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Petzold"],"technology_type":["Mass Spectrometry","Bottom-up proteomics","Data-independent acquisition"],"software":[""],"submitter_keywords":["Catabolite repression","Isoprenyl acetate","T-brsc","Lignocellulosic feedstocks","Esterase knockout","Isoprenol","Advanced aviation fuel"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD067010"],"sample_protocol":["Protein was extracted from cell pellets and tryptic peptides were prepared by following established proteomic sample preparation protocol (dx.doi.org/10.17504/protocols.io.6qpvr6xjpvmk/v1). Briefly, cell pellets were resuspended in Qiagen P2 Lysis Buffer (Qiagen, Germany) to promote cell lysis. Proteins were precipitated with addition of 1 mM NaCl and 4 x vol acetone, followed by two additional washes with 80% acetone in water. The recovered protein pellet was homogenized by pipetting mixing with 100 mM ammonium bicarbonate in 20% methanol. Protein concentration was determined by the DC protein assay (BioRad, USA). Protein reduction was accomplished using 5 mM tris 2-(carboxyethyl)phosphine (TCEP) for 30 min at room temperature, and alkylation was performed with 10 mM iodoacetamide (IAM; final concentration) for 30 min at room temperature in the dark. Overnight digestion with trypsin was accomplished with a 1:50 trypsin:total protein ratio."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["DIA raw data files were analyzed by an integrated software suite DIA-NN. The databases used in the DIA-NN search (library-free mode)is P. putida KT2440 latest Uniprot proteome FASTA sequences plus the protein sequences of the heterologous proteins and common proteomic contaminants. DIA-NN determines mass tolerances automatically based on first pass analysis of the samples with automated determination of optimal mass accuracies. The retention time extraction window was determined individually for all MS runs analyzed via the automated optimization procedure implemented in DIA-NN. Protein inference was enabled, and the quantification strategy was set to Robust LC = High Accuracy. Output main DIA-NN reports were filtered with a global false discovery rate set at 0.01 (FDR <= 0.01) on both the precursor level and protein group level. The Top3 method, which is the average MS signal response of the three most intense tryptic peptides of each identified protein, was used to plot the quantity of the targeted proteins in the samples"],"omics_type":["Proteomics"],"labhead":["Christopher Petzold"],"instrument_platform":[""],"submission_type":["PARTIAL"],"labhead_affiliation":["Staff Scientist Biological Systems & Engineering Division Lawrence Berkeley National Laboratory Berkeley CA 94720"],"species":["Pseudomonas Putida Kt2440"],"publication":["41858775 Kang CW, Carruthers DN, McCauley J, Chen Y, Gin JW, Petzold CJ, Simmons BA, Lee TS. Multi-layered metabolic remodeling of &lt;i&gt;Pseudomonas putida&lt;/i&gt; for efficient conversion of lignocellulosic sugars to the precursors of advanced aviation fuel. Metab Eng Commun. 2026 22:e00274 10.1016/j.mec.2026.e00274"],"submitter_mail":["cjpetzold@lbl.gov"],"submitter_affiliation":["Lawrence Berkeley National Laboratory"],"submitter_country":["United States"],"additional_accession":[]},"is_claimable":false,"name":"Multi-layered metabolic remodeling of Pseudomonas putida for efficient conversion of lignocellulosic sugars to a precursor of advanced aviation fuel","description":"Isoprenyl acetate, a volatile ester derived from isoprenol, is a key biosynthetic intermediate for a candidate for next-generation aviation fuel, 1,4-dimethylcyclooctane. Here, we engineered Pseudomonas putida KT2440 for the production of isoprenyl acetate from mixed sugar substrates. We first generated isoprenyl acetate by introducing a heterologous alcohol acetyltransferase (ATF1) and deleting three promiscuous native esterases to reduce product degradation. Then, we engineered efficient glucose and xylose co-utilization by integrating a heterologous xylose isomerase pathway and deleting global regulators crc and hexR to alleviate catabolite repression. Additionally, intracellular acetyl-CoA flux was reinforced through the expression of auxiliary carbon-conserving routes, including non-oxidative glycolysis and acetate assimilation. Culture conditions were systematically optimized by adjusting media composition, induction, and overlay solvent to maximize product yield. These cumulative efforts led to isoprenyl acetate titers of 1.5 g/L in shake flasks and 1.9 g/L in fed-batch fermentor cultures from mixed sugars, corresponding to a yield of 0.067 g/g of total sugar consumed. Our work demonstrates the potential of P. putida as a robust microbial chassis for scalable biosynthesis of ester-based biofuels from lignocellulosic feedstocks.","dates":{"publication":"2026-03-23","submission":"2025-08-06"},"accession":"PXD067010","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:1073882","NEWT:6819","NEWT:475174","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:1590","NEWT:10034","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:1589","NEWT:135622","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656064","NEWT:36630","NEWT:383379","NCBITaxon:12721","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:7719","NEWT:868565","NEWT:135674","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:224308","NEWT:3347","NEWT:160621","NEWT:212790","NEWT:110368","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:410658","NEWT:44688","NEWT:44689","NEWT:498211","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:44685","NEWT:161934","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:410661","NEWT:1140","NCBITaxon:2157","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:3714","NEWT:2065263","NEWT:177437","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:244366","NEWT:1246791","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:33548","NEWT:52299","NEWT:8165","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:485","NEWT:56636","NEWT:2853422","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:1502","NEWT:128017","NEWT:376686","NEWT:95486","NEWT:1883446","NEWT:1233435","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:491","NEWT:460523","NEWT:235443","NEWT:1757312","NEWT:85694","NEWT:108458","NEWT:5936","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:1255228","NEWT:410289","NEWT:373153","NEWT:1640278","NEWT:472","NEWT:940825","NEWT:529507","NEWT:1071661","NEWT:470","NEWT:5911","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:445","NEWT:153009","NEWT:261756","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:215402","NEWT:1547","NEWT:906914","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:150475","NEWT:267872","NEWT:381666","NEWT:172269","NEWT:9534","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:2903","NEWT:185579","NEWT:13076","NEWT:1006581","NEWT:33940","NEWT:550","NEWT:554","NEWT:451516","NEWT:1325291","NEWT:36185","NEWT:1054211","NEWT:1225786","NEWT:771870","NEWT:51338","NEWT:575412","NEWT:28112","NEWT:6493","NEWT:10832","NEWT:6494","NEWT:6491","NEWT:507601","NEWT:520","NEWT:186441","NEWT:643680","NEWT:214092","NCBITaxon:6157","NEWT:13095","NEWT:162425","NEWT:104105","NEWT:216257","NEWT:9986","NEWT:8654","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:5147","NEWT:28108","NEWT:28104","NEWT:407821","NEWT:568703","NCBITaxon:2","NEWT:568708","NEWT:986","NEWT:52641","NEWT:28532","NEWT:353152","NEWT:298176","NEWT:227377","NEWT:1774284","NEWT:40674","NEWT:1194669","NEWT:443906","NEWT:519","NEWT:2510939","NEWT:7394","NEWT:6063","NEWT:1328388","NEWT:105487","NEWT:1548728","NEWT:667127","NEWT:9557","NEWT:377586","NEWT:300641","NEWT:39655","NEWT:9554","NEWT:38323","NEWT:475932","NEWT:256318","NEWT:206411","NCBITaxon:6191","NEWT:229533","NEWT:214053","NEWT:28550","NEWT:80863","NEWT:90675","NEWT:52638","NEWT:57075","NEWT:1666905","NEWT:8697","NEWT:8695","NEWT:884204","NEWT:1123869","NEWT:9544","NEWT:9545","NEWT:979","NEWT:7370","NEWT:83906","NEWT:99488","NEWT:1134506","NEWT:255470","NEWT:38783","NEWT:6426","NEWT:33090","NEWT:9935","NEWT:287889","NEWT:305959","NEWT:13489","NEWT:29829","NEWT:92867","NEWT:92866","NCBITaxon:3055","NEWT:51750","NEWT:202950","NEWT:441772","NEWT:48106","NEWT:295027","NCBITaxon:11320","NEWT:632957","NEWT:9925","NCBITaxon:9606","NEWT:90690","NEWT:1436183","NEWT:300273","NEWT:4232","NEWT:416348","NEWT:11298","NEWT:4236","NEWT:7742","NEWT:196627","NEWT:200308","NEWT:200302","NEWT:870435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