Characterization of the polyhydroxyalkanoate (PHA) synthase from Ideonella sakaiensis, a bacterium that is capable of degrading and assimilating poly(ethylene terephthalate)

Tan, Hua; Chek, Min Fey; Neoh, Soon Zher; Ang, Shaik Ling; Yoshida, Shosuke; Hakoshima, Toshio; Sudesh, Kumar

doi:10.1016/j.polymdegradstab.2022.110160

Cited by 7 publications

(5 citation statements)

References 47 publications

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“…The columns were TSK guard column H HR -H and TSKgel GMH HR -H (Tosoh, Japan). The flow rate was set to 0.8 mL/min and the column temperature was maintained at 40 °C [ 42 ].…”

Section: Methodsmentioning

confidence: 99%

N-terminal truncation of PhaCBP-M-CPF4 and its effect on PHA production

Neoh,

Tan,

Trakunjae

et al. 2024

Microb Cell Fact

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Background Among the polyhydroxyalkanoate (PHA), poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] [P(3HB-co-3HHx)] is reported to closely resemble polypropylene and low-density polyethylene. Studies have shown that PHA synthase (PhaC) from mangrove soil (PhaCBP-M-CPF4) is an efficient PhaC for P(3HB-co-3HHx) production and N-termini of PhaCs influence its substrate specificity, dimerization, granule morphology, and molecular weights of PHA produced. This study aims to further improve PhaCBP-M-CPF4 through N-terminal truncation. Results The N-terminal truncated mutants of PhaCBP-M-CPF4 were constructed based on the information of the predicted secondary and tertiary structures using PSIPRED server and AlphaFold2 program, respectively. The N-terminal truncated PhaCBP-M-CPF4 mutants were evaluated in C. necator mutant PHB−4 based on the cell dry weight, PHA content, 3HHx molar composition, molecular weights, and granule morphology of the PHA granules. The results showed that most transformants harbouring the N-terminal truncated PhaCBP-M-CPF4 showed a reduction in PHA content and cell dry weight except for PhaCBP-M-CPF4 G8. PhaCBP-M-CPF4 G8 and A27 showed an improved weight-average molecular weight (Mw) of PHA produced due to lower expression of the truncated PhaCBP-M-CPF4. Transformants harbouring PhaCBP-M-CPF4 G8, A27, and T74 showed a reduction in the number of granules. PhaCBP-M-CPF4 G8 produced higher Mw PHA in mostly single larger PHA granules with comparable production as the full-length PhaCBP-M-CPF4. Conclusion This research showed that N-terminal truncation had effects on PHA accumulation, substrate specificity, Mw, and granule morphology. This study also showed that N-terminal truncation of the amino acids that did not adopt any secondary structure can be an alternative to improve PhaCs for the production of PHA with higher Mw in mostly single larger granules.

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“…The columns were TSK guard column H HR -H and TSKgel GMH HR -H (Tosoh, Japan). The flow rate was set to 0.8 mL/min and the column temperature was maintained at 40 °C [ 42 ].…”

Section: Methodsmentioning

confidence: 99%

N-terminal truncation of PhaCBP-M-CPF4 and its effect on PHA production

Neoh,

Tan,

Trakunjae

et al. 2024

Microb Cell Fact

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“…After optimization of culture conditions, I. sakaiensis accumulated PHA from PET up to 48% of dry cell weight, which corresponds to PHA titer of 0.75 g/L (Table 1 ). In addition, a subsequent study revealed substrate specificity of PHA synthase from I. sakaiensis (Tan et al 2022 ). Recently, a protocol for gene manipulation of I. sakaiensis was established (Hachisuka et al 2021 ), enabling metabolic engineering of I. sakaiensis for the production of various chemicals from PET waste.…”

Section: Ethylene Glycol Catabolism In the Pet Assimilating Bacterium...mentioning

confidence: 99%

Novel aspects of ethylene glycol catabolism

Shimizu,

Inui

2024

Appl Microbiol Biotechnol

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Ethylene glycol (EG) is an industrially important two-carbon diol used as a solvent, antifreeze agent, and building block of polymers such as poly(ethylene terephthalate) (PET). Recently, the use of EG as a starting material for the production of bio-fuels or bio-chemicals is gaining attention as a sustainable process since EG can be derived from materials not competing with human food stocks including CO2, syngas, lignocellulolytic biomass, and PET waste. In order to design and construct microbial process for the conversion of EG to value-added chemicals, microbes capable of catabolizing EG such as Escherichia coli, Pseudomonas putida, Rhodococcus jostii, Ideonella sakaiensis, Paracoccus denitrificans, and Acetobacterium woodii are candidates of chassis for the construction of synthetic pathways. In this mini-review, we describe EG catabolic pathways and catabolic enzymes in these microbes, and further review recent advances in microbial conversion of EG to value-added chemicals by means of metabolic engineering. Key points • Ethylene glycol is a potential next-generation feedstock for sustainable industry. • Microbial conversion of ethylene glycol to value-added chemicals is gaining attention. • Ethylene glycol-utilizing microbes are useful as chassis for synthetic pathways.

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“…46 Afterwards, I. sakaiensis was investigated to accumulate PHA at high levels from PET. 158 A recent study by Tan et al 159 explores in vivo and in vitro characterization of PHA synthase gene of I. sakaiensis (phaC Is ). Indeed, phaC Is was cloned and heterologously expressed in a PHAnegative mutant strain Cupriavidus necator PHBff4, the finding revealed that the strain produced P(3HB) accumulation of up to 71 wt% of the cell dry weight.…”

Section: ) Bioconversion Of Pet To Phamentioning

confidence: 99%

Sustainable bioconversion of synthetic plastic wastes to polyhydroxyalkanoate (PHA) bioplastics: recent advances and challenges

Neifar¹,

Hammami²,

Souissi³

et al. 2023

MOJABB

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Millions of tons of chemical plastics are accumulated annually worldwide in terrestrial and marine environments due to inadequate recycling plants and facilities and low circular use. Their continuous accumulation and contamination of soil and water pose a severe threat to the environment and to human, animal and plant health. There is therefore an urgent need to develop effective eco-environmental strategies to overcome the significant environmental impacts of traditional plastic waste management practises (incineration, landfilling, and recycling). In recent years, reports on microbial strains equipped with the potential of degrading plastic materials, which can further be converted into usable products such as PHA bioplastics have sprung up, and these offer a possibility to develop microbial and enzymatic technologies for plastic waste treatment and then progressing plastics circularity. In this chapter, an overview of the reported microbial and enzymatic degradations of petroleum-based synthetic plastics, specifically polyethylene, polystyrene, polypropylene, polyvinyl chloride, polyurethane and polyethylene terephthalate, is detailed. Furthermore, the harvesting of depolymerization products to produce new PHA materials with high added industrial value can be considered as an innovative solution, helping to increase synthetic plastic recycling rate and creating new circular economy opportunities. Finally, the challenge of ending plastic pollution is still difficult, but sustainable, renewable, bio-based and completely biodegradable, PHA will hold enormous promise for replacing plastics made from petrochemicals.

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Characterization of the polyhydroxyalkanoate (PHA) synthase from Ideonella sakaiensis, a bacterium that is capable of degrading and assimilating poly(ethylene terephthalate)

Cited by 7 publications

References 47 publications

N-terminal truncation of PhaCBP-M-CPF4 and its effect on PHA production

N-terminal truncation of PhaCBP-M-CPF4 and its effect on PHA production

Novel aspects of ethylene glycol catabolism

Sustainable bioconversion of synthetic plastic wastes to polyhydroxyalkanoate (PHA) bioplastics: recent advances and challenges

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