Sequential <scp>One‐Pot Three‐Enzyme</scp> Synthesis of the Tetrasaccharide Repeating Unit of Group B <i>Streptococcus</i> Serotype <scp>VIII</scp> Capsular Polysaccharide

Liang, Min; Gong, Wei; Sun, Chongzhen; Zhao, Jielin; Wang, Hong; Chen, Zonggang; Xiao, Min; Gu, Guofeng

doi:10.1002/cjoc.202100822

Cited by 3 publications

(6 citation statements)

References 33 publications

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“…As depicted in Figure 5 , the reaction velocity catalyzed by WciN was dramatically reduced as the concentration of Glcα-PP-(CH 2 ) 11 -OPh 1 was greater than 0.25 mM, indicating the activity of WciN could be easily inhibited even by a slightly higher concentration of acceptor substrate. This finding was radically different from those reported glycosyltransferases that could well accept Glcα-PP-(CH 2 ) 11 -OPh 1 as substrate acceptor ( Wang et al, 2019 ; Liang et al, 2022 ). Bioinformatics analysis of the amino acid sequence of WciN revealed that there was no transmembrane domain in WciN, lacking a domain that interacts with the membrane.…”

Section: Resultscontrasting

confidence: 99%

“…We have recently reported the optimized synthesis of the glycolipid Glcα-PP-(CH 2 ) 11 -OPh 1 ( Liang et al, 2022 ) and utilized it as an acceptor substrate to characterize several bacterial glycosyltransferases related to CPS RU biosynthesis ( Wang et al, 2019 ; Wang et al, 2021 ; Liang et al, 2022 ). Therefore, using it as an enzymatic substrate, the detailed biochemical properties of WciN were then investigated.…”

Section: Resultsmentioning

confidence: 99%

“…Sugar nucleotide donors UDP-Gal and UDP-Glc were prepared as previously described ( Li et al, 2020 ). Enzymatic acceptor substrate Glcα-PP-(CH 2 ) 11 -OPh 1 was synthesized followed the protocol reported previously ( Liang et al, 2022 ). Ni 2+ Sepharose high performance was the product of GE healthcare.…”

Section: Methodsmentioning

confidence: 99%

“…Reaction mixtures were performed for 10 min and then terminated by boiling at 100°C for 30 s. After centrifuging for 10 min under 12,000 rpm, the supernatant was analyzed with HPLC (DionexCarboPac TM PA-100 column, 4 × 250 mm, 0–1 M ammonium acetate buffer eluent). The byproduct UDP was monitored to assess the reaction process owing to its strong UV absorption at 260 nm and its convenience to be quantitated by HPLC ( Wang et al, 2021 ; Liang et al, 2022 ).…”

Section: Methodsmentioning

confidence: 99%

“…Thus, the more in vitro evidence to confirm such serotype evolution are further worthy of exploring. In this study, we presented in detail the biochemical properties of WciN derived from pneumococcus type 6B using the synthesized glycolipid Glcα-PP-(CH 2 ) 11 -OPh 1 ( Wang et al, 2019 ; Wang et al, 2021 ; Liang et al, 2022 ) ( Figure 1B ) as acceptor substrate, and carried out the single or dual amino acid substitution of D38 and/or A150 residue of WciN to verify the different glycosylation functions of the resultant glycosyltransferase mutants.…”

Section: Introductionmentioning

confidence: 99%

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Biochemical Characterization and Synthetic Application of WciN and Its Mutants From Streptococcus pneumoniae Serotype 6B

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The biochemical properties of α-1,3-galactosyltransferase WciN from Streptococcus pneumoniae serotype 6B were systemically characterized with the chemically synthesized Glcα-PP-(CH2)11-OPh as an acceptor substrate. The in vitro site-directed mutation of D38 and A150 residues of WciN was further investigated, and the enzymatic activities of those WciN mutants revealed that A150 residue was the pivotal residue responsible for nucleotide donor recognition and the single-site mutation could completely cause pneumococcus serotype switch. Using WciNA150P and WciNA150D mutants as useful tool enzymes, the disaccharides Galα1,3Glcα-PP-(CH2)11-OPh and Glcα1,3Glcα-PP-(CH2)11-OPh were successfully prepared in multi-milligram scale in high yields.

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Section: Resultscontrasting

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Biochemical Characterization and Synthetic Application of WciN and Its Mutants From Streptococcus pneumoniae Serotype 6B

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Chemical Synthesis of an Octasaccharide Derivative Related to Group B StreptococcusCell‐Wall Polysaccharide

et al. 2022

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Comprehensive Summary Group B Streptococcus (GBS) is the major pathogen that causes invasive infectious diseases in neonates and infants. The development of preventive and therapeutic strategies against GBS infection has been becoming the most pressing subject worldwide. Group B carbohydrate (GBC), the group B‐specific polysaccharide that distinguishes GBS with other streptococci species, has been identified as an attractive antigen for diagnosis and vaccine development because of its highly conservative tetra‐antennary structure. In this paper, a highly convergent [3 + 5] glycosylation strategy for efficient synthesis of an octasaccharide derivative related to GBC oligosaccharide unit II has been developed. In this synthesis, each glycosylation reaction was efficiently constructed with glycosyl imidates, especially trifluoroacetimidate, as donors, and each glycosidic bond was stereoselectively controlled via the neighboring group participation effect of acyl group on the 2‐O‐position of imidate donors or the solvent effect of Et2O. Furthermore, the aminoethylphosphate group was smoothly installed on the 6‐O‐position of d‐glucitol residue using the phosphoramidite method. After global deprotection, the target octasaccharide was successfully obtained from d‐glucitol in 29 steps with an overall yield of 1.37%. The free amino group installed on the aminoethylphosphate spacer of the target molecule enables its modification with functionalized biomolecules for further biological studies.

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Rhamnose-Containing Compounds: Biosynthesis and Applications

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et al. 2022

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Rhamnose-associated molecules are attracting attention because they are present in bacteria but not mammals, making them potentially useful as antibacterial agents. Additionally, they are also valuable for tumor immunotherapy. Thus, studies on the functions and biosynthetic pathways of rhamnose-containing compounds are in progress. In this paper, studies on the biosynthetic pathways of three rhamnose donors, i.e., deoxythymidinediphosphate-L-rhamnose (dTDP-Rha), uridine diphosphate-rhamnose (UDP-Rha), and guanosine diphosphate rhamnose (GDP-Rha), are firstly reviewed, together with the functions and crystal structures of those associated enzymes. Among them, dTDP-Rha is the most common rhamnose donor, and four enzymes, including glucose-1-phosphate thymidylyltransferase RmlA, dTDP-Glc-4,6-dehydratase RmlB, dTDP-4-keto-6-deoxy-Glc-3,5-epimerase RmlC, and dTDP-4-keto-Rha reductase RmlD, are involved in its biosynthesis. Secondly, several known rhamnosyltransferases from Geobacillus stearothermophilus, Saccharopolyspora spinosa, Mycobacterium tuberculosis, Pseudomonas aeruginosa, and Streptococcus pneumoniae are discussed. In these studies, however, the functions of rhamnosyltransferases were verified by employing gene knockout and radiolabeled substrates, which were almost impossible to obtain and characterize the products of enzymatic reactions. Finally, the application of rhamnose-containing compounds in disease treatments is briefly described.

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Sequential One‐Pot Three‐Enzyme Synthesis of the Tetrasaccharide Repeating Unit of Group B Streptococcus Serotype VIII Capsular Polysaccharide

Cited by 3 publications

References 33 publications

Biochemical Characterization and Synthetic Application of WciN and Its Mutants From Streptococcus pneumoniae Serotype 6B

Biochemical Characterization and Synthetic Application of WciN and Its Mutants From Streptococcus pneumoniae Serotype 6B

Chemical Synthesis of an Octasaccharide Derivative Related to Group B StreptococcusCell‐Wall Polysaccharide

Rhamnose-Containing Compounds: Biosynthesis and Applications

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