利用固定化重组大肠杆菌细胞生产D-塔格糖

中国生物工程杂志

2011, Vol. 31

Issue (7): 85-90

研究报告

利用固定化重组大肠杆菌细胞生产D-塔格糖

付凤根, 徐铮, 李贵祥, 李莎, 冯小海, 徐虹

南京工业大学食品与轻工学院材料化学工程国家重点实验室南京 210009

D-tagatose Production Utilizing Immobilized Recombinant Escherichia coli cells

FU Feng-gen, XU Zheng, LI Gui-xiang, LI Sha, FENG Xiao-hai, XU Hong

State Key Laboratory of Materials-Oriented Chemical Engineering,College of Food Science and Light Industry, Ningjing University of Technology,Nanjing 210009,China

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摘要：

利用经海藻酸钙包埋的重组大肠杆菌细胞催化D-半乳糖生产D-塔格糖,考察了细胞包埋量、反应条件对固定化细胞催化效率以及对D-塔格糖生产稳定性的影响。确定的最优转化条件为:温度65℃,pH 6.5,添加终浓度为1 mmol/L Mn²⁺,底物(D-半乳糖)浓度100 g/L,重组大肠杆菌细胞用量40 g/L。固定化小球在0.3%戊二醛溶液中交联30 min可以显著提高其在高温下的机械强度。考察了异构化反应体系中硼酸与底物间的摩尔比对产率的影响。研究结果表明,添加适量的硼酸可以改变原有的化学反应平衡,实现D-塔格糖的高产。利用D-半乳糖为底物在最优的反应条件下催化24 h,固定化细胞对D-半乳糖的转化率最高,可达65.8%,连续转化8批次的平均转化率为60.6%,为工业化生产D-塔格糖奠定了基础。

关键词： D-塔格糖; 固定化细胞; 重组大肠杆菌; 硼酸; L-阿拉伯糖异构酶

Abstract:

Recombinant E.coli cells were immobilized with calcium alginate for the isomerization of D-galactose to D-tagatose. The influences of immobilization process and reaction conditions on the catalytic efficiency and D-tagatose production stability of the immobilized cells are investigated. The optimal conditions were as follows: the temperature, pH and mental ion were 65℃,6.5 and 1mmol/L Mn²⁺, respectively. The optimal substrate (D-galactose) and cell concentrations of 100 g/L and 40 g/L were selected, respectively. Stability of the alginate beads under high temperatures was enhanced after cross-linked by 0.3% glutaraldehyde for 30min. The effect of molar ratio between borate and substrate on isomerization was studied, it was found that borate could change the initial chemical equilibrium of D-galactose isomerization and lead to a high production of D-tagatose. After conversion for 24h, the highest conversion rate for D-tagatose using D-galactose as the substrate reached to 65.8% by immobilized recombinant E.coli cells, and the average conversion rate of eight repeated batch conversions was 60.6%, which laid the foundation for industrial production of D-tagatose.

Key words: D-tagatose Immobilized cells Recombinant E.coli Borate L-arabinose isomerase

收稿日期: 2011-03-17 出版日期: 2011-07-25

ZTFLH:

Q784

基金资助:

中央级公益性科研院所基本科研业务费专项资金项目(B2F100306)

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引用本文:

付凤根, 徐铮, 李贵祥, 李莎, 冯小海, 徐虹. 利用固定化重组大肠杆菌细胞生产D-塔格糖[J]. 中国生物工程杂志, 2011, 31(7): 85-90.

FU Feng-gen, XU Zheng, LI Gui-xiang, LI Sha, FENG Xiao-hai, XU Hong. D-tagatose Production Utilizing Immobilized Recombinant Escherichia coli cells. China Biotechnology, 2011, 31(7): 85-90.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2011/V31/I7/85

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