A Process Engineering Approach to Improve Production of P(3HB) by <i>Cupriavidus necator</i> from Used Cooking Oil

Cruz, Madalena V.; Gouveia, Ana Rosa; Dionísio, Madalena; Freitas, Filomena; Reis, Maria A. M.

doi:https://doi.org/10.1155/2019/2191650

International Journal of Polymer Science

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Abstract Introduction Methods Results and Discussion Conclusions Data Availability Conflicts of Interest Acknowledgments References Copyright Related Articles

Research Article | Open Access

Volume 2019 | Article ID 2191650 | https://doi.org/10.1155/2019/2191650

A Process Engineering Approach to Improve Production of P(3HB) by Cupriavidus necator from Used Cooking Oil

Madalena V. Cruz,¹Ana Rosa Gouveia,¹Madalena Dionísio,²Filomena Freitas,¹and Maria A. M. Reis¹

Academic Editor: Luc Averous

Received30 Apr 2018

Accepted12 Nov 2018

Published13 Jan 2019

Abstract

Different feeding strategies, namely, exponential feeding and DO-stat mode, were implemented for the production of poly(3-hydroxybutyrate), P(3HB), by Cupriavidus necator DSM 428 with used cooking oil (UCO) as the sole carbon source. With the exponential feeding strategy, a cell dry mass of 21.3 ± 0.9 g L⁻¹ was obtained, with a polymer content of 84.0 ± 4.5 wt.%, giving an overall volumetric productivity of 4.5 ± 0.2 g L⁻¹ day⁻¹. However, the highest P(3HB) volumetric productivity, 12.6 ± 0.8 g L⁻¹ day⁻¹, was obtained when the DO-stat mode was implemented together with the use of ammonium hydroxide for pH control, which served as an additional nitrogen source and allowed to reach higher cell dry mass (7.8 ± 0.6 g L⁻¹). The P(3HB) obtained in all experiments had a high molecular mass, ranging from 0.6 × 10⁵ to 2.6 × 10⁵ g mol⁻¹, with low polydispersity indexes of 1.2-1.6. Melting and glass transition temperatures were also similar for the polymer produced with both cultivation strategy, 174°C and 3.0-4.0°C, respectively. The polymer exhibited a crystallinity ranging from 52 to 65%. The DO-stat strategy to feed oil containing substrates as the sole carbon sources was reported for the first time in this study, and the preliminary results obtained show that it is a promising strategy to improve P(3HB) production. Nevertheless, the process requires further optimization in order to make it economically viable.

1. Introduction

The cultivation strategy used for the production of polyhydroxyalkanoates (PHA) is an important factor to be taken into account for bioprocess optimization and improvement. Although the batch mode is the simplest and primary strategy for any bioprocess [1, 2], and it has been the most extensively used strategy for PHA production [3], the fed-batch mode is considered more efficient to achieve high cell density cultures with high volumetric productivities [1, 3]. However, the process set-up requires the selection of a suitable substrate feeding strategy to properly control the concentration of the carbon source throughout the fed-batch phase. Several strategies are available to feed the cultures and have been tested for PHA production from different substrates, including, among others, pulse feeding [4, 5], continuous feeding with defined feed rate [6, 7], exponential feeding [8, 9], control of nutrient feed through dissolved oxygen (DO) concentration (DO-stat mode) [10–12], and pH control (pH-stat mode) [13, 14].

The use of feeding profiles designed to match the maximum specific cell growth rate of the microorganism during the growth phase and linear or decaying linear feed rates during the nongrowth-associated production phase are often used [15]. The exponential feeding strategy has been successfully used for PHA production by Cupriavidus necator DSM 545 [9] and Pseudomonas putida KT2440 [15], using glucose as carbon source. Rathinasabapathy et al. [16] also tested an exponential feeding strategy for cultivation of C. necator in fructose and canola oil. However, adjusting the feeding profile to the culture’s needs in terms of substrate is a difficult task and the defined profile may result in over- or underfeeding [9].

Feeding may also be based on physiology as in pH-stat and DO-stat modes where it depends on acid production or oxygen utilization, respectively [15, 17]. Since this strategy is based on a parameter that is measured online (i.e., the DO concentration or the pH value), it allows for the substrate to be automatically fed to the culture. The DO-stat strategy has been tested for cultivation of different PHA producers, such as recombinant Escherichia coli strains [18], Alcaligenes latus DSM1123 [19], Cupriavidus sp. USMAA2-4 [20], and Cupriavidus sp. DSM19416 [11]. A strategy combining DO- and pH-stat modes was also reported to feed oleic acid to Aeromonas hydrophila [21] and Pseudomonas putida KT2442 [22] to overcome the fact that, for high cell densities, the DO concentration did not respond to substrate depletion. There are some reports on the cultivation of C. necator using the DO-stat mode, either alone or in combination with the pH-stat mode, to feed spent coffee grounds oil [10] or a mixture of acetic, propionic, and butyric acids [22] to the culture in the fed-batch phase. However, this strategy has never been reported for the cultivation of C. necator using used cooking oil (UCO) as sole substrate.

In the previous work, UCO was demonstrated to be a suitable substrate for the cultivation of C. necator DSM 428 for production of P(3HB) [23, 24]. However, process optimization conditions were not explored. In this study, two different fed-batch strategies were tested for the first time, namely, exponential feeding and DO-stat mode, to feed UCO to the culture, aiming at improving polymer productivity and yield on a substrate basis. The impact of the tested cultivation strategies on the polymer’s composition and molecular mass distribution was evaluated.

2. Material and Methods

2.1. Microorganism and Media

C. necator DSM 428 was reactivated from stock cultures kept at −80°C by inoculation in solid Luria Bertani (LB) medium (15 g L⁻¹ agar), as described by Cruz et al. [10]. The mineral medium used for inoculum preparation and the bioreactor experiments had the following composition (per liter): (NH₄)₂HPO₄, 3.3 g; K₂HPO₄, 5.8 g; KH₂PO₄, 3.7 g; 10 mL of a 100 mM MgSO₄ solution; and 1 mL of a micronutrient solution. The micronutrient solution had the following composition (per liter of 1 N HCl): FeSO₄·7H₂O, 2.78 g; MnCl₂·4H₂O, 1.98 g; CoSO₄·7H₂O, 2.81 g; CaCl₂·2H₂O, 1.67 g; CuCl₂·2H₂O, 0.17 g; and ZnSO₄·7H₂O, 0.29 g [25]. The mineral medium was supplemented with 20 g L⁻¹ UCO as sole carbon source. The UCO used in this study was supplied by the University canteen, and it was mainly composed of triglycerides (83.4 ± 9.1 wt.%), with minor amounts of di- and monoglycerides (6.7 ± 0.4 and 0.4 ± 0.1 wt.%, respectively). Oleic and linoleic acids were the major constituent fatty acids of UCO (37.5 ± 0.6 and 49.8 ± 0.5 wt.%, respectively), with minor contents of palmitic and stearic acids (9.0 ± 0.1 and 3.4 ± 0.6 wt.%, respectively) [26].

2.2. Bioreactor Cultivation

Bioreactor cultivation experiments were performed in 2 L bioreactors (BIOSTAT B-Plus, Sartorius, Germany), with an initial working volume of 1.5 L. The inoculum was 10% (v/v) of the initial reactor working volume. It was prepared by inoculating a single C. necator colony into the LB medium and incubation in an orbital shaker, at 30°C and 200 rpm, for 24 hours. The culture thus obtained was then transferred into the mineral medium supplemented with 20 g L⁻¹ UCO as sole carbon source, further incubated for 48 hours, as described above, and used as inoculum for the bioreactor experiments.

In all experiments, the temperature was maintained at 30 ± 1°C and the pH was controlled at 6.8 ± 0.2 by the automatic addition of NaOH 2 M and/or 25% (v/v) NH₄OH. The dissolved oxygen concentration (DO) was maintained at 30% air saturation. The experiments comprised an initial batch phase (18-20 hours), followed by the fed-batch phase, wherein the UCO was supplied to the culture.

In experiment A (exponential feeding), UCO was fed according to the following profile: where is the feeding rate (g UCO h⁻¹ L⁻¹), (g_s g_x⁻¹ h⁻¹) is the specific substrate uptake rate, (g L⁻¹) is the active biomass concentration at the end of the exponential phase, (h⁻¹) is the specific growth rate, (h) is the initial time of feeding, and (h) is the end of batch time, respectively. The pH was controlled by the addition of NaOH 2 M throughout the entire experiment.

In experiment B (DO-stat mode), the UCO feeding flow rate was automatically controlled as a function of DO concentration (under a constant stirring of 500 rpm). The pH was initially controlled by the addition of NH₄OH to prolong the exponential growth phase by serving also as a nitrogen source. Afterwards, the pH was controlled with NaOH 2 M to impose nitrogen-limiting conditions.

Samples (15 ± 5 mL) were periodically withdrawn from the bioreactor for determination of the cell dry mass (CDM), UCO concentration, P(3HB) content in the biomass, and polymer composition.

2.3. Polymer Extraction and Purification

At the end of the cultivation runs, the broth (100 mL) was washed with n-hexane (1 : 1, v/v) to remove residual oil and centrifuged (7012 × g, 20 min). The biomass was further washed twice with deionized water (200 mL) and freeze-dried. The polymer was recovered from the dried biomass by Soxhlet extraction with chloroform (~10 g dry biomass extracted with 250 mL chloroform), at 70°C, for 24 hours. The solution thus obtained was filtered with 0.45 μm pore size filters (GxF, GHP membrane, PALL) to remove cell debris and precipitated in cold ethanol (1 : 10, v/v) under strong stirring. The polymer was collected by centrifugation (7012 × g, 20 min), dried at room temperature, and stored at 4°C.

2.4. Analytical Techniques

For CDM and residual oil quantification, 4-5 mL broth samples were mixed with n-hexane (1 : 1, v/v) and centrifuged (15,777 × g, 10 min). The biomass pellet was collected, washed with deionized water, and lyophilized for the gravimetric CDM quantification. The upper hexane layer (2-3 mL) containing the residual UCO was transferred to preweighed tubes and placed in a fume hood at room temperature for 24 h, for solvent evaporation and oil quantification. All analyses were performed in duplicate.

Polymer content in the biomass was determined as described by Cruz et al. [26]. Briefly, 2-3 mg dried cells were hydrolyzed with 1 mL 20% (v/v) sulphuric acid in methanol and 1 mL benzoic acid (internal standard) in chloroform (1 g L⁻¹), at 100°C, during 3.5 hours. The resulting methyl esters were analyzed by gas chromatography (GC), as described by Cruz et al. [26]. Poly(3-hydroxybutyrate-co-3-hydroxyvalerate), P(3HB-co-3HV), poly(3-hydroxyhexanoate-co-3-hydroxyoctanoate), P(3HHx-co-HO), and poly(3-hydroxyocatonate-co-3-decanoate-co-3-dodecanoate), P(3HO-co-3HD-co-3HDd), in concentrations ranging from 0.325 to 5 mg mL⁻¹ were used as standards.

2.5. Calculations

The active biomass was determined by where (g L⁻¹) and (g L⁻¹) are the cell dry mass and the concentration of polymer at time (h). The overall volumetric productivity (, g L⁻¹ day⁻¹) was calculated by the following equation: where (g L⁻¹) is the polymer produced during cultivation time (day). The growth (, g_x g_s⁻¹) and storage (, g_p g_s⁻¹) yields were calculated, respectively, by where (g L⁻¹) and (g L⁻¹) are the active biomass and the polymer, respectively, produced during the run, and (g L⁻¹) is the UCO consumed for the same period of time.

2.6. Polymer Characterization

Polymer composition and purity were evaluated by GC analysis, as described above. Weight average () and number average () molecular mass were determined using a size exclusion chromatography (SEC) apparatus (Waters), equipped with a solvent delivery system composed of a model 510 pump, a Rheodyne injector, and a refractive index detector (Waters 2410), according to the procedure described by Cruz et al. [26]. The polydispersity index (PDI) was given by the ratio between () and ().

The thermal properties of the polymers were determined by differential scanning calorimetry (DSC), as described by Morais et al. [27]. The glass transition temperatures (, °C) were taken as the midpoint of the heat flux step transition; melting (, °C) temperature and enthalpy (, J g⁻¹) were estimated, respectively, from the center and area of the endothermic peaks. The crystallinity (, %) of the PHA samples was estimated as the ratio between associated with the detected melting peak and the melting enthalpy of 100% crystalline poly-3-hydroxybutyrate, P(3HB), estimated as 146 J g⁻¹ [27].

3. Results and Discussion

3.1. Fed-Batch Cultivation with Exponential Feeding Profile

The results obtained in experiment A, wherein the exponential feeding strategy was tested, are presented in Figure 1 and Table 1. During the initial batch phase of the experiment, the culture grew with a maximum specific cell growth rate of 0.14 ± 0.02 h⁻¹, which is similar to the values obtained in previous studies for cultivation of C. necator under batch mode, 0.12-0.14 h⁻¹ [23, 24].

The fed-batch phase was initiated after 20 hours of cultivation (Figure 1) by supplying the culture with UCO under the defined exponential profile. The exponential feeding profile was designed aiming at maintaining the specific cell growth rate observed during the batch phase (0.14 ± 0.02 h⁻¹) for an extended period of time during the fed-batch phase, thus increasing the overall biomass production. However, the oil feeding was stopped at 55 hours of cultivation since accumulation of unconsumed UCO was noticed. The experiment was further prolonged up to 96 hours to allow the culture to use the accumulated UCO. This UCO accumulation in the bioreactor might have been caused by an overestimation of the feeding profile that was based on data obtained in previous batch experiments [23]. This dependency of the designed feeding profile on previously determined data is a disadvantage of this strategy since it is more prone to result in inadequate feeding of the culture.

A final polymer production of 17.9 ± 0.9 g L⁻¹ was obtained, corresponding to an overall volumetric productivity of 4.5 ± 0.2 g L⁻¹ day⁻¹ (Table 1). These values are considerably higher than the ones obtained under batch cultivation of C. necator with UCO, namely, a polymer production of 3.8-7.4 g L⁻¹ and a volumetric productivity of 3.4-3.6 g L⁻¹ day⁻¹ [23, 24]. The improved polymer production and productivity were a result of the higher polymer content in the biomass that reached 84.0 ± 4.5 wt.% at the end of the cultivation, while under the batch mode it had reached only 38-63 wt% [23, 24]. The polymer content in the biomass obtained in experiment A is also slightly higher than the values (72-81 wt.%) reported for the fed-batch cultivation of C. necator using soybean oil, with pulse feeding [5, 28].

There was an overall oil consumption of 28.0 ± 1.3 g L⁻¹ during the 96 hours of experiment A. The growth and storage yields were 0.11 ± 0.01 g_x g_s⁻¹ and 0.65 ± 0.03 g_p g_s⁻¹, respectively. The storage yield obtained in experiment A was within the values obtained under batch mode (0.29-0.70 g_p g_s⁻¹) [23, 24] and close to the values reported for the fed-batch cultivation of C. necator with pulse feeding of soybean oil (0.72-0.85 g_p g_s⁻¹) [5, 28].

The results obtained with experiment A show that the exponential feeding strategy tested for the fed-batch cultivation of C. necator with UCO allowed for improvement of P(3HB) production compared to the batch mode operation. However, it was difficult to adjust the feeding profile to the actual culture’s substrate consumption and an overfeeding resulted in UCO accumulation in the bioreactor. Mozumder et al. [9] also reported the difficulty in maintaining the exponential glucose profile feeding adequate to the culture’s needs in terms of substrate, and the defined profile resulted in over- or underfeeding. Nevertheless, the exponential feeding strategy might be more successful when used in combination with other feeding strategies, for example, based on the monitoring of other parameters, such as suggested by Mozumder et al. [9].

3.2. Fed-Batch Cultivation under a DO-Stat Mode

In an attempt to have a feeding strategy that more accurately matched the culture’s requirements for cell growth and polymer synthesis, the DO-stat mode was tested in experiment B. This strategy is based on the online measurement of the DO concentration, which tends to increase upon substrate depletion, thus signaling the automatic feeding of UCO to the culture. Figure 2 presents the results obtained for the fed-batch cultivation of C. necator under the DO-stat mode. The culture grew with a specific growth rate of 0.21 ± 0.01 h⁻¹ and reached an active biomass of 5.9 ± 1.1 g L⁻¹ within 7 hours of cultivation. The higher cell growth rate compared to experiment A was due to the fact that the pH was controlled with ammonium hydroxide, which served as an additional nitrogen source. The nitrogen availability during the initial batch phase changed the carbon-to-nitrogen ratio and promoted a faster cell growth.

The DO-stat mode was implemented at 17 hours of cultivation by starting the substrate feeding as a function of the DO concentration that was set at 30% of air saturation. At that time, NH₄OH was changed to NaOH for pH control so that nitrogen-limiting conditions were imposed. The culture continued to grow, though at a lower rate, and reached a maximum active biomass of 9.6 g L⁻¹, at 27 hours of cultivation (Figure 2). No further cell growth was noticed afterwards as a result of nitrogen exhaustion.

The automatic substrate feeding rate was very high during the first 5 hours after initiating the DO-stat control mode, as a response to the rise in DO concentration (Figure 2). Overall, 116 g L⁻¹ of UCO entered the reactor. Afterwards, the DO concentration remained rather constant (26-30%) and no further substrate feeding occurred. Polymer accumulation was initiated during the batch phase, but increased production occurred during the fed-batch phase (Figure 2). Maximum polymer concentration (19.8 ± 1.8 g L⁻¹) was reached at 37 hours of cultivation, corresponding to an overall volumetric productivity of 12.6 ± 0.8 g L⁻¹ day⁻¹ (Table 1). This value is considerably higher than that obtained in experiment A (4.5 ± 0.2 g L⁻¹ day⁻¹) and within the wide range of values (6.3-25 g L⁻¹ day⁻¹) reported for fed-batch cultivations of the same strain with pulse feeding of soybean oil [28] or jatropha oil [29].

A lower volumetric productivity (4.7 g L⁻¹ day⁻¹) was reported in a previous study, in which C. necator was cultivated using spent coffee grounds (SCG) oil as substrate, under the DO-stat mode [10]. The observed differences are probably related to the substrate used in each study. In fact, although both oils were rich in linoleic acid (49.8 and 38.4 wt.%, respectively), UCO had a higher content of oleic acid (37.5 wt.%) [26], while SCG oil was richer in palmitic acid (39.4 wt.%) [10]. Cell growth and polymer synthesis may have been stimulated in experiment B by the presence of higher contents of linoleic and oleic acids in UCO, since both fatty acids are known to be preferred carbon sources for C. necator cultivation [10, 28, 29]. Moreover, the presence of other components in SCG oil, such as sterols, tocopherols, and esters, may have also impacted on the culture’s performance.

Although the overall consumption of UCO in experiment B (38.0 ± 2.0 g L⁻¹) was higher than that observed for experiment A (28.0 ± 1.3 g L⁻¹), a lower storage yield was observed (0.52 ± 0.07 g g⁻¹) (Table 1). It is likely that more carbon has been deviated for cell growth and maintenance, which is in accordance with the higher growth yield (0.21 ± 0.02 g g⁻¹) obtained in experiment B. In fact, by controlling the pH with ammonium hydroxide, cell growth was faster and a higher cell density was reached, which required more substrate to fulfill the cell metabolism needs.

These results demonstrate that the DO-stat strategy was successful for feeding UCO to the culture, resulting in improved volumetric productivity. A huge advantage of this strategy is that it allows for the substrate to be automatically fed to the culture as a function of parameter that is measured online (i.e., the DO concentration). This strategy has been reported in some studies for the production of different PHA polymers. Faezah et al. [11] reported the use of the DO-stat mode to feed oleic acid and/or γ-butyrolactone to Cupriavidus sp. USMAA1020, as a strategy to regulate the molar fraction of 4-hydroxybutyrate in the P(3HB-co-4HB) polymer. The same strategy was tested by Kim et al. [12] to feed fructose and/or γ-butyrolactone to C. necator ATCC 17699. A combination of pH-stat with DO-stat strategies was also proposed by Huschner et al. [30] for feeding a mixture of sodium salts of acetic, propionic, and butyric acids to C. necator H16. There are no reports on the use of the DO-stat strategy to feed UCO or other oils containing substrates as the sole carbon sources for the fed-batch cultivation of C. necator.

3.3. PHA Characterization

The polymer produced from UCO in this study was a 3-hydroxybutyrate homopolymer, poly(3-hydroxybutyrate), P(3HB) (Table 2), which is in accordance with the literature reports for polymers produced by C. necator when cultivated in oil-containing substrates as sole carbon sources, under different cultivation modes [5, 27–29, 31].

Fed-batch cultivation under the DO-stat mode (experiment B) resulted in a polymer with a molecular weight of 2.6 × 10⁵ g mol⁻¹ (Table 2). Values of the same order of magnitude were reported for P(3HB) synthesized by C. necator from UCO under batch conditions (2.6 × 10⁵ g mol⁻¹) [24] or pulse feeding (2.0 × 10⁵–20 × 10⁵ g mol⁻¹) [32]. The values are also similar to the polymer produced by the same culture from SCG oil under pulse feeding (2.3 × 10⁵–4.7 × 10⁵ g mol⁻¹) [33] and with a DO-stat mode [10].

A lower molecular weight value (0.6 × 10⁵ g mol⁻¹) was obtained for the P(3HB) produced in experiment A, with UCO exponential feeding (Table 2). This low value might be related to the prolonged cultivation time (96 hours). According to Budde et al. [34], PHA is continuously turned over by C. necator, which is accompanied by a decrease in average polymer molecular mass. This might justify the observed decrease in the average molecular weight given the long cultivation time of the experiment. On the other hand, the polydispersity index (PDI) of the polymers was found to be lower (1.2-1.6) than those reported for P(3HB) obtained from SCG oil (2.2-2.5) [33], meaning that the P(3HB) produced in this study was highly homogeneous.

Thermal properties were also assessed after polymer extraction and purification. A melting temperature of 174°C was determined for the polymers produced in experiments A and B (Table 2). Similar values (172-173°C) were reported for P(3HB) produced from UCO [24], margarine waste [27], and SCG oil [10]. The glass transition temperatures (3.0-4.0°C) were also within the range of values reported for P(3HB), 3.0-8.4°C. According to Laycock et al. [35], melting and glass transition temperatures of P(3HB) typically range between 162-181 and −4 and 18°C, respectively, depending on the bacterial strain, carbon source, polymer extraction and purification procedures, etc.

The polymer’s crystallinity was 65 and 52% for experiments A and B, respectively. The simultaneous detection of a glass transition and melting, which are thermal events associated, respectively, with the amorphous and crystalline regions in the polymer, shows that the P(3HB) produced in both experiments is a semicrystalline material. The crystallinity of the polymer depends on many factors, including its chemical structure, intermolecular interactions, and processing conditions, but it is commonly between 55 and 80% [35], roughly including the crystalline degrees determined for the produced P(3HB).

4. Conclusions

Improved P(3HB) volumetric productivity was obtained with the implementation of a DO-stat strategy to feed UCO to C. necator DSM 428 during the fed-batch phase of a bioreactor cultivation. This strategy was tested for the first time, and it resulted in adequate substrate feeding for cell growth and polymer synthesis. A high molecular weight homogeneous homopolymer, with thermal properties similar to the P(3HB) synthesized by C. necator under different cultivation conditions and substrates, was obtained. The results of these preliminary studies show that the DO concentration can be used as a reliable online parameter for the automatic feeding of UCO to C. necator for P(3HB) production and can be used to improve the overall economic viability of the process.

Data Availability

The data used to support the findings of this study are available from the corresponding author upon request.

Conflicts of Interest

The authors declare that there is no conflict of interest regarding the publication of this paper.

Acknowledgments

This work was supported by the Unidade de Ciências Biomoleculares Aplicadas (UCIBIO) and Associated Laboratory for Sustainable Chemistry - Clean Processes and Technologies (LAQV), which are financed by national funds from FCT/MEC (UID/Multi/04378/2013 and UID/QUI/50006/2013, respectively) and co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728 and POCI-01-0145-FEDER-007265, respectively), exploratory project grant IF/00589/2015 attributed within the 2015 FCT Researcher Program, and fellowship SFRH/BD/72142/2010.

References

G. Kaur, A. K. Srivastava, and S. Chand, “Advances in biotechnological production of 1,3-propanediol,” Biochemical Engineering Journal, vol. 64, pp. 106–118, 2012.
View at: Publisher Site | Google Scholar
G. Kaur and I. Roy, “Strategies for large-scale production of polyhydroxyalkanoates,” Chemical and Biochemical Engineering Quarterly, vol. 29, no. 2, pp. 157–172, 2015.
View at: Publisher Site | Google Scholar
C. Peña, T. Castillo, A. García, M. Millán, and D. Segura, “Biotechnological strategies to improve production of microbial poly-(3-hydroxybutyrate): a review of recent research work,” Microbial Biotechnology, vol. 7, no. 4, pp. 278–293, 2014.
View at: Publisher Site | Google Scholar
J. M. B. T. Cavalheiro, M. C. M. D. de Almeida, C. Grandfils, and M. M. R. da Fonseca, “Poly(3-hydroxybutyrate) production by Cupriavidus necator using waste glycerol,” Process Biochemistry, vol. 44, no. 5, pp. 509–515, 2009.
View at: Publisher Site | Google Scholar
J. G. da Cruz Pradella, J. L. Ienczak, C. R. Delgado, and M. K. Taciro, “Carbon source pulsed feeding to attain high yield and high productivity in poly(3-hydroxybutyrate) (PHB) production from soybean oil using Cupriavidus necator,” Biotechnology Letters, vol. 34, no. 6, pp. 1003–1007, 2012.
View at: Publisher Site | Google Scholar
E. Akaraonye, C. Moreno, J. C. Knowles, T. Keshavarz, and I. Roy, “Poly(3-hydroxybutyrate) production by Bacillus cereus SPV using sugarcane molasses as the main carbon source,” Biotechnology Journal, vol. 7, no. 2, pp. 293–303, 2012.
View at: Publisher Site | Google Scholar
A. Hafuka, K. Sakaida, H. Satoh, M. Takahashi, Y. Watanabe, and S. Okabe, “Effect of feeding regimens on polyhydroxybutyrate production from food wastes by Cupriavidus necator,” Bioresource Technology, vol. 102, no. 3, pp. 3551–3553, 2011.
View at: Publisher Site | Google Scholar
E. Grothe and Y. Chisti, “Poly(β-hydroxybutyric acid) thermoplastic production by Alcaligenes latus: behavior of fed-batch cultures,” Bioprocess Engineering, vol. 22, no. 5, pp. 441–449, 2000.
View at: Publisher Site | Google Scholar
M. S. I. Mozumder, H. De Wever, E. I. P. Volcke, and L. Garcia-Gonzalez, “A robust fed-batch feeding strategy independent of the carbon source for optimal polyhydroxybutyrate production,” Process Biochemistry, vol. 49, no. 3, pp. 365–373, 2014.
View at: Publisher Site | Google Scholar
M. V. Cruz, A. Paiva, P. Lisboa et al., “Production of polyhydroxyalkanoates from spent coffee grounds oil obtained by supercritical fluid extraction technology,” Bioresource Technology, vol. 157, pp. 360–363, 2014.
View at: Publisher Site | Google Scholar
A. N. Faezah, A. Rahayu, S. Vigneswari, M. I. A. Majid, and A. A. Amirul, “Regulating the molar fraction of 4-hydroxybutyrate in poly(3-hydroxybutyrate-co-4-hydroxybutyrate) by biological fermentation and enzymatic degradation,” World Journal of Microbiology and Biotechnology, vol. 27, no. 10, pp. 2455–2459, 2011.
View at: Publisher Site | Google Scholar
J. S. Kim, B. H. Lee, and B. S. Kim, “Production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) by Ralstonia eutropha,” Biochemical Engineering Journal, vol. 23, no. 2, pp. 169–174, 2005.
View at: Publisher Site | Google Scholar
W. S. Ahn, S. J. Park, and S. Y. Lee, “Production of poly(3-hydroxybutyrate) by fed-batch culture of recombinant Escherichia coli with a highly concentrated whey solution,” Applied and Environmental Microbiology, vol. 66, no. 8, pp. 3624–3627, 2000.
View at: Publisher Site | Google Scholar
S. Kulpreecha, A. Boonruangthavorn, B. Meksiriporn, and N. Thongchul, “Inexpensive fed-batch cultivation for high poly(3-hydroxybutyrate) production by a new isolate of Bacillus megaterium,” Journal of Bioscience and Bioengineering, vol. 107, no. 3, pp. 240–245, 2009.
View at: Publisher Site | Google Scholar
Z. Sun, J. A. Ramsay, M. Guay, and B. A. Ramsay, “Automated feeding strategies for high-cell-density fed-batch cultivation of Pseudomonas putida KT2440,” Applied Microbiology and Biotechnology, vol. 71, no. 4, pp. 423–431, 2006.
View at: Publisher Site | Google Scholar
A. Rathinasabapathy, B. A. Ramsay, J. A. Ramsay, and F. Pérez-Guevara, “A feeding strategy for incorporation of canola derived medium-chain-length monomers into the PHA produced by wild-type Cupriavidus necator,” World Journal of Microbiology and Biotechnology, vol. 30, no. 4, pp. 1409–1416, 2014.
View at: Publisher Site | Google Scholar
J. Lee, S. Y. Lee, S. Park, and A. P. J. Middelberg, “Control of fed-batch fermentations,” Biotechnology Advances, vol. 17, no. 1, pp. 29–48, 1999.
View at: Publisher Site | Google Scholar
S. J. Park, W. S. Ahn, P. R. Green, and S. Y. Lee, “Production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by metabolically engineered Escherichia coli strains,” Biomacromolecules, vol. 2, no. 1, pp. 248–254, 2001.
View at: Publisher Site | Google Scholar
F. Wang and S. Y. Lee, “Poly(3-hydroxybutyrate) production with high productivity and high polymer content by a fed-batch culture of Alcaligenes latus under nitrogen limitation,” Applied Environmental Microbiology, vol. 63, no. 9, pp. 3703–3706, 1997.
View at: Google Scholar
K. Shantini, A. R. M. Yahya, and A. A. Amirul, “Influence of feeding and controlled dissolved oxygen level on the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer by Cupriavidus sp. USMAA2-4 and its characterization,” Applied Biochemistry and Biotechnology, vol. 176, no. 5, pp. 1315–1334, 2015.
View at: Publisher Site | Google Scholar
S. H. Lee, D. H. Oh, W. S. Ahn, Y. Lee, J.-i. Choi, and S. Y. Lee, “Production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by high-cell-density cultivation of Aeromonas hydrophila,” Biotechnology and Bioengineering, vol. 67, no. 2, pp. 240–244, 2000.
View at: Publisher Site | Google Scholar
S. Y. Lee, H. H. Wong, J. Choi, S. H. Lee, S. C. Lee, and C. S. Han, “Production of medium-chain-length polyhydroxyalkanoates by high-cell-density cultivation of Pseudomonas putida under phosphorus limitation,” Biotechnology and Bioengineering, vol. 68, no. 4, pp. 466–470, 2000.
View at: Publisher Site | Google Scholar
M. V. Cruz, M. C. Sarraguça, F. Freitas, J. A. Lopes, and M. A. M. Reis, “Online monitoring of P(3HB) produced from used cooking oil with near-infrared spectroscopy,” Journal of Biotechnology, vol. 194, pp. 1–9, 2015.
View at: Publisher Site | Google Scholar
L. Martino, M. V. Cruz, A. Scoma et al., “Recovery of amorphous polyhydroxybutyrate granules from Cupriavidus necator cells grown on used cooking oil,” International Journal of Biological Macromolecules, vol. 71, pp. 117–123, 2014.
View at: Publisher Site | Google Scholar
F. Freitas, V. D. Alves, J. Pais et al., “Production of a new exopolysaccharide (EPS) by Pseudomonas oleovorans NRRL B-14682 grown on glycerol,” Process Biochemistry, vol. 45, no. 3, pp. 297–305, 2010.
View at: Publisher Site | Google Scholar
M. V. Cruz, F. Freitas, A. Paiva et al., “Valorization of fatty acids-containing wastes and byproducts into short- and medium-chain length polyhydroxyalkanoates,” New Biotechnology, vol. 33, no. 1, pp. 206–215, 2016.
View at: Publisher Site | Google Scholar
C. Morais, F. Freitas, M. V. Cruz, A. Paiva, M. Dionísio, and M. A. M. Reis, “Conversion of fat-containing waste from the margarine manufacturing process into bacterial polyhydroxyalkanoates,” International Journal of Biological Macromolecules, vol. 71, pp. 68–73, 2014.
View at: Publisher Site | Google Scholar
P. Kahar, T. Tsuge, K. Taguchi, and Y. Doi, “High yield production of polyhydroxyalkanoates from soybean oil by Ralstonia eutropha and its recombinant strain,” Polymer Degradation and Stability, vol. 83, no. 1, pp. 79–86, 2004.
View at: Publisher Site | Google Scholar
K. S. Ng, W. Y. Ooi, L. K. Goh, R. Shenbagarathai, and K. Sudesh, “Evaluation of jatropha oil to produce poly(3-hydroxybutyrate) by Cupriavidus necator H16,” Polymer Degradation and Stability, vol. 95, no. 8, pp. 1365–1369, 2010.
View at: Publisher Site | Google Scholar
F. Huschner, E. Grousseau, C. J. Brigham et al., “Development of a feeding strategy for high cell and PHA density fed-batch fermentation of Ralstonia eutropha H16 from organic acids and their salts,” Process Biochemistry, vol. 50, no. 2, pp. 165–172, 2015.
View at: Publisher Site | Google Scholar
B. Füchtenbusch, D. Wullbrandt, and A. Steinbüchel, “Production of polyhydroxyalkanoic acids by Ralstonia eutropha and Pseudomonas oleovorans from an oil remaining from biotechnological rhamnose production,” Applied Microbiology and Biotechnology, vol. 53, no. 2, pp. 167–172, 2000.
View at: Publisher Site | Google Scholar
R. A. Verlinden, D. J. Hill, M. A. Kenward, C. D. Williams, Z. Piotrowska-Seget, and I. K. Radecka, “Production of polyhydroxyalkanoates from waste frying oil by Cupriavidus necator,” AMB Express, vol. 1, no. 1, p. 11, 2011.
View at: Publisher Site | Google Scholar
S. Obruca, S. Petrik, P. Benesova, Z. Svoboda, L. Eremka, and I. Marova, “Utilization of oil extracted from spent coffee grounds for sustainable production of polyhydroxyalkanoates,” Applied Microbiology and Biotechnology, vol. 98, no. 13, pp. 5883–5890, 2014.
View at: Publisher Site | Google Scholar
C. F. Budde, S. L. Riedel, L. B. Willis, C. Rha, and A. J. Sinskey, “Production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) from plant oil by engineered Ralstonia eutropha strains,” Applied and Environmental Microbiology, vol. 77, no. 9, pp. 2847–2854, 2011.
View at: Publisher Site | Google Scholar
B. Laycock, P. Halley, S. Pratt, A. Werker, and P. Lant, “The chemomechanical properties of microbial polyhydroxyalkanoates,” Progress in Polymer Science, vol. 38, no. 3-4, pp. 536–583, 2013.
View at: Publisher Site | Google Scholar

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Copyright © 2019 Madalena V. Cruz et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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