Co-expression of chaperones from P. furiosus enhanced the soluble expression of the recombinant hyperthermophilic α-amylase in E. coli

Peng, Shuaiying; Chu, Zema; Lu, Junhui; Li, Dongxiao; Wang, Yonghong; Yang, Shengli; Zhang, Yi

doi:10.1007/s12192-016-0675-7

Cited by 30 publications

(18 citation statements)

References 40 publications

(35 reference statements)

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“…After solubilization, a total of 58,000 U/g wet cells yield was obtained. In another study, Peng et al ( 2016 ) co-expressed PFA with chaperones in E. coli . Both chaperonin and a small heat shock protein (sHSP) increased the solubility of PFA to a certain degree, while pre-folding seemed to be the most efficient that increased the enzyme activity to about 60,000 U g −1 wet weight over that of 5,000 U g −1 wet weight without chaperone.…”

Section: Cloning and Expression Of Acid-stable α-Amylase Encoding Genmentioning

confidence: 99%

An Insight Into Ameliorating Production, Catalytic Efficiency, Thermostability and Starch Saccharification of Acid-Stable α-Amylases From Acidophiles

Parashar

Satyanarayana

2018

Front. Bioeng. Biotechnol.

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Most of the extracellular enzymes of acidophilic bacteria and archaea are stable at acidic pH with a relatively high thermostability. There is, however, a dearth of information on their acid stability. Although several theories have been postulated, the adaptation of acidophilic proteins to low pH has not been explained convincingly. This review highlights recent developments in understanding the structure and biochemical characteristics, and production of acid-stable and calcium-independent α-amylases by acidophilic bacteria with special reference to that of Bacillus acidicola.

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Section: Cloning and Expression Of Acid-stable α-Amylase Encoding Genmentioning

confidence: 99%

An Insight Into Ameliorating Production, Catalytic Efficiency, Thermostability and Starch Saccharification of Acid-Stable α-Amylases From Acidophiles

Parashar

Satyanarayana

2018

Front. Bioeng. Biotechnol.

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“…As shown by Peng et. al., re‐hosting the P. furiosus chaperone systems to E. coli cells indeed increases the soluble expression of a recombinant α‐amylase from P. furiosus . In addition, cellular proteases are apparently incapable of degrading the highly stable aberrant LamA species, as we isolate these in considerable quantities from E. coli .…”

Section: Discussionmentioning

confidence: 95%

Chaotropic heat treatment resolves native‐like aggregation of a heterologously produced hyperthermostable laminarinase

Westphal

Geerke-Volmer

Mierlo

et al. 2017

Biotechnology Journal

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Production of hyperthermostable enzymes in mesophilic hosts frequently causes undesired aggregation of these proteins. During production of Pyrococcus furiosus endo-β-1,3 glucanase (LamA) in Escherichia coli, soluble and insoluble species form. Here, the authors address the composition of this mixture, including the nature of LamA conformers, and establish a method to increase the yield of native monomer. With gel electrophoresis, size-exclusion chromatography, light scattering, circular dichroism and enzyme kinetics the authors show that approximately 50 % of heterologously produced LamA is soluble, and that 40 % of this fraction constitutes native-like oligomers and non-native monomers. Soluble oligomers display, like native LamA monomer, substrate inhibition, although with poor activity. Treatment of soluble oligomers with 3 M guanidinium hydrochloride at 80 °C yields up to 75 % properly active monomer. Non-native monomer shows low specific activity without substrate inhibition. Incubating non-native monomer with 3 M guanidinium hydrochloride at 80 °C causes formation of 25 % native LamA. Also, a large amount of insoluble LamA aggregates can be converted into soluble native monomer by application of this procedure. Thus, chaotropic heat treatment can improve the yield and quality of hyperthermostable proteins that form aberrant species during production in E. coli.

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“…The sarcosine oxidase from Thermomicrobium roseum, which was codon-optimized, was functionally expressed in E. coli, and the co-expression of GroE and GroE with the HSP70 complex significantly increased the solubility of expressed sarcosine oxidase [76]. Another example is that the expression of α-amylase from the hyper-thermophilic archaeum Pyrococcus furiosus could be improved via co-expression of the autologous chaperones [77].…”

Section: Improving the Temperature Tolerancementioning

confidence: 99%

The Role of GroE Chaperonins in Developing Biocatalysts for Biofuel and Chemical Production

Xia¹,

Turner²,

Jayakody³

2016

Enz Eng

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As the need and interest for producing renewable biofuels and biochemical has grown, new avenues to improve product yields and productivity have been explored. Specifically, improving the tolerance of host microbes towards stressors, such as heat shock or the presence of harmful solvents, has been an especially important route to improve industrial-scale chemical production. In this review, we discuss recent advances in microbial engineering for renewable chemical production through the introduction and expression of chaperonins, especially the bacterial GroE complex. The GroE complex provides a closed-off environment and allows vital proteins to enter and engage in post-translational folding or refolding in a more-ideal environment, allowing the microbe to possess increased survival rates in low/high temperatures or in high concentrations of otherwise harmful end-products. Overall, we highlighted how chaperonin systems such as the GroE complex could have many industrially-relevant uses in the coming years.

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Co-expression of chaperones from P. furiosus enhanced the soluble expression of the recombinant hyperthermophilic α-amylase in E. coli

Cited by 30 publications

References 40 publications

An Insight Into Ameliorating Production, Catalytic Efficiency, Thermostability and Starch Saccharification of Acid-Stable α-Amylases From Acidophiles

An Insight Into Ameliorating Production, Catalytic Efficiency, Thermostability and Starch Saccharification of Acid-Stable α-Amylases From Acidophiles

Chaotropic heat treatment resolves native‐like aggregation of a heterologously produced hyperthermostable laminarinase

The Role of GroE Chaperonins in Developing Biocatalysts for Biofuel and Chemical Production

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