全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

自诱导系统在酶促合成2’-脱氧胞苷中的应用

, PP. 225-232

Keywords: 生物催化,N-脱氧核糖转移酶,2’-脱氧胞苷,自诱导系统,核苷类似物

Full-Text   Cite this paper   Add to My Lib

Abstract:

旨在高效合成2’-脱氧胞苷(dCyd)。DCyd作为一类重要的药物中间体,能够用于合成吉西他滨(dFdC)、扎西他滨(ddC)等多种抗病毒或抗肿瘤的药物。采用ZYM-Fe-5052自诱导培养基对含有N-脱氧核糖转移酶II(NDT)的大肠杆菌BL21(DE3)进行诱导,所得完整菌体直接用于催化合成dCyd。结果表明,自诱导系统不仅无需额外添加诱导剂,还能够达到较高的菌体密度,且与LB诱导系统所得菌体在合成dCyd方面具有同样高效的催化活性。2’-脱氧胸苷(dThd)和胞嘧啶的浓度比为1∶3时,产物转化率可高达86.5%。合成dCyd的最适反应条件为:2’-脱氧胸苷(dThd)和胞嘧啶的浓度比为1∶1.5,pH6.5的磷酸缓冲液,1mg/mL的菌体量,终体系10mL,30℃条件下反应1h,产物转化率可达71.9%。此方法还可用于制备其他核苷类似物,具有广泛的适用性和应用前景。

References

[1]  Mikhailopulo IA, Miroshnikov AI. New trends in nucleoside Biotech-nology[J]. Acta Naturae, 2010, 2:36-58.
[2]  严琳, 孔军, 侯莉莉.核苷类似物的设计合成及其抗肿瘤活性[J].河南大学学报:自然科学版, 2011, 41(3):257-261.
[3]  Galmarini CM, Mackey JR, Dumontet C. Nucleoside analogues and nucleobases in cancer treatment[J]. Lancet Oncol, 2002, 3:415-424.
[4]  Mathe C, Gosselin G. L-nucleoside enantiomers as antivirals drugs:a mini-review[J]. Antivir Res, 2006, 71:276-281.
[5]  Zhang J, Visser F, King KM, et al. The role of nucleoside transporters in cancer chemotherapy with nucleoside drugs[J]. Cancer Metast Rev, 2007, 26:85-110.
[6]  Zou Z, Ding Q, Ou L, Yan B. Efficient production of deoxynucleoside-5’-monophosphates using deoxynucleoside kinase coupled with a GTP-regeneration system[J]. Appl Microbiol Biotechnol, 2013, 97:9389-9395.
[7]  徐渊, 王鸿, 易喻, 等.核苷合成技术的研究进展[J].化工生产与技术, 2009, 16(2):32-39.
[8]  王丽慧, 丁庆豹, 欧伶, 等.酶法合成5-杂氮-2’-脱氧胞苷[J].华东理工大学学报:自然科学报, 2011, 37(7):325-329.
[9]  樊华伟, 傅绍军, 邵志宇, 朱利民.核苷类药物酶法合成研究进展[J].生物技术通讯, 2005, 16(6):690-692.
[10]  邱蔚然, 丁庆豹.酶法合成核苷类抗病毒药物[J].中国医药工业杂志, 1999, 30(10):474-478.
[11]  Macnutt WS. The enzymically catalysed transfer of the deoxyribosyl group from one purine or pyrimidine to another[J]. Biochem J, 1951, 50:384-397.
[12]  Holguin J, Cardinaud R. Trans-N-Deoxyribosylase:Purification by affinity chromatography and characterization[J]. Eur J Biochem, 1975, 54:505-514.
[13]  Holguin J, Cardinaud R. Trans-N-Deoxyribosylase:Substrate specificity studies purine bases as acceptors[J]. Eur J Biochem, 1975, 54:515-520.
[14]  Kaminski PA. Functional cloning, heterologous expression, and purification of two different N-deoxyribosyltransferases from Lactobacillus helveticus[J]. J Biol Chem, 2002, 277:14400-14407.
[15]  Carson DA, Wasson DB. Synthesis of 2’, 3’-dideoxynucleosides by enzymatic trans-glycosylation[J]. Biochem Bioph Res Co, 1988, 155:829-834.
[16]  Freeman GA, Shaver SR, Rideout JL, Short SA. 2-amino-9-(3-azido-2, 3-dideoxy-β-D- erythro-pentofuranosyl)-6-substituted-9H-purines:Synthesis and anti-HIV activity[J]. Bioorgan Med Chem, 1995, 3:447-458.
[17]  Anand R, Kaminski PA, Ealick SE. Structures of purine 2¢-Deoxy-ribosyltransferase, substrate complexes, and the ribosylated enzyme intermediate at 2.0 ? resolution[J]. Biochemistry, 2004, 43:2384-2393.
[18]  Kiyoshi O, Toshitada N. Molecular cloning and expression of the nucleoside deoxyribosyltransferase-II Gene from lactobacillus helveticus[J]. Biosci Biotechnol Biochem, 2000, 64:2243-2245.
[19]  Gombert AK, Kilikian BV. Recombinant gene expression in Escher-ichia colicultivation using lactose as inducer[J]. J Biotechnol, 1998, 60:47-54.
[20]  Donovan RS, Robinson CW, Glick BR. Review:Optimizing inducer and culture conditions for expression of foreign proteins under the control of the lac promoter[J]. J Ind Microbiol, 1996, 16:145-154.
[21]  Neubauer P, Hofmann K, Holst O, et al. Maximizing the expression of a recombinant gene in Escherichia coli by manipulation of indu-ction time using lactose as inducer[J]. Appl Microbiol Biote-chnol, 1992, 36:739-744.
[22]  Studier FW. Protein production by auto-induction in high-density shaking cultures[J]. Protein Expres Purif, 2005, 41:207-234.
[23]  Xiong J, Zhang W, Su J, et al. Improved synthesis of 2’-deoxyaden-osine and 5-methyluridine by Escherichia coli using an auto-induct-ion system[J]. World J Microbiol Biotechnol, 2012, 28:721-727.
[24]  Lin Y, Zhang W, Zhu F, et al. Subcellular localization of N-deoxyribosyltransferase in Lactobacillus fermentum:cell surface association of an intracellular nucleotide metabolic enzyme[J]. FEMS Microbiol Lett, 2011, 323:132-141.
[25]  Fernandez-Lucas J, Acebal C, Sinisterra JV, et al. Lactobacillus reuteri 2’-deoxyribosyltransferase, a novel biocatalyst for tailoring of nucleosides[J]. Appl Environ Microb, 2010, 76:1462-1470.
[26]  Britos CN, Cappa VA, Rivero CW, et al. Biotransformation of halo-genated 2’-deoxyribosides by immobilized lactic acid bacteria[J]. J Mol Catal B-Enzym, 2012, 79:49-53.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133