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. 2023 Aug;107(15):4803-4813.
doi: 10.1007/s00253-023-12628-8. Epub 2023 Jun 19.

Efficient production of α-monoglucosyl hesperidin by cyclodextrin glucanotransferase from Bacillus subtilis

Jiawei Zhou  1   2 , Yuan Shi  1   2 , Jingyi Fang  1   2 , Tian Gan  1   2 , Yuele Lu  1   2 , Linjiang Zhu  3   4 , Xiaolong Chen  1   2
Affiliations

Efficient production of α-monoglucosyl hesperidin by cyclodextrin glucanotransferase from Bacillus subtilis

Jiawei Zhou et al. Appl Microbiol Biotechnol. 2023 Aug.

Abstract

α-Monoglucosyl hesperidin is a promising food additive with various activities. However, there are a few reports about the production of α-monoglucosyl hesperidin. Here, to develop a practical and safe process for α-monoglucosyl hesperidin synthesis, we used nonpathogenic Bacillus subtilis as a host to express cyclodextrin glucanotransferase (CGTase) from Bacillus sp. A2-5a. The promoters and signal peptides were screened to optimize the transcription and secretion of CGTase in B. subtilis. The results of optimization showed that the best signal peptide and promoter were YdjM and PaprE, respectively. Finally, the enzyme activity increased to 46.5 U mL-1, 8.7 times that of the enzyme expressed from the strain containing pPHpaII-LipA, and the highest yield of α-monoglucosyl hesperidin was 2.70 g L-1 by enzymatic synthesis using the supernatant of the recombinant B. subtilis WB800 harboring the plasmid pPaprE-YdjM. This is the highest α-monoglucosyl hesperidin production level using recombinant CGTase to date. This work provides a generally applicable method for the scaled-up production of α-monoglucosyl hesperidin. KEY POINTS: • A three-step procedure was created for high throughput signal peptide screening. • YdjM and PaprE were screened from 173 signal peptides and 13 promoters. • α-Monoglucosyl hesperidin was synthesized by CGTase with a yield of 2.70 g L-1.

Keywords: Bacillus subtilis; Cyclodextrin glucanotransferase; Enzymatic synthesis; Optimization; α-Monoglucosyl hesperidin.

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