凯发一触即发

至今,GenScript的服务及产品已被Cell, Nature, Science, PNAS等1300多家生物医药类杂志引用近万次,处于行业领先水平。NIH、哈佛、耶鲁、斯坦福、普林斯顿、杜克大学等约400家全球著名机构使用GenScript的基因合成、多肽服务、抗体服务和蛋白服务等成功地发表科研成果,再次证明GenScript 有能力帮助业内科学家Make research easy.

Overexpression of epoxide hydrolase in Rhodococcus ruber with high robustness for the synthesis of chiral epichlorohydrin

Process Biochemistry. 2019; 
YouxiangLiangabSongJiaoabMiaomiaoWangabHuiminYuabcZhongyaoShenab
Products/Services Used Details Operation
Gene Synthesis Restriction enzymes, DNA polymerases, T4 DNA ligases and a site-directed mutagenesis kit were purchased from Takara (Dalian, China) and Vazyme (Nanjing, China). DNA purification kits, gel extraction kits, BCA-protein quantification assay kits, and plasmid extraction kits were purchased from Biomega (Shanghai, China). Protein marker (MM1397-500) was purchased from GenScript (Nanjing, China). ArEH gene synthesis was completed by Qinglan (Wuxi, China) and DNA sequencingwas performed by GENEWIZ (Suzhou, China). DNA manipulations including PCR, agarose gel electrophoresis, restriction enzyme digestion, ligation and transformation were performed following standard procedures from the manufacturers’ protocols. Get A Quote

摘要

Epoxide hydrolases (EHs) are attractive enzymes for producing enantiopure epoxides and diols, but do not display enough stability when lysates or Escherichia coli whole cells are used as biocatalysts. In this work, an organic-solvent tolerant strain Rhodococcus ruber THdAdN was utilized to overexpress an epoxide hydrolase from Agrobacterium radiobacter (ArEH), using E. coli BL21(DE3) as a control. The proportion of ArEH in all soluble proteins of R. ruber THdAdN(ArEH) reached 30.3%, which was comparable to that of E. coli BL21(DE3)(ArEH). Due to a higher cell density in flask cultivation, the maximum ArEH activity of R. ruber THdAdN(ArEH) toward epichlorohydrin (ECH) reached 5.4 U/mL, approximatel... More

关键词

Rhodococcus ruberWhole-cell biocatalystsEpoxide hydrolaseStability and organic-solvent toleranceChiral epoxides