scholarly journals Single amino acid substitutions on the surface of Escherichia coli maltose-binding protein can have a profound impact on the solubility of fusion proteins

2001 ◽  
Vol 10 (3) ◽  
pp. 622-630 ◽  
Author(s):  
J. D. Fox
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Fusion Proteins ◽  
Binding Protein ◽  
Maltose Binding Protein ◽  
Single Amino Acid ◽  
Amino Acid Substitutions ◽  
Profound Impact

Structural and functional consequences of single amino acid substitutions in the pyrimidine base binding pocket of Escherichia coli CMP kinase

FEBS Journal ◽  
2007 ◽  
Vol 274 (13) ◽  
pp. 3363-3373 ◽  
Author(s):  
Augustin Ofiteru ◽  
Nadia Bucurenci ◽  
Emil Alexov ◽  
Thomas Bertrand ◽  
Pierre Briozzo ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Binding Pocket ◽  
Single Amino Acid ◽  
Amino Acid Substitutions ◽  
Pyrimidine Base ◽  
Functional Consequences

scholarly journals Active Expression of Membrane-Bound L-Amino Acid Deaminase from Proteus mirabilis in Recombinant Escherichia coli by Fusion with Maltose-Binding Protein for Enhanced Catalytic Performance

Catalysts ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 215
Author(s):  
Dan-Ping Zhang ◽  
Xiao-Ran Jing ◽  
An-Wen Fan ◽  
Huan Liu ◽  
Yao Nie ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Proteus Mirabilis ◽  
Binding Protein ◽  
Catalytic Efficiency ◽  
Maltose Binding Protein ◽  
Catalytic Performance ◽  
Cell Lysate ◽  
Active Expression ◽  
Membrane Bound

L-amino acid deaminases (LAADs) are membrane flavoenzymes that catalyze the deamination of neutral and aromatic L-amino acids to α-keto acids and ammonia. LAADs can be used to develop many important biotechnological applications. However, the transmembrane α-helix of LAADs restricts its soluble active expression and purification from a heterologous host, such as Escherichia coli. Herein, through fusion with the maltose-binding protein (MBP) tag, the recombinant E. coli BL21 (DE3)/pET-21b-MBP-PmLAAD was constructed and the LAAD from Proteus mirabilis (PmLAAD) was actively expressed as a soluble protein. After purification, the purified MBP-PmLAAD was obtained. Then, the catalytic activity of the MBP-PmLAAD fusion protein was determined and compared with the non-fused PmLAAD. After fusion with the MBP-tag, the catalytic efficiency of the MBP-PmLAAD cell lysate was much higher than that of the membrane-bound PmLAAD whole cells. The soluble MBP-PmLAAD cell lysate catalyzed the conversion of 100 mM L-phenylalanine (L-Phe) to phenylpyruvic acid (PPA) with a 100% yield in 6 h. Therefore, the fusion of the MBP-tag not only improved the soluble expression of the PmLAAD membrane-bound protein, but also increased its catalytic performance.

scholarly journals NDM-4 Metallo-β-Lactamase with Increased Carbapenemase Activity from Escherichia coli

2012 ◽  
Vol 56 (4) ◽  
pp. 2184-2186 ◽  
Author(s):  
Patrice Nordmann ◽  
Anne E. Boulanger ◽  
Laurent Poirel
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Amino Acid Substitution ◽  
Active Sites ◽  
Hydrolytic Activity ◽  
Single Amino Acid Substitution ◽  
Single Amino Acid ◽  
Amino Acid Substitutions ◽  
Content Type

ABSTRACTA clinicalEscherichia coliisolate resistant to all β-lactams, including carbapenems, expressed a novel metallo-β-lactamase (MBL), NDM-4, differing from NDM-1 by a single amino acid substitution (Met154Leu). NDM-4 possessed increased hydrolytic activity toward carbapenems and several cephalosporins compared to that of NDM-1. This amino acid substitution was not located in the known active sites of NDM-1, indicating that remote amino acid substitutions might also play a role in the extended activity of this MBL.

Low plasma concentrations of retinol-binding protein in individuals with mutations affecting position 84 of the transthyretin molecule

1995 ◽  
Vol 41 (9) ◽  
pp. 1288-1291 ◽  
Author(s):  
R P Waits ◽  
T Yamada ◽  
T Uemichi ◽  
M D Benson
Keyword(s):  
Amino Acid ◽  
Vitamin A ◽  
Binding Protein ◽  
Plasma Concentrations ◽  
Substantial Reduction ◽  
Point Mutations ◽  
Gene Mutations ◽  
Retinol Binding Protein ◽  
Single Amino Acid ◽  
Amino Acid Substitutions

Abstract Retinol-binding protein (RBP), the principal carrier for vitamin A, is known to form a complex with transthyretin (TTR) for transport in plasma. Individuals from a kindred with the amino acid substitution of serine for isoleucine at position 84 (Ser84) of the TTR molecule show substantial reduction in plasma concentrations of RBP. In the present study, we measured plasma RBP in individuals from several kindreds, demonstrating 17 different point mutations within the TTR gene. In each case, these mutations caused single amino acid substitutions at various positions throughout the TTR molecule. Of all the individuals examined, only those with mutations causing amino acid substitutions at position 84 of the TTR molecule (Ser84 and Asn84) demonstrated substantial decreases in plasma concentrations of RBP. These results suggest that the isoleucine at position 84 on the TTR molecule may be critically involved in mediating RBP binding. Further, these findings demonstrate the importance of considering TTR gene mutations when clinically evaluating patients with low RBP.

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