Modification of quaternary structure of Candida albicans GlcN-6-P synthase and its desensitization to inhibition by UDP-GlcNAc by site-directed mutagenesis

Kwiatkowska-Semrau, Karolina; Wojciechowski, Marek; Gabriel, Iwona; Crucho, Sara; Milewski, Sławomir

doi:10.1016/j.bbapap.2018.08.003

Cited by 6 publications

(3 citation statements)

References 26 publications

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“…UDP-GlcNAc is a common metabolic intermediate, which can be endogenously synthesized in E. coli for anabolism of chondroitin-like polysaccharides , and peptidoglycan . UDP-GlcNAc can be synthesized from Fru-6-P, the most basic sugar intermediate of the glycolytic pathway, using three consecutive biocatalysis processes by GlmS, GlmM, and GlmU.…”

Section: Resultsmentioning

confidence: 99%

“…11 UDP-GlcNAc is a common metabolic intermediate, which can be endogenously synthesized in E. coli for anabolism of chondroitin-like polysaccharides 11,23 and peptidoglycan. 24 UDP-GlcNAc can be synthesized from Fru-6-P, the most basic sugar intermediate of the glycolytic pathway, using three consecutive biocatalysis processes by GlmS, GlmM, and GlmU. Therefore, to construct an efficient pathway for LNT II production, LgtA was introduced and the UDP-GlcNAc synthesis pathway was strengthened to improve the precursor supply in the engineered E. coli (Figure 1).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

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Metabolic Engineering of Escherichia coli for Lacto-N-triose II Production with High Productivity

Zhu

Wan

Meng

et al. 2021

J. Agric. Food Chem.

111

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Lacto-N-triose II (LNT II), a core structural unit of human milk oligosaccharides (HMOs), has attracted substantial attention for its nutraceutical potentials and applications in the production of complex HMOs. In this study, Escherichia coli was metabolically engineered to efficiently produce LNT II using glycerol as a carbon source and lactose as a substrate. The UDP-Nacetylglucosamine (UDP-GlcNAc) biosynthesis pathway was strengthened, and β-1,3-N-acetylglucosaminyltransferase (LgtA) was introduced to construct an LNT II-producing base strain. To increase the titer and yield of LNT II, combinatorial optimization of the copy number and the ribosomal binding site sequence was performed to tune the gene expression strength and translation rates of the pathway enzymes. Next, multipoint mutations were introduced to glucosamine-6-phosphatesynthase (GlmS) to relieve the feedback inhibition. Then, a series of engineered strains were constructed by blocking the futile pathways by the deletion of the relevant genes. Finally, the culture conditions were optimized. LNT II titer was improved step-by-step from 0.53 to 5.52 g/L in shake-flask cultivations. Fed-batch culture of the final engineered strain produced 46.2 g/L of LNT II, with an LNT II productivity and content of 0.77 g/(L•h) and 0.95 g/g dry cell weight, respectively.

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Section: Resultsmentioning

confidence: 99%

Section: ■ Results and Discussionmentioning

confidence: 99%

Metabolic Engineering of Escherichia coli for Lacto-N-triose II Production with High Productivity

Zhu

Wan

Meng

et al. 2021

J. Agric. Food Chem.

111

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“…According to the method described by Kwiatkowska-Semrau et al [16], discontinuous urea SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gelelectrophoresis) electrophoresis was used to determine the molecular weight of endogenous LPL in yak milk. The separating gel concentration was 15%, and the stacking gel concentration was 5%.…”

Section: Lpl Molecular Weight Measurementmentioning

confidence: 99%

HS-SPME-GC-MS Combined with Orthogonal Partial Least Squares Identification to Analyze the Effect of LPL on Yak Milk’s Flavor under Different Storage Temperatures and Times

Zhang,

Zhong,

Wang

et al. 2024

Foods

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Flavor is a crucial parameter for assessing the sensory quality of yak milk. However, there is limited information regarding the factors influencing its taste. In this study, the effects of endogenous lipoprotein lipase (LPL) on the volatile flavor components of yak milk under storage conditions of 4 °C, 18 °C and 65 °C were analyzed via headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS) combined with orthogonal partial least-squares (OPSL) discrimination, and the reasons for the changes in yak milk flavors were investigated. Combined with the difference in the changes in volatile flavor substance before and after the action of LPL, LPL was found to have a significant effect on the flavor of fresh yak milk. Fresh milk was best kept at 4 °C for 24 h and pasteurized for more than 24 h. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were employed to characterize the volatile components in yak milk under various treatment conditions. Twelve substances with significant influence on yak milk flavor were identified by measuring their VIP values. Notably, 2-nonanone, heptanal, and ethyl caprylate exhibited OAV values greater than 1, indicating their significant contribution to the flavor of yak milk. Conversely, 4-octanone and 2-heptanone displayed OAV values between 0.1 and 1, showing their important role in modifying the flavor of yak milk. These findings can serve as monitoring indicators for assessing the freshness of yak milk.

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Phosphate solubilization and plant growth properties are promoted by a lactic acid bacterium in calcareous soil

Li,

Chen,

Wang

et al. 2023

Appl Microbiol Biotechnol

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Modification of quaternary structure of Candida albicans GlcN-6-P synthase and its desensitization to inhibition by UDP-GlcNAc by site-directed mutagenesis

Cited by 6 publications

References 26 publications

Metabolic Engineering of Escherichia coli for Lacto-N-triose II Production with High Productivity

Metabolic Engineering of Escherichia coli for Lacto-N-triose II Production with High Productivity

HS-SPME-GC-MS Combined with Orthogonal Partial Least Squares Identification to Analyze the Effect of LPL on Yak Milk’s Flavor under Different Storage Temperatures and Times

Phosphate solubilization and plant growth properties are promoted by a lactic acid bacterium in calcareous soil

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