{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T21:51:41Z","timestamp":1776289901565,"version":"3.50.1"},"reference-count":32,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2016,8,8]],"date-time":"2016-08-08T00:00:00Z","timestamp":1470614400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["11264004"],"award-info":[{"award-number":["11264004"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31460035"],"award-info":[{"award-number":["31460035"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004607","name":"Guangxi Natural Science Foundation","doi-asserted-by":"publisher","award":["2012GXNSFCA053001"],"award-info":[{"award-number":["2012GXNSFCA053001"]}],"id":[{"id":"10.13039\/501100004607","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004607","name":"Guangxi Natural Science Foundation","doi-asserted-by":"publisher","award":["2014GXNSFAA118193"],"award-info":[{"award-number":["2014GXNSFAA118193"]}],"id":[{"id":"10.13039\/501100004607","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>Poly-beta-hydroxybutyrate (PHB) can be formed in large amounts in Cupriavidus necator and is important for the industrial production of biodegradable plastics. In this investigation, laser tweezers Raman spectroscopy (LTRS) was used to characterize dynamic changes in PHB content\u2014as well as in the contents of other common biomolecule\u2014in C. necator during batch growth at both the population and single-cell levels. PHB accumulation began in the early stages of bacterial growth, and the maximum PHB production rate occurred in the early and middle exponential phases. The active biosynthesis of DNA, RNA, and proteins occurred in the lag and early exponential phases, whereas the levels of these molecules decreased continuously during the remaining fermentation process until the minimum values were reached. The PHB content inside single cells was relatively homogenous in the middle stage of fermentation; during the late growth stage, the variation in PHB levels between cells increased. In addition, bacterial cells in various growth phases could be clearly discriminated when principle component analysis was performed on the spectral data. These results suggest that LTRS is a valuable single-cell analysis tool that can provide more comprehensive information about the physiological state of a growing microbial population.<\/jats:p>","DOI":"10.3390\/s16081257","type":"journal-article","created":{"date-parts":[[2016,8,8]],"date-time":"2016-08-08T10:14:38Z","timestamp":1470651278000},"page":"1257","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Probing the Kinetic Anabolism of Poly-Beta-Hydroxybutyrate in Cupriavidus necator H16 Using Single-Cell Raman Spectroscopy"],"prefix":"10.3390","volume":"16","author":[{"given":"Zhanhua","family":"Tao","sequence":"first","affiliation":[{"name":"Guangxi Academy of Sciences, Nanning 530007, Guangxi, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lixin","family":"Peng","sequence":"additional","affiliation":[{"name":"Guangxi Academy of Sciences, Nanning 530007, Guangxi, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pengfei","family":"Zhang","sequence":"additional","affiliation":[{"name":"Optical Imaging Laboratory at Washington University in St. Louis, One Brookings Drive, St Louis, MO 63130, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yong-Qing","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Physics, East Carolina University, Greenville, NC 27858, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Guiwen","family":"Wang","sequence":"additional","affiliation":[{"name":"Guangxi Academy of Sciences, Nanning 530007, Guangxi, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2016,8,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Gao, S., Feng, J., Sun, L.H., and Tian, H. (2015, January 27\u201328). The Evolvement of Co-Production of Biodegradable Biological Material PHB. Proceedings of the 2015 4th International Conference on Sensors, Measurement and Intelligent Materials, Shenzhen, China.","DOI":"10.2991\/icsmim-15.2016.136"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"578","DOI":"10.1038\/nrmicro2354","article-title":"Bacterial polymers: Biosynthesis, modifications and applications","volume":"8","author":"Rehm","year":"2010","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2434","DOI":"10.1039\/b812677c","article-title":"A microbial polyhydroxyalkanoates (PHA) based bio- and materials industry","volume":"38","author":"Chen","year":"2009","journal-title":"Chem. Soc. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1128\/mr.54.4.450-472.1990","article-title":"Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates","volume":"54","author":"Anderson","year":"1990","journal-title":"Microbiol. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1007\/s002530051357","article-title":"Factors affecting the economics of polyhydroxyalkanoate production by bacterial fermentation","volume":"51","author":"Choi","year":"1999","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3788","DOI":"10.1128\/JB.185.13.3788-3794.2003","article-title":"Ralstonia eutropha H16 encodes two and possibly three intracellular poly[d-(\u2212)-3-hydroxybutyrate] depolymerase genes","volume":"185","author":"York","year":"2003","journal-title":"J. Bacteriol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3814","DOI":"10.1128\/JB.187.11.3814-3824.2005","article-title":"Kinetic studies of polyhydroxybutyrate granule formation in wautersia eutropha H16 by transmission electron microscopy","volume":"187","author":"Tian","year":"2005","journal-title":"J. Bacteriol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1257","DOI":"10.1038\/nbt1244","article-title":"Genome sequence of the bioplastic-producing \u201cknallgas\u201d bacterium ralstonia eutropha H16","volume":"24","author":"Pohlmann","year":"2006","journal-title":"Nat. Biotechnol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1227","DOI":"10.1038\/nbt1006-1227","article-title":"Deciphering bioplastic production","volume":"24","author":"Lee","year":"2006","journal-title":"Nat. Biotechnol."},{"key":"ref_10","first-page":"38","article-title":"Genomic view of energy metabolism in ralstonia eutropha H16","volume":"16","author":"Cramm","year":"2009","journal-title":"J. Mol. Microbiol. Biotechnol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"943","DOI":"10.1007\/s00449-013-1066-4","article-title":"Biosynthesis of poly(3-hydroxybutyrate) (PHB) by cupriavidus necator H16 from jatropha oil as carbon source","volume":"37","author":"Batcha","year":"2014","journal-title":"Bioprocess Biosyst. Eng."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"15298","DOI":"10.1016\/S0021-9258(19)84825-1","article-title":"Poly-beta-hydroxybutyrate (PHB) biosynthesis in alcaligenes eutrophus H16. Identification and characterization of the PHB polymerase gene (phbC)","volume":"264","author":"Peoples","year":"1989","journal-title":"J. Biol. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1016\/S1389-1723(00)88950-1","article-title":"Construction and selection of the novel recombinant Escherichia coli strain for poly(beta-hydroxybutyrate) production","volume":"89","author":"Yu","year":"2000","journal-title":"J. Biosci. Bioeng."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1043","DOI":"10.1002\/bit.260431107","article-title":"Cybernetic model for synthesis of poly-beta-hydroxybutyric acid in alcaligenes-eutrophus","volume":"43","author":"Yoo","year":"1994","journal-title":"Biotechnol. Bioeng."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1339","DOI":"10.1128\/aem.46.6.1339-1344.1983","article-title":"Analysis of poly-beta-hydroxybutyrate in rhizobium-japonicum bacteroids by ion-exclusion high-pressure liquid-chromatography and UV detection","volume":"46","author":"Karr","year":"1983","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1002\/cphc.200390004","article-title":"Raman spectroscopy\u2014A prospective tool in the life sciences","volume":"4","author":"Petry","year":"2003","journal-title":"ChemPhysChem"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"4452","DOI":"10.1021\/ac049753k","article-title":"Raman microscopic analysis of single microbial cells","volume":"76","author":"Huang","year":"2004","journal-title":"Anal. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"632","DOI":"10.1111\/j.1472-765X.2010.02941.x","article-title":"Intracellular ethanol accumulation in yeast cells during aerobic fermentation: A raman spectroscopic exploration","volume":"51","author":"Peng","year":"2010","journal-title":"Lett. Appl. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2155","DOI":"10.1021\/ac702185d","article-title":"Monitoring poly(3-hydroxybutyrate) production in Cupriavidus necator DSM 428 (H16) with raman spectroscopy","volume":"80","author":"Gelder","year":"2008","journal-title":"Anal. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1129","DOI":"10.1039\/c0an00526f","article-title":"Observation of content and heterogeneity of poly-beta-hydroxybutyric acid (PHB) in legionella bozemanii by vibrational spectroscopy","volume":"136","author":"Hermelink","year":"2011","journal-title":"Analyst"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1364\/OL.27.000249","article-title":"Near-infrared raman spectroscopy of single optically trapped biological cells","volume":"27","author":"Xie","year":"2002","journal-title":"Opt. Lett."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1002\/jbio.201200143","article-title":"Recent advances in laser tweezers Raman spectroscopy (LTRS) for label-free analysis of single cells","volume":"6","author":"Chan","year":"2013","journal-title":"J. Biophotonics"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"19021","DOI":"10.3390\/s150819021","article-title":"Raman spectroscopy of optically trapped single biological micro-particles","volume":"15","author":"Redding","year":"2015","journal-title":"Sensors"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2620","DOI":"10.1128\/JB.06543-11","article-title":"Mutations derived from the thermophilic polyhydroxyalkanoate synthase PhaC enhance the thermostability and activity of PhaC from cupriavidus necator H16","volume":"194","author":"Sheu","year":"2012","journal-title":"J. Bacterio."},{"key":"ref_25","unstructured":"La, R. (2005). Encyclopedia of Soil Science, CRC Press."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3132","DOI":"10.1016\/j.polymer.2006.03.010","article-title":"Raman microspectroscopy study of structure, dispersibility, and crystallinity of poly(hydroxybutyrate)\/poly(l-lactic acid) blends","volume":"47","author":"Furukawa","year":"2006","journal-title":"Polymer"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1002\/bip.10378","article-title":"Spectroscopic study of human lung epithelial cells (A549) in culture: Living cells versus dead cells","volume":"72","author":"Notingher","year":"2003","journal-title":"Biopolymers"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/j.vibspec.2010.07.011","article-title":"Ft-raman investigation of biodegradable polymers: Poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate)","volume":"54","author":"Izumi","year":"2010","journal-title":"Vib. Spectrosc."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1080\/05704920701551530","article-title":"Raman spectroscopy of biological tissues","volume":"42","author":"Movasaghi","year":"2007","journal-title":"Appl. Spectrosc. Rev."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2173","DOI":"10.1016\/j.procbio.2004.08.011","article-title":"Statistical media optimization studies for growth and PHB production by ralstonia eutropha","volume":"40","author":"Khanna","year":"2005","journal-title":"Process Biochem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1351\/pac198961030303","article-title":"Poly-(R)-3-hydroxybutyrate (PHB) biosynthesis\u2014Mechanistic studies on the biological claisen condensation catalyzed by beta-ketoacyl thiolase","volume":"61","author":"Masamune","year":"1989","journal-title":"Pure Appl. Chem."},{"key":"ref_32","unstructured":"Klug, W.S., Cummings, M.R., Spencer, C., and Palladino, M.A. (2011). Concepts of Genetics, Benjamin Cummings."}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/16\/8\/1257\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T19:28:00Z","timestamp":1760210880000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/16\/8\/1257"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,8,8]]},"references-count":32,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2016,8]]}},"alternative-id":["s16081257"],"URL":"https:\/\/doi.org\/10.3390\/s16081257","relation":{},"ISSN":["1424-8220"],"issn-type":[{"value":"1424-8220","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,8,8]]}}}