The NT11, a novel fusion tag for enhancing protein expression in Escherichia coli

Nguyen, Thi Khoa My; Ki, Mi Ran; Son, Ryeo Gang; Pack, Seung Pil

doi:10.1007/s00253-018-09595-w

Cited by 55 publications

(32 citation statements)

References 55 publications

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“…In the area of small peptide tags, Ojima‐Kato et al reported that the addition of the sequence coding for serine‐lysine‐isoleucine‐lysine (SKIK tag) after the initial N‐terminal methionine codon markedly improves the expression of recombinant proteins . Nguyen et al devised the NT‐11 tag, derived from the first 11 amino acid residues of a duplicated carbonic anhydrase from Dunaliella . The NT‐11 tag enhanced the soluble production of all tested proteins, which were also His‐tagged.…”

Section: Advances In Plasmid Designmentioning

confidence: 99%

New tools for recombinant protein production in Escherichia coli: A 5‐year update

2019

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The production of proteins in sufficient amounts is key for their study or use as biotherapeutic agents. Escherichia coli is the host of choice for recombinant protein production given its fast growth, easy manipulation, and cost-effectiveness. As such, its protein production capabilities are continuously being improved. Also, the associated tools (such as plasmids and cultivation conditions) are subject of ongoing research to optimize product yield. In this work, we review the latest advances in recombinant protein production in E. coli.

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Section: Advances In Plasmid Designmentioning

confidence: 99%

New tools for recombinant protein production in Escherichia coli: A 5‐year update

2019

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“…So the conversion rate from St to Reb A by TrxA-UGT76G1 showed at least 30% higher than that by UGT76G1 (pET22b-UGT76G1) with the same volume dosage of the crude extract enzymes. And previous research also concluded that fusion partners could enhance the soluble expression of heterologous enzymes in E. coli cells [ 21 , 23 , 24 ]. Thus, we selected other five fusion partners (Fh8, MBP, Smt3, DsbA and DsbC) to find a more suitable fusion partner for the soluble expression of UGT76G1 ( Figure 1 A).…”

Section: Resultsmentioning

confidence: 99%

Enhanced Heterologous Production of Glycosyltransferase UGT76G1 by Co-Expression of Endogenous prpD and malK in Escherichia coli and Its Transglycosylation Application in Production of Rebaudioside

Shu

Zheng

et al. 2020

IJMS

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Steviol glycosides (SGs) with zero calories and high-intensity sweetness are the best substitutes of sugar for the human diet. Uridine diphosphate dependent glycosyltransferase (UGT) UGT76G1, as a key enzyme for the biosynthesis of SGs with a low heterologous expression level, hinders its application. In this study, a suitable fusion partner, Smt3, was found to enhance the soluble expression of UGT76G1 by 60%. Additionally, a novel strategy to improve the expression of Smt3-UGT76G1 was performed, which co-expressed endogenous genes prpD and malK in Escherichia coli. Notably, this is the first report of constructing an efficient E. coli expression system by regulating prpD and malK expression, which remarkably improved the expression of Smt3-UGT76G1 by 200% as a consequence. Using the high-expression strain E. coli BL21 (DE3) M/P-3-S32U produced 1.97 g/L of Smt3-UGT76G1 with a yield rate of 61.6 mg/L/h by fed-batch fermentation in a 10 L fermenter. The final yield of rebadioside A (Reb A) and rebadioside M (Reb M) reached 4.8 g/L and 1.8 g/L, respectively, when catalyzed by Smt3-UGT76G1 in the practical UDP-glucose regeneration transformation system in vitro. This study not only carried out low-cost biotransformation of SGs but also provided a novel strategy for improving expression of heterologous proteins in E. coli.

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“…The strategy of using protein fusion partners has been effective for increasing expression of target proteins and inhibiting IB formation [67,68]. There are numerous solubility tags reported in the literature; the majority of recent work has focussed on a few tags, notably maltose-binding protein, N-utilization substance A, thioredoxin and GST [68].…”

Section: Expression Of Pgex-4t-1-bpfaementioning

confidence: 99%

Soluble expression of a novel feruloyl esterase from Burkholderia pyrrocinia B1213 in Escherichia coli and optimization of production conditions

Fan

Zhu

et al. 2020

Biotechnology & Biotechnological Equipment

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This report examines the soluble expression of a novel feruloyl esterase, BpFae, from Burkholderia pyrrocinia in Escherichia coli overexpression systems using three expression vectors, pET28a, pCold-TF and pGEX-4T-1. BpFae was overexpressed as insoluble inclusion bodies when pET28a was used as the expression vector. BpFae was overexpressed in the soluble form when using pCold-TF; however, the overexpressed protein showed negligible activity. Recombinant BpFae was produced in the soluble form and active when E. coli cells were transformed with the pGEX-4T-1-BpFae vector. Optimal conditions for soluble overexpression of recombinant BpFae were determined, and the highest activity of recombinant BpFae was 2.54 U mL À1 . Multiple sequence alignment, phylogenetic analysis and construction of a three-dimensional model indicated that BpFae is a unique FAE from B. pyrrocinia with underlying research value.

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The NT11, a novel fusion tag for enhancing protein expression in Escherichia coli

Cited by 55 publications

References 55 publications

New tools for recombinant protein production in Escherichia coli: A 5‐year update

New tools for recombinant protein production in Escherichia coli: A 5‐year update

Enhanced Heterologous Production of Glycosyltransferase UGT76G1 by Co-Expression of Endogenous prpD and malK in Escherichia coli and Its Transglycosylation Application in Production of Rebaudioside

Soluble expression of a novel feruloyl esterase from Burkholderia pyrrocinia B1213 in Escherichia coli and optimization of production conditions

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