Production of medium-chain-length polyhydroxyalkanoates by high-cell-density cultivation of Pseudomonas putida under phosphorus limitation

Lee, Sang Yup; Wong, Heng Ho; Choi, Jong Il; Lee, Seung Hwan; Lee, Sang Cheol; Han, Chul Soo

doi:10.1002/(sici)1097-0290(20000520)68:4<466::aid-bit12>3.3.co;2-k

Cited by 34 publications

(56 citation statements)

References 19 publications

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“…PHAs consist of about 120 diVerent (R)-3-hydroxyacid (3HA) monomeric units with the most common being poly( -hydroxybutyrate)(PHB) [19] and poly( -hydroxyvalerate) (PHV) [5]. Several bacteria use various carbon substrates to produce PHAs under limiting growth conditions [1,13,24,[26][27][28]50]. Pseudomonas oleovorans, for example, employs substrates requiring -oxidation, but not those requiring fatty acid de novo synthesis to produce medium chain length (mcl) PHAs [11,12,14,17,23,36,43,49,51].…”

Section: Introductionmentioning

confidence: 99%

Biosynthesis and characterization of copolymer poly(3HB-co-3HV) from saponified Jatropha curcas oil by Pseudomonas oleovorans

Allen

Anderson

Ayorinde

et al. 2010

J Ind Microbiol Biotechnol

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Polyhydroxyalkanoates (PHAs) are naturally occurring biodegradable polymers with promising application in the formulation of plastic materials. PHAs are produced by numerous bacteria as energy/carbon storage materials from various substrates, including sugars and plant oils. Since these substrates compete as food sources, their use as raw material for industrial-scale production of PHA is limited. Therefore, efforts have been focused on seeking alternative sources for bacterial production of PHA. One substrate that seems to have great potential is the seed oil of Jatropha curcas plant. Among other favorable properties, J. curcas seed oil is non-edible, widely available, and can be cheaply produced. In this study, Pseudomonas oleovorans (ATCC 29347) was grown in a mineral salt medium supplemented with saponified J. curcas seed oil as the only carbon source under batch fermentation. Optimum PHA yield of 26.06% cell dry weight was achieved after 72 h. The PHA had a melting point (T(m)) between 150 and 160 degrees C. Results of polymer analyses by gas chromatography/mass spectrometry (GC/MS) identified only the methyl 3-hydroxybutanoate monomeric unit. However, electrospray ionization-time of flight mass spectroscopy (ESI-TOF MS) confirmed that the PHA was a copolymer with the characteristic HB/HV peaks at m/z 1155.49 (HB) and 1,169, 1,184-1,194 (HV). The data were further supported by 1H and 13C NMR analysis. Polymer analysis by gel permeation chromatography (GPC) indicated a peak molecular weight (MP) of 179,797, molecular weight (M(W)) of 166,838, weight number average mass (M(n)) of 131,847, and polydispersity (M(w)/M (n)) of 1.3. The data from this study indicate that J. curcas seed oil can be used as a substrate to produce the copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate), poly(3HB-co-3HV).

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Section: Introductionmentioning

confidence: 99%

Biosynthesis and characterization of copolymer poly(3HB-co-3HV) from saponified Jatropha curcas oil by Pseudomonas oleovorans

Allen

Anderson

Ayorinde

et al. 2010

J Ind Microbiol Biotechnol

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“…The maximum cell density ever obtained with P. putida KT2442 was 141 g CDM/liter containing 51% (wt/wt) of PHA; however, this was achieved when oleic acid was used as the carbon source (41). When octanoic acid was used as the carbon source, cells could be grown only to densities of about 55 g CDM/liter, with a maximum PHA content of 75% (wt/wt) (35); it has to be mentioned that these experiments were done at the 2.5-liter scale only (36).…”

Section: Discussionmentioning

confidence: 99%

Large-Scale Production of Poly(3-Hydroxyoctanoic Acid) by Pseudomonas putida GPo1 and a Simplified Downstream Process

Elbahloul

Steinbüchel

2009

Appl Environ Microbiol

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The suitability of Pseudomonas putida GPo1 for large-scale cultivation and production of poly(3-hydroxyoctanoate) (PHO) was investigated in this study. Three fed-batch cultivations of P. putida GPo1 at the 350-or 400-liter scale in a bioreactor with a capacity of 650 liters were done in mineral salts medium containing initially 20 mM sodium octanoate as the carbon source. The feeding solution included ammonium octanoate, which was fed at a relatively low concentration to promote PHO accumulation under nitrogen-limited conditions. During cultivation, the pH was regulated by addition of NaOH, NH 4 OH, or octanoic acid, which was used as an additional carbon source. Partial O 2 pressure (pO 2 ) was adjusted to 20 to 40% by controlling the airflow and stirrer speed. Under the optimized conditions, P. putida GPo1 was able to grow to cell densities as high as 18, 37, and 53 g cells (dry mass) (CDM) per liter containing 49, 55, and 60% (wt/wt) of PHO, respectively. The resulting 40 kg CDM from these three cultivations was used directly for extraction of PHO. Three different methods of extraction of PHO were applied. From these, only acetone extraction showed better performance and resulted in 94% recovery of the PHO contents of cells. A novel mixture of precipitation solvents composed of 70% (vol/vol) methanol and 70% (vol/vol) ethanol was identified in this study. The ratio of PHO concentrate to the mixture was 0.2:1 (vol/vol) and allowed complete precipitation of PHO as white flakes. However, at a ratio of 1:1 (vol/vol) of the solvent mixture to PHO concentrate, a highly purified PHO was obtained. Precipitation yielded a dough-like polymeric material which was cast into thin layers and then shredded into small strips to allow evaporation of the remaining solvents. Gas chromatographic analysis revealed a purity of about 99% ؎ 0.2% (wt/wt) of the polymer, which consisted mainly of 3-hydroxyoctanoic acid (96 mol%).

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“…In our studies, P(3HB-co-3HV) content (wt%) was obtained as the percentage of the ratio of PHA concentration to CDW [6].…”

Section: Quantitative Analysismentioning

confidence: 99%

A fed-batch fermentation process for poly (3-hydroxybutyrate-co-3-hydroxyvalerate) production by Yangia sp. ND199 using molasses as substrate

Thược¹,

Phong²,

Khuong³

2016

V J Bio

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A locally isolated halophilic bacterium, Yangia sp. ND199 was able to use molasses as substrate for copolymers poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] production. Cell dry weight (CDW) of 6.37 g/l, P(3HB-co-3HV) content of 43.1 wt% and P(3HB-co-3HV) concentration of 2.75 g/l were obtained by Yangia sp. after 60 h of cultivation in flask. In a batch cultivation mode in a fermentor, the CDW was increased to 9.1 g/l but P(3HB-co-3HV) content was decreased to 37 wt%. Fed-batch fermentation with two different nutrient feeding strategies was used. High CDW of 54.8 g/l was obtained after 54 h of cultivation but P(3HB-co-3HV) content was still low (39.8 wt%). Two-step fed-batch fermentation with two different nutrient feeding strategies was then designed. High CDW of 50 g/l and P(3HB-co-3HV) content of 52.9 wt% were obtained after 54 h of cultivation. The two-step fed-batch process designed here for the production of P(3HB-co-3HV) by Yangia sp. ND199 can be developed and used for further studies.

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Production of medium-chain-length polyhydroxyalkanoates by high-cell-density cultivation of Pseudomonas putida under phosphorus limitation

Cited by 34 publications

References 19 publications

Biosynthesis and characterization of copolymer poly(3HB-co-3HV) from saponified Jatropha curcas oil by Pseudomonas oleovorans

Biosynthesis and characterization of copolymer poly(3HB-co-3HV) from saponified Jatropha curcas oil by Pseudomonas oleovorans

Large-Scale Production of Poly(3-Hydroxyoctanoic Acid) by Pseudomonas putida GPo1 and a Simplified Downstream Process

A fed-batch fermentation process for poly (3-hydroxybutyrate-co-3-hydroxyvalerate) production by Yangia sp. ND199 using molasses as substrate

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