Recovery of low-molecular-weight γ-PGA by metal cation from the fermentation broth

Yuan, Zhenhong; Ran, Qiong; Chang, Zhongyi; Gao, Hongliang; Chen, Jia

doi:10.1016/j.procbio.2019.04.001

Cited by 6 publications

(5 citation statements)

References 23 publications

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“…45 Given the favourable adsorption of metal ions at pH 6 for the PA-g-PGA layer, a drop in pH to an acidic value is advantageous for the desorption of metal ion. 33,34,40 Harnessing this characteristic, we can achieve remarkable repeatability in detecting Ni 2+ by utilizing PA-g-PGA-modied asymmetric nanochannels. Fig.…”

Section: Resultsmentioning

confidence: 98%

“…Consequently, the modied PA-g-PGA layer can adsorb heavy metal ions at pH 6 and desorb at pH 3. 33,34,40,41 Fig. 2a depicts a schematic diagram of the modied PA-g-PGA layer adsorbing metal ions in nanochannels at pH 6.…”

Section: Resultsmentioning

confidence: 99%

“…Notably, the oxygen atom of the free carboxyl group and the nitrogen atom of the amide group of g-PGA can form coordination bonds with several metal ions, leading to the formation of stable complexes at pH = 6. 33,34 This unique property enables the constructed PA-g-PGA-modied multinanochannels to facilitate label-free and ultra-trace detection of metal ions in water. The experimental results underscore the capability of these modied nanochannels to achieve specic recognition and detection of nickel ions, and the detection limit can reach 1 × 10 −8 M. Consequently, we are condent that this system holds the potential for rapid and efficient recognition and detection of Ni 2+ , offering key advantages such as userfriendly operation, swi response, high sensitivity, portability, and miniaturization.…”

Section: Introductionmentioning

confidence: 99%

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Layer-by-layer assembly of homopolypeptide polyelectrolytes on asymmetric nanochannels for the detection of nickel ions

An,

Yang,

Wang

et al. 2024

Anal. Methods

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Section: Resultsmentioning

confidence: 98%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Layer-by-layer assembly of homopolypeptide polyelectrolytes on asymmetric nanochannels for the detection of nickel ions

An,

Yang,

Wang

et al. 2024

Anal. Methods

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“…B. subtilis strains were also grown in similar conditions, except without the addition of IPTG and antibiotic. The PGA produced in the supernatant was either measured directly or purified by a modified version of the copper precipitation method described by ( Manocha and Margaritis, 2010 ; Yuan et al., 2019 ). In brief, 0.5 M CuSO 4 was added to the supernatant and the solution was mixed by inversion and let stand for 1 h at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Characterization of a membrane enzymatic complex for heterologous production of poly-γ-glutamate in E. coli

Nascimento

Nair

2020

Metabolic Engineering Communications

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Poly-γ-glutamic acid (PGA) produced by many Bacillus species is a polymer with many distinct and desirable characteristics. However, the multi-subunit enzymatic complex responsible for its synthesis, PGA Synthetase (PGS), has not been well characterized yet, in native nor in recombinant contexts. Elucidating structural and functional properties are crucial for future engineering efforts aimed at altering the catalytic properties of this enzyme. This study focuses on expressing the enzyme heterologously in the Escherichia coli membrane and characterizing localization, orientation, and activity of this heterooligomeric enzyme complex. In E. coli , we were able to produce high molecular weight PGA polymers with minimal degradation at titers of approximately 13 mg/L in deep-well microtiter batch cultures. Using fusion proteins, we observed, for the first time, the association and orientation of the different subunits with the inner cell membrane. These results provide fundamental structural information on this poorly studied enzyme complex and will aid future fundamental studies and engineering efforts.

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“…Cultures were incubated at 30 ºC, 250 rpm, for 24 h. B. subtilis strains were also grown in similar conditions, except without the addition of IPTG and antibiotic. The PGA produced in the supernatant was either measured directly or purified by a modified version of the copper precipitation method described by (Manocha and Margaritis, 2010;Yuan et al, 2019). In brief, 0.5 M CuSO 4 was added to the supernatant and the solution was mixed by inversion and let stand for 1 h at room temperature.…”

Section: Pga Productionmentioning

confidence: 99%

Characterization of a membrane enzymatic complex for heterologous production of poly-γ-glutamate inE. coli

Nascimento

Nair

2020

Preprint

View full text Add to dashboard Cite

Poly-γ-glutamic acid (PGA) produced by many Bacillus species is a polymer with many distinct and desirable characteristics. However, the multi-subunit enzymatic complex responsible for its synthesis, PGA Synthetase (PGS), has not been well characterized yet, in native nor in recombinant contexts. Elucidating structural and functional properties are crucial for future engineering efforts aimed at altering the catalytic properties of this enzyme. This study focuses on expressing the enzyme heterologously in the Escherichia coli membrane and characterizing localization, orientation, and activity of this heterooligomeric enzyme complex. In E. coli, we were able to produce high molecular weight PGA polymers with minimal degradation at titers of approximately 13 mg/L in deep-well microtiter batch cultures. Using fusion proteins, we observed, for the first time, the association and orientation of the different subunits with the inner cell membrane. These results elucidate provide fundamental structural information on this poorly studied enzyme complex and will aid future fundamental studies and engineering efforts.

show abstract

Recovery of low-molecular-weight γ-PGA by metal cation from the fermentation broth

Cited by 6 publications

References 23 publications

Layer-by-layer assembly of homopolypeptide polyelectrolytes on asymmetric nanochannels for the detection of nickel ions

Layer-by-layer assembly of homopolypeptide polyelectrolytes on asymmetric nanochannels for the detection of nickel ions

Characterization of a membrane enzymatic complex for heterologous production of poly-γ-glutamate in E. coli

Characterization of a membrane enzymatic complex for heterologous production of poly-γ-glutamate inE. coli

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