Chemo-enzymatic synthesis of Neu5Gc-containing sialylated lactulose

Zeng, Jie; Tian, Jia; Hu, Yajie; Zhang, Ruiyao; Sun, Junliang; Li, Bangbang; Gao, Haiyan

doi:10.4314/tjpr.v16i7.26

Cited by 2 publications

(1 citation statement)

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“…As described in detail previously [ 21 ], the assays were performed in a total volume of 10 μL in Tris-HCl buffer (100 mM) with pH values varying from 7.0 to 9.0; the reaction contained lactulose (10 mM), Neu5Ac (15 mM), CTP (15 mM), MgCl 2 (20 mM), NmCSS (5 μg), and PmST1_M144D or Pd26ST (5 μg). The reactions were allowed to proceed at 32, 37 and 42°C and monitored by thin-layer chromatography (TLC) every 30 min.…”

Section: Methodsmentioning

confidence: 99%

Chemoenzymatic synthesis of sialylated lactuloses and their inhibitory effects on Staphylococcus aureus

Zeng

Tian

et al. 2018

PLoS ONE

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BackgroundSialylated glycoconjugates play important roles in physiological and pathological processes. However, available sialylated oligosaccharides source is limited which is a barrier to study their biological roles. This work reports an efficient approach to produce sialic acid-modified lactuloses and investigates their inhibitory effects on Staphylococcus aureus (S. aureus).MethodsA one-pot two-enzyme (OPTE) sialylation system was used to efficiently synthesize sialylated lactuloses. Silica gel flash chromatography column was employed to purify the sialylated products. The purity and identity of the product structures were confirmed with mass spectrometry (MS) and nuclear magnetic resonance (NMR). The inhibitory effect of sialylated lactuloses against S. aureus was evaluated by using microplate assay, fluorescence microscopy, DAPI (4',6-diamidino-2-phenylindole) fluorescence staining and protein leakage quantification.ResultsNeu5Ac-containing sialylated lactuloses with either α2,3- or α2,6-linkages were efficiently synthesized via an efficient OPTE sialylation system using α-2,3-sialyltransferase or α-2,6-sialyltransferase, respectively. Neu5Ac-α2,3-lactulose and Neu5Ac-α2,6-lactulose significantly inhibited the growth of S. aureus. Fluorescence microscopy and DAPI fluorescence staining indicated that the sialylated lactuloses might disrupt nucleic acid synthesis of S. aureus.ConclusionsNeu5Ac-containing sialylated lactuloses had higher antibacterial activity against S. aureus than non-sialylated lactulose. The inhibitory effect of Neu5Ac-α2,3-lactulose was superior to that of Neu5Ac-α2,6-lactulose. The sialylated lactuloses might inhibit S. aureus by causing cell membrane leakage and disrupting nucleic acid synthesis.

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Section: Methodsmentioning

confidence: 99%