Abstract
Polyhydroxyalkanoates (PHAs) are biological plastics that are sustainable alternative to synthetic ones. Numerous microorganisms have been identified as PHAs producers. They store PHAs as cellular inclusions to use as an energy source backup. They can be produced in shake flasks and in bioreactors under defined fermentation and physiological culture conditions using suitable nutrients. Their production at bioreactor scale depends on various factors such as carbon source, nutrients supply, temperature, dissolved oxygen level, pH, and production modes. Once produced, PHAs find diverse applications in multiple fields of science and technology particularly in the medical sector. The present review covers some recent developments in sustainable bioreactor scale production of PHAs and identifies some areas in which future research in this field might be focused.
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History
Polyhydroxyalkanoates (PHAs) comprise a range of different biodegradable biopolymers—bioplastics or bio-polyesters synthesized by various microorganisms [1]. They were discovered, in 1888, by Beijerinck as cellular inclusions of bacteria and other microorganisms [2]. In 1926 Maurice Lemoigne, a French scientist, was the first to synthesize a PHA (named poly-3-hydroxybutyric acid) using a bacterial strain of Bacillus megaterium [3]. Later on, in 1958, Macrae and Wilkinson claimed that the bacterial cells play a role as cellular factories for PHAs synthesis and storage [4]. Imperial Chemical Industries (ICI), UK, initiated commercial production of PHAs in 1961. In 1980, ICI synthesized a copolymer of PHA via fermentation, while in 1983, De Smet et al. [5] reported synthesis of a medium chain length polyhydroxyalkanoate (mcl-PHAs) which in turn became a milestone in the synthesis of PHAs. Also in 1980, the researchers had started investigations on the physiochemical properties of PHAs [6] and later on in 1990, scientists used them for the first time in the biomedical field. By 2010, the PHAs market had reached up to 1010 US$ with a growth of 170–180 Kt, annually [7].
Introduction
Plants have always been used as a major source of polymers such as starch, cellulose, flax and rubber for thousands of years. During the past few decades, plant polymers had extensively been replaced by petrochemical based polymers on mass scale production [8]. At the end of the twentieth century, it was realized that these synthetic polymers are non-biodegradable, persistent, and continually accumulate in the environment which ultimately represents a threat to life on Earth. During the past two decades environmental protection agencies had started awareness programs for both the public, in general, and manufacturers, in specific, on the importance of preserving and protecting the nature from toxic products and hazardous wastes [9]. These concerns had compelled scientists and researchers to further explore new approaches to synthesize and use eco-friendly bioplastics such as PHAs as alternative materials to plastics [10].
PHAs are biodegradable linear thermoplastic polyesters, which can be used as alternative polymers to synthetic ones. They are synthesized by different bacterial strains (both Gram positive and Gram negative) cultivated on different carbon sources like sugars, alkanoic acids, alkanes, alkenes and other renewable carbon sources [11]. During the last two decades, PHAs have attained much attention because of their diverse features such as hydrophobicity, elastomeric nature and biodegradability under a wide range of environmental conditions. They also constitute a natural part of renewable carbon cycle and are used as alternatives to synthetic polyesters. Nevertheless, there are still some limitations in their bulk scale production including low yields and high production costs [12].
Some 150 different types of PHAs congeners with different structures (varying side chains or functional groups) and properties had been synthesized using different microbial strains. PHAs can be synthesized though microbial fermentations both at laboratory and pilot plant scales followed by appropriate downstream processing [13]. They are classified according to their chemical unit structure. The polymers containing repeating units with 3–5 carbon atoms are known as short chain length PHA (scl-PHA) and a polymer with repeating units of 6–13 carbon atoms are known as medium chain length PHA (mcl-PHA), while those with more than 13 carbon atoms are known as long chain length PHA (lcl-PHA). A copolymer like poly(3-hydroxybutyrate-co-4-hydroxyhexanoate) could be produced by simply mixing poly(3-hydroxybutyrate), a scl-PHAs, and poly(4-hydroxyhexanoate), a mcl-PHAs monomers under desirable fermentation conditions [14]. A general structure of PHAs is shown in Fig. 1.
PHAs find a large number of applications in different fields of science and technology. For instance, a blend of poly(hydroxybutyrate) P(HB) and poly(hydroxyoctanoate) P(HO) is used as food additive which had been approved by the US food and drug authority (FDA) [15]. PHB has also been used as a component in making plastic accessories, coating paper, biodegradable bottles, electronic accessories, pharmaceuticals, garments, upholstery, and packaging materials [16]. In the field of medicine, PHAs allow us to control the drug release and targeted drug action whereas PHB had been used to produce different medical devices [17]. A drug delivery system can be a sustainable way to administrate certain drugs by minimizing their toxic effects. An mcl-PHA had been used to administrate drugs through a transdermal drug delivery system. There are three basic components of a drug delivery system i.e., a carrier, a target moiety and an active part of the drug. Genetically engineered bacterial strains produce PHAs and allow us to utilize them in vascular grafting, blood vessel and heart valve development. P(3-HAs) has also been suggested for use as a component of biofuels to reduce global warming [18]. The wide range of applications of PHAs in different fields of life is listed in Table 1.
In this review, we present recent developments in the bacterial production of PHAs on various carbon sources. We focused on the challenges in PHAs production in different types of bioreactors and operational regime using various bacterial strains. A particular emphasis is dedicated to current advantages and limitations of different bioreactor scale setup while concentrating on the efforts devoted to upgrade these processes to industrial scales. Finally, we endeavour to identify the most suitable downstream processing technology.
Financial obstacles in PHAs commercialization
Commercialization of a product mainly depends on its production cost, potential applications and market requirements. In the past decades, magnificent efforts had been made to improve PHAs yield both at laboratory and pilot plant scales [7]. The main challenges in the commercialization of PHAs had been their high production cost and low product yields. PHAs produced from different sources require different tools like fermenter, autoclave etc. with different fermentation approaches which increase the production cost ultimately limiting their commercialization. The inability to achieve optimal bacterial growth conditions and production yields for PHAs had been a major disadvantage in this context [19]. In addition, PHAs have some disadvantages in their mechanical properties including; incompatibility with conventional thermal processing, limited functionalities, susceptibility to thermal degradation and production cost which limits their competitiveness and application as a feasible biomaterial when compared with synthetic plastics. The possibility of commercialization of PHAs might be enhanced using mixed microbial culture (MMC) with cheap carbon sources like waste frying oils and crude oils. The main obstacle in the commercialization of PHAs had been their production cost which could be reduced using edible oils as carbon sources with non-sterilized fermentation conditions [11, 20].
Sustainable solutions for PHAs production
The economics of PHA production mainly depends upon substrate cost and compatibility to use product in similar to those produced using expensive growth substrate, efficiency of production, and downstream processing [21]. Recent studies on PHAs production not only consider the sustainability of polymer, but also its cost-effectiveness. Nitrogen and carbon sources get significant share in the production cost. Using cheap nitrogen sources such as urea and sodium nitrate reduces the cost of production significantly [10]. The utilization of industrial wastes as carbon source to produce a competitive cost effective PHA has also been reported as a promising approach for decreasing overall costs [2]. The economics of whole process is based on three approaches; large scale aerobic batch fermentation, use of cheap carbon source soybean and other waste frying oils, and simplification of the downstream recovery process to achieve an economical PHAs production approach [22]. PHAs bacterial sources and their chemical structures are listed in Table 2.
Other ways to meet financial barrier is though value addition by inserting new functionalities in PHAs to overcome their inherent limitations, improve their desirable properties and enhance potential applications [20]. This is usually achieved by blending the bio-polyesters with other natural biodegradable polymers, including cellulose derivatives, poly lactic acid (PLA), starch, lignin and poly(caprolactone). The introduction of different functional groups via chemical routes has been described with regard to the two main synthesis approaches, graft co-polymerization and block co-polymerization [23]. Grafting of different functional groups on side chains of linear bio-polyesters introduces many additional properties, which makes them sustainable with desired properties and controlled polymer structures. For instance, grafting of chitosan on PHA produces a copolymer P(H-co-chitosan) which is a sustainable biopolymer with some important applications in medical field. A cost effective production has been reported, carried out using MMC, under unsterilized conditions [20, 24].
PHA production in shake flasks
Shake flask fermentations are normally carried out in laboratories mainly to explore feasibility. Here, the information is often vogue and the experimental expenditures are high with large number of trails typically required to identify a set of optimum fermentation conditions. Such fermentations are usually carried out in Erlenmeyer flasks provided with minimal media, carbon source and any precursors at specific pH levels and incubated at certain temperature and shaking speed for a certain period of time. During shake flask fermentations, the process is followed up based on certain parameters such as carbon source or any other nutrients utilization, biomass formation and fermentation kinetics. In the previous studies, bacterial production of PHAs had mostly been carried out using glucose as carbon source. Later on, Pseudomonas sp. grown on MMC had been employed with volatile fatty acids (VFAs) as carbon source and also used as precursor for PHAs production with higher productivity [25]. The optimized fermentation conditions had been used to produce PHAs with efficient recovery. Different ratios of carbon and nitrogen sources had been used to optimize product yields. An ammonia free fermentation media were used to accumulate PHAs with VFAs. During the first phase of fermentation, glucose was used as carbon source after inoculating the media in Erlenmeyer flask and cell biomass was harvested via centrifugation at 6000 rpm and 4 °C with 20–40% g/g of product yield. The shake flask study was used to determine the feasibility of fermentation process with different parameters. The main disadvantage of shake flask investigations is the wide variation in the quality of produced product [26].
Kinetics of PHA production
The fermentation kinetics were previously investigated by determining the product yields with respect to substrate consumption. The kinetics of production determines how much PHAs cell dry mass (CDM) are produced with respect to time and expressed as g/L/h. A linear increase in PHAs yield was observed following a zero order reaction kinetics until the carbon source growth substrate was depleted [27]. The kinetics of PHAs production for different bacterial strains with large number of substrates had been established e.g., Azotobacter beijerinckii with glucose as growth substrate produced 0.09 g/L/h [28]. Burkholderia cepacia bacterial strain was grown on glycerol as energy substrate produced 0.103 g/L/h [29], Ralstonia eutropha with butyrate, propanoic, lactic and acetate acid as growth substrates produced 0.001–0.037 g/L/h [30]. Alcaligenes eutrophus grown on potato starch and saccharified wastes as growth substrates produced 1.5 g/L/h [31]. Hydrogenophaga pseudoflava with sucrose and lactose as growth substrates produced 0.02–0.12 g/L/h [32]. Pseudomonas frederiksbergensis used terephthalic acid as growth substrate and produced 0.004 g/L/h [33], Pseudomonas putida KT 224 produced 0.006 g/L/h of PHAs using glucose as growth substrate [34]. Haloferax mediterranei produced 0.05–0.2 g/L/h of PHAs using vinasse as growth substrate [35] and the same bacterial strain with glycerol produced 0.12 g/L/h [36]. Further kinetics of PHAs production with different bacterial sources, substrates and production scales are presented in Table 3.
Bioreactor scale production of PHAs
Bioreactor scale production of biopolymers is the most suitable production technique on industrial scale using large working volume and avoiding many restrictions encountered under shake flask conditions. Because, PHAs biosynthesis constitutes a multiple phase process, both the feeding strategy and bioreactor operation mode need to be adapted via optimization of operation conditions. Industrial bioreactor scale production of PHAs can also be operated under both fed batch and continuous fed batch feeding strategies. In the following text, various types of bioreactors and their conditions have been described and analyzed for enhanced PHA production opportunities based on the available literature.
Single batch production
During batch production, desirable amounts of different nutrients are added in the fermentation media. The single batch production process is simple but gives low productivity. The maximum allowed concentration of both carbon and nitrogen sources are added at the beginning of batch fermentation processes which restrict nutrient addition during PHAs production. In batch productions of PHAs, the amount of nitrogen source are typically added at 0.2–5 g/L and carbon source at 1–30 g/L at the beginning of fermentation process [37]. Moreover, the excess amount of carbon source added in the presence of other growth limiting nutrients such as N, P, S, and K. The single batch fermentation process is analogous to other PHAs production processes with minimum conversion toward biomass, CO2, PHAs and other metabolites; thus resulting in low overall conversion of growth substrate carbon source to PHAs cell biomass yield. Considering hypothetical biotransformation of biogenic carbon to PHAs, the yield was calculated to be below 0.4 g/g which indicates that such processes are not economically feasible. The single batch fermentation scale production of PHAs, therefore, suffers limitations which need to be addressed. The main set back of batch scale production is that the nutrients are not further provided which inhibited the bacterial growth, thus restricting the production of PHAs. If the fermentation period is extended above the optimum period (e.g., 7 days) then the degradation of PHAs starts. A list of single batch PHAs production processes using different bacterial strains is shown in Table 4.
Sequence batch fermentation production processes for PHAs
In sequence batch bioreactor experiments, two or more reactors are connected in series to perform different functions simultaneously to produce PHAs. Albuquerque et al. [38] reported a study of two-configurations system used to produce PHAs from organic activated sludge waste as substrate. In the first configuration, nitrogen (as ammonium) was limited by simple conversion to nitrate and bacteria started to respond these conditions and produced PHAs. Both aerobic feasting stage (ammonia conversion to nitrate) and anoxic famine stage (denitrifying) conditions drive internally stored PHAs as carbon source [38].
First configuration: In the first configuration, continuous feeding with fix interval of 5 min had been carried throughout the fermentation process until desired cell mass was achieved. The fermentation medium was allowed to settle down for 15 min and then fermentation process was further run under aerobic conditions for an hour which was followed by anaerobic fermentation condition to produce PHAs [38].
Second configuration: Here, the fermentation process had been carried in two bioreactors which were connected in a series. In one of the bioreactors, nitration took place and in second bioreactor, biomass is optimized. The nitration was carried out under aerobic fermentation conditions. After that, the fermentation medium was further processed in the second reactor under aerobic feast and anoxic famine reaction conditions to accumulate high cell mass for PHAs inclusions [38].
Fed batch production of PHAs
During fed batch cultivation, the substrate is added through a pulse feeding when its concentration drops below its optimum value without removing any culture media. In the case of PHAs production, both carbon and nitrogen sources are added at periodic intervals according to their consumption by microorganisms. In fed batch fermentation process, these substrates are added to avoid any depletion during the production process until the desired cell biomass yield is achieved. Some researchers used fed batch process to produce 0.41 g/g of PHAs using Pseudomonas putida KT 2440 through feeding octanoic acid as a carbon source [39].
The fed batch production had also been used to examine the effect of different carbon to nitrogen (C/N) ratios on product yield. For that purpose, nutrient enrich media had been prepared using the following recipe: MgCl2·6H2O, KCl, Fe2+, NaHCO3, C6H12O6, NaCl, CaCl2, MgSO4·7H2O, NH4Cl, KH2PO4, NaBr and 1 mL of trace elements solution. An aliquot amount of 60 mL of inoculum was added in the nutrient enrich media of sequence batch reactor after 48 h of cultivation. The pH of fermentation media was maintained at 7.5–7.8 using 1 M NaOH and 1 M HCl solutions. The amount of dissolved oxygen in the nutrients enriched media was 100% at 40 °C for 72 h. The maximal substrate consumption, CDM and PHAs yield decreased with increase in C/N ratio. The yields of 0.970, 0.955, and 0.890 g/L of CDM were reported, respectively, using 0.47, 1.95, and 7.64 g/L of nitrogen concentrations [13].
Continuous production process
The fermentative production processes are typically carried out in continuous stirred tank bioreactors (CSTR) where process parameters like agitation speed, air flow, pH, substrate, product concentration and temperature are controlled to investigate PHAs yield. The PHAs production using MMC under continuous feeding strategies had been established. Such a process produced PHAs with improved composition on monomeric level and results in a more sustainable process. Continuous PHAs production process is clearly better than the batch production in terms of composition, flexibility and productivity of bio-polyesters [38]. Pseudomonas putida had also been reported to produce 80% (g/g) PHAs, at both shake flask and bioreactor scale fermentations. The pH of fermentation media at lab scale were kept neutral using 2M NaOH and 4M H2SO4 solutions at 30 °C, 250 rpm and 20% of dissolved oxygen (DO) throughout fermentation process. At pilot scale, CSTR fermentation with different working volumes has been carried out to produce PHAs. In CSTR, the fermentation parameters were kept at pH 7 (using 30% H2SO4and 2 M NaOH solutions), temperature 30 °C, rate of agitation 100–1000 rpm and aeration 0–30 L/min to produce 0.53 g/g of PHAs. The continuous production of PHAs had been carried out at single, double and multiple stages to facilitate the bacteria during fermentation process. In continuous production, each and every aspect regarding the fermentation process proceeded more accurately than fed batch mode. In fed batch process, nutrients were further added once their depletion inside fermention media to produce higher PHAs yields [40].
One stage continuous stirred tank reactor
The efficiency of a single stage continuous stirred tank reactor in terms of flexibility in polyester composition and productivity. The schematic diagram used is shown in Fig. 2a. Single stage continuous stirred tank reactors (single SCSTRs) are not productive for PHAs cell biomass production, because intercellular products of secondary metabolism only boost PHAs production under limited nutrients growth conditions. Hence, it is not possible to continuously supply these nutrients throughout the fermentation process, that’s why optimization is needed for both intracellular PHAs mass and high biomass formation. Atlic et al. [41] reported on the validity of single SCSTR using propionic acid as growth substrate, Cupriavidus necator bacterial strain and glucose as carbon source producing 0.33 g/g P(3-HB-co-3-HV) co-polyesters. The amount of cell biomass and cell dry mass produced in the single SCSTR was not competitive with fed batch process using the same bacterial production strain [41].
Two-stage continuous stirred tank reactor
The two-stage continuous stirred tank reactor fermentation process is used to produce PHAs with maximum yield which is not possible under shake flask conditions and with single SCSTR. A schematic diagram for this is shown in Fig. 2b. During two SCSTR fermentation process, different functions are carried out in two bioreactors, simultaneously. Regarding the cell biomass accumulation for the bacterial strain and substrate ratio, it had been observed that autocatalytic growth was much higher than the PHAs accumulation process. Due to this reason, it is impossible to carry out such process with sufficient productivity at single SCSTR scale. Therefore, it was consequently demonstrated that two-stage continuous stirred tank reactor (two SCSTR) process is better for such type of production. Here, in the first CSTR, a higher amount of cell biomass could be produced with higher density and continuously transferred to second CSTR, where large amount of carbon source are provided continuously, under limiting nitrogen substrate to stimulate PHA production [42].
The efficiency of two SCSTR has been demonstrated in the literature. For instance, the bacterial strain of Alcaligenes lata had been used to produce 0.38 g/g of P(3-HB) and 0.55 g/g of P(3-HB-co-3-HV) a copolymer using propionic acid (as precursor), residual sucrose (as carbon source) in the minimal nutrients media. In the first CSTR, the nitrogen source and precursor were completely utilized by the bacterial cells. The fermented broth containing residual sucrose was continuously transferred to the second CSTR, but no additional nutrients were added to facilitate the conversion of sucrose as growth substrate to increase PHAs content in the bacterial cell biomass [43]. Hence, in this phase, bacterial cell was allowed to complete the fermentation process to maximize the PHAs yield while depleting the sucrose (i.e., as carbon source). In the context of two SCSTR, the optimized bacterial strain and substrates were used to produce different scl-PHA, mcl-PHA as homo and hetero polymers. Some researchers cultivated the bacterial strain of P. putida GPo1 in two SCSTR with gaseous substrate n-octane as carbon source and produced a blend of two different polymers with block structures [21]. The block copolymer of poly(3-hydroxy-10-undecenoate-co-3-hydroxy-8-nonenoate-co-3-hydroxy-6-heptenoate) was produced at 0.63 g/g. This observation is in contrast to other reports, which claimed that block PHAs copolymers were only produced when adding sufficient amount of growth substrate during the intracellular accumulation of PHAs [44].
Multi stage continuous stirred tank reactor
In the case of multi stage CSTR, three or more bioreactors are connected with each other using plastic tubing. The schematic diagram of continuous stirred tank bioreactor is shown in Fig. 2c. PHAs can be produced with exponential growth rate under continuous growth conditions as used in two SCSTR. The culture condition can be used throughout the process until the desired cell biomass is achieved. The concentrated substrate was loaded to avoid any dilution of nutrients during continuous production condition in three SCSTR [45].
Factors affecting the production of PHAs
The bacterial production of PHAs through fermentation is influenced by different factors such as, strain used, growth substrate, C/N ratio, pH, DO and sodium chloride which are further elaborated in the following subsections.
Bacterial strains for PHAs production
More than 300 types of different Gram positive, Gram negative and archaea bacteria had been reported to produce PHAs. Different carbon source and CDM, PHAs yields and production kinetics are shown have been reported in Table 3.
Growth substrates
The carbon source is the most crucial and major factor during bacterial production of cellular metabolites like PHAs. Several types of carbon sources such as alkanes, polysaccharides, glycerides, acids, edible oils, gases, industrial waste, agricultural waste, carbohydrates, alcohols, petroleum products, animal waste, benzene and its derivatives had been investigated for PHAs production. The carbon source is used as basic food component to fulfil energy requirements and to act as biogenetic substrate for microbial growth and precursor for PHAs biosynthesis. In the above context, there are three basic functions for the carbon source within the bacterial cells such as cell maintenance, cell biomass synthesis and polymerization of PHAs molecules [46]. The small amount of growth substrate was used to restrict the molecular size of polymer during fermentation. Because bacteria have thick and rigid cell wall, it does not allow large polymeric molecule to be transported into cells. Different concentrations of glucose (as carbon source) had been used to optimize cellular growth and product yield. The results demonstrated a proportional increase up to a point after which glucose exhibited an inverse effect on microbial growth. The effect of growth substrates on PHAs yield is entirely based on nutrients media [47].
The selectivity of growth substrate not only depends upon their cost but also on their feasibility in the fermentation process. The selectivity of carbon source as growth substrate depends also on the bacterial strain used. The bacterial strain of P. aeruginosa shows compatibility for PHAs production with large number of growth substrates like oils, polysaccharide’s and acids [48].
Carbon sources as growth substrates generally belong to three main groups viz. triglycerol, sugars and hydrocarbons. The growth substrate used include carbohydrates such as sucrose [49], lactose [50], starch [51] and lignocellulose [52]. Other substrates such as ethyl alcohol [11], methanol [53], methane [54] and triglycerol containing compounds such as animal fat [55], plant oils [11], fatty acid [56], glycerol [57] and waste frying oil [58] had also been reported. Hydrocarbons had shown the highest affinity to produce PHAs including alkane, alkene, alkyne and a host of other hydrocarbons [59]. The use of fats, salts, ashes, whey as carbon sources had also been reported to have a large impact on PHAs yield hence decreasing the production costs up to 50%. Both culture conditions and the substrate have shown significant effects on both quality and quantity of the PHAs produced [60]. A number of different carbon sources with specific bacterial strains used in PHAs production are listed in Table 3.
Nitrogen sources and limitation
Several organic and inorganic nitrogen sources had been investigated for suitability and improvement of PHAs yield including different nitrogen sources such as amides [61], ammonium sulphate [26, 40], ammonium nitrate and ammonia nitrogen [40, 62], ammonium bicarbonate [63], ammonium carbonate, ammonium chloride [13], polyamide poly-γ-glutamate (PGA) [64], urea, nitrates [65] and sodium nitrate [27]. The amount of nitrogen sources (like organic and inorganic ones) had also shown linear relation with the bacterial growth, i.e., the number of bacterial cells increased with increasing nitrogen contents in the culture media. The PHAs stored inside bacterial cell and nitrogen source concentration had an inverse effect on cell biomass yield [66]. Lower concentration of nitrogen source showed better PHAs accumulation inside the bacterial cell, but higher values increased the biocatalyst activity to increase the number of bacterial cell in the fermentation media [38]. The effect of organic and inorganic nitrogen sources on PHAs production are listed in Table 5.
Carbon to nitrogen ratio
The effect of C/N ratio on PHA yield had also been reported elsewhere [68]. The concept of C/N ratio originated from the biological law according to which limitation of carbon and nitrogen sources controls the molecular size of PHAs and the number of bacterial cells. A continuous increase in C/N ratio promotes the accumulation of bacterial cell mass with inverse effect on growth kinetics (yield with respect to time) of PHAs [11]. The depletion of nitrogen in nutrient media promotes the accumulation of PHAs. The highest percentage of PHAs within cell dry mass was produced as 47.22% at 35 C/N ratio with Haloferax mediterranei [13]. In another study using C. necator, it was reported that the C/N ratio caused a prominent effect on PHAs composition and accumulation where most of the cell mass formation occurred during first 12 h of incubation [69]. The effect of C/N ratio on PHAs is determined from production kinetics of PHAs cell mass [68]. Increasing the nitrogen concentration had a positive effect on growth rate, however, negative on PHA yield. During fermentative production of PHAs, the excess amount of carbon source is typically used with limited nitrogen source. When investigating growth-associated production of PHAs using A. lata, it was observed that PHAs cell biomass yield weren’t enough until sufficient amount of both carbon and nitrogen sources were provided. This C/N ratio generates high active biomass productivity with sufficient PHAs yield [13].
Effect of pH
The pH value of extracellular environment greatly affects the bacterial enzymatic activity. A fermentation medium at neutral pH produces more PHAs than both under acidic and basic environments. In the case of pure culture media, any fluctuation in neutral pH slows down the fermentative process, with a reduction in cellular activities of enzyme, i.e., Pha C and Pha Z, thereby affecting the growth rate, and ultimately the cell biomass. The overall, the effect of pH on PHAs production was totally depended on the composition of media. A general trend of fluctuations in pH value is usually observed for neutral fermentation media. After loading with a substrate, the pH of a fermentation medium increases in the early stages of incubation, and then according reinstates [70]. A similar effect of pH fluctuation had also been observed both in continuous and fed batch fermentations [71].
Dissolved oxygen demand
The gaseous requirement in most of the cells is atmospheric oxygen, which is essential for the bio-oxidative respiration process. Oxygen plays a vital role in adenosine triphosphate (ATP) formation and produces energy which is utilizable in vital cellular activities. The dissolved oxygen (DO) acts as carrier gas in the fermentation medium during PHAs production [72]. Under the anaerobic conditions, the bacterial cells which lack the enzyme CoA for the respiration. Both metabolic activity and the amount of DO were reported to increase during continuous feeding and pulse feeding fermentation as process proceeded to PHAs production. In an MMC media, the amount of DO had reverse effect on metabolic activity in the bacterial cells [73]. This reversal effect of both pH and DO was observed in MMC when pH of the fermentative media was less than 8 [74].
Concentration of phosphorus
Phosphorus in the fermentation media plays an important role in the synthesis of proteins, carbohydrate and fats with some additional roles in cells repair and maintenance of growth rate. The amount of phosphorus present in the nutrients enrich media also acts as buffer to resist any change in neutral pH throughout the fermentation process. Various concentrations of phosphorus had been used to optimize the fermentation condition with respect to product yield. The optimized concentration of phosphorus should be used because at higher concentration a reverse effect on product yield had been reported [75, 76].
Production with PHAs precursor addition
The PHA co-polyesters are biosynthesized via metabolic pathways using a variety of monomers as precursors. Over 150 different types of precursors have been reported for PHA co-polyesters formation. However, most of them were expensive, which limit their utilization at industrial scale. The chemical nature and concentration of these precursors have to be controlled carefully to avoid any inhibitions, toxicity and to ensure the formation desired product. For instance, butyrate, citrate and acetate species can act as precursors for PHAs production. Likewise, R. eutropha showed compatibility with specific precursors of propionate for poly-3-hydroxybutyrate-co-3-hydroxyvalerate, and ϒ-butyrolactone, 1,4-butanediol and 4-hydroxybutyrate for poly(3-hydroxybutyrate-co-4-hydroxybutyrate). Fatty acids with odd number carbon atoms have also been used as precursors in PHA co-polymers production [77]. Different organic acids such as propionic acid and alcohols like methanol had also been used as precursors in the biosynthesis of PHA copolymers. For example, the late addition of propionic acid to the fermentation media while using glucose as main substrate produced poly-3-hydroxybutyrate-co-3-hydroxyvalerate as co-polyester. Lignocellulosic carbon sources mainly present in the liquefied wood have also been used as precursor in PHAs production [78]. The effect of these nutrients on PHA production was studied individually through optimization. The addition of these precursors increased the PHAs yield 30–35% on using butyrate, citrate and acetate as precursors [8]. The addition of precursor facilitates the formation of co-polymer PHAs with remarkable properties which allow using them in different field of life. A co-polymer of PHAs showed different properties from the parent polymers in different aspect like drug delivery, durability, and many others [79].
Addition of sodium chloride
The external stress factors such as increased temperature or ionic species (NaCl) cause a stress response leading to enhanced PHAs production. The amount of NaCl as 9 g/L was added additionally into nutrient enriched media. The NaCl had been used as a cheap, non-toxic, sustainable, simple and nonreactive external stress factor which had been reported to increase PHAs productivity up to 30%(w/w) with C. necator. The results also demonstrated that the nutrients media supplemented with NaCl produced 6% (w/w) more cell biomass as compare to negative control. The product yield increased at a rate of 65–77% (w/w) in the presence of NaCl. This might be attributed to lower DO in the media at higher salinity stress [80].
Production of PHAs using gases
In recent years, PHAs had been produced from industrial by-product gases such as methane CH4 or CO2. These greenhouse gases can be fixed and used as carbon sources with additional advantages of cost-effectiveness and sustainability [81]. A pretreatment usually assists the utilization of these gases as carbon source to produce PHAs. Continuous fed batch production process also accounts for PHAs production using gaseous carbon sources, CH4 (by a methanotrophic bacterial strain) and CO2 (autotrophs bacteria) [82]. In the case of Cyanobacteria sp., the availability of substrate to bacteria is limited by the solubility of these greenhouse gases, in the aqueous fermentation media. Both the solubility and availability of substrate under these conditions are influenced by parameters such as pH value, size of gas bubbles, and the temperature. A sufficient supply of these gaseous growth substrates to cell biomass is possible in a continuous mode [83]. The plants are natural sources to fix the anthropogenic gases like CO2. In recent days different approaches have been considered to fix them using metal organic framework (MOF).
Downstream recovery processes
The downstream recovery process (DsRP) should be an economical eco-friendly process in which toxic solvents are not used [11]. In DsRP, organic solvent such as methylene, chloroform, carbonate, propylene and dichloroethane are used for ultimately degrading the cell wall and cell membrane to recover the PHAs [67]. Polymers recovery methods also contribute to production costs of polymers produced. The main purpose of DsRP is to minimize production cost, increase the purity of product, obtain greater yield, avoid cell disruption and minimize the use of toxic solvent. In the case of DsRP, different parameters like purity of product, substrate used and properties required must be taken into consideration before a PHAs recovery [84]. A solvent free recovery method (like sodium hypochlorite recovery method) could also be used to recover PHAs by simple dissolving the non-PHAs cellular mass; however, this method reduced the molecular mass of polymer. The polymers used in medical field must be of high purity with no impurities [85, 86]. Some downstream recovery processes and their advantage in purity and recovery of polymer are listed in Table 6.
Future trends
In the early stages, PHAs production had been limited due to high costs which restricted their production. Researchers, therefore, had always been looking for low cost substrates to reduce overall production cost. In the recent years, large number of cheaper carbon sources such as waste frying oil, vegetable oil, whey and organic substrate etc., have been investigated to minimize the production cost. A mutant strain of P. aeruginosa produced high yields of PHAs using cheap carbon sources. The strain showed good synthesis of PHAs in preliminary investigation. The future development in PHAs production would be based on two factors; (a) lower production cost with higher yield (b) wider applications in different fields. Synthetic biology and genetic engineering techniques are expected to produce higher yield PHAs strain with high growing density under optimizing fermentation conditions in short period of time. The purification techniques with controllable lysis can be used to accumulate large size granules and reduce the cost of production. Continuous fermentation with MMC can be used to minimize the production cost operated in two SCSTR and three SCSTR configurations using cheap substrates to obtain higher yield. Different functional groups are attached to the side chain of basic structure of PHAs to give them different properties. The molecular evolution technique should be used to produce the expected structure product of short chain length or medium chain length PHAs. Downstream processing is needed to obtain the desired purity of product for specific application in the mechanical sector and many other fields of science and engineering. The mutant bacterial strain P. aeruginosa was used with soybean oil always produces mcl-PHA like P(3-HB). P. aeruginosa synthase has a good ability to produce mcl-PHA monomers efficiently utilizing vegetable oil as growth substrate to produce PHA with low production of by-products. Moreover, there is need to work on the production of desirable congeners of PHAs under controlled fermentation and to elucidate the physico-chemical and biological properties of each individual congener.
Conclusions
The present review exemplifies a number of different bioreactors (fed batch, continuous fed batch and multiple stages continuous stirring reactors) systems and bacterial strains with large number of feeding regimes to produce PHAs of different molecular mass. It became clear that a number of different combinations of bacterial strain and substrates demand different fermentation schemes, bioreactor facilities and feeding regimes. The cultivation time and process design have to be accommodated to the physiological kinetics of the system to optimize the final cell biomass productivity and product quality. As a concluding result of recent studies or experiments, it could be expected that future PHAs production would be combined with continuous stirring method. Robust extremophilic bacterial strain would be used to produce PHAs with minimum energy input. The utilization of cheap carbon enriched feed stocks should be applied to minimize substrate cost and develop sustainable DsRP method for recovery and purification of PHAs. Only the combination of these different techniques would allow for economic and sustainable production of PHAs biopolyesters. A downstream recovery process allows us to obtain PHAs with high purity to be used in for medical application.
References
Mozejko-Ciesielska J, Kiewisz K (2016) Bacterial polyhydroxyalkanoates: still fabulous. Microbiol Res 192:271–282
Khanna S, Srivastava AK (2005) Recent advances in microbial polyhydroxyalkanoates. Process Biochem 40:607–619
Lemoigne M (1926) Products of dehydration and of polymerization of β-hydroxybutyric acid. Bull Soc Chem Biol 8:770–782
Macrae R, Wilkinson J (1958) Poly-β-hyroxybutyrate metabolism in washed suspensions of bacillus cereus and bacillus megaterium. Microbiology 19:210–222
De Smet M, Eggink G, Witholt B, Kingma J, Wynberg H (1983) Characterization of intracellular inclusions formed by Pseudomonas oleovorans during growth on octane. J Bacteriol 154:870–878
Volova TG, Zhila NO, Shishatskaya EI, Mironov PV, Vasil’ev AD, Sukovatyi AG, Sinskey AG (2013) The physicochemical properties of polyhydroxyalkanoates with different chemical structures. Poly Sci Ser A 55:427–437
Virov P (2013) Polyhydroxyalkanoates: biodegradable polymers and plastics from renewable resources. Mater Technol 47:5–12
Colombo B, Favini F, Scaglia B, Sciarria TP, D’Imporzano G, Pognani M, Alekseeva A, Eisele G, Cosentino C, Adani F (2017) Enhanced polyhydroxyalkanoate (PHA) production from the organic fraction of municipal solid waste by using mixed microbial culture. Biotechnol Biofuels 10:201–215
Dietrich K, Dumont MJ, Del Rio LF, Orsat V (2017) Producing PHAs in the bioeconomy-towards a sustainable bioplastic. Sustain Prod Consum 9:58–70
Rai R, Roy I (2011) Polyhydroxyalkanoates: the emerging new green polymers of choice. In: Sharma SK, Mudhoo A (eds) A handbook of applied biopolymer technology. Royal Society of Chemistry, Cambridge, pp 79–101
Raza ZA, Abid S, Banat IM (2018) Polyhydroxyalkanoates: characteristics, production, recent developments and applications. Int Biodeterior Biodegrad 126:45–56
Tan GYA, Chen C-L, Li L, Ge L, Lin W, Mutiara I, Razaad N, Li Y, Lei Z, Mo Yu, Wang J-Y (2014) Start a research on biopolymer polyhydroxyalkanoate (PHA): a review. Polymers 6:706–754
Cui YW, Shi YP, Gong XY (2017) Effects of C/N in the substrate on the simultaneous production of polyhydroxyalkanoates and extracellular polymeric substances by Haloferax mediterranei via kinetic model analysis. J R Soc Chem 7:18953–18961
Kaur G, Roy I (2015) Strategies for large-scale production of polyhydroxyalkanoates. Chem Biochem Eng 29:157–172
Clarinval AM, Halleux J (2005) Classification of biodegradable polymers in biodegradable polymers for industrial applications. CRIF, France
Chen GQ (2009) A microbial polyhydroxyalkanoates (PHA) based bio-and materials industry. Chem Soc Rev 38:2434–2446
Greene J (2013) PHA biodegradable blow-molded bottles: compounding and performance. Plast Eng 69:16–21
Chen GQ, Wu Q (2005) The application of polyhydroxyalkanoates as tissue engineering materials. Biomaterials 26:6565–6578
Philip S, Keshavarz T, Roy I (2007) Polyhydroxyalkanoates: biodegradable polymers with a range of applications. J Chem Technol Biotechnol 82:233–247
Li Z, Yang J, Loh XJ (2016) Polyhydroxyalkanoates: opening doors for a sustainable future. NPG Asia Mater 8:265–330
Koller M, Sandholzer D, Salerno A, Braunegg G, Narodoslawsky M (2013) Biopolymer from industrial residues: life cycle assessment of poly(hydroxyalkanoates) from whey. Resour Conserv Recycl 73:64–71
Bohlmann GM (2006) Polyhydroxyalkanoate production in crops. J Am Chem Soc 921:253–270
Raza ZA, Riaz S, Banat IM (2018) Polyhydroxyalkanoates: properties and chemical modification approaches for their functionalization. Biotechnol Prog 34:29–41
Kleerebezem R, Van Loosdrecht RL (2007) Mixed culture biotechnology for bioenergy production. Curr Opin Biotechnol 18:207–212
Majone M, Massanisso P, Carucci A, Lindrea K, Tandoi V (1996) Influence of storage on kinetic selection to control aerobic filamentous bulking. Water Sci Technol 34:223–232
Martinez GA, Rebecchi S, Decorti D, Domingos JMB, Rio DD, Bertin L, Porto CD, Fava F (2016) Towards multi-purpose biorefinery platforms for the valorisation of red grape pomace: production of polyphenols, volatile fatty acids, polyhydroxyalkanoates and biogas. Green Chem 18:261–270
Raza ZA, Abid S, Rehman A, Hussain T (2016) Synthesis kinetics of poly(3-hydroxybutyrate) by using a Pseudomonas aeruginosa mutant strain grown on hexadecane. Int Biodeterior Biodegrad 115:171–178
Lasemi Z, Darzi GN, Baei MS (2013) Media optimization for poly (β-hydroxybutyrate) production using Azotobacter beijerinckii. Int J Polym Mater Polym Biomater 62:265–269
Zhu C, Nomura CT, Perrotta JA, Stipanovic AJ, Nakas JP (2010) Production and characterization of poly-3-hydroxybutyrate from biodiesel-glycerol by Burkholderia cepacia ATCC 17759. Biotechnol Prog 26:424–430
Chakraborty P, Gibbons W, Muthukumarappan K (2009) Conversion of volatile fatty acids into polyhydroxyalkanoate by Ralstonia eutropha. J Appl Microbiol 106:1996–2005
Haas R, Jin B, Zepf FT (2008) Production of poly(3-hydroxybutyrate) from waste potato starch. Biosci Biotechnol Biochem 72:253–256
Povolo S, Romanelli MG, Basaglia M, Ilieva VI, Corti A, Morelli A, Chiellini E, Casella S (2013) Polyhydroxyalkanoate biosynthesis by Hydrogenophaga pseudoflava DSM1034 from structurally unrelated carbon sources. New Biotechnol 30:629–634
Kenny ST, Kaminsky W, Wood T, Babu RP, Keely CM, Blau W, O’Connor KE (2008) Up-cycling of PET (polyethylene terephthalate) to the biodegradable plastic PHA (polyhydroxyalkanoate). Environ Sci Technol 42:7696–7701
Davis R (2013) Conversion of grass biomass into fermentable sugars and its utilization for medium chain length polyhydroxyalkanoate (mcl-PHA) production by Pseudomonas strains. Bioresour Technol 150:202–209
Bhattacharyya A, Saha J, Haldar S, Bhowmic A, Mukhopadhyay UK, Mukherjee J (2014) Production of poly-3-(hydroxybutyrate-co-hydroxyvalerate) by Haloferax mediterranei using rice-based ethanol stillage with simultaneous recovery and re-use of medium salts. Extremophiles 18:463–470
Hermann-Krauss C, Koller M, Muhr A, Fasl H, Stelzer F, Braunegg G (2013) Archaeal production of polyhydroxyalkanoate (PHA) co and ter-polyesters from biodiesel industry-derived by-products. Archaea 2013:1–10
Koller M, Bona R, Chiellini E, Fernandes EG, Horvat P, Kutschera C, Hesse P, Braunegg G (2008) Polyhydroxyalkanoate production from whey by Pseudomonas hydrogenovora. Bioresour Technol 99:4854–4863
Albuquerque M, Torres C, Reis M (2010) Polyhydroxyalkanoate (PHA) production by a mixed microbial culture using sugar molasses: effect of the influent substrate concentration on culture selection. Water Res 44:3419–3433
Follonier S, Riesen R, Zinn M (2015) Pilot-scale production of functionalized mcl-PHA from grape pomace supplemented with fatty acids. Chem Biochem Eng 29:113–121
Kellerhals MB, Kessler B, Witholt B, Tchouboukov A, Brandl H (2000) Renewable long-chain fatty acids for production of biodegradable medium-chain-length polyhydroxyalkanoates (mcl-PHAs) at laboratory and pilot plant scales. Macromolecules 33:4690–4698
Atlic A, Koller M, Scherzer D, Kutschera C, Grillo-Fernandes E, Horvat P, Chiellini E, Braunegg G (2011) Continuous production of poly (R-3-hydroxybutyrate) by Cupriavidus necator in a multistage bioreactor cascade. Appl Microbiol Biotechnol 91:295–304
Durner R, Witholt B, Egli T (2000) Accumulation of poly[(R)-3-hydroxyalkanoates] in Pseudomonas oleovorans during growth with octanoate in continuous culture at different dilution rates. Appl Environ Microbiol 66:3408–3414
Heinrich D, Raberg M, Fricke P, Kenny ST, Morales-Gamez L, Babu RP, O’Connor KE, Steinbuchel A (2016) Synthesis gas (Syngas)-derived medium-chain-length polyhydroxyalkanoate synthesis in engineered Rhodospirillum rubrum. Appl Environ Microbiol 82:6132–6140
Pagliano G, Ventorino V, Panico A, Pepe O (2017) Integrated systems for biopolymers and bioenergy production from organic waste and by-products: a review of microbial processes. Biotechnol Biofuels 10:113–136
Ienczak JL, Schmidell W, De Aragao GMAF (2013) High-cell-density culture strategies for polyhydroxyalkanoate production: a review. J Ind Microbiol Biotechnol 40:275–286
Israni N, Shivakumar S (2015) Evaluation of upstream process parameters influencing the growth associated PHA accumulation in Bacillus sp. J Sci Ind Res 74:290–295
Masood F, Abdul-Salam M, Yasin T, Hameed A (2017) Effect of glucose and olive oil as potential carbon sources on production of PHAs copolymer and tercopolymer by Bacillus cereus FA11. 3 Biotech 7:87–101
Shahid S, Mosrati R, Ledauphin J, Amiel C, Fontaine P, Gaillard JL, Corroler D (2013) Impact of carbon source and variable nitrogen conditions on bacterial biosynthesis of polyhydroxyalkanoates: evidence of an atypical metabolism in Bacillus megaterium DSM 509. J Biosci Bioeng 116:302–308
Park SJ, Jang YA, Noh W, Oh YH, Lee H, David Y, Baylon MG, Shin J, Yang JE, Choi SY, Lee SH, Lee SY (2015) Metabolic engineering of Ralstonia eutropha for the production of polyhydroxyalkanoates from sucrose. Biotechnol Bioeng 112:638–643
Eggink G, Steinbuchel A, Poirier A, Witholt B (1997) International symposium on bacterial polyhydroxyalkanoates. NRC Research Press, Toulouse
Byrom D (1992) Production of poly-β-hydroxybutyrate: poly-β-hydroxyvalerate copolymers. FEMS Microbiol Lett 103:247–250
Pandian SR, Deepak V, Kalishwaralal K, Rameshkumar N, Jeyaraj M, Gurunathan S (2010) Optimization and fed-batch production of PHB utilizing dairy waste and sea water as nutrient sources by Bacillus megaterium SRKP-3. Bioresour Technol 101:705–711
Kim SW, Kim P, Lee HS, Kim JH (1996) High production of poly-β-hydroxybutyrate (PHB) from Methylobacterium organophilum under potassium limitation. Biotechnol Lett 18:25–30
Khosravi-Darani K, Mokhtari ZB, Amai T, Tanaka K (2013) Microbial production of poly(hydroxybutyrate) from C1 carbon sources. Appl Microbiol Biotechnol 97:1407–1424
Muhr A (2013) Biodegradable latexes from animal-derived waste: biosynthesis and characterization of mcl-PHA accumulated by Ps. citronellolis. React Funct Polym 73:1391–1398
Akiyama M, Taima Y, Doi Y (1992) Production of poly(3-hydroxyalkanoates) by a bacterium of the genus Alcaligenes utilizing long-chain fatty acids. Appl Microbiol Biotechnol 37:698–701
Shay EG (1993) Diesel fuel from vegetable oils: status and opportunities. Biomass Bioenerg 4:227–242
Rincon J, Camarillo R, Rodriguez L, Ancillo V (2010) Fractionation of used frying oil by supercritical CO2 and cosolvents. Ind Eng Chem Res 49:2410–2418
Jiang G (2016) Carbon sources for polyhydroxyalkanoates and an integrated biorefinery. Int J Mol Sci 17:1157
Gumel A, Annuar M, Heidelberg T (2014) Growth kinetics, effect of carbon substrate in biosynthesis of mcl-PHA by Pseudomonas putida Bet001. Braz J Microbiol 45:427–438
Kumar M, Singhal A, Verma PK, Thakur IS (2017) Production and characterization of polyhydroxyalkanoate from lignin derivatives by Pandoraea sp. ISTKB. J Am Chem Soc 2:9156–9163
Hao J, Wang X, Wang H (2017) Overall process of using a valerate-dominant sludge hydrolysate to produce high-quality polyhydroxyalkanoates (PHA) in a mixed culture. J Nat 7:6939–6943
Yu J, Si Y (2001) A dynamic study and modeling of the formation of polyhydroxyalkanoates combined with treatment of high strength wastewater. Environ Sci Technol 35:3584–3588
Begun G, Palko A, Brown L (1956) The ammonia-ammonium carbonate system for the concentration of nitrogen-15. J Phy Chem 60:48–51
Hu D (2011) Biosynthesis and characterization of polyhydroxyalkanoate block copolymer P-3-HB-b-P-4-HB. Biomacromolecule 12:3166–3173
Rehm BH (2010) Bacterial polymers: biosynthesis, modifications and applications. Nat Rev Microbiol 8:578
Ivanov V, Stabnikov V, Ahmed Z, Dobrenko S, Saliuk A (2015) Production and applications of crude polyhydroxyalkanoate-containing bioplastic from the organic fraction of municipal solid waste. Int J Biotechnol Technol 12:725–738
Sreekanth M, Vijayendra S, Joshi G, Shamala T (2013) Effect of carbon and nitrogen sources on simultaneous production of α-amylase and green food packaging polymer by Bacillus sp. CFR 67. J Food Sci Technol 50:404–408
Zhao D (2013) Improving polyhydroxyalkanoate production by knocking out the genes involved in exopolysaccharide biosynthesis in Haloferax mediterranei. Appl Microbiol Biotechnol 97:3027–3036
Xu Z, Dai X, Chai X (2018) Effect of influent pH on biological denitrification using biodegradable PHBV/PLA blends as electron donor. Biochem Eng J 131:24–30
Liu C, Luo G, Wang W, He Y, Zhang R, Liu G (2018) The effects of pH and temperature on the acetate production and microbial community compositions by syngas fermentation. Fuel 224:537–544
Poblete-Castr I, Escapa IE, Jager C, Puchalka J, Lam JMC, Schomburg D, Prieto MP, Dos Santos VAP (2012) 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. Microb Cell Fact 11:1–21
Kang M, Peng S, Tian Y, Zhang H (2018) Effects of dissolved oxygen and nutrient loading on phosphorus fluxes at the sediment–water interface in the hai river estuary, China. Mar Pollut Bull 130:132–139
Third KA, Newland M, Cord-Ruwisch R (2003) The effect of dissolved oxygen on PHB accumulation in activated sludge cultures. Biotechnol Bioeng 82:238–250
De Almeida A, Giordano AM, Nikel PI, Pettinari MJ (2010) Effects of aeration on the synthesis of poly(3-hydroxybutyrate) from glycerol and glucose in recombinant Escherichia coli. Appl Environ Microbiol 76:2036–2040
Korkakaki E, Van Loosdrecht MC, Kleerebezem R (2017) Impact of phosphate limitation on PHA production in a feast-famine process. Water Res 126:472–480
Koller M (2015) Novel precursors for production of 3-hydroxyvalerate-containing poly[(R)-hydroxyalkanoate]s. Biocatal Biotransform 32:161–167
Koller M (2015) Liquefied wood as inexpensive precursor-feedstock for bio-mediated incorporation of (R)-3-hydroxyvalerate into polyhydroxyalkanoates. Materials 8:6543–6557
De Paula FC, Kakazu S, De Paula CBC, Gomez JGC, Contiero J (2017) Polyhydroxyalkanoate production from crude glycerol by newly isolated Pandoraea sp. J King Saud Univ Sci 29:166–173
Passanha P, Kedia G, Dinsdale RM, Guwy AJ, Esteves SR (2014) The use of NaCl addition for the improvement of polyhydroxyalkanoate production by Cupriavidus necator. Bioresour Technol 163:287–294
Ghysels S, Mozumder MSI, De Wever H, Volcke EI, Garcia-Gonzalez L (2018) Targeted poly(3-hydroxybutyrate-co-3-hydroxyvalerate) bioplastic production from carbon dioxide. Bioresour Technol 249:858–868
Garcia-Perez T, Lopez JC, Passos F, Lebrero R, Revah S, Munoz R (2018) Simultaneous methane abatement and PHB production by methylocystis hirsuta in a novel gas-recycling bubble column bioreactor. Chem Eng J 334:691–697
Koller M (2015) Cyanobacterial polyhydroxyalkanoate production: status quo and quo vadis. Curr Biotechnol 4:464–480
Ishak K, Annuar M, Heidelberg H, Gumel A (2016) Ultrasound-assisted rapid extraction of bacterial intracellular medium-chain-length poly(3-hydroxyalkanoates)(mcl-PHAs) in medium mixture of solvent/marginal non-solvent. Arabia J Sci Eng 41:33–44
Koller M, Niebelschütz H, Braunegg G (2013) Strategies for recovery and purification of poly[R-3-hydroxyalkanoates] (PHA) biopolyesters from surrounding biomass. Eng Life Sci 13:549–562
Pathak VM (2017) Review on the current status of polymer degradation: a microbial approach. Bioresour Bioprocess 4:15
Li ZJ, Cai L, Wu Q, Chen GQ (2009) Overexpression of NAD kinase in recombinant Escherichia coli harboring the phbCAB operon improves poly(3-hydroxybutyrate) production. Appl Microbiol Biotechnol 83:939–947
Chen GQ, Wu Q (2005) Microbial production and applications of chiral hydroxyalkanoates. Appl Microbiol Biotechnol 67:592–599
Massieu L, Haces M, Montiel T, Hernandez-Fonseca K (2003) Acetoacetate protects hippocampal neurons against glutamate-mediated neuronal damage during glycolysis inhibition. Neuroscience 120:365–378
Kashiwaya Y, Takeshima T, Mori N, Nakashima K, Clarke K, Veech RL (2000) d-β-hydroxybutyrate protects neurons in models of alzheimer’s and prkinson’s disease. Proc Natl Acad Sci 97:5440–5444
Yao YC (2015) A specific drug targeting system based on polyhydroxyalkanoate granule binding protein PhaP fused with targeted cell ligands. Biomaterials 29:4823–4830
Ahmed T, Marcal H, Lawless M, Wanandy NS, Chiu A, Foster LJR (2010) Polyhydroxybutyrate and its copolymer with polyhydroxyvalerate as biomaterials: influence on progression of stem cell cycle. Biomacromolecules 11:2707–2715
Gadgil BST, Killi N, Rathna GV (2017) Polyhydroxyalkanoates as biomaterials. Med Chem Commun 8:1774–1787
Furutate S, Nakazaki H, Maejima K, Hiroe A, Abe H, Tsuge T (2017) Biosynthesis and characterization of novel polyhydroxyalkanoate copolymers consisting of 3-hydroxy-2-methylbutyrate and 3-hydroxyhexanoate. J Polym Res 24:221
Fukui T, Doi Y (1998) Efficient production of polyhydroxyalkanoates from plant oils by Alcaligenes eutrophus and its recombinant strain. Appl Microbiol Biotechnol 49:333–336
Yu P, Chua H, Huang A, Lo W, Chen G (1998) Conversion of food industrial wastes into bioplastics. Appl Biochem Biotechnol 70:603–614
Yamane T, Fukunaga M, Lee YW (1996) Increased PHB productivity by high-cell-density fed-batch culture of Alcaligenes latus, a growth-associated PHB producer. Biotechnol Bioeng 50:197–202
Aditi S, Souza Shalet NMD, Pranesh R, Katyayini T (2015) Microbial production of polyhydroxyalkanoates (PHA) from novel sources: a review. Int J RBS 4:16–28
Gomez J (1996) Evaluation of soil gram-negative bacteria yielding polyhydroxyalkanoic acids from carbohydrates and propionic acid. Appl Microbiol Biotechnol 45:785–791
Pan W, Perrotta JA, Stipanovic AJ, Nomura CT, Nakas JP (2012) Production of polyhydroxyalkanoates by Burkholderia cepacia ATCC 17759 using a detoxified sugar maple hemicellulosic hydrolysate. J Ind Microbiol Biotechnol 39:459–469
Chee JY, Tan Y, Samian MR, Sudesh K (2010) Isolation and characterization of a Burkholderia sp. USM (JCM15050) capable of producing polyhydroxyalkanoate (PHA) from triglycerides, fatty acids and glycerols. J Polym Environ 18:584–592
Qi Q, Rehm BH (2001) Polyhydroxybutyrate biosynthesis in caulobacter crescentus: molecular characterization of the polyhydroxybutyrate synthase. Microbiology 147:3353–3358
Valentin HE, Lee EY, Choi CY, Steinbüchel A (1994) Identification of 4-hydroxyhexanoic acid as a new constituent of biosynthetic polyhydroxyalkanoic acids from bacteria. Appl Microbiol Biotechnol 40:710–716
Sonnleitner B, Heinzle E, Braunegg G, Lafferty R (1979) Formal kinetics of poly-β-hydroxybutyric acid (PHB) production in Alcaligenes eutrophus H 16 and Mycoplana rubra R 14 with respect to the dissolved oxygen tension in ammonium-limited batch cultures. Eur J Appl Microbiol Biotechnol 7:1–10
Ishizaki A, Tanaka K (1991) Production of poly-β-hydroxybutyric acid from carbon dioxide by Alcaligenes eutrophus ATCC 17697T. J Ferment Bioeng 71:254–257
Kim BS, Lee SC, Lee SY, Chang HN, Chang YK, Woo SI (1994) Production of poly (3-hydroxybutyric acid) by fed-batch culture of Alcaligenes eutrophus with glucose concentration control. Biotechnol Bioeng 43:892–898
Beaulieu M, Beaulieu Y, Melinard J, Pandian S, Goulet J (1995) Influence of ammonium salts and cane molasses on growth of Alcaligenes eutrophus and production of polyhydroxybutyrate. Appl Environ Microbiol 61:165–169
Cavalheiro JM, De Almeida MCM, Grandfils C, Da Fonseca M (2009) Poly(3-hydroxybutyrate) production by Cupriavidus necator using waste glycerol. Process Biochem 44:509–515
Van-Thuoc D, Quillaguaman J, Mamo G, Mattiasson B (2008) Utilization of agricultural residues for poly(3-hydroxybutyrate) production by Halomonas boliviensis LC1. J Appl Microbiol 104:420–428
Koller M, Hesse P, Bona R, Kutschera C, Atlic A, Braunegg G (2007) Potential of various archae-and eubacterial strains as industrial polyhydroxyalkanoate producers from whey. Macromol Biosci 7:218–226
Bourque D, Pomerleau Y, Groleau D (1995) High-cell-density production of poly-β-hydroxybutyrate (PHB) from methanol by Methylobacterium extorquens: production of high-molecular-mass PHB. Appl Microbiol Biotechnol 44:367–376
Wendlandt KD, Jechorek M, Helm J, Stottmeister U (1998) Production of PHB with a high molecular mass from methane. Polym Degrad Stab 59:191–194
Smit AM, Strabala TJ, Peng L, Rawson P, Lloyd-Jones G, Jordan TW (2012) Proteomic phenotyping of novosphingobium nitrogenifigens reveals a robust capacity for simultaneous nitrogen fixation, polyhydroxyalkanoate production, and resistance to reactive oxygen species. Appl Environ Microbiol 78:4802–4815
Yamane T, Chen X, Ueda S (1996) Growth associated production of Poly(3-hydroxyvalerate) from n-pentanol by a Methylotrophic bacterium, Paracoccus denitrificans. Appl Environ Microbiol 62:380–384
Tripathi AD, Yadav A, Jha A, Srivastava S (2012) Utilizing of sugar refinery waste (cane molasses) for production of bio-plastic under submerged fermentation process. J Poly Environ 20:446–453
Lee E, Jendrossek D, Schirmer A, Choi C, Steinbüchel A (1995) Biosynthesis of copolyesters consisting of 3-hydroxybutyric acid and medium-chain-length 3-hydroxyalkanoic acids from 1, 3-butanediol or from 3-hydroxybutyrate by Pseudomonas sp. A33. Appl Microbiol Biotechnol 42:901–909
Ward PG, Goff M, Donner M, Kaminsky W, Connor KEQ (2006) A two step chemo-biotechnological conversion of polystyrene to a biodegradable thermoplastic. Environ Sci Technol 40:2433–2437
Sun Z, Ramsay JA, Guay M, Ramsay BA (2007) Carbon-limited fed-batch production of medium-chain-length polyhydroxyalkanoates from nonanoic acid by Pseudomonas putida KT2440. Appl Microbiol Biotechnol 74:69–77
Nikodinovic J, Kenny ST, Babu RP, Woods T, Blau WJ, O’Connor KE (2008) The conversion of BTEX compounds by single and defined mixed cultures to medium-chain-length polyhydroxyalkanoate. Appl Microbiol Biotechnol 80:665–673
Haywood GW, Anderson AJ, Williams DR, Dawes EA, Ewing DF (1991) Accumulation of a poly(hydroxyalkanoate) copolymer containing primarily 3-hydroxyvalerate from simple carbohydrate substrates by Rhodococcus sp. NCIMB 40126. Int J Biol Macromol 13:83–88
Akar A (2006) Accumulation of polyhydroxyalkanoates by Microlunatus phosphovorus under various growth conditions. J Ind Microbiol Biotechnol 33:215–220
Koller M, Atlic A, Gonzalez-Garcia Y, Kutschera C, Braunegg G (2008) Polyhydroxyalkanoate (PHA) biosynthesis from whey lactose. Macromol Symp 272:87–92
Cui B, Huang S, Xu F, Zhang R, Zhang Y (2015) Improved productivity of poly(3-hydroxybutyrate)(PHB) in thermophilic Chelatococcus daeguensis TAD1 using glycerol as the growth substrate in a fed-batch culture. Appl Microbiol Biotechnol 99:6009–6019
Ng KS, Wong YM, Tsuge T, Sudesh K (2011) Biosynthesis and characterization of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) copolymers using jatropha oil as the main carbon source. Process Biochem 46:1572–1578
Sindhu R, Silviya N, Binod P, Pandey A (2013) Pentose-rich hydrolysate from acid pretreated rice straw as a carbon source for the production of poly-3-hydroxybutyrate. Biochem Eng J 78:67–72
Gomaa EZ (2014) Production of polyhydroxyalkanoates (PHAs) by Bacillus subtilis and Escherichia coli grown on cane molasses fortified with ethanol. Braz Arch Biol Technol 57:145–154
Negi S, Banerjee R (2010) Optimization of culture parameters to enhance production of amylase and protease from Aspergillus awamori in a single fermentation. Afr J Biochem Res 4:73–80
Shamala T, Vijayendra S, Joshi G (2012) Agro-industrial residues and starch for growth and co-production of polyhydroxyalkanoate copolymer and α-amylase by Bacillus sp. CFR-67. Braz J Microbiol 43:1094–1102
Rinnan R, Baath E (2009) Differential utilization of carbon substrates by bacteria and fungi in tundra soil. Appl Environ Microbiol 75:3611–3620
Ryu HW, Cho KS, Goodrich PR, Park CH (2008) Production of polyhydroxyalkanoates by Azotobacter vinelandii UWD using swine wastewater: effect of supplementing glucose, yeast extract, and inorganic salts. Biotechnol Bioprocess Eng 13:651–658
Wei YH (2011) Screening and evaluation of polyhydroxybutyrate-producing strains from indigenous isolate Cupriavidus taiwanensis strains. Int J Mol Sci 12:252–265
Lopez-Abelairas M, García-Torreiro M, Lu-Chau T, Lema J, Steinbuchel A (2015) Comparison of several methods for the separation of poly(3-hydroxybutyrate) from Cupriavidus necator H16 cultures. Biochem Eng J 93:250–259
Kapritchkoff FM (2006) Enzymatic recovery and purification of polyhydroxybutyrate produced by Ralstonia eutropha. J Biotechnol 122:453–462
Kosseva M, Webb C (2013) Food industry wastes: assessment and recuperation of commodities. Academic Press, Cambridge
Divyashree M, Shamala T (2010) Extractability of polyhydroxyalkanoate synthesized by Bacillus flexus cultivated in organic and inorganic nutrient media. India J Microbiol 50:63–69
Khosravi-Darani K (2010) Research activities on supercritical fluid science in food biotechnology. Crit Rev Food Sci Nutr 50:479–488
Murugan P, Han L, Gan CY, Maurer FH, Sudesh K (2016) A new biological recovery approach for PHA using mealworm, Tenebrio molitor. J Biotechnol 239:98–105
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Authors acknowledge the financial support of Higher Education Commission of Pakistan for this study.
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Raza, Z.A., Tariq, M.R., Majeed, M.I. et al. Recent developments in bioreactor scale production of bacterial polyhydroxyalkanoates. Bioprocess Biosyst Eng 42, 901–919 (2019). https://doi.org/10.1007/s00449-019-02093-x
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DOI: https://doi.org/10.1007/s00449-019-02093-x