Protein aggregation is commonly observed in genetically engineered bacteria over-expressing foreign genes, in the context of protein production processes. Very often, recombinant polypeptides deposit as insoluble protein clusters named inclusion bodies, whose formation is driven by stereo-specific cross-molecular interactions between partially folded polypeptide chains. The formation of inclusion bodies has been historically considered as the main bottleneck in industrial production processes of proteins, since a wide diversity of protein species tend to aggregate in bacteria. As the formation of inclusion bodies can be eventually minimized but rarely prevented, aggregated polypeptides of industrial interest need to be refolded in vitro before use. However, the progressive understanding of the molecular and physiological mechanisms regulating aggregation has revealed that inclusion bodies contain significant amounts of biologically active protein species making them suitable for the straightforward use in different in vitro processes, as functional, particulate entities. Therefore, when formed by enzymes, inclusion bodies are catalytic particles ready for industrial use. As discussed here, the genetic background of the host bacteria and the protein production conditions can be adjusted to tune the biological and biophysical properties of bacterial inclusion bodies, to gain manipulability_x000D_ and to make them more biotechnologically friendly.

Inclusion Bodies Biosynthesis Proteases Chaperones E. coli

Prouty WF, Karnovsky MJ, Goldberg AL, J. Biol. Chem., 250, 1112 (1975)
Marston FA, Biochem. J., 240, 1 (1986)
Gasser B, Saloheimo M, Rinas U, Dragosits M, Rodriguez-Carmona E, Baumann K, Giuliani M, Parrilli E, Branduardi P, Lang C, Porro D, Ferrer P, Tutino ML, Mattanovich D, Villaverde A, Microb. Cell Fact., 7, 11 (2008)
Baneyx F, Mujacic M, Nat. Biotechnol., 22, 1399 (2004)
Carrio MM, Villaverde A, Febs Lett., 489, 29 (2001)
Carrio MM, Corchero JL, Villaverde A, FEMS Microbiol. Lett., 169, 9 (1998)
Rinas U, Bailey JE, Appl. Environ. Microbiol., 59, 561 (1993)
Rinas U, Boone TC, Bailey JE, J. Biotechnol., 28, 313 (1993)
Margreiter G, Messner P, Caldwell KD, Bayer K, J. Biotechnol., 138, 67 (2008)
Peternel S, Jevsevar S, Bele M, Gaberc-Porekar V, Menart V, Biotechnol. Appl. Biochem., 49, 239 (2008)
Bowden GA, Paredes AM, Georgiou G, Biotechnology (NY), 9, 725 (1991)
Carrio MM, Cubarsi R, Villaverde A, Febs Lett., 471, 7 (2000)
Garcia-Fruitos E, Rodriguez-Carmona E, Diez-Gil C, Ferraz RM, Vazquez E, Corchero JL, Cano-Sarabia M, Ratera I, Ventosa N, Vaciana J, Villaverde A, Adv. Mat., 21, 4249 (2009)
Arie JP, Miot M, Sassoon N, Betton JM, Mol. Microbiol., 62, 427 (2006)
Miot M, Betton JM, Microb. Cell Fact., 3, 4 (2004)
Hunke S, Betton JM, Mol. Microbiol., 50, 1579 (2003)
Villaverde A, Carrio MM, Biotechnol. Lett., 25(17), 1385 (2003)
Carrio MM, Villaverde A, J. Biotechnol., 96, 3 (2002)
Rinas U, Bailey JE, Appl. Microbiol. Biotechnol., 37, 609 (1992)
Carrio MM, Villaverde A, J. Bacteriol., 187, 3599 (2005)
Taylor G, Hoare M, Gray DR, Marston FAO, Biotechnology, 4, 553 (1986)
Carrio M, Gonzalez-Montalban N, Vera A, Villaverde A, Ventura S, J. Mol. Biol., 347, 1025 (2005)
de Groot NS, Sabate R, Ventura S, Trends Biochem. Sci., 34, 408 (2009)
Gonzalez-Montalban N, Villaverde A, Aris A, Biochem. Biophys. Res. Commun., 355(3), 637 (2007)
Sabate R, Espargaro A, Saupe SJ, Ventura S, Microb. Cell Fact., 8, 56 (2009)
Wang L, Maji SK, Sawaya MR, Eisenberg D, Riek R, PLoS Biol., 6, e195 (2008)
Ami D, Natalello A, Taylor G, Tonon G, Maria DS, Biochim. Biophys. Acta, 1764, 793 (2006)
Doglia SM, Ami D, Natalello A, Gatti-Lafranconi P, Lotti M, Biotechnol. J., 3, 193 (2008)
Gonzalez-Montalban N, Natalello A, Garcia-Fruitos E, Villaverde A, Doglia SM, Biotechnol. Bioeng., 100(4), 797 (2008)
Ami D, Natalello A, Gatti-Lafranconi P, Lotti M, Doglia SM, Febs Lett., 579, 3433 (2005)
Wasmer C, Benkemoun L, Sabate R, Steinmetz MO, Coulary-Salin B, Wang L, Riek R, Saupe SJ, Meier BH, Angew. Chem. Int. Ed. Engl., 48, 4858 (2009)
Lewandowska A, Matuszewska M, Liberek K, J. Mol. Biol., 371, 800 (2007)
Morell M, Bravo R, Espargaro A, Sisquella X, Aviles FX, Fernandez-Busquets X, Ventura S, Biochim. Biophys. Acta, 1783, 1815 (2008)
Speed MA, Wang DI, King J, Nat. Biotechnol., 14, 1283 (1996)
Gonzalez-Montalban N, Garcia-Fruitos E, Ventura S, Aris A, Villaverde A, Microb. Cell. Fact., 5, 26 (2006)
Rinas U, Hoffmann F, Betiku E, Estape D, Marten S, J. Biotechnol., 127, 244 (2007)
Vera A, Aris A, Carrio M, Gonzalez-Montalban N, Villaverde A, J. Biotechnol., 119, 163 (2005)
Carrio MM, Villaverde A, Febs Lett., 537, 215 (2003)
Espargaro A, Sabate R, Ventura S, Febs Lett., 582, 3669 (2008)
Georgiou G, Valax P, Methods Enzymol., 309, 48 (1999)
Vallejo LF, Rinas U, Microb. Cell Fact., 3, 11 (2004)
Ferrer-Miralles N, Domingo-Espin J, Corchero JL, Vazquez E, Villaverde A, Microb. Cell Fact., 8, 17 (2009)
Peternel S, Grdadolnik J, Gaberc-Porekar V, Komel R, Microb. Cell Fact., 7, 34 (2008)
Jevsevar S, Gaberc-Porekar V, Fonda I, Podobnik B, Grdadolnik J, Menart V, Biotechnol. Prog., 21(2), 632 (2005)
Vera A, Gonzalez-Montalban N, Aris A, Villaverde A, Biotechnol. Bioeng., 96(6), 1101 (2007)
Oberg K, Chrunyk BA, Wetzel R, Fink AL, Biochemistry, 33, 2628 (1994)
Natalello A, Santarella R, Doglia SM, de Marco A, Protein Expr. Purif., 58(2), 356 (2008)
Ami D, Bonecchi L, Cali S, Orsini G, Tonon G, Doglia SM, Biochim. Biophys. Acta, 1624, 6 (2003)
Garcia-Fruitos E, Martinez-Alonso M, Gonzalez-Montalban N, Valli M, Mattanovich D, Villaverde A, J. Mol. Biol., 374, 195 (2007)
Ventura S, Villaverde A, Trends Biotechnol., 24, 179 (2006)
Garcia-Fruitos E, Gonzalez-Montalban N, Morell M, Vera A, Ferraz RM, Aris A, Ventura S, Villaverde A, Microb. Cell Fact., 4, 27 (2005)
Garcia-Fruitos E, Aris A, Villaverde A, Appl. Environ. Microbiol., 73, 289 (2007)
Nahalka J, J. Ind. Microbiol. Biotechnol., 35, 219 (2008)
Nahalka J, Gemeiner P, Bucko M, Wang PG, Artif. Cells Blood Substit. Immobil. Biotechnol., 34, 515 (2006)
Nahalka J, Nidetzky B, Biotechnol. Bioeng., 97(3), 454 (2007)
Nahalka J, Patoprsty V, Org. Biomol. Chem., 7, 1778 (2009)
Nahalka J, Vikartovska A, Hrabarova E, J. Biotechnol., 134, 146 (2008)
Nahalka J, Mislovicova D, Kavcova H, Mol. Biosyst., 5, 819 (2009)
Thomas JG, Baneyx F, J. Bacteriol., 180, 5165 (1998)
Han MJ, Lee JW, Lee SY, J. Proteome Res., 4, 2429 (2005)
Mogk A, Deuerling E, Vorderwulbecke S, Vierling E, Bukau B, Mol. Microbiol., 50, 585 (2003)
Gonzalez-Montalban N, Garcia-Fruitos E, Villaverde A, Nat. Biotechnol., 25, 718 (2007)
de Marco A, Deuerling E, Mogk A, Tomoyasu T, Bukau B, BMC Biotechnol., 7, 32 (2007)
de Marco A, Schroedel A, BMC Biochem., 6, 10 (2005)
Martinez-Alonso M, Gonzalez-Montalban N, Garcia-Fruitos E, Villaverde A, Appl. Environ. Microbiol., 101, 1353 (2008)
Martinez-Alonso M, Gonzalez-Montalban N, Garcia-Fruitos E, Villaverde A, Microb. Cell Fact., 8, 4 (2009)
Martinez-Alonso M, Vera A, Villaverde A, FEMS Microbiol. Lett., 273, 187 (2007)
Martinez-Alonso M, Garcia-Fruitos E, Villaverde A, Biotechnol. Bioeng., 101(6), 1353 (2008)
Martinez-Alonso M, Toledo-Rubio V, Noad R, Unzueta U, Ferrer-Miralles N, Roy P, Villaverde A, Appl. Environ. Microbiol., 75, 7850 (2009)
Martinez-Alonso M, Gomez-Sebastian S, Escribano JM, Saiz JC, Ferrer-Miralles N, Villaverde A, Appl. Microbiol. Biotechnol., Oct. 30, [Epub ahead of print] (2009)
Roessl U, Nahalka J, Nidetzky B, Biotechnol. Lett., Nov. 27, [Epub ahead of print] (2009)
de Marco A, Sevastsyanovich YR, Cole JA, N. Biotechnol., 25, 170 (2009)
de Marco A, Microb. Cell Fact., 7, 20 (2008)
Weibezahn J, Schlieker C, Tessarz P, Mogk A, Bukau B, Biol. Chem., 386, 739 (2005)
Mogk A, Bukau B, Curr. Biol., 14, R78 (2004)
Weibezahn J, Bukau B, Mogk A, Microb. Cell Fact., 3, 1 (2004)
Schlieker C, Tews I, Bukau B, Mogk A, Febs Lett., 578, 351 (2004)