Biosynthesis of 2-deoxysugars using whole-cell catalyst expressing 2-deoxy-d-ribose 5-phosphate aldolase
文献类型：期刊论文
| 作者 | Li, Jitao1; Yang, Jiangang1; Men, Yan1; Zeng, Yan1; Zhu, Yueming1; Dong, Caixia2; Sun, Yuanxia1; Ma, Yanhe1 |
| 刊名 | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
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| 出版日期 | 2015-10-01 |
| 卷号 | 99期号:19页码:7963-7972 |
| 关键词 | 2-Deoxy-D-ribose 5-phosphate aldolase (DERA) Deoxysugars Whole-cell transformation Directed mutagenesis |
| 英文摘要 | 2-Deoxy-d-ribose 5-phosphate aldolase (DERA) accepts a wide variety of aldehydes and is used in de novo synthesis of 2-deoxysugars, which have important applications in drug manufacturing. However, DERA has low preference for non-phosphorylated substrates. In this study, DERA from Klebsiella pneumoniae (KDERA) was mutated to increase its enzyme activity and substrate tolerance towards non-phosphorylated polyhydroxy aldehyde. Mutant KDERA(K12) (S238D/F200I/Delta Y259) showed a 3.15-fold improvement in enzyme activity and a 1.54-fold increase in substrate tolerance towards d-glyceraldehyde compared with the wild type. Furthermore, a whole-cell transformation strategy using resting cells of the BL21(pKDERA12) strain, containing the expressed plasmid pKDERA12, resulted in increase in 2-deoxy-d-ribose yield from 0.41 mol/mol d-glyceraldehyde to 0.81 mol/mol d-glyceraldehyde and higher substrate tolerance from 0.5 to 3 M compared to in vitro assays. With further optimization of the transformation process, the BL21(pKDERA12) strain produced 2.14 M (287.06 g/L) 2-deoxy-d-robose (DR), with a yield of 0.71 mol/mol d-glyceraldehyde and average productivity of 0.13 mol/L center dot h (17.94 g/L center dot h). These results demonstrate the potential for large-scale production of 2-deoxy-d-ribose using the BL21(pKDERA12) strain. Furthermore, the BL21(pKDERA12) strain also exhibited the ability to efficiently produce 2-deoxy-d-altrose from d-erythrose, as well as 2-deoxy-l-xylose and 2-deoxy-l-ribose from l-glyceraldehyde. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biotechnology & Applied Microbiology |
| 研究领域[WOS] | Biotechnology & Applied Microbiology |
| 关键词[WOS] | ORGANIC-SYNTHESIS ; ESCHERICHIA-COLI ; ENZYMES ; PATHWAYS |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000360839800014 |
| 源URL | [http://124.16.173.210/handle/834782/1568]  |
| 专题 | 天津工业生物技术研究所_功能糖与天然活性物质研究组 孙媛霞 _期刊论文 |
| 作者单位 | 1.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Natl Engn Lab Ind Enzymes, Tianjin 300308, Peoples R China 2.Tianjin Med Univ, Sch Pharm, Tianjin Key Lab Technol Enabling Dev Clin Therape, Tianjin 300070, Peoples R China |
| 推荐引用方式 GB/T 7714 | Li, Jitao,Yang, Jiangang,Men, Yan,et al. Biosynthesis of 2-deoxysugars using whole-cell catalyst expressing 2-deoxy-d-ribose 5-phosphate aldolase[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2015,99(19):7963-7972. |
| APA | Li, Jitao.,Yang, Jiangang.,Men, Yan.,Zeng, Yan.,Zhu, Yueming.,...&Ma, Yanhe.(2015).Biosynthesis of 2-deoxysugars using whole-cell catalyst expressing 2-deoxy-d-ribose 5-phosphate aldolase.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,99(19),7963-7972. |
| MLA | Li, Jitao,et al."Biosynthesis of 2-deoxysugars using whole-cell catalyst expressing 2-deoxy-d-ribose 5-phosphate aldolase".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 99.19(2015):7963-7972. |
入库方式： OAI收割
来源：天津工业生物技术研究所
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