β-半乳糖苷酶催化合成半乳糖基氯苯那敏及其性质研究。

β-galactosidase-catalyzed synthesis of galactosyl chlorphenesin and its characterization.

机构信息

Department of Biotechnology, Korea National University of Transportation, Jeungpyung, Chungbuk, 368-701, Republic of Korea.

出版信息

Appl Biochem Biotechnol. 2013 Nov;171(6):1299-312. doi: 10.1007/s12010-013-0213-3. Epub 2013 Apr 7.

DOI:10.1007/s12010-013-0213-3

PMID:23564435

Abstract

We synthesized galactosyl chlorphenesin (CPN-G) using β-gal-containing Escherichia coli (E. coli) cells in which the conversion yield of chlorphenesin (CPN) to CPN-G reached about 64 % during 12 h. CPN-G was identified and characterized using high-performance liquid chromatography, liquid chromatography-mass spectrometry, Fourier transform-infrared spectrometry, and nuclear magnetic resonance analysis ((1)H and (13)C). We verified that a galactose was covalently bound to a CPN alcohol group during CPN-G synthesis throughout these analyses. In particular, by the hydrolysis of CPN-G using β-gal, it was confirmed that a galactose was bound to CPN. The minimal inhibitory concentration (MIC) results showed that the CPN-G MICs were fairly similar to those of CPN. HACAT cell viability was significantly higher in CPN-G-treated cells than in CPN-treated cells at concentrations of 0.0-20.0 mM. Finally, we accomplished the synthesis of less toxic CPN-G, compared with CPN, using β-gal-containing E. coli cells as whole cells without changes in the MICs against microorganisms.

摘要

我们使用含有β-半乳糖苷酶的大肠杆菌（E. coli）细胞合成了半乳糖基氯苯那敏（CPN-G），在 12 小时内，CPN 到 CPN-G 的转化率约为 64%。通过高效液相色谱、液质联用、傅里叶变换红外光谱和核磁共振分析（1H 和 13C）对 CPN-G 进行了鉴定和表征。通过这些分析，我们验证了在 CPN-G 合成过程中，半乳糖与 CPN 的醇基发生了共价结合。特别是，通过β-半乳糖苷酶对 CPN-G 的水解，证实了半乳糖与 CPN 的结合。最小抑菌浓度（MIC）结果表明，CPN-G 的 MIC 与 CPN 的 MIC 相当。在 0.0-20.0 mM 的浓度下，CPN-G 处理的 HACAT 细胞的细胞活力明显高于 CPN 处理的细胞。最后，我们使用含有β-半乳糖苷酶的大肠杆菌细胞作为全细胞，在不改变对微生物的 MIC 的情况下，完成了比 CPN 毒性更小的 CPN-G 的合成。