Optimized soluble expression of a novel endoglucanase from Burkholderia pyrrocinia in Escherichia coli

Hu, Xiaoqing; Fan, Guangsen; Liao, Hui; Fu, Zhaohui; Ni, Hui

doi:10.1007/s13205-020-02327-w

Cited by 3 publications

(5 citation statements)

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“…During Daqu production, the raw materials are colonized by networks of microorganisms from the environment. Thus, Daqu used is a natural microbial library of bacteria, fungi, and yeasts that play important roles during the Baijiu fermentation (Hu et al, 2020). Because Daqu is produced using a range of different preparation methods and in different environments, the microbial composition of Daqu produced in different places can be quite different (Zuo et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

Isolation and Identification of a High-Yield Ethyl Caproate-Producing Yeast From Daqu and Optimization of Its Fermentation

Fan

Liu

Xu³

et al. 2021

Front. Microbiol.

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Baijiu is an important fermented product in China. A yeast named YX3307 that is capable of producing a large amount of ethyl caproate (EC) was isolated from Daqu, a crude fermentation starter for Baijiu. This yeast was identified as Clavispora lusitaniae on the basis of its morphological properties, physiological and biochemical characteristics, and 26S rDNA sequence. Single-factor experiments were conducted to obtain the optimum fermentation conditions for EC production by YX3307. The highest EC yield (62.0 mg/L) from YX3307 was obtained with the following culture conditions: inoculum size 7.5% (v/v), seed cell age 30 h, sorghum hydrolysate medium (SHM) with a sugar content of 10 Brix and an initial pH of 6.0; incubation at 28°C with shaking at 180 rpm for 32 h; addition of 10% (v/v) anhydrous ethanol and 0.04% (v/v) caproic acid at 32 and 40 h, respectively, static culture at 20°C until 72 h. YX3307 synthesized more EC than ethyl acetate, ethyl lactate, ethyl butyrate, and ethyl octanoate. An intracellular enzyme or cell membrane enzyme was responsible for EC synthesis. YX3307 can produce many flavor compounds that are important for high-quality Baijiu. Thus, it has potential applications in improving the flavor and quality of Baijiu.

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Section: Introductionmentioning

confidence: 99%

Isolation and Identification of a High-Yield Ethyl Caproate-Producing Yeast From Daqu and Optimization of Its Fermentation

Fan

Liu

Xu³

et al. 2021

Front. Microbiol.

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“…The Escherichia coli Rosetta-gami B (DE3) producer-strain was designed with many modifications to improve the activity and solubility of recombinant proteins, including enhancement of correct disulfide bridge formation, through mutations to improve glutathione reductase and thioredoxin reductase [ 20 ]. Unlike the commonly used pET series plasmids, pCold was designed as a cold-shock vector that includes a translation-enhancing element, a highly efficient Shine–Dalgarno sequence (GAGG) and a trigger factor (TF) chaperone, which facilitates co-translational folding of newly expressed polypeptides and high-yield protein expression [ 21 ].…”

Section: Introductionmentioning

confidence: 99%

Enhanced Thermostability of D-Psicose 3-Epimerase from Clostridium bolteae through Rational Design and Engineering of New Disulfide Bridges

Zhao

Chen

Wang

et al. 2021

IJMS

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D-psicose 3-epimerase (DPEase) catalyzes the isomerization of D-fructose to D-psicose (aka D-allulose, a low-calorie sweetener), but its industrial application has been restricted by the poor thermostability of the naturally available enzymes. Computational rational design of disulfide bridges was used to select potential sites in the protein structure of DPEase from Clostridium bolteae to engineer new disulfide bridges. Three mutants were engineered successfully with new disulfide bridges in different locations, increasing their optimum catalytic temperature from 55 to 65 °C, greatly improving their thermal stability and extending their half-lives (t1/2) at 55 °C from 0.37 h to 4–4.5 h, thereby greatly enhancing their potential for industrial application. Molecular dynamics simulation and spatial configuration analysis revealed that introduction of a disulfide bridge modified the protein hydrogen–bond network, rigidified both the local and overall structures of the mutants and decreased the entropy of unfolded protein, thereby enhancing the thermostability of DPEase.

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“…The commercial cspA promoter-TF system directs the expression of a trigger-factor (TF), a major ribosome-associated chaperone of E. coli involved in protein folding. The fusion-associated protein, TF, increases the solubility of the target protein, thus facilitating the co-translational folding of the expressed protein [ 55 , 159 , 160 ]. Hu et al [ 55 ] contrasted the endoglucanase (EG) expression by the T7 expression system with that of the cspA expression system fused to a TF partner by cloning the BpEG01790 from Burkholderia pyrrocinia B1213.…”

Section: Cold Shock Promoters Used For the Expression Of Recombinant ...mentioning

confidence: 99%

“…The fusion-associated protein, TF, increases the solubility of the target protein, thus facilitating the co-translational folding of the expressed protein [ 55 , 159 , 160 ]. Hu et al [ 55 ] contrasted the endoglucanase (EG) expression by the T7 expression system with that of the cspA expression system fused to a TF partner by cloning the BpEG01790 from Burkholderia pyrrocinia B1213. Although BpEG01790 was cloned into the conventional pET28a(+) vector under the control of the T7 promoter, no EG expression was detected with the E. coli BL21 (DE3) host system.…”

Section: Cold Shock Promoters Used For the Expression Of Recombinant ...mentioning

confidence: 99%

“…Even so, the unique commercial genetic platform and its variants, available today to overexpress recombinant proteins under cold-shock conditions, have provided remarkable results in E. coli . That is, reduced inclusion bodies formation, the production of difficult-to-express proteins in a soluble and active form, and the expression of toxic proteins to cells [ 55 , 155 ]. Initial research about cold shock-directed expression platforms for recombinant protein production suggests a promising future for protein biotechnology and the production of proteins that have not been readily expressed so far.…”

Section: Perspectivesmentioning

confidence: 99%

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The potential of cold-shock promoters for the expression of recombinant proteins in microbes and mammalian cells

Bartolo-Aguilar

Chávez-Cabrera

Flores-Cotera

et al. 2022

Journal of Genetic Engineering and Biotechnology

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Background Low-temperature expression of recombinant proteins may be advantageous to support their proper folding and preserve bioactivity. The generation of expression vectors regulated under cold conditions can improve the expression of some target proteins that are difficult to express in different expression systems. Main body of the abstract The cspA encodes the major cold-shock protein from Escherichia coli (CspA). The promoter of cspA has been widely used to develop cold shock-inducible expression platforms in E. coli. Moreover, it is often necessary to employ expression systems other than bacteria, particularly when recombinant proteins require complex post-translational modifications. Currently, there are no commercial platforms available for expressing target genes by cold shock in eukaryotic cells. Consequently, genetic elements that respond to cold shock offer the possibility of developing novel cold-inducible expression platforms, particularly suitable for yeasts, and mammalian cells. Conclusions This review covers the importance of the cellular response to low temperatures and the prospective use of cold-sensitive promoters to direct the expression of recombinant proteins. This concept may contribute to renewing interest in applying white technologies to produce recombinant proteins that are difficult to express. Graphical Abstract

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Optimized soluble expression of a novel endoglucanase from Burkholderia pyrrocinia in Escherichia coli

Cited by 3 publications

References 58 publications

Isolation and Identification of a High-Yield Ethyl Caproate-Producing Yeast From Daqu and Optimization of Its Fermentation

Isolation and Identification of a High-Yield Ethyl Caproate-Producing Yeast From Daqu and Optimization of Its Fermentation

Enhanced Thermostability of D-Psicose 3-Epimerase from Clostridium bolteae through Rational Design and Engineering of New Disulfide Bridges

The potential of cold-shock promoters for the expression of recombinant proteins in microbes and mammalian cells

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