利用重组海藻糖合酶在 中通过发酵和生物催化生产海藻糖。
Trehalose Production Using Recombinant Trehalose Synthase in by Integrating Fermentation and Biocatalysis.
机构信息
State Key Laboratory of Biobased Material and Green Papermaking (LBMP) , Qilu University of Technology (Shandong Academy of Sciences) , Jinan , Shandong 250353 , People's Republic of China.
Key Laboratory of Shandong Microbial Engineering, School of Bioengineering , Qilu University of Technology (Shandong Academy of Sciences) , Jinan , Shandong 250353 , People's Republic of China.
出版信息
J Agric Food Chem. 2019 Aug 21;67(33):9314-9324. doi: 10.1021/acs.jafc.9b03402. Epub 2019 Aug 8.
Trehalose, a stable nonreducing disaccharide, protects biomolecules against environmental stress. However, trehalose production using secretory trehalose synthase (TreS) by has not been well studied. In this study, a mutant TreS was successfully secreted and expressed in WB800N. The extracellular enzyme activity of TreS regulated by the P promoter and SP signal peptide in recombinant WB800N reached 23080.6 ± 1119.4 U/L in a 5-L fermenter after optimizing the culture medium, while , , , and were knocked out. To reduce maltose consumption, and corresponding to maltose transporters were further deleted. To simplify the trehalose production process, we invented a fermentation-coupling biocatalysis process involving recombinant bacteria fermentation to secrete TreS and simultaneous conversion of maltose to trehalose by TreS and found that the conversion rate of maltose to trehalose reached 75.5%, suggesting that this is an efficient strategy for large-scale trehalose production using recombinant .
海藻糖是一种稳定的非还原二糖，能够保护生物分子免受环境压力的影响。然而，利用分泌型海藻糖合酶（TreS）生产海藻糖的研究还不够完善。在本研究中，成功地在 WB800N 中分泌和表达了突变型 TreS。通过优化培养基，在 5-L 发酵罐中，重组 WB800N 中受 P 启动子和 SP 信号肽调控的 TreS 胞外酶活性达到 23080.6±1119.4 U/L，同时敲除了 、 、 、 。为了降低麦芽糖的消耗，进一步缺失了麦芽糖转运蛋白 和 。为了简化海藻糖的生产工艺，我们发明了一种发酵-偶联生物催化过程，涉及重组菌发酵分泌 TreS 和同时通过 TreS 将麦芽糖转化为海藻糖，发现麦芽糖转化为海藻糖的转化率达到 75.5%，表明这是一种利用重组 进行大规模生产海藻糖的有效策略。
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