[21] Thioredoxin as a fusion partner for production of soluble recombinant proteins in Escherichia coli

LaVallie, Edward R.; Lu, Zhijian; DiBlasio-Smith, Elizabeth A.; Collins-Racie, Lisa A.; McCoy, John

doi:10.1016/s0076-6879(00)26063-1

Cited by 159 publications

(87 citation statements)

References 23 publications

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“…One of the E. coli thioredoxins, TrxA, is an 11.6 kDa protein that demonstrates high solubility in the E. coli cytoplasm and inherent thermal stability, which may be conferred to TrxA fusion proteins. For example, TrxA has been used as an N-or C-terminal fusion protein [38,39] to increase soluble protein expression of recombinant proteins [40]. In addition to enhancing the solubility of fusion proteins, TrxA has been shown to aid in the crystallization of proteins.…”

Section: Thioredoxin Amentioning

confidence: 99%

Recombinant protein expression and purification: A comprehensive review of affinity tags and microbial applications

Young

Britton

Robinson

2012

Biotechnology Journal

410

267

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Protein fusion tags are indispensible tools used to improve recombinant protein expression yields, enable protein purification, and accelerate the characterization of protein structure and function. Solubility-enhancing tags, genetically engineered epitopes, and recombinant endoproteases have resulted in a versatile array of combinatorial elements that facilitate protein detection and purification in microbial hosts. In this comprehensive review, we evaluate the most frequently used solubility-enhancing and affinity tags. Furthermore, we provide summaries of well-characterized purification strategies that have been used to increase product yields and have widespread application in many areas of biotechnology including drug discovery, therapeutics, and pharmacology. This review serves as an excellent literature reference for those working on protein fusion tags.

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Section: Thioredoxin Amentioning

confidence: 99%

Recombinant protein expression and purification: A comprehensive review of affinity tags and microbial applications

Young

Britton

Robinson

2012

Biotechnology Journal

410

267

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“…Although EngE and ExgS could be expressed as soluble proteins by the pET-22b vector, EngH was expressed as an inclusion body by this vector (data not shown). To express EngH as a soluble protein, EngH was expressed as a fused protein with thioredoxin by using the pBAD/Thio vector (Invitrogen), since fusion with thioredoxin is known to help solubilize expressed proteins (23). As a result, EngH was also expressed as a soluble protein.…”

Section: Preparation Of Recombinant Cellulosomal Subunitsmentioning

confidence: 99%

Synergistic Effects on Crystalline Cellulose Degradation between Cellulosomal Cellulases fromClostridium cellulovorans

2002

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Clostridium cellulovorans produces a multienzyme cellulose-degrading complex called the cellulosome. In this study, we determined the synergistic effects on crystalline cellulose degradation by three different recombinant cellulosomes containing either endoglucanase EngE, endoglucanase EngH, or exoglucanase ExgS bound to mini-CbpA, a part of scaffolding protein CbpA. EngE, EngH, and ExgS are classified into the glycosyl hydrolase families 5, 9, and 48, respectively. The assembly of ExgS and EngH with mini-CbpA increased the activity against insoluble cellulose 1.5-to 3-fold, although no effects on activity against soluble cellulose were observed. These results indicated that mini-CbpA could help cellulase components degrade insoluble cellulose but not soluble cellulose. The mixture of the cellulosomes containing ExgS and EngH showed higher activity and synergy degrees than the other cellulosome mixtures, indicating the synergistic effect between EngH and ExgS was the most dominant effect among the three mixtures for crystalline cellulose degradation. Reactions were also performed by adding different cellulosomes in a sequential manner. When ExgS was used for the initial reaction followed by EngE and EngH, almost no synergistic effect was observed. On the other hand, when EngE or EngH was used for the first reaction followed by ExgS, synergistic effects were observed. These results indicated that the initial reactions by EngH and/or EngE promoted cellulose degradation by ExgS.

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“…The essential requirement in this case is the process of synthesizing membrane proteins together with natural or synthetic lipids, as well as detergents, that can help solubilize the membrane's protein content. CF type of aquaporin production has been illustrated at analytical levels [31,[56][57][58], and recent tests showed high expression of properly folded AqpZ. Furthermore, plant aquaporin has been achieved with E. coli CF protocols and implementing various fusion vectors [59,60].…”

Section: Production Of Aquaporinsmentioning

confidence: 99%

Aquaporin Biomimetic Membranes

Abdelrasoul¹,

Doan²,

Lohi³

2017

Biomimetic and Bioinspired Membranes for New Frontiers in Sustainable Water Treatment Technology

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Recent research looked at an array of aquaporin protein structures, their unique functions, and potential applications in the research and industrial sectors. This chapter focuses on the specific functional features of aquaporin biomimetic membranes to interrogate their permeability properties in relation to various biomimetic water-transporting membranes. This chapter discusses in detail functional characteristics of aquaporin, how to produce it, and the status of aquaporin development.

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[21] Thioredoxin as a fusion partner for production of soluble recombinant proteins in Escherichia coli

Cited by 159 publications

References 23 publications

Recombinant protein expression and purification: A comprehensive review of affinity tags and microbial applications

Recombinant protein expression and purification: A comprehensive review of affinity tags and microbial applications

Synergistic Effects on Crystalline Cellulose Degradation between Cellulosomal Cellulases fromClostridium cellulovorans

Aquaporin Biomimetic Membranes

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