Engineering Glucosamine-6-Phosphate Synthase to Achieve Efficient One-Step Biosynthesis of Glucosamine.
ACS Chem Biol
; 19(6): 1237-1242, 2024 Jun 21.
Article
in En
| MEDLINE
| ID: mdl-38723147
ABSTRACT
As an important functional monosaccharide, glucosamine (GlcN) is widely used in fields such as medicine, food nutrition, and health care. Here, we report a distinct GlcN biosynthesis method that utilizes engineered Bacillus subtilis glucosamine-6-phosphate synthase (BsGlmS) to convert D-fructose to directly generate GlcN. The best variant obtained by using a combinatorial active-site saturation test/iterative saturation mutagenesis (CAST/ISM) strategy was a quadruple mutant S596D/V597G/S347H/G299Q (BsGlmS-BK19), which has a catalytic activity 1736-fold that of the wild type toward D-fructose. Upon using mutant BK19 as a whole-cell catalyst, D-fructose was converted into GlcN with 65.32% conversion in 6 h, whereas the wild type only attained a conversion rate of 0.31% under the same conditions. Molecular docking and molecular dynamics simulations were implemented to provide insights into the mechanism underlying the enhanced activity of BK19. Importantly, the BsGlmS-BK19 variant specifically catalyzes D-fructose without the need for phosphorylated substrates, representing a significant advancement in GlcN biosynthesis.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Bacillus subtilis
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Protein Engineering
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Glucosamine
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Glutamine-Fructose-6-Phosphate Transaminase (Isomerizing)
Language:
En
Journal:
ACS Chem Biol
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ACS chem. biol. (Online)
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ACS chemical biology (Online)
Year:
2024
Document type:
Article
Country of publication: