Metabolic improvements and use of inexpensive carbon sources in microbial production of polyhydroxyalkanoates

doi:10.1016/S1389-1723(02)80198-0

Journal of Bioscience and Bioengineering

Volume 94, Issue 6, December 2002, Pages 579-584

https://doi.org/10.1016/S1389-1723(02)80198-0 Get rights and content

Abstract

This paper deals with the microbial production of polyhydroxyalkanoates (PHAs), biodegradable thermoplastics which perform excellently as a material, from inexpensive renewable carbon sources. To date, with the help of genetic engineering techniques, it has become possible to design several types of PHAs with different compositions and to enhance the productivities of PHAs. In addition, molecular breeding of PHA biosynthesis enzymes has been demonstrated to improve polymer production. Mutant PHA synthases generated by an in vitro evolution technique have allowed the enhanced production and quality alteration of PHAs. Furthermore, use of inexpensive renewable carbon sources, such as plant oils, waste materials, and carbon dioxide, would be a key for a reduction in PHA production cost.

References (32)

K Sudesh et al.
Synthesis, structure and properties of polyhydroxyalkanoates: biological polyesters
Prog. Polym. Sci.
(2000)
S Kusaka et al.
Properties and biodegradability of ultra-high-molecular-weight poly[(R)-hydroxybutyrate] produced by a recombinant Escherichia coli
Int. J. Biol. Macromol.
(1999)
B.H Rehm et al.
A new metabolic link between fatty acid de novo synthesis and polyhydroxyalkanoic acid synthesis
The phaG gene from Pseudomonas putida KT2440 encodes a 3-hydroxyacyl-acyl carrier protein-coenzyme A transferase
J. Biol. Chem.
(1998)
K Taguchi et al.
Co-expression of 3-ketoacyl-ACP reductase and polyhydroxyalkanoate synthase genes induces PHA production in Escherichia coli HB101 strain
FEMS Microbiol. Lett.
(1999)
T Tsuge et al.
Molecular cloning of two (R)-specific enoyl-CoA hydratase genes from Pseudomonas aeruginosa and their use for polyhydroxyalkanoate synthesis
FEMS Microbiol. Lett.
(2000)
Y Modis et al.
Crystallographic analysis of the reaction pathway of Zoogloea ramigera biosynthetic thiolase
J. Mol. Biol.
(2000)
S.Y Lee et al.
Recent advances in polyhydroxyalkanoate production by bacterial fermentation
Int. J. Biol. Macromol.
(1999)
T Yamane
Cultivation engineering of microbial bioplastics production
FEMS Microbiol. Rev.
(1992)
T Tsuge et al.
Development of a novel method for feeding a mixture of l-lactic acid and acetic acid in fed-batch culture of Ralstonia eutropha for poly-d-3-hydroxybutyrate production
J. Biosci. Bioeng.
(2001)
T Tsuge et al.
Optimization of l-lactic acid feeding for production of poly-d-3-hydroxybutyric acid by Alcaligenes eutrophus in fed-batch culture
J. Biosci. Bioeng.
(1999)

M Miyake et al.

A thermophilic cyanobacterium, Synechococcus sp. MA19, capable of accumulating poly-β-hydroxybutyrate

J. Ferment. Bioeng.

(1996)

A Ishizaki et al.

Production of poly-β-hydroxybutyric acid from carbon dioxide by Alcaligenes eutrophus ATCC17697^T

J. Ferment. Bioeng.

(1991)

Y Doi

(1990)

L.L Madison et al.

Metabolic engineering of poly(3-hydroxyalkanoates): from DNA to plastic

Microbiol. Mol. Biol. Rev.

(1999)

T Urakami

Biogreen

Plastics

(1999)

Cited by (190)

Properties, production, and modification of polyhydroxyalkanoates
2024, Resources, Conservation and Recycling Advances
Polyhydroxyalkanoates (PHAs) are polymers synthesized by diverse bacteria for carbon and energy storage applications. PHAs are biodegradable and nontoxic. They also exhibit properties similar to those of petroleum-based polymers. Therefore, these materials are promising alternatives to conventional plastics. This review aims to provide a comprehensive overview of PHA research, from production to application. This review summarizes the thermal and mechanical properties of various PHA homopolymers and copolymers, and compares them with those of common petroleum-based polymers. This comparison indicates that elongation at break is a major weakness of many PHAs. Different organic wastes used in PHA microbial production by mixed culture fermentation are summarized in this review. Important parameters of feedstock fermentation, culture selection, and PHA accumulation were compared. The pH and organic loading rate significantly affected the overall PHA yield, and various feedstocks led to different PHA compositions. Physical (blending and fiber reinforcement) and biological (cofeeding) modifications to improve the mechanical properties of PHA are elaborated in this review. Tensile properties are the major improvements among the mechanical properties after modification. Current applications of PHA and its derivatives are also presented in this work. They are primarily applied in the medicine, agriculture, and packaging industries. The widespread application of PHA faces challenges such as high production costs and limited mechanical properties. This study intends to stimulate further research into cost-effective methods for PHA production and to explore additional modification techniques.
Biosynthesis, characterization, and biodegradation of elastomeric polyhydroxyalkanoates consisting of α-dimethylated monomer units
2023, Materials Today Sustainability
Although poly[(R)-3-hydroxybutyrate)] [P(3HB)] is a microbial polyester that can be used as a sustainable and biodegradable plastic, its use has been limited because of poor material properties due to its stiff and brittle nature. To overcome this issue, it is critical to diversify the monomeric composition of 3HB-based polymers through the introduction of new repeating units that can improve material properties. In this study, a new 3HB-based copolymer was biosynthesized with 3-hydroxypivalate (3HPi) units containing signature dimethyl groups on the α-carbon. To biosynthesize poly[(R)-3-hydroxybutyrate-co-3-hydroxypivalate] {P(3HB-co-3HPi)}, recombinant Escherichia coli LSBJ was cultured with glucose and 3-hydroxypivalic acid (3HPiA) as the carbon source and 3HPi precursor, respectively. This resulted in the successful synthesis of P(3HB-co-3HPi) copolymers with a 3HPi fraction in the range of 2–50 mol% by adjusting the precursor concentration to the microbial catalyst. Interestingly, the biosynthesized polymers exhibited an increasing molecular weight with increasing 3HPi fractions, up to 424 × 10⁴ in weight average molecular weight. The thermal properties of P(3HB-co-3HPi) were similar to those of conventional 3HB-based copolymers, but the copolymers displayed extraordinary mechanical performance compared to other 3HB homo- and copolymers. P(3HB-co-32 mol% 3HPi) displayed an elongation at break of 1919 %, the highest value ever reported for bacterial polyesters. Given its high flexibility, P(3HB-co-3HPi) copolymers exhibited reversible deformation during tension testing, unlike other 3HB-based copolymers. Furthermore, P(3HB-co-3HPi) copolymers and monomeric 3HPiA were both biodegradable. This study has demonstrated that the incorporation of 3HPi monomers into 3HB-based copolymers results in superior mechanical properties and has introduced a promising new class of materials for use as sustainable and biodegradable plastics.
High production of poly(3-hydroxybutyrate) in Escherichia coli using crude glycerol
2023, Bioresource Technology
Poly(3-hydroxybutyrate) (PHB) is a prominent bio-plastic and recognized as the potential replacement of petroleum-derived plastics. To make PHB cost-effective, the production scheme based on crude glycerol was developed using Escherichia coli. The heterogeneous synthesis pathway of PHB was introduced into the E. coli strain capable of efficiently utilizing glycerol. The central metabolism that links to the synthesis of acetyl-CoA and NADPH was further reprogrammed to improve the PHB production. Key genes were targeted for manipulation, involving those in glycolysis, the pentose phosphate pathway, and the tricarboxylic cycle. As a result, the engineered strain gained a 22-fold increase in the PHB titer. Finally, the fed-batch fermentation was conducted with the producer strain to give the PHB titer, content, and productivity reaching 36.3 ± 3.0 g/L, 66.5 ± 2.8%, and 1.2 ± 0.1 g/L/h, respectively. The PHB yield on crude glycerol accounts for 0.3 g/g. The result indicates that the technology platform as developed is promising for the production of bio-plastics.
Polyhydroxyalkanoates production from ethanol- and lactate-rich fermentate of confectionary industry effluents
2023, International Journal of Biological Macromolecules
Polyhydroxyalkanoate (PHA) production has been the focus of considerable research to increase productivities and reduce production costs. In this study, a fermented confectionary industry wastewater was used as feedstock for mixed microbial culture PHA production. The feedstock was dominated by lactate and ethanol (60–90 % of all soluble fermentation products). The culture selection reactor was inoculated with municipal activated sludge and was operated at an organic loading rate (OLR) of 100 Cmmol·L⁻¹·d⁻¹, achieving a robust PHA-accumulating enrichment, which produced up to 52.6 ± 0.4 wt% of PHA in accumulation assays. An OLR increase in the culture selection stage to 150 Cmmol·L⁻¹·d⁻¹ led to a PHA content of 59.1 ± 0.6, a yield of 0.93 ± 0.01 Cmol-PHA·Cmol-S⁻¹ and a productivity of 0.93 ± 0.01 g-PHA L⁻¹·h⁻¹. A correlation analysis of the impact of ethanol concentrations from 3.19 to 20.3 Cmmol·L⁻¹ in the reactor showed that ethanol inhibited PHA production rate and yield and the consumption of other carbon sources available. Microbial community analysis revealed the increase of Amaricoccus genus during the bioreactor operation time, a known PHA accumulator. The produced polymer was poly(3-hydroxybutyrate) with an average molecular weight of 4.3 × 10⁵ Da and a polydispersity index of 1.88.
Oxidation of methionine-derived 2-hydroxyalkanoate unit in biosynthesized polyhydroxyalkanoate copolymers
2023, International Journal of Biological Macromolecules
In this study, 2-hydroxy-4-methylthiobutyrate (2H4MTB)-containing polyhydroxyalkanoate (PHA) copolymers were biosynthesized using methionine as the 2H4MTB precursor. 2H4MTB is a novel monomer unit that contains a sulfur atom in its side chain. The 2H4MTB-containing PHA was biosynthesized by functionalizing the leucine degradation and PHA synthesis pathways in recombinant Escherichia coli. The 2H4MTB fraction in the PHA copolymer was increased up to 30.6 mol% by increasing the methionine concentration in the medium. Using purified polymers containing 10.9 mol% 2H4MTB unit, the sulfide group of the 2H4MTB side chain was oxidized with hydrogen peroxide or peracetic acid, which resulted in the conversion of sulfide to sulfoxide and sulfone groups. The oxidized polymer was relatively hydrophilic, as revealed by water contact angle measurements, and swelled slightly when soaked in water. These results suggest that the 2H4MTB unit can be used as an oxidation site to impart hydrophilicity to the PHA copolymers.
A review on biorefining of palm oil and sugar cane agro-industrial residues by bacteria into commercially viable bioplastics and biosurfactants
2022, Fuel
The growing concerns on the rising population, global economy, food safety and environmental pollution have given impetus to the production of polyhydroxyalkanoates (PHAs) and biosurfactant, biodegradable alternatives to petrochemical plastics and surfactants, respectively. However, the application of these compounds is limited due to processing costs. Carbon sources represent half of the PHA and biosurfactant fermentation cost, therefore it is of interest to recycle agro-industrial waste as renewable carbon sources, such as palm oil and sugar cane production waste. This review aimed to provide insight on the utilisation of molasses and sweetwater from sugar cane and palm oil as carbon substrates for PHA and biosurfactant production, as well as providing examples of the application of such compounds. sugar cane and palm oil by-products are feasible as inexpensive feed for the PHA and biosurfactant production. Moreover, since sugar cane and palm oil plants are cultivated in tropical and subtropical areas, utilising these wastes can instil the circular economy spirit in line with the Sustainable Development Goals proposed by the United Nations.

View all citing articles on Scopus

^∗: phone: +81-(0)45-924-5286 fax: +81-(0)45-924-5426

View full text

Article preview

Journal of Bioscience and Bioengineering

Abstract

References (32)

Synthesis, structure and properties of polyhydroxyalkanoates: biological polyesters

Prog. Polym. Sci.

Properties and biodegradability of ultra-high-molecular-weight poly[(R)-hydroxybutyrate] produced by a recombinant Escherichia coli

Int. J. Biol. Macromol.

A new metabolic link between fatty acid de novo synthesis and polyhydroxyalkanoic acid synthesis

The phaG gene from Pseudomonas putida KT2440 encodes a 3-hydroxyacyl-acyl carrier protein-coenzyme A transferase

J. Biol. Chem.

Co-expression of 3-ketoacyl-ACP reductase and polyhydroxyalkanoate synthase genes induces PHA production in Escherichia coli HB101 strain

FEMS Microbiol. Lett.

Molecular cloning of two (R)-specific enoyl-CoA hydratase genes from Pseudomonas aeruginosa and their use for polyhydroxyalkanoate synthesis

FEMS Microbiol. Lett.

Crystallographic analysis of the reaction pathway of Zoogloea ramigera biosynthetic thiolase

J. Mol. Biol.

Recent advances in polyhydroxyalkanoate production by bacterial fermentation

Int. J. Biol. Macromol.

Cultivation engineering of microbial bioplastics production

FEMS Microbiol. Rev.

Development of a novel method for feeding a mixture of l-lactic acid and acetic acid in fed-batch culture of Ralstonia eutropha for poly-d-3-hydroxybutyrate production

J. Biosci. Bioeng.

Optimization of l-lactic acid feeding for production of poly-d-3-hydroxybutyric acid by Alcaligenes eutrophus in fed-batch culture

J. Biosci. Bioeng.

A thermophilic cyanobacterium, Synechococcus sp. MA19, capable of accumulating poly-β-hydroxybutyrate

J. Ferment. Bioeng.

Production of poly-β-hydroxybutyric acid from carbon dioxide by Alcaligenes eutrophus ATCC17697^T

J. Ferment. Bioeng.

Metabolic engineering of poly(3-hydroxyalkanoates): from DNA to plastic

Microbiol. Mol. Biol. Rev.

Biogreen

Plastics

Cited by (190)

Properties, production, and modification of polyhydroxyalkanoates

Biosynthesis, characterization, and biodegradation of elastomeric polyhydroxyalkanoates consisting of α-dimethylated monomer units

High production of poly(3-hydroxybutyrate) in Escherichia coli using crude glycerol

Polyhydroxyalkanoates production from ethanol- and lactate-rich fermentate of confectionary industry effluents

Oxidation of methionine-derived 2-hydroxyalkanoate unit in biosynthesized polyhydroxyalkanoate copolymers

A review on biorefining of palm oil and sugar cane agro-industrial residues by bacteria into commercially viable bioplastics and biosurfactants

ReviewMetabolic improvements and use of inexpensive carbon sources in microbial production of polyhydroxyalkanoates

Abstract

Prog. Polym. Sci.

Int. J. Biol. Macromol.

J. Biol. Chem.

FEMS Microbiol. Lett.

FEMS Microbiol. Lett.

J. Mol. Biol.

Int. J. Biol. Macromol.

FEMS Microbiol. Rev.

J. Biosci. Bioeng.

J. Biosci. Bioeng.

J. Ferment. Bioeng.

J. Ferment. Bioeng.

Metabolic engineering of poly(3-hydroxyalkanoates): from DNA to plastic

Microbiol. Mol. Biol. Rev.

Biogreen

Plastics

Review
Metabolic improvements and use of inexpensive carbon sources in microbial production of polyhydroxyalkanoates