Active protein aggregates induced by terminally attached self-assembling peptide ELK16 in Escherichia coli
Tóm tắt
In recent years, it has been gradually realized that bacterial inclusion bodies (IBs) could be biologically active. In particular, several proteins including green fluorescent protein, β-galactosidase, β-lactamase, alkaline phosphatase, D-amino acid oxidase, polyphosphate kinase 3, maltodextrin phosphorylase, and sialic acid aldolase have been successfully produced as active IBs when fused to an appropriate partner such as the foot-and-mouth disease virus capsid protein VP1, or the human β-amyloid peptide Aβ42(F19D). As active IBs may have many attractive advantages in enzyme production and industrial applications, it is of considerable interest to explore them further. In this paper, we report that an ionic self-assembling peptide ELK16 (LELELKLK)2 was able to effectively induce the formation of cytoplasmic inclusion bodies in Escherichia coli (E. coli) when attached to the carboxyl termini of four model proteins including lipase A, amadoriase II, β-xylosidase, and green fluorescent protein. These aggregates had a general appearance similar to the usually reported cytoplasmic inclusion bodies (IBs) under transmission electron microscopy or fluorescence confocal microscopy. Except for lipase A-ELK16 fusion, the three other fusion protein aggregates retained comparable specific activities with the native counterparts. Conformational analyses by Fourier transform infrared spectroscopy revealed the existence of newly formed antiparallel beta-sheet structures in these ELK16 peptide-induced inclusion bodies, which is consistent with the reported assembly of the ELK16 peptide. This has been the first report where a terminally attached self-assembling β peptide ELK16 can promote the formation of active inclusion bodies or active protein aggregates in E. coli. It has the potential to render E. coli and other recombinant hosts more efficient as microbial cell factories for protein production. Our observation might also provide hints for protein aggregation-related diseases.
Tài liệu tham khảo
Martinez-Alonso M, Gonzalez-Montalban N, Garcia-Fruitos E, Villaverde A: Learning about protein solubility from bacterial inclusion bodies. Microb Cell Fact. 2009, 8: 4-10.1186/1475-2859-8-4.
Ventura S, Villaverde A: Protein quality in bacterial inclusion bodies. Trends Biotechnol. 2006, 24: 179-185. 10.1016/j.tibtech.2006.02.007.
Gonzalez-Montalban N, Garcia-Fruitos E, Villaverde A: Recombinant protein solubility - does more mean better?. Nat Biotechnol. 2007, 25: 718-720. 10.1038/nbt0707-718.
Garcia-Fruitos E: Inclusion bodies: a new concept. Microb Cell Fact. 2010, 9: 80-10.1186/1475-2859-9-80.
Worrall DM, Goss NH: The formation of biologically active beta-galactosidase inclusion bodies in Escherichia coli. Aust J Biotechnol. 1989, 3: 28-32.
Peternel S, Grdadolnik J, Gaberc-Porekar V, Komel R: Engineering inclusion bodies for non denaturing extraction of functional proteins. Microb Cell Fact. 2008, 7: 9-10.1186/1475-2859-7-34.
Tokatlidis K, Dhurjati P, Millet J, Beguin P, Aubert JP: High activity of inclusion bodies formed in Escherichia coli overproducing Clostridium thermocellum endoglucanase D. FEBS Letters. 1991, 282: 205-208. 10.1016/0014-5793(91)80478-L.
Garcia-Fruitos E, Gonzalez-Montalban N, Morell M, Vera A, Ferraz RM, Aris A, Ventura S, Villaverde A: Aggregation as bacterial inclusion bodies does not imply inactivation of enzymes and fluorescent proteins. Microb Cell Fact. 2005, 4: 6-10.1186/1475-2859-4-27.
Arie JP, Miot M, Sassoon N, Betton JM: Formation of active inclusion bodies in the periplasm of Escherichia coli. Mol Microbiol. 2006, 62: 427-437. 10.1111/j.1365-2958.2006.05394.x.
Nahalka J, Nidetzky B: Fusion to a pull-down domain: A novel approach of producing Trigonopsis variabilis D-amino acid oxidase as insoluble enzyme aggregates. Biotechnol Bioeng. 2007, 97: 454-461. 10.1002/bit.21244.
Nahalka J, Vikartovska A, Hrabarova E: A crosslinked inclusion body process for sialic acid synthesis. J Biotechnol. 2008, 134: 146-153. 10.1016/j.jbiotec.2008.01.014.
Nahalka J: Physiological aggregation of maltodextrin phosphorylase from Pyrococcus furiosus and its application in a process of batch starch degradation to alpha-D-glucose-1-phosphate. J Ind Microbiol Biotechnol. 2008, 35: 219-223. 10.1007/s10295-007-0287-4.
Nahalka J, Mislovicova D, Kavcova H: Targeting lectin activity into inclusion bodies for the characterisation of glycoproteins. Mol Biosyst. 2009, 5: 819-821. 10.1039/b900526a.
Nahalka J, Patoprsty V: Enzymatic synthesis of sialylation substrates powered by a novel polyphosphate kinase (PPK3). Org Biomol Chem. 2009, 7: 1778-1780. 10.1039/b822549b.
Roessl U, Nahalka J, Nidetzky B: Carrier-free immobilized enzymes for biocatalysis. Biotechnol Lett. 2010, 32: 341-350. 10.1007/s10529-009-0173-4.
Garcia-Fruitos E, Rodriguez-Carmona E, Diez-Gil C, Ferraz RM, Vazquez E, Corchero JL, Cano-Sarabia M, Ratera I, Ventosa N, Veciana J, Villaverde A: Surface cell growth engineering assisted by a novel bacterial nanomaterial. Adv Mater. 2009, 21: 4249-4253. 10.1002/adma.200900283.
Anantharamaiah GM, Jones JI, Brouillette CG, Schmidt CF, Chung BH, Hughes TA, Bhown AS, Segrest JP: Studies of synthetic peptide analogs of the amphipathic helix - structure of complexes with dimyristoyl phosphatidylcholine. J Biol Chem. 1985, 260: 248-255.
Lazar KL, Miller-Auer H, Getz GS, Orgel J, Meredith SC: Helix-turn-helix peptides that form alpha-helical fibrils: Turn sequences drive fibril structure. Biochemistry. 2005, 44: 12681-12689. 10.1021/bi0509705.
Zhang SG, Holmes T, Lockshin C, Rich A: Spontaneous assembly of a self-complementary oligopeptide to form a stable macroscopic membrane. Proc Natl Acad Sci USA. 1993, 90: 3334-3338. 10.1073/pnas.90.8.3334.
Zhang SG, Lockshin C, Herbert A, Winter E, Rich A: Zuotin, a putative Z-DNA binding-protein in Saccharomyces-cerevisiae. Embo J. 1992, 11: 3787-3796.
van Pouderoyen G, Eggert T, Jaeger KE, Dijkstra BW: The crystal structure of Bacillus subtilis lipase: A minimal alpha/beta hydrolase fold enzyme. J Mol Biol. 2001, 309: 215-226. 10.1006/jmbi.2001.4659.
Williams DC, Vanfrank RM, Muth WL, Burnett JP: Cytoplasmic inclusion-bodies in Escherichia coli producing biosynthetic human insulin proteins. Science. 1982, 215: 687-689. 10.1126/science.7036343.
Seshadri S, Khurana R, Fink AL: Fourier transform infrared spectroscopy in analysis of protein deposits. Amyloid, Prions, and Other Protein Aggregates. 1999, San Diego: Academic Press Inc, 309: 559-576. full_text. Methods in Enzymology,
Mitraki A: Protein aggregation: from inclusion bodies to amyloid and biomaterials. Advances in Protein Chemistry and Structural Biology. 2010, San Diego: Elsevier Academic Press Inc, 79: 89-125. full_text. Volume 79. Advances in Protein Chemistry and Structural Biology
Pan KM, Baldwin M, Nguyen J, Gasset M, Serban A, Groth D, Mehlhorn I, Huang ZW, Fletterick RJ, Cohen FE, Prusiner SB: Conversion of alpha-helices into beta-sheets features in the formation of the scrapie prion proteins. Proc Natl Acad Sci USA. 1993, 90: 10962-10966. 10.1073/pnas.90.23.10962.
Collard F, Zhang J, Nemet I, Qanungo KR, Monnier VM, Yee VC: Crystal structure of the deglycating enzyme fructosamine oxidase (Amadoriase II). J Biol Chem. 2008, 283: 27007-27016. 10.1074/jbc.M804885200.
Nilsson MR: Techniques to study amyloid fibril formation in vitro. Methods. 2004, 34: 151-160. 10.1016/j.ymeth.2004.03.012.
Carrio M, Gonzalez-Montalban N, Vera A, Villaverde A, Ventura S: Amyloid-like properties of bacterial inclusion bodies. J Mol Biol. 2005, 347: 1025-1037. 10.1016/j.jmb.2005.02.030.
Ami D, Natalello A, Taylor G, Tonon G, Doglia SM: Structural analysis of protein inclusion bodies by Fourier transform infrared microspectroscopy. BBA-Proteins Proteomics. 2006, 1764: 793-799. 10.1016/j.bbapap.2005.12.005.
Yang YL, Khoe U, Wang XM, Horii A, Yokoi H, Zhang SG: Designer self-assembling peptide nanomaterials. Nano Today. 2009, 4: 193-210. 10.1016/j.nantod.2009.05.001.
Morell M, Bravo R, Espargaro A, Sisquella X, Aviles FX, Fernandez-Busquets X, Ventura S: Inclusion bodies: Specificity in their aggregation process and amyloid-like structure. Biochim Biophys Acta-Mol Cell Res. 2008, 1783: 1815-1825. 10.1016/j.bbamcr.2008.06.007.
Wei QD, Kim YS, Seo JH, Jang WS, Lee IH, Cha HJ: Facilitation of expression and purification of an antimicrobial peptide by fusion with baculoviral polyhedrin in Escherichia coli. Appl Environ Microbiol. 2005, 71: 5038-5043. 10.1128/AEM.71.9.5038-5043.2005.
Pazgier M, Lubkowski J: Expression and purification of recombinant human alpha-defensins in Escherichia coli. Protein Expr Purif. 2006, 49: 1-8. 10.1016/j.pep.2006.05.004.
Zorko M, Japelj B, Hafner-Bratkovic I, Jerala R: Expression, purification and structural studies of a short antimicrobial peptide. Biochim Biophys Acta-Biomembr. 2009, 1788: 314-323. 10.1016/j.bbamem.2008.10.015.
Wood DW, Wu W, Belfort G, Derbyshire V, Belfort M: A genetic system yields self-cleaving inteins for bioseparations. Nat Biotechnol. 1999, 17: 889-892. 10.1038/12879.
Serpell LC: Alzheimer's amyloid fibrils: structure and assembly. Biochim Biophys Acta-Mol Basis Dis. 2000, 1502: 16-30.
Zheng J, Guan H, Xu LH, Yang R, Lin ZL: Engineered amadoriase II exhibiting expanded substrate range. Appl Microbiol Biotechnol. 2010, 86: 607-613. 10.1007/s00253-009-2319-7.
Chen TJ, Zhang JQ, Liang L, Yang R, Lin ZL: An in vivo, label-free quick assay for xylose transport in Escherichia coli. Anal Biochem. 2009, 390: 63-67. 10.1016/j.ab.2009.03.048.
Winkler UK, Stuckmann M: Glycogen, hyaluronate, and some other polysaccharides greatly enhance the formation of exolipase by Serratia-marcescens. J Bacteriol. 1979, 138: 663-670.