{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T23:38:48Z","timestamp":1777505928670,"version":"3.51.4"},"reference-count":58,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2018,10,27]],"date-time":"2018-10-27T00:00:00Z","timestamp":1540598400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000038","name":"Natural Sciences and Engineering Research Council of Canada","doi-asserted-by":"publisher","award":["RGPIN-5961-2016"],"award-info":[{"award-number":["RGPIN-5961-2016"]}],"id":[{"id":"10.13039\/501100000038","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100008762","name":"Genome Canada","doi-asserted-by":"publisher","award":["FiCoGen"],"award-info":[{"award-number":["FiCoGen"]}],"id":[{"id":"10.13039\/100008762","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Polymers"],"abstract":"<jats:p>Pseudomonas chlororaphis PA23 was isolated from the rhizosphere of soybeans and identified as a biocontrol bacterium against Sclerotinia sclerotiorum, a fungal plant pathogen. This bacterium produces a number of secondary metabolites, including phenazine-1-carboxylic acid, 2-hydroxyphenazine, pyrrolnitrin (PRN), hydrogen cyanide, proteases, lipases and siderophores. It also synthesizes and accumulates polyhydroxyalkanoate (PHA) polymers as carbon and energy storage compounds under nutrient-limited conditions. Pseudomonads like P. chlororaphis metabolize glucose via the Entner-Doudoroff and Pentose Phosphate pathways, which provide precursors for phenazine production. Mutants defective in phenazine (PHZ; PA23-63), PRN (PA23-8), or both (PA23-63-1) accumulated higher concentrations of PHAs than the wild-type strain (PA23) when cultured in Ramsay\u2019s Minimal Medium with glucose or octanoic acid as the carbon source. Expression levels of six pha genes, phaC1, phaZ, phaC2, phaD, phaF, and phaI, were compared with wild type PA23 by quantitative real time polymerase chain reaction (qPCR). The qPCR studies indicated that there was no change in levels of transcription of the PHA synthase genes phaC1 and phaC2 in the phz- (PA23-63) and phz- prn- (PA23-63-1) mutants in glucose medium. There was a significant increase in expression of phaC2 in octanoate medium. Transcription of phaD, phaF and phaI increased significantly in the phz- prn- (PA23-63-1) mutant. Mutations in regulatory genes like gacS, rpoS, and relA\/spoT, which affect PHZ and PRN production, also resulted in altered gene expression. The expression of phaC1, phaC2, phaF, and phaI genes was down-regulated significantly in gacS and rpoS mutants. Thus, it appears that PHZ, PRN, and PHA production is regulated by common mechanisms. Higher PHA production in the phz- (PA23-63), prn- (PA23-8), and phz- prn- (PA23-63-1) mutants in octanoic medium could be correlated with higher expression of phaC2. Further, the greater PHA production observed in the phz- and prn- mutants was not due to increased transcription of PHA synthase genes in glucose medium, but due to more accessibility of carbon substrates and reducing power, which were otherwise used for the synthesis of PHZ and PRN.<\/jats:p>","DOI":"10.3390\/polym10111203","type":"journal-article","created":{"date-parts":[[2018,10,29]],"date-time":"2018-10-29T11:10:41Z","timestamp":1540811441000},"page":"1203","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Polyhydroxyalkanoate (PHA) Polymer Accumulation and pha Gene Expression in Phenazine (phz-) and Pyrrolnitrin (prn-) Defective Mutants of Pseudomonas chlororaphis PA23"],"prefix":"10.3390","volume":"10","author":[{"given":"Parveen K.","family":"Sharma","sequence":"first","affiliation":[{"name":"Department of Biosystems Engineering, University of Manitoba, Winnipeg, MB R3T 5V6, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Riffat I.","family":"Munir","sequence":"additional","affiliation":[{"name":"Department of Biosystems Engineering, University of Manitoba, Winnipeg, MB R3T 5V6, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jocelyn","family":"Plouffe","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University of Manitoba, Winnipeg, MB R3T 5V6, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nidhi","family":"Shah","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University of Manitoba, Winnipeg, MB R3T 5V6, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Teresa","family":"De Kievit","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University of Manitoba, Winnipeg, MB R3T 5V6, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David B.","family":"Levin","sequence":"additional","affiliation":[{"name":"Department of Biosystems Engineering, University of Manitoba, Winnipeg, MB R3T 5V6, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2018,10,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.cropro.2006.04.007","article-title":"Biological control of Sclerotinia sclerotiorum (Lib.) de Bary by Pseudomonas and Bacillus species on canola petals","volume":"26","author":"Fernando","year":"2007","journal-title":"Crop Prot."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1016\/j.femsec.2004.04.014","article-title":"Effect of timing of application and population dynamics on the degree of biological control of Sclerotinia sclerotiorum by bacterial antagonists","volume":"49","author":"Savchuk","year":"2004","journal-title":"FEMS Microbiol. Ecol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.1139\/w06-079","article-title":"A GacS deficiency does not affect Pseudomonas chlororaphis PA23 fitness when growing on canola, in aged batch culture or as a biofilm","volume":"52","author":"Poritsanos","year":"2006","journal-title":"Can. J. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"476","DOI":"10.1139\/w05-152","article-title":"Detection of antibiotic-related genes from bacterial biocontrol agents with polymerase chain reaction","volume":"52","author":"Zhang","year":"2006","journal-title":"Can. J. Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1659","DOI":"10.1007\/s00253-010-2509-3","article-title":"Metabolism and function of phenazines in bacteria: Impacts on the behavior of bacteria in the environment and biotechnological processes","volume":"86","author":"Pierson","year":"2010","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1111\/j.1462-2920.2012.02846.x","article-title":"Recent insights into the diversity, frequency and ecological roles of phenazines in fluorescent Pseudomonas spp.","volume":"15","author":"Mavrodi","year":"2013","journal-title":"Environ. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Chincholkar, S., and Thomashow, L. (2013). Phenazines in the environment: Microbes, habitats, and ecological relevance. Microbial Phenazines, Springer.","DOI":"10.1007\/978-3-642-40573-0"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Chincholkar, S., and Thomashow, L. (2013). Genomic features and regulation of phenazine biosynthesis in the rhizosphere strain Pseudomonas aeruginosa M18. Microbial Phenazines, Springer.","DOI":"10.1007\/978-3-642-40573-0"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1094\/MPMI.2004.17.5.557","article-title":"Influence of environmental conditions on the production of phenazine-1-carboxamide by Pseudomonas chlororaphis PCL1391","volume":"17","author":"Wesselink","year":"2004","journal-title":"Mol. Plant-Microbe Interact."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3142","DOI":"10.1128\/AEM.66.8.3142-3150.2000","article-title":"Controlling instability in gacS-gacA regulatory genes during inoculant production of Pseudomonas fluorescens biocontrol strains","volume":"66","author":"Duffy","year":"2000","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Chincholkar, S., and Thomashow, L. (2013). The Biosynthesis of phenazines. Microbial phenazines: Biosynthesis, Agriculture and Health, Springer.","DOI":"10.1007\/978-3-642-40573-0"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1111\/j.1574-6941.2009.00792.x","article-title":"Phenazines are not essential for Pseudomonas chlororaphis PA23 biocontrol of Sclerotinia sclerotiorum, but do play a role in biofilm formation","volume":"71","author":"Selin","year":"2010","journal-title":"FEMS Microbiol. Ecol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"896","DOI":"10.1099\/mic.0.054254-0","article-title":"The PhzI\/PhzR quorum-sensing system is required for pyrrolnitrin and phenazine production, and exhibits cross-regulation with RpoS in Pseudomonas chlororaphis PA23","volume":"158","author":"Selin","year":"2012","journal-title":"Microbiology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2159","DOI":"10.1099\/mic.0.000391","article-title":"The global regulator ANR is essential for Pseudomonas chlororaphis strain PA23 biocontrol","volume":"162","author":"Nandi","year":"2016","journal-title":"Microbiology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"544","DOI":"10.1016\/j.biortech.2018.08.120","article-title":"Valorization of polyhydroxyalkanoates production process by co-synthesis of value-added products","volume":"269","author":"Kumar","year":"2018","journal-title":"Bioresour. Technol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1111\/j.1574-6968.1996.tb08034.x","article-title":"Phenazine antibiotic production in Pseudomonas aureofaciens: Role in rhizosphere ecology and pathogen suppression","volume":"136","author":"Pierson","year":"1996","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_17","first-page":"64","article-title":"Characterization of a tacky poly[3-hydroxyalkanoate] produced by Pseudomonas chlororaphis HS21 from palm kernel oil","volume":"13","author":"Yun","year":"2003","journal-title":"J. Microbiol. Biotechnol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.jbiotec.2013.02.003","article-title":"Novel description of mcl-PHA biosynthesis by Pseudomonas chlororaphis from animal-derived waste","volume":"165","author":"Muhr","year":"2013","journal-title":"J. Biotechnol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.bcab.2013.09.009","article-title":"Rhamnolipid and poly[hydroxyalkanoate] biosynthesis in 3-hydroxyacyl-ACP: CoA transacylase [phaG]-knockouts of Pseudomonas chlororaphis","volume":"3","author":"Solaiman","year":"2014","journal-title":"Biocatal. Agric. Biotechnol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"982","DOI":"10.1139\/w2012-074","article-title":"Kinetics of medium-chain-length polyhydroxyalkanoate production by a novel isolate of Pseudomonas putida LS46","volume":"58","author":"Sharma","year":"2012","journal-title":"Can. J. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"123","DOI":"10.15255\/CABEQ.2014.2252","article-title":"Plant oils and products of their hydrolysis as substrates for polyhydroxyalkanoate synthesis","volume":"29","author":"Walsh","year":"2015","journal-title":"Chem. Biochem. Eng. Q."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1186\/s13568-014-0037-8","article-title":"Genome features of Pseudomonas putida LS46, a novel polyhydroxyalkanoate producer and its comparison with other P. putida strains","volume":"4","author":"Sharma","year":"2014","journal-title":"AMB Express"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1009","DOI":"10.1139\/cjm-2017-0412","article-title":"Synthesis of polyhydroxyalkanoates [PHAs] from vegetable oils and free fatty acids by wild-type and mutant strains of Pseudomonas chlororaphis","volume":"63","author":"Sharma","year":"2017","journal-title":"Can. J. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.jbiotec.2010.10.072","article-title":"Metabolic relationship between polyhydroxyalkanoic acid and rhamnolipid synthesis in Pseudomonas aeruginosa: Comparative 13C NMR analysis of the products in wild-type and mutants","volume":"151","author":"Choi","year":"2011","journal-title":"J. Biotechnol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1099\/mic.0.053082-0","article-title":"Stringent response mutants of Pseudomonas chlororaphis PA23 exhibit enhanced antifungal activity against Sclerotinia sclerotiorum in vitro","volume":"158","author":"Manuel","year":"2012","journal-title":"Microbiology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1099\/mic.0.28284-0","article-title":"Regulatory roles of psrA and rpoS in phenazine-1-carboxamide synthesis by Pseudomonas chlororaphis PCL1391","volume":"152","author":"Girard","year":"2006","journal-title":"Microbiology"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.biocontrol.2013.10.015","article-title":"Expression of the Pseudomonas chlororaphis strain PA23 Rsm system is under control of GacA, RpoS, PsrA, quorum sensing and the stringent response","volume":"69","author":"Selin","year":"2014","journal-title":"Biol. Control"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1002\/mbo3.90","article-title":"Roles of the Gac-Rsm pathway in the regulation of phenazine biosynthesis in Pseudomonas chlororaphis 30\u201384","volume":"2","author":"Wang","year":"2013","journal-title":"MicrobiologyOpen"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1111\/j.1574-6968.2006.00562.x","article-title":"RpoS as an intermediate in RsmA-dependent regulation of secondary antifungal metabolites biosynthesis in Pseudomonas sp. M18","volume":"268","author":"Ge","year":"2006","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1795","DOI":"10.1128\/AEM.02962-12","article-title":"GacS-dependent regulation of polyhydroxyalkanoate synthesis in Pseudomonas putida CA-3","volume":"79","author":"Ryan","year":"2013","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1111\/j.1574-6968.2008.01203.x","article-title":"The alternative sigma factor, rpoS affects polyhydroxyalkanoate metabolism in Pseudomonas putida","volume":"284","author":"Ruiz","year":"2008","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1007\/s00284-003-4183-5","article-title":"rpoS gene expression in carbon-starved cultures of the polyhydroxyalkanoate-accumulating species Pseudomonas oleovorans","volume":"48","author":"Ruiz","year":"2004","journal-title":"Curr. Microbiol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1128\/AEM.00755-17","article-title":"Absence of ppGpp leads to increased mobilization of intermediately accumulated poly [3-hydroxybutyrate] in Ralstonia eutropha H16","volume":"83","author":"Juengert","year":"2017","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"8033","DOI":"10.1128\/AEM.01693-12","article-title":"Whole-genome microarray and gene deletion studies reveal regulation of the polyhydroxyalkanoate production cycle by the stringent response in Ralstonia eutropha H16","volume":"78","author":"Brigham","year":"2012","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1186\/s13568-017-0396-z","article-title":"Medium-chain-length polyhydroxyalkanoates synthesis by Pseudomonas putida KT2440 relA\/spoT mutant: Bioprocess characterization and transcriptome analysis","volume":"7","author":"Dabrowska","year":"2017","journal-title":"AMB Express"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"e00689-14","DOI":"10.1128\/genomeA.00689-14","article-title":"Genome Sequence of Pseudomonas chlororaphis strain PA23","volume":"2","author":"Loewen","year":"2014","journal-title":"Genome Annoc."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1007\/BF00500854","article-title":"A rapid gas chromatographic method for the determination of polyhydroxybutyric acid in microbial biomass","volume":"6","author":"Braunegg","year":"1978","journal-title":"Eur. J. Appl. Microbiol. Biotechnol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of Relative gene expression data using real time quantitative PCR and the 2\u2212\u03b4\u03b4CT method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1111\/1462-2920.12760","article-title":"A holistic view of polyhydroxyalkanoate metabolism in Pseudomonas putida","volume":"18","author":"Prieto","year":"2016","journal-title":"Environ. Microbiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1111\/j.1574-6968.2004.tb09538.x","article-title":"Polyhydroxyalkanoate synthases PhaC1 and PhaC2 from Pseudomonas stutzeri 1317 had different substrate specificities","volume":"234","author":"Chen","year":"2004","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1146\/annurev.phyto.44.013106.145710","article-title":"Phenazine compounds in fluorescent Pseudomonas spp. biosynthesis and regulation","volume":"44","author":"Mavrodi","year":"2006","journal-title":"Ann. Rev. Phytopathol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1591","DOI":"10.1111\/j.1462-2920.2010.02199.x","article-title":"The PhaD regulator controls the simultaneous expression of the pha genes involved in polyhydroxyalkanoate metabolism and turnover in Pseudomonas putida KT2442","volume":"12","author":"Escapa","year":"2010","journal-title":"Environ. Microbiol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.1007\/s00253-006-0373-y","article-title":"Regulation of polyhydroxyalkanoate synthases (phaC1 and phaC2) gene expression in Pseudomonas corrugata","volume":"72","author":"Conte","year":"2006","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1038\/nchembio764","article-title":"Rethinking \u2018secondary\u2019 metabolism: Physiological roles for phenazine antibiotics","volume":"2","author":"Newman","year":"2006","journal-title":"Nat. Chem. Biol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1186\/1475-2859-13-40","article-title":"Metabolic flux responses to genetic modification for shikimic acid production by Bacillus subtilis strains","volume":"13","author":"Liu","year":"2014","journal-title":"Microb. Cell Fact."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1016\/j.ymben.2003.09.001","article-title":"Metabolic engineering for microbial production of shikimic acid","volume":"5","author":"Bongaerts","year":"2003","journal-title":"Metab. Eng."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.ymben.2015.09.003","article-title":"Engineering the central biosynthetic and secondary metabolic pathways of Pseudomonas aeruginosa strain PA1201 to improve phenazine-1-carboxylicacid production","volume":"32","author":"Jin","year":"2015","journal-title":"Metab. Eng."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.sbi.2014.08.013","article-title":"The structural biology of phenazine biosynthesis","volume":"29","author":"Blankenfeldt","year":"2014","journal-title":"Curr. Opin. Struct. Biol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1007\/s11274-017-2356-9","article-title":"Engineering Pseudomonas for phenazine biosynthesis, regulation, and biotechnological applications: A review","volume":"33","author":"Bilal","year":"2017","journal-title":"World J. Microbiol. Biotechnol."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Shen, X., Wang, Z., Huang, X., Hu, H., Wang, W., and Zhang, X. (2017). Developing genome-reduced Pseudomonas chlororaphis strains for the production of secondary metabolites. BMC Genom., 18.","DOI":"10.1186\/s12864-017-4127-2"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"6372","DOI":"10.1128\/JB.00505-07","article-title":"Pyocyanin alters redox homeostasis and carbon flux through central metabolic pathways in Pseudomonas aeruginosa PA14","volume":"189","author":"Dietrich","year":"2007","journal-title":"J. Bacteriol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.procbio.2017.04.028","article-title":"Microaerophilic environments improve the productivity of medium chain length polyhydroxyalkanoate biosynthesis from fatty acids in Pseudomonas putida LS46","volume":"59","author":"Blunt","year":"2017","journal-title":"Process Biochem."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1186\/1475-2859-11-34","article-title":"The metabolic response of P. putida KT2442 producing high levels of polyhydroxyalkanoate under single and multiple-nutrient-limited growth: Highlights from a multi-level omics approach","volume":"11","author":"Escapa","year":"2012","journal-title":"Microb. Cell Fact."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1186\/1475-2859-13-88","article-title":"Production of medium chain length polyhydroxyalkanoate in metabolic flux optimized Pseudomonas putida","volume":"13","author":"Bielecka","year":"2014","journal-title":"Microb. Cell Fact."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1002\/(SICI)1097-0290(19961220)52:6<707::AID-BIT8>3.0.CO;2-S","article-title":"Regulatory effects of cellular nicotinamide nucleotides and enzyme activities on poly(3-hydroxybutyrate) synthesis in recombinant Escherichia coli","volume":"52","author":"Lee","year":"1996","journal-title":"Biotechnol. Bioeng."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1049","DOI":"10.1111\/j.1462-2920.2011.02684.x","article-title":"The polyhydroxyalkanoate metabolism controls carbon and energy spillage in Pseudomonas putida","volume":"14","author":"Escapa","year":"2012","journal-title":"Environ. Microbiol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"916","DOI":"10.1021\/bm801431c","article-title":"Simultaneous accumulation and degradation of polyhydroxyalkanoates: Futile cycle or clever regulation?","volume":"10","author":"Ren","year":"2009","journal-title":"Biomacromolecules"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1016\/j.synbio.2017.11.001","article-title":"Rapid generation of recombinant Pseudomonas putida secondary metabolite producers using yTREX","volume":"2","author":"Weihmann","year":"2017","journal-title":"J. Synth. Syst. Biotechnol."}],"container-title":["Polymers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4360\/10\/11\/1203\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:26:36Z","timestamp":1760196396000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4360\/10\/11\/1203"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,10,27]]},"references-count":58,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2018,11]]}},"alternative-id":["polym10111203"],"URL":"https:\/\/doi.org\/10.3390\/polym10111203","relation":{},"ISSN":["2073-4360"],"issn-type":[{"value":"2073-4360","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,10,27]]}}}