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用海藻酸钙固定化表达 L-阿拉伯糖异构酶的重组大肠杆菌细胞生产 L-核酮糖。

Alginate immobilization of recombinant Escherichia coli whole cells harboring L-arabinose isomerase for L-ribulose production.

机构信息

Department of Chemical Engineering, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul, 143-701, Korea.

出版信息

Bioprocess Biosyst Eng. 2010 Aug;33(6):741-8. doi: 10.1007/s00449-009-0397-7. Epub 2009 Nov 28.

Abstract

Recombinant Escherichia coli whole cells harboring Bacillus licheniformis L-arabinose isomerase (BLAI) were immobilized with alginate. The operational conditions for immobilization were optimized with response surface methodology. Optimal alginate concentration, Ca(2+) concentration, and cell mass loading were 1.8% (w/v), 0.1 M, and 44.5 g L(-1), respectively. The interactions between Ca(2+) concentration, alginate concentration, and initial cell mass were significant. After immobilization of BLAI, cross-linking with 0.1% glutaraldehyde significantly reduced cell leakage. The half-life of immobilized whole cells was 150 days, which was 50-fold longer than that of free cells. In seven repeated batches for L-ribulose production, the productivity was as high as 56.7 g L(-1) h(-1) at 400 g L(-1) substrate concentration. The immobilized cells retained 89% of the initial yield after 33 days of reaction. Immobilization of whole cells harboring BLAI, therefore, makes a suitable biocatalyst for the production of L-ribulose, particularly because of its high stability and low cost.

摘要

含地衣芽孢杆菌 L-阿拉伯糖异构酶(BLAI)的重组大肠杆菌全细胞用藻酸盐固定化。用响应面法优化了固定化的操作条件。最佳藻酸钠浓度、Ca2+浓度和细胞质量负载分别为 1.8%(w/v)、0.1 M 和 44.5 g/L。Ca2+浓度、藻酸钠浓度和初始细胞质量之间的相互作用是显著的。BLAI 固定化后,用 0.1%戊二醛交联可显著降低细胞渗漏。固定化全细胞的半衰期为 150 天,是游离细胞的 50 倍。在 400 g/L 底物浓度下进行了 7 次重复批次的 L-核酮糖生产,产率高达 56.7 g/L/h。反应 33 天后,固定化细胞保留了初始产率的 89%。因此,固定化 BLAI 全细胞是生产 L-核酮糖的合适生物催化剂,特别是因为它具有高稳定性和低成本。

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