 201(10):4719-4729.
[7]王中旺,许梦川,杨雯茜,等.槲皮素对2型糖尿病患者机体铁代谢的影响[J].河北师范大学学报:自然科学版, 2017, 41(3):272-276.
[8]汤喜兰,刘建勋,董伟,等.槲皮素对大鼠乳鼠心脏成纤维细胞炎症分泌的干预作用[J].中国中药杂志, 2014, 39(12):2314-2317.
[9]Byun EB, Yang MS, Choi HG, et al. Quercetin negatively regulates TLR4 signaling induced by lipopolysaccharide through Tollip expression[J]. Biochem Biophys ResCommun, 2013, 431(4):698-705.
[10]万巧凤,吴莉,杨美玲,等.槲皮素对甲型H1N1流感病毒诱导的A549细胞凋亡效应酶Caspase-3的影响[J].中国中医药信息杂志, 2011, 18(10):42-44.
[11]张丹丹,方建国,陈娟娟,等.连翘及其主要有效成分槲皮素体外抗人巨细胞病毒的实验研究[J].中国中药杂志, 2010,35(8):1055-1059.
[12]Hui L, Xiao LG, Yi C, et al. Heart protective effects and mechanism of quercetinpreconditioning onanti-myocardialischemia reperfusion(IR)injuries in rats[J]. Gene, 2014, 4(43):1-7.
[13]吴薛明,许婷婷,储建林,等.黄酮类化合物酶法糖基化修饰的研究进展[J].中国天然药物, 2010, 8(5):389-400.
[14]Ko JH, Kim BG, Ahn JH. Glycosylation of flavonoids with a glycosyltransferase from Bacillus cereus[J]. FEMS Microbiology Letters, 2006, 258(2):263-268.
[15]Lim EK, Ashford DA, Hou BK, et al. Arabidopsis glycosyltransferases as biocatalysts in fermentation for regioselective synthesis of diverse quercetin glucosides[J]. Biotechnol Bioeng, 2004, 87(5):623-631.
[16]Son MH, Kim BG, Kim DH, et al. Production of flavonoid O-glucoside using sucrose synthase and flavonoid O-glucosyltransferasefusionprotein[J].JournalofMicrobiologyand Biotechnology, 2009, 19(7):709-712.
[17]Xu JQ, Fan N, Yu BY, et al. Biotransformation of quercetin by Gliocladium deliquescens NRRL 108[J]. Chinese Journal of Nature Medicines, 2017, 15(8):615-424.
[18]De Bruyn F, Van Brempt M, Maertens J. et al. Metabolic engineering of Escherichia coli into a versatile glycosylation platform:production of bio-active quercetin glycosides[J].Microbial Cell Factories, 2015, 14:1-12.
[19]焦洁.考马斯亮蓝G-250染色法测定苜蓿中可溶性蛋白含量[J].农业工程技术, 2016, 36(17):33-34.
[20]李侠,臧学丽,徐祎博,等. AB-8大孔树脂纯化绿豆皮黄酮工艺优化及纯化前后抗氧化能力比较[J].食品科学, 2018, 39(10):283-290.
[21]Aramsangtienchai P, Chavasiri W, Ito K, et al. Synthesis of epicatechin glucosides by a cyclodextrin glycosyltransferase[J].Molecular Catalysis B:Enzymatic, 2011, 73:27-34.
[22]Chiu HH, Hsieh YC, Chen YH, et al. Three important amino acids control the regioselectivity of flavonoid glucosidation in glycosyltransferase-1 from Bacillus cereus[J]. Applied Microbiology and Biotechnology, 2016, 100(9):8411-8424.
[2]Gachon CMM, Langlois-Meurinne M, Saindrenan P. Plant secondary metabolismglycosyltransferases:theemergingfunctional analysis[J]. Trends Plant Sci, 2005, 10(11):542-549.
[3]Coutinho PM, Deleury E, Davies GJ, et al. An evolving hierarchical family classification for glycosyltransferases[J]. Mol Biol, 2003,328(2):307-317.
[4]尹恒,王文霞,赵小明,等.植物糖生物学研究进展[J].植物学报, 2010, 45(5):521-529.
[5]Khorsandi L, Orazizadeh M, Niazvand F, et al. Quercetin induces apoptosis and necroptosis in MCF-7 breast cancer cells[J]. Bratisl Lek Listy, 2017, 118(2):123-128.
[6]Yu D, Ye T, Xiang Y, et al. Quercetin inhibits epithelialmesenchymal transition, decreases invasiveness and metastasis, and reverses IL-6induced epithelial-mesenchymal transition, expression of MMP by inhibiting STAT3 signaling in pancreatic cancer cells[J]. Onco Targets Ther, 2017, 201(10):4719-4729.
[7]王中旺,许梦川,杨雯茜,等.槲皮素对2型糖尿病患者机体铁代谢的影响[J].河北师范大学学报:自然科学版, 2017, 41(3):272-276.
[8]汤喜兰,刘建勋,董伟,等.槲皮素对大鼠乳鼠心脏成纤维细胞炎症分泌的干预作用[J].中国中药杂志, 2014, 39(12):2314-2317.
[9]Byun EB, Yang MS, Choi HG, et al. Quercetin negatively regulates TLR4 signaling induced by lipopolysaccharide through Tollip expression[J]. Biochem Biophys ResCommun, 2013, 431(4):698-705.
[10]万巧凤,吴莉,杨美玲,等.槲皮素对甲型H1N1流感病毒诱导的A549细胞凋亡效应酶Caspase-3的影响[J].中国中医药信息杂志, 2011, 18(10):42-44.
[11]张丹丹,方建国,陈娟娟,等.连翘及其主要有效成分槲皮素体外抗人巨细胞病毒的实验研究[J].中国中药杂志, 2010,35(8):1055-1059.
[12]Hui L, Xiao LG, Yi C, et al. Heart protective effects and mechanism of quercetinpreconditioning onanti-myocardialischemia reperfusion(IR)injuries in rats[J]. Gene, 2014, 4(43):1-7.
[13]吴薛明,许婷婷,储建林,等.黄酮类化合物酶法糖基化修饰的研究进展[J].中国天然药物, 2010, 8(5):389-400.
[14]Ko JH, Kim BG, Ahn JH. Glycosylation of flavonoids with a glycosyltransferase from Bacillus cereus[J]. FEMS Microbiology Letters, 2006, 258(2):263-268.
[15]Lim EK, Ashford DA, Hou BK, et al. Arabidopsis glycosyltransferases as biocatalysts in fermentation for regioselective synthesis of diverse quercetin glucosides[J]. Biotechnol Bioeng, 2004, 87(5):623-631.
[16]Son MH, Kim BG, Kim DH, et al. Production of flavonoid O-glucoside using sucrose synthase and flavonoid O-glucosyltransferasefusionprotein[J].JournalofMicrobiologyand Biotechnology, 2009, 19(7):709-712.
[17]Xu JQ, Fan N, Yu BY, et al. Biotransformation of quercetin by Gliocladium deliquescens NRRL 108[J]. Chinese Journal of Nature Medicines, 2017, 15(8):615-424.
[18]De Bruyn F, Van Brempt M, Maertens J. et al. Metabolic engineering of Escherichia coli into a versatile glycosylation platform:production of bio-active quercetin glycosides[J].Microbial Cell Factories, 2015, 14:1-12.
[19]焦洁.考马斯亮蓝G-250染色法测定苜蓿中可溶性蛋白含量[J].农业工程技术, 2016, 36(17):33-34.
[20]李侠,臧学丽,徐祎博,等. AB-8大孔树脂纯化绿豆皮黄酮工艺优化及纯化前后抗氧化能力比较[J].食品科学, 2018, 39(10):283-290.
[21]Aramsangtienchai P, Chavasiri W, Ito K, et al. Synthesis of epicatechin glucosides by a cyclodextrin glycosyltransferase[J].Molecular Catalysis B:Enzymatic, 2011, 73:27-34.
[22]Chiu HH, Hsieh YC, Chen YH, et al. Three important amino acids control the regioselectivity of flavonoid glucosidation in glycosyltransferase-1 from Bacillus cereus[J]. Applied Microbiology and Biotechnology, 2016, 100(9):8411-8424.