3HV Fraction Research Articles

Enrichment of mixed methanotrophic cultures producing polyhydroxyalkanoates (PHAs) from various environmental sources

Methanotrophic bacteria can use atmospheric methane (CH4) as a sole carbon source for the growth and production of polyhydroxyalkanoates (PHA). The development of CH4 bioconversion processes relies heavily on the selection of an efficient methanotrophic culture. This research assessed the effect of selected growth conditions, such as nitrogen sources on the enrichment of methanotrophic cultures from various environments for PHA accumulation. Nitrate-based medium favoured the culture growth and selection for PHA-producing methanotrophic cultures with Methylocystis sp. as a major genus and accumulation of up to 27 % polyhydroxybutyrate (PHB) in the biomass. Three PHB-producing cultures: enriched from waste activated sludge (AS), peat bog soil (PB) and landfill biocover soil (LB) were then tested for their ability to produce PHA copolymer at different CH4:O2 ratios. All enriched cultures were able to utilise valeric acid as a cosubstrate for the accumulation of PHA with a 3-hydroxyvaleric (3HV) fraction of 21–41 mol% depending on the inoculum source and CH4 concentration. The process performance of selected cultures was evaluated and compared to the culture of reference strain Methylocystis hirsuta DSM 18500. All mixed cultures irrespective of their inoculum source had similar levels of 3HV fraction in the PHA (38 ± 2 mol%). The highest poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) production was observed for AS culture at 10 % CH4 with an accumulation of 27 ± 3 % of dry cell weight (DCW), 3HV fraction of 39 ± 2 mol% and yield of 0.42 ± 0.02 g-PHA/g-substrate.

Relation of 3HV fraction and thermomechanical properties of poly(3–hydroxybutyrate–co–3–hydroxyvalerate) produced by Azotobacter vinelandii OP

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) has attracted substantial attention as a promising material for industrial applications. In this study, different PHBV films with distinct 3-hydroxyvalerate (3HV) contents produced by Azotobacter vinelandii OP were evaluated. The 3HV fraction ranged from 18.6 to 36.7 mol%, and the number-average molecular weight (Mn) was between 238 and 434 kDa. In the bioreactor, a 3HV fraction (36.7 mol%) and an Mn value of 409 kDa were obtained with an oxygen transfer rate (OTR) of 12.5 mmol L−1 h−1. Thermal analysis measurements showed decreased melting (Tm) and glass transition (Tg) temperatures, and values with relatively high 3HV fractions indicated improved thermomechanical properties. The incorporation of the 3HV fraction in the PHBV chain improved the thermal stability of the films, reduced the polymer Tm, and affected the tensile strength. PHBV film with 36.7 mol% 3HV showed an increase in its tensile strength (51.8 MPa) and a decrease in its Tm (170.61 °C) compared with PHB. Finally, scanning electron microscopy (SEM) results revealed that the PHBV film with 32.8 mol% 3HV showed a degradation upon contact with soil, water, or soil bacteria, showing more porous surfaces after degradation. The latter phenomenon indicated that thermomechanical properties played an important role in biodegradation.

Enhanced poly(3-hydroxybutyrateco-3-hydroxyvalerate) production from high-concentration propionate by a novel halophile Halomonas sp. YJ01: Detoxification of the 2-methylcitrate cycle

As a carbon substrate, propionate can be used to synthesize poly(3-hydroxybutyrateco-3-hydroxyvalerate) [PHBV] biopolymer, but high concentrations can inhibit PHBV production. Therefore, novel PHBV producers that can utilize high propionate concentrations are needed. Here, a novel halophile, Halomonas sp. YJ01 was applied to PHBV production via a propionate-dependent pathway, and optimal culture growth conditions were determined. The maximum poly(3-hydroxybutyrate) [PHB] content and yield in the presence of glucose were 89.5 wt% and 5.7 g/L, respectively. This strain utilizes propionate and volatile fatty acids (VFAs) for PHBV accumulation. Multiple genes related to polyhydroxyalkanoate (PHA) synthesis were identified using whole-genome annotation. The PHBV yield and 3HV fraction obtained by strain YJ01 utilizing 15 g/L propionate were 0.86 g/L and 29 mol%, respectively, but in cultures with glucose-propionate, it decreased its copolymer dry weight. This indicates that propionyl-CoA was converted to pyruvate through the 2-methylcitrate cycle (2MCC), which reduced propionate detoxification for the strain.

Inexpensive Production of Poly (3‑hydroxybutyrate‑co‑ 3‑hydroxyvalerate) from Bacillus megaterium PP-10 Using Pineapple Peel Waste

Pineapple peel waste has recently been interested in being utilized as a low-cost carbon source in PHA biosynthesis to reduce the production cost of PHA. The production of copolymer Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [PHBV] by the new Bacillus megaterium PP-10 was investigated. The bacteria were grown in a mixture of pineapple peel hydrolysate (PPH) and 3-hydroxyvalerate (3HV) precursor such as sodium propionate or sodium valerate at ratios of 1:1. Remarkably, the microbial growth and PHBV production in a mixture of PPH and sodium valerate exhibited higher biomass and higher PHA amount than that of sodium propionate, accounted about 2.40 ± 0.07 g/L of DCW and 0.71 ± 0.03 g/L of PHA concentration (PHA content of 29.6%DCW). Moreover, to control the 3HV molar fraction in PHBV, various sodium valerate concentration from 2 to 18 g/L was supplemented with PPH, and the result showed that the 3HV fraction increased linear trend with an increase in valerate concentration and was in the range between 6-35 mol%HV. In contrast, a maximum PHA concentration of 1.65 ± 0.04 g/L content (about 49%DCW) was obtained when B. megaterium PP-10 was cultivated in 18 g/L of total reducing sugar in PPH with 2 g/L of sodium valerate at 12 h of cultivation. Finally, the produced PHBV containing 20 mol%HV was further determined by some thermal properties and found that it possessed the melting and glass transition temperatures of 148°C and -10°C, respectively. Therefore, PHBV synthesized by B. megaterium PP-10 with various 3HV fractions was an excellent choice for biopolymer production.

Enhanced production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) with modulated 3-hydroxyvalerate fraction by overexpressing acetolactate synthase in Cupriavidus necator H16

The elastomeric properties of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), a biodegradable copolymer, strongly depend on the molar composition of 3-hydroxyvalerate (3HV). This paper reports an improved artificial pathway for enhancing the 3HV component during PHBV biosynthesis from a structurally unrelated carbon source by Cupriavidus necator H16. To increase the intracellular accumulation of propionyl-CoA, a key precursor of the 3HV monomer, we developed a recombinant strain by genetically manipulating the branched-chain amino acid (e.g., valine, isoleucine) pathways. Overexpression of the heterologous feedback-resistant acetolactate synthase (alsS), (R)-citramalate synthase (leuA), homologous 3-ketothiolase (bktB), and the deletion of 2-methylcitrate synthase (prpC) resulted in biosynthesis of 42.5 % (g PHBV/g dry cell weight) PHBV with 64.9 mol% 3HV monomer from fructose as the sole carbon source. This recombinant strain also accumulated the highest PHBV content of 54.5 % dry cell weight (DCW) with 24 mol% 3HV monomer from CO2 ever reported. The lithoautotrophic cell growth and PHBV production by the recombinant C. necator were promoted by oxygen stress. The thermal properties of PHBV showed a decreasing trend of the glass transition and melting temperatures with increasing 3HV fraction. The average molecular weights of PHBV with modulated 3HV fractions were between 20 and 26 × 104 g/mol.

Electrospun poly(3-hydroxybutyrate-co-3-hydroxyvalerate) scaffolds – a step towards ligament repair applications

ABSTRACT The incidence of anterior cruciate ligament (ACL) ruptures is approximately 50 per 100,000 people. ACL rupture repair methods that offer better biomechanics have the potential to reduce long term osteoarthritis. To improve ACL regeneration biomechanically similar, biocompatible and biodegradable tissue scaffolds are required. Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), with high 3-hydroxyvalerate (3HV) content, based scaffold materials have been developed, with the advantages of traditional tissue engineering scaffolds combined with attractive mechanical properties, e.g., elasticity and biodegradability. PHBV with 3HV fractions of 0 to 100 mol% were produced in a controlled manner allowing specific compositions to be targeted, giving control over material properties. In conjunction electrospinning conditions were altered, to manipulate the degree of fibre alignment, with increasing collector rotating speed used to obtain random and aligned PHBV fibres. The PHBV based materials produced were characterised, with mechanical properties, thermal properties and surface morphology being studied. An electrospun PHBV fibre mat with 50 mol% 3HV content shows a significant increase in elasticity compared to those with lower 3HV content and could be fabricated into aligned fibres. Biocompatibility testing with L929 fibroblasts demonstrates good cell viability, with the aligned fibre network promoting fibroblast alignment in the axial fibre direction, desirable for ACL repair applications. Dynamic load testing shows that the 50 mol% 3HV PHBV material produced can withstand cyclic loading with reasonable resilience. Electrospun PHBV can be produced with low batch variability and tailored, application specific properties, giving these biomaterials promise in tissue scaffold applications where aligned fibre networks are desired, such as ACL regeneration.

Conversion of Short and Medium Chain Fatty Acids into Novel Polyhydroxyalkanoates Copolymers by Aeromonas sp. AC_01.

Polyhydroxyalkanoates (PHAs) production by Aeromonas sp. AC_01 was investigated using synthetic and waste derived short and medium chain fatty acids (SMCFAs). The obtained results revealed that the analyzed bacterial strain was able to grow and synthesize PHAs using SMCFAs. The highest PHA productivity was observed in the cultivation supplemented with a mixture of acetic acid and butyric acid (3.89 mg/L·h). Furthermore, SMCFAs-rich stream, derived from acidogenic mixed culture fermentation of acid whey, was found to be less beneficial for PHA productivity than its synthetic mixture, however the PHA production was favored by the nitrogen-limited condition. Importantly, Aeromonas sp. AC_01 was capable of synthesizing novel scl-mcl copolymers of 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), 3-hydroxytridecanoate (3HtriD) and/or 3-hydroxytetradecaonate (3HTD) with high 3HB and 3HV fractions. They were identified with alterable monomers composition depending on the culture conditions used. Moreover, in-depth thermal analyses proved that they are highly resistant to thermal degradation regardless of their monomeric composition. The obtained results confirm that Aeromonas sp. AC_01 is a promising candidate for the biotechnological production of PHAs from SMCFAs with thermal properties that can be tuned together with their chemical composition by the corresponding adjustment of the cultivation process.

Heterologous phasin expression in Rhodopseudomonas palustris CGA009 for bioplastic production from lignocellulosic biomass

Rhodopseudomonas palustris CGA009 is a metabolically robust microbe that can utilize lignin breakdown products to produce polyhydroxyalkanoates (PHAs), biopolymers with the potential to replace conventional plastics. Our recent efforts suggest PHA granule formation is a limiting factor for maximum production of the bioplastic poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) by R. palustris. The Phap1 phasin (phaP1) from the PHB-producing model bacterium Cupriavidus necator H16 was expressed in R. palustris with the aim of overproducing PHBV from the lignin breakdown product p-coumarate by fostering smaller and more abundant granules. Expression of phaP1 yielded PHBV production from R. palustris aerobically (0.7 g/L), which does not occur in the wild-type strain, and led to a significantly higher PHBV titer than wild-type anaerobic production (0.41 g/L). The 3HV fractions were also significantly increased under both anaerobic and aerobic conditions, which boosts thermomechanical properties and potential for application. Thus, heterologous phasin expression in R. palustris provides flexibility for industrial processing and could foster compositional changes in copolymers with better thermomechanical properties compared to PHB alone.

Composition control of poly(3‐hydroxybutyrate‐co‐3‐hydroxyvalerate) copolymerization by oxygen transfer rate (OTR) in Azotobacter vinelandii OPNA

AbstractBACKGROUNDThe aim of this study was to evaluate changes in the composition of poly‐3‐hydoxybutyrate‐co‐3‐hydroxyvalerate (PHBV) by the polyhydroxybutyrate (PHB) overproducing mutant OPNA of Azotobacter vinelandii cultured under different conditions.RESULTSThe oxygen transfer rate (OTR) significantly affected PHBV composition in cultures in shaken flasks; the highest content of 3‐hydroxyvalerate (3HV; 28%) was achieved from cultures developed at an OTRmax of 5.87 mmol L−1 h−1. In the bioreactor, cultures were grown under oxygen‐limited conditions and different OTRs, varying the agitation rate. The highest PHBV accumulation (85%) was achieved at a low OTRmax (4.96 mmol L−1 h −1), whereas the maximum content of 3HV (8.5 mol%) was obtained at a higher OTRmax (20.3 mmol L−1 h−1). The OPNA strain produced a copolymer of ultra‐high molecular weight.CONCLUSIONSOur results reveal that increasing the OTRmax raises the molar fraction of 3HV in the PHBV copolymer. Therefore, the manipulation of OTR could be a feasible strategy to produce PHBV copolymers with different monomeric compositions at industrial level. © 2021 Society of Chemical Industry (SCI).

Introducing the Newly Isolated Bacterium Aneurinibacillus sp. H1 as an Auspicious Thermophilic Producer of Various Polyhydroxyalkanoates (PHA) Copolymers-2. Material Study on the Produced Copolymers.

Aneurinibacillus sp. H1 is a promising, moderately thermophilic, novel Gram-positive bacterium capable of the biosynthesis of polyhydroxyalkanoates (PHA) with tunable monomer composition. In particular, the strain is able to synthesize copolymers of 3-hydroxybutyrate (3HB), 4-hydroxybutyrate (4HB) and 3-hydroxyvalerate (3HV) with remarkably high 4HB and 3HV fractions. In this study we performed an in-depth material analysis of PHA polymers produced by Aneurinibacillus sp. H1 in order to describe how the monomer composition affects fundamental structural and physicochemical parameters of the materials in the form of solvent-casted films. Results of infrared spectroscopy, X-ray diffractometry and thermal analysis clearly show that controlling the monomer composition enables optimization of PHA crystallinity both qualitatively (the type of the crystalline lattice) and quantitatively (the overall degree of crystallinity). Furthermore, resistance of the films against thermal and/or enzymatic degradation can also be manipulated by the monomer composition. Results of this study hence confirm Aneurinibacillus sp. H1 as an auspicious candidate for thermophilic production of PHA polymers with material properties that can be tuned together with their chemical composition by the corresponding adjustment of the cultivation process.

Accumulation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by Azotobacter vinelandii with different 3HV fraction in shake flasks and bioreactor.

In the present study, the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) by Azotobacter vinelandii was evaluated in shake flasks and bioreactors, utilizing different precursors and oxygen transfer rates (OTRs). In shake flask cultures, the highest PHBV yield from sucrose (0.16gg-1) and 3-hydroxyvalerate (3HV) fraction in the PHA chain (27.4mol%) were obtained with valerate (1.0g L-1). In the bioreactor, the cultures were grown under oxygen-limited conditions, and the maximum OTR (OTRmax) was varied by adjusting the agitation rate. In the cultures grown at low OTRmax (4.3mmol L-1h-1), the intracellular content of PHBV (73% w w-1) was improved, whereas a maximum 3HV fraction (35mol %) was obtained when a higher OTRmax (17.2mmol L-1h-1, to 600rpm) was employed. The findings obtained suggest that the PHBV production and the content of 3HV incorporated into the polymer were affected by the OTR. Based on the evidence, it is possible to produce PHBV with a different composition by varying the OTR of the culture; thus, the approach in this study could be used to scale up PHBV production.

The effect of methane and odd-chain fatty acids on 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) synthesis by a Methylosinus-dominated mixed culture

A methanotrophic community was enriched in a semi-continuous reactor under non-aseptic conditions with methane and ammonia as carbon and nitrogen source. After a year of operation, Methylosinus sp., accounted for 80% relative abundance of the total sequences identified from potential polyhydroxyalkanoates (PHAs) producers, dominated the methane-fed enrichment. Prior to induction of PHA accumulation, cells harvested from the parent reactor contained low level of PHA at 4.0 ± 0.3 wt%. The cells were later incubated in the absence of ammonia with various combinations of methane, propionic acid, and valeric acid to induce biosynthesis of poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). Previous studies reported that methanotrophic utilization of odd-chain fatty acids for the production of PHAs requires reducing power from methane oxidation. However, our findings demonstrated that the PHB-containing methanotrophic enrichment does not require methane availability to generate 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV)—when odd-chain fatty acids are presented. The enrichment yielded up to 14 wt% PHA with various mole fractions of 3HV monomer depending on the availability of methane and odd-fatty acids. Overall, the addition of valeric acid resulted in a higher PHA content and a higher 3HV fraction. The highest 3HV fraction (up to 65 mol%) was obtained from the methane–valeric acid experiment, which is higher than those previously reported for PHA-producing methanotrophic mixed microbial cultures.

Production of polyhydroxyalkanoates on waste frying oil employing selected Halomonas strains

The aim of this work was to study the potential of selected Halomonas species for conversion of waste frying oil into polyhydroxyalkanoates (PHA). In total nine Halomonas strains were experimentally screened for their capability of PHA production. Among them, Halomonas neptunia and Halomonas hydrothermalis were identified as potent PHA producers. Initial concentration of NaCl was identified as parameter influencing PHA yields as well as molecular weight of the polymer. In addition, H. hydrothermalis was capable of biosynthesis of a copolymer of 3-hydroxybutyrate and 3-hydroxyvalerate P(3HB-co-3HV). When valerate was utilized as a precursor, the 3HV fraction in the copolymer reached high values of 50.15 mol.%. PHA production on lipid substrates by Halomonas has not been reported so far. Bearing in mind all the positive aspects of employing extremophiles in industrial biotechnology, H. hydrothermalis seems to be a very interesting halophilic strain for production of PHA using lipid substrates.

Polyhydroxybutyrate production in halophilic marine bacteria Vibrio proteolyticus isolated from the Korean peninsula.

Polyhydroxybutyrates (PHB) are biodegradable polymers that are produced by various microbes, including Ralstonia, Pseudomonas, and Bacillus species. In this study, a Vibrio proteolyticus strain, which produces a high level of polyhydroxyalkanoate (PHA), was isolated from the Korean marine environment. To determine optimal growth and production conditions, environments with different salinity, carbon sources, and nitrogen sources were evaluated. We found that the use of a medium containing 2% (w/v) fructose, 0.3% (w/v) yeast extract, and 5% (w/v) sodium chloride (NaCl) in M9 minimal medium resulted in high PHA content (54.7%) and biomass (4.94g/L) over 48h. Addition of propionate resulted in the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (P(HB-co-HV)) copolymer as propionate acts as a precursor for the HV unit. In these conditions, the bacteria produced poly(3-hydroxybutyrate-co-3-hydroxyvalerate) containing a 15.8% 3HV fraction with 0.3% propionate added as the substrate. To examine the possibility of using unsterilized media with high NaCl content for PHB production, V. proteolyticus was cultured in sterilized and unsterilized conditions. Our results indicated a higher growth, leading to a dominant population in unsterilized conditions and higher PHB production. This study showed the conditions for halophilic PHA producers to be later implemented at a larger scale.

Acetic Acid as an Indirect Sink of CO2 for the Synthesis of Polyhydroxyalkanoates (PHA): Comparison with PHA Production Processes Directly Using CO2 as Feedstock

White biotechnology is promising to transform CO2 emissions into a valuable commodity chemical such as the biopolymer polyhydroxyalkanaotes (PHA). Our calculations indicated that the indirect conversion of acetic acid from CO2 into PHA is an interesting alternative for the direct production of PHA from CO2 in terms of CO2 fixation, H2 consumption, substrate cost, safety and process performance. An alternative cultivation method using acetic acid as an indirect sink of CO2 was therefore developed and a proof-of-concept provided for the synthesis of both the homopolymer poly(3-hydroxybutyrate) (PHB) and the copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). The aim was to compare key performance parameters with those of existing cultivation methods for direct conversion of CO2 to PHA. Fed-batch cultivations for PHA production were performed using a pH-stat fed-batch feeding strategy in combination with an additional Dissolved Oxygen (DO)-dependent feed. After 118 h of fermentation, 60 g/L cell dry matter (CDM) containing 72% of PHB was obtained, which are the highest result values reported so far. Fed-batch cultivations for PHBV production resulted in 65 g/L CDM and 48 g/L PHBV concentration with a 3HV fraction of 27 mol %. Further research should be oriented towards process optimisation, whole process integration and design, and techno-economic assessment.

CRISPR/Cas9 editing genome of extremophile Halomonas spp.

Extremophiles are suitable chassis for developing the next generation industrial biotechnology (NGIB) due to their resistance to microbial contamination. However, engineering extremophiles are not an easy task. Halomonas, an industrially interesting halophile able to grow under unsterile and continuous conditions in large-scale processes, can only be engineered using suicide plasmid-mediated two-step homologous recombination which is very laborious and time-consuming (up to half a year). A convenient approach for the engineering of halophiles that can possibly be extended to other extremophiles is therefore urgently required. To meet this requirement, a rapid, efficient and scarless method via CRISPR/Cas9 system was developed in this study for genome editing in Halomonas. The method achieved the highest efficiency of 100%. When eight different mutants were constructed via this special CRISPR/Cas9 method to study the combinatorial influences of four different genes on the glucose catabolism in H. bluephagenesis TD01, it took only three weeks to complete the deletion and insertion of up to 4.5 kb DNA. H. bluephagenesis was designed to produce a microbial copolymer P(3HB-co-3HV) consisting of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV). The CRISPR/Cas9 was employed to delete the prpC gene in H. bluephagenesis TD01. Shake flask studies showed that the 3HV fraction in the copolymers increased approximately 16-folds, demonstrating enhanced effectiveness of the ΔprpC mutant to synthesize PHBV. This genome engineering strategy significantly speeds up the studies on Halomonas engineering, opening up a wide area for developing NGIB.

Biosynthesis and Characterization of Polyhydroxyalkanoates with Controlled Composition and Microstructure.

Volatile fatty acids (VFA) C2:0 to C6:0 were used as the sole carbon source for poly(3-hydroxybutyrate- co-3-hydroxyvalerate) (PHBV) production with controllable composition and microstructure in Haloferax mediterranei. Feeding carbon-even VFA gave >90 mol % poly(3-hydroxybutyrate) (3HB) PHBV, while carbon-odd VFA generated >87 mol % poly(3-hydroxyvalerate) (3HV) PHBV. Bespoke random, block, and blend copolymers with 0-100 mol % 3HV were synthesized using C4:0/C5:0 mixtures. The copolymer 3HV fraction is proportional to the %C5:0 in the feed mixture, allowing control over copolymer composition. Microstructure depends on the substrate addition order: cofeeding generated random copolymers, while sequential feeding created block and blend copolymers. On average, the PHBV had an ultrahigh molecular weight of 3 × 106 g/mol. 3HV rich copolymers showed lower melting temperatures, enhanced elasticity, and ductility. H. mediterranei is ideal for large-scale production of PHBV due to its inherent bioprocessing advantages, while control over the composition and microstructure of PHBV will facilitate the production of biopolymers capable of meeting industrial criteria for specific applications.

Engineering Escherichia coli for efficient coproduction of polyhydroxyalkanoates and 5-aminolevulinic acid.

Single-cell biorefineries are an interesting strategy for using different components of feedstock to produce multiple high-value biochemicals. In this study, a strategy was applied to refine glucose and fatty acid to produce 5-aminolevulinic acid (ALA) and polyhydroxyalkanoates (PHAs). To express the ALA and PHAs dual-production system efficiently and stably, multiple copies of the poly-β-3-hydroxybutyrate (PHB) synthesis operon were integrated into the chromosome of Escherichia coli DH5αΔpoxB. The above strain harboring the ALA C5 synthesis pathway genes hemA and hemL resulted in coproduction of 38.2% PHB (cell dry weight, CDW) and 3.2g/L extracellular ALA. To explore coproduction of ALA and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), the PHBV synthetic pathway was also integrated into engineered E. coli and coexpressed with hemA and hemL; cells produced 38.9% PHBV (CDW) with 10.3mol% 3HV fractions and 3.0g/L ALA. The coproduction of ALA with PHB and PHBV can improve the utilization of carbon sources and maximize the value derived from the feedstock.

Productivity of Marine Archaea Haloferax mediterranei Culture in Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Production with Yeast Extract as Nutrient Source

Currently, most of household goods are made from polymer or plastic. The polymer manufacturing industry has been relying on fossil fuel-based raw materials which is non-biodegradable. Plastics derived from these non-biodegradable sources will be difficult to degrade in nature, which in turn will pollute the environment and harm the ecosystem. One way to reduce the ecosystem damages caused by plastic made from petroleum-based plastic is to use biodegradable materials as plastic industry raw materials, such as Polyhydroxyalkanoates (PHA). Poly (3-hydroxybutyrate-co-3-hydroxyvalerate) or PHBV is an example of PHA that can be produced by several types of microorganisms, such as marine archaea Haloferax mediterranei. This objective of this study is to determine the productivity of marine archaea cultures H. mediterranei to produce PHBV with yeast extract as main nutrient sources. H. mediterranei will produce PHBV at extreme conditions with excess carbon and limited nutrients. Experiments and analysis were conducted in triplicate and batch operating systems. During the fermentation process, Optical Density, glucose levels, phosphorus levels, pH, and Dry Cell Weight (DCW) were observed. The final product then analysed using GC-MS to determine the PHBV content and 3HB (3-hydroxybutyrate) and 3HV (3-hydroxyvalerate) fraction composition in the PHBV. The results show that the maximum specific growth rate and doubling time (Td) of the H. mediterranei were 0.1258 hours-1 and 5.51 hours. H. mediterranei biomass were increasing until it reached stationary phase at 95 hours. The results show PHBV content, dry biomass and yield PHA to substrate were 2.62 g/L, 29.1% and 0.27 g/g.

Production of Poly(3-Hydroxybutyrate-Co-3-Hydroxyvalerate) by Marine Archaea Haloferax mediterranei Dsm 1411 with Yeast Extract As Nutrient Source

In many countries, most of household goods are made from polymer or plastic. The polymer manufacturing industry has been relying on fossil fuel-based raw materials which are non-biodegradable. Plastics derived from these non-biodegradable sources will be difficult to degrade in nature, which in turn will pollute the environment and harm the ecosystem. One way to reduce the ecosystem damages caused by petroleum-based plastic is by using biodegradable materials for plastic industry, such as Polyhydroxyalkanoates (PHA). Poly (3-hydroxybutyrate-co-3-hydroxyvalerate) or PHBV is an example of PHA that can be produced by several types of microorganism, one of them is marine archaea Haloferax mediterranei. The objective of this study is to determine the productivity of marine archaea H. mediterranei cultures to produce PHBV with yeast extract as main nutrient sources. Experiments and analysis were conducted in triplicate and batch operating system. During the fermentation process, optical density, glucose levels, phosphorus levels, pH, and dry cell weight (DCW) were observed. The final product was then analysed using GC-MS to determine 3HB (3-hydroxybutyrate) and 3HV (3-hydroxyvalerate) fraction composition in the PHBV. The results showed that the maximum specific growth rate (m) and doubling time (Td) of the H. mediterranei were 0.1258 hours-1 and 5.51 hours, respectively. H. mediterranei biomass increased until it reached stationary phase after 95 hours incubation. The PHBV content, dry biomass and yield of PHBV to substrate were 2.62 g/L, 29.1% and 0.27 g/g, respectively.