Biogenesis and secretion of overproduced protein in recombinant strains of Escherichia coli

Badyakina, A. O.; Nesmeyanova, M. A.

doi:10.1016/j.procbio.2004.02.012

Cited by 7 publications

(5 citation statements)

References 52 publications

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“…Detailed knowledge of E. coli at genetic and metabolic levels has been exploited by the recombinant DNA technology for an economic production of a wide variety of biochemicals. However, the heterologous proteins produced by this organism are not typically excreted due to the presence of the cell envelope that consists of a cytoplasmic membrane and an outer cell wall (1). The outer cell wall forms a barrier to the periplasmic proteins.…”

Section: Introductionmentioning

confidence: 99%

Study of Physical and Biological Factors Involved in the Disruption of E. coli by Hydrodynamic Cavitation

Balasundaram

Harrison

2006

Biotechnol. Prog.

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Hydrodynamic cavitation results in flow restriction in a flow system causing rapid pressure fluctuations and significant fluid forces. These can be harnessed to mediate microbial cell damage. Hydrodynamic cavitation was studied for the partial disruption of E. coli and selective release of specific proteins relative to the total soluble protein. The effects of the cavitation number, the number of passes, and the specific growth rate of E. coli on the release of periplasmic and cytoplasmic proteins were studied. At the optimum cavitation number of 0.17 for this experimental configuration, 48% of the total soluble protein, 88% of acid phosphatase, and 67% of beta-galactosidase were released by hydrodynamic cavitation in comparison with the maximum release attained using multiple passes through the French Press. The higher release of the acid phosphatase over the total soluble protein suggested preferred release of periplasmic compounds. This was supported by SDS-PAGE analysis. The absence of micronization of cell material resulting in the potential for ease of solid-liquid separation downstream of the cell disruption operation was confirmed by TEM microscopy. E. coli cells cultivated at a higher specific growth rate (0.36 h(-1)) were more easily disrupted than slower grown cells (0.11 h(-1)). The specific activity of the enzyme of interest released by hydrodynamic cavitation, defined as the units of enzyme in solution per milligram of total soluble protein, was greater than that obtained on release by the French Press, high-pressure homogenization, osmotic shock, and EDTA treatment. The selectivity offered indicates the potential of enzyme release by hydrodynamic cavitation to ease the purification in the subsequent downstream processing.

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Section: Introductionmentioning

confidence: 99%

Study of Physical and Biological Factors Involved in the Disruption of E. coli by Hydrodynamic Cavitation

Balasundaram

Harrison

2006

Biotechnol. Prog.

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“…The polyextremophilic bacterium, Bacillus halodurans TSEV1 was isolated from the effluent sample collected from the Century Pulp and Paper Industry, Lalkuan (Uttarakhand State, India) [12]. The genomic DNA of B. halodurans TSEV1 was used as the source of xylanase gene.…”

Section: Strains and Plasmids For Dna Manipulationsmentioning

confidence: 99%

“…The recovery of substantial yields of correctly folded proteins is, however, a problem. Most of the proteins synthesized in the cytoplasm of E. coli are not secreted into the culture medium because the cell envelope of this gram-negative bacterium has both cytoplasmic and outer membranes, which is a barrier to permeability for cytoplasmic proteins [10,12]. One approach to resolve these problems is to have recombinant proteins secreted into culture medium by membrane permeabilization.…”

Section: Introductionmentioning

confidence: 99%

“…Some strategies for the permeabilization of the outer membrane of E. coli have been developed to enhance the secretion of periplasmic proteins into the medium. These approaches mainly include the use of E. coli mutants with a defect in the cell envelope components (lky-and exc-mutants) [13,14] and the addition of chemicals such as glycine and Triton X-100 [12,15]. Most of these approaches are, however, non-selective, and thus, result in the release of other periplasmic proteins into the medium along with the protein of interest.…”

Section: Introductionmentioning

confidence: 99%

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Secretion of recombinant thermo-alkali-stable endoxylanase of polyextremophilic Bacillus halodurans TSEV1 and its utility in generating xylooligosaccharides from renewable agro-residues

Kumar

Satyanarayana

2014

Process Biochemistry

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“…changing of pH-optimum, temperature stability, substrate spectrum, etc.) through modification of primary and secondary protein structure 25 . cytochrome P450 aromatics... Table 3.…”

Section: Recombinant Dna-technology and Protein-engineeringmentioning

confidence: 99%

Towards a ‘tailor made’ biocatalysis

Cerdobbel¹,

Vandamme²,

Soetaert³

2008

Microbiol. Aust.

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Microbial enzymes are used today in the most diverse industrial sectors. However, it is only in recent years that fundamental knowledge has been acquired that allows for a rational ? rather than an empirical ? development and application of microbial enzyme processes. These enzymes are primarily used as an end product or a processing aid in the agricultural sector, food and feed industry, the detergent sector, the textile industry and health care sector. In these sectors they replace or adjust the familiar (bio)chemical processes, or plant or animal enzymes. But they can also perform totally new ?chemical? reactions, with the principal aim of ?improving? and/or modifying the ?substrate?.

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Biogenesis and secretion of overproduced protein in recombinant strains of Escherichia coli

Cited by 7 publications

References 52 publications

Study of Physical and Biological Factors Involved in the Disruption of E. coli by Hydrodynamic Cavitation

Study of Physical and Biological Factors Involved in the Disruption of E. coli by Hydrodynamic Cavitation

Secretion of recombinant thermo-alkali-stable endoxylanase of polyextremophilic Bacillus halodurans TSEV1 and its utility in generating xylooligosaccharides from renewable agro-residues

Towards a ‘tailor made’ biocatalysis

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