Amino Acid Accumulation Limits the Overexpression of Proteins in Lactococcus lactis 英文参考文献.docVIP
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Amino Acid Accumulation Limits the Overexpression of Proteins in Lactococcus lactis 英文参考文献
AminoAcidAccumulationLimitstheOverexpressionof
ProteinsinLactococcuslactis
RaviK.R.Marreddy1,EricR.Geertsma1¤,HjalmarP.Permentier1,JoaoP.C.Pinto2,JanKok2 ,Bert
Poolman1*
1DepartmentofBiochemistry,GroningenBiomolecularSciencesandBiotechnologyInstitute,NetherlandsProteomicsCentre,ZernikeInstituteforAdvancedMaterials,
UniversityofGroningen,Groningen,TheNetherlands,2DepartmentofMolecularGenetics,GroningenBiomolecularSciencesandBiotechnologyInstitute,Universityof
Groningen,Haren,TheNetherlands
Abstract
Background:Understandingthebiogenesispathwaysforthefunctionalexpressionofrecombinantproteins,inparticular
membraneproteinsandcomplexmultidomainassemblies,isafundamentalissueincellbiologyandofhighimportancefor
futureprogressinstructuralgenomics.Inthisstudy,weemployedaproteomicapproachtounderstandthedifferencein
expressionlevelsforvariousmultidomainmembraneproteinsinL.lactiscellsgrownincomplexandsyntheticmedia.
Methodology/PrincipalFindings:Theproteomicprofilesofcellsgrowinginmediainwhichtheproteinswereexpressedto
highorlowlevelssuggestedalimitationintheavailabilityofbranched-chainaminoacids,morespecificallyatoolimited
capacitytoaccumulatethesenutrients.BysupplyingthecellswithanalternativepathforaccumulationofIle,Leuand/or
Val,i.e.,amediumsupplementoftheappropriatedipeptides,orbyengineeringthetransportcapacityforbranched-chain
aminoacids,theexpressionlevelscouldbeincreasedseveralfold.
Conclusions: We show that the availability of branched chain amino acids is a critical factor for the (over)expression of
proteinsinL.lactis.Theforwardengineeringofcellsforfunctionalproteinproductionrequiredfine-tuningofco-expression
ofthebranchedchainaminoacidtransporter.
Citation: Marreddy RKR, Geertsma ER, Permentier HP, Pinto JPC, Kok J, et al. (2010) Amino Acid Accumulation Limits the Overexpression of Proteins in
Lactococcuslactis.PLoSONE5(4):e10317.doi:10.1371/journal.pone.0010317
Editor:ChristopheHerman,BaylorCollegeofMedicine,UnitedStatesofAmerica
ReceivedFebruary26,2010;AcceptedMarc
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