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Fulltext Biosynthesized Poly(3-Hydroxybutyrate) on Coated Pineapple Leaf Fiber Papers for Biodegradable Packaging Application

Vaithanomsat P, Kongsin K, Trakunjae C, Boonyarit J, Jarerat A, Sudesh K, et al.

Polymers (Basel), 2021 May 26;13(11).
PMID: 34073164 DOI: 10.3390/polym13111733

This paper is aimed at investigating the usage of biosynthesized poly(3-hydroxybutyrate) (P(3-HB)) for a coating on pineapple leaf fiber paper (PLFP). For this purpose, (P(3-HB)) was produced by Rhodococcus pyridinivorans BSRT1-1, a highly potential P(3-HB) producing bacterium, with a weight-average molecular weight (Mw) of 6.07 × 10 -5 g/mol. This biosynthesized P(3-HB) at 7.5% (w/v) was then coated on PLFP through the dip-coating technique with chloroform used as a solvent. The respective coated PLFP showed that P(3-HB) could be well coated all over on the PLFP surface as confirmed by scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectroscopy. The brightness and mechanical properties of PLFP could be improved by coating with biosynthesized P(3-HB) in comparison to commercially available P(3-HB) and non-coated PLFP. Furthermore, coating of P(3-HB) significantly increased the water drop penetration time on the surface of PLFP and was similar to that of the commercial P(3-HB) with the same content. The results showed that all the coated PLPF samples can be degraded under the soil burial test conditions. We have demonstrated that the P(3-HB) coated PLFP paper has the ability to prevent water drop penetration and could undergo biodegradation. Taken together, the P(3-HB) coated PLFP can be applied as a promising biodegradable paper packaging.
Fulltext Enhanced polyhydroxybutyrate (PHB) production by newly isolated rare actinomycetes Rhodococcus sp. strain BSRT1-1 using response surface methodology

Trakunjae C, Boondaeng A, Apiwatanapiwat W, Kosugi A, Arai T, Sudesh K, et al.

Sci Rep, 2021 01 21;11(1):1896.
PMID: 33479335 DOI: 10.1038/s41598-021-81386-2

Poly-β-hydroxybutyrate (PHB) is a biodegradable polymer, synthesized as carbon and energy reserve by bacteria and archaea. To the best of our knowledge, this is the first report on PHB production by a rare actinomycete species, Rhodococcus pyridinivorans BSRT1-1. Response surface methodology (RSM) employing central composite design, was applied to enhance PHB production in a flask scale. A maximum yield of 3.6 ± 0.5 g/L in biomass and 43.1 ± 0.5 wt% of dry cell weight (DCW) of PHB were obtained when using RSM optimized medium, which was improved the production of biomass and PHB content by 2.5 and 2.3-fold, respectively. The optimized medium was applied to upscale PHB production in a 10 L stirred-tank bioreactor, maximum biomass of 5.2 ± 0.5 g/L, and PHB content of 46.8 ± 2 wt% DCW were achieved. Furthermore, the FTIR and 1H NMR results confirmed the polymer as PHB. DSC and TGA analysis results revealed the melting, glass transition, and thermal decomposition temperature of 171.8, 4.03, and 288 °C, respectively. In conclusion, RSM can be a promising technique to improve PHB production by a newly isolated strain of R. pyridinivorans BSRT1-1 and the properties of produced PHB possessed similar properties compared to commercial PHB.
Fulltext Statistical optimization of P(3HB-co-3HHx) copolymers production by Cupriavidus necator PHB-4/pBBR_CnPro-phaCRp and its properties characterization

Trakunjae C, Boondaeng A, Apiwatanapiwat W, Janchai P, Neoh SZ, Sudesh K, et al.

Sci Rep, 2023 Jun 02;13(1):9005.
PMID: 37268758 DOI: 10.1038/s41598-023-36180-7

Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] is a bacterial copolymer in the polyhydroxyalkanoates (PHAs) family, a next-generation bioplastic. Our research team recently engineered a newly P(3HB-co-3HHx)-producing bacterial strain, Cupriavidus necator PHB-4/pBBR_CnPro-phaCRp. This strain can produce P(3HB-co-2 mol% 3HHx) using crude palm kernel oil (CPKO) as a sole carbon substrate. However, the improvement of P(3HB-co-3HHx) copolymer production by this strain has not been studied so far. Thus, this study aims to enhance the production of P(3HB-co-3HHx) copolymers containing higher 3HHx monomer compositions using response surface methodology (RSM). Three significant factors for P(3HB-co-3HHx) copolymers production, i.e., CPKO concentration, sodium hexanoate concentration, and cultivation time, were studied in the flask scale. As a result, a maximum of 3.6 ± 0.4 g/L of P(3HB-co-3HHx) with 4 mol% 3HHx compositions was obtained using the RSM optimized condition. Likewise, the higher 3HHx monomer composition (5 mol%) was obtained when scaling up the fermentation in a 10L-stirrer bioreactor. Furthermore, the produced polymer's properties were similar to marketable P(3HB-co-3HHx), making this polymer suitable for a wide range of applications.
N-terminal truncation of PhaCBP-M-CPF4 and its effect on PHA production

Neoh SZ, Tan HT, Trakunjae C, Chek MF, Vaithanomsat P, Hakoshima T, et al.

Microb Cell Fact, 2024 Feb 15;23(1):52.
PMID: 38360657 DOI: 10.1186/s12934-024-02329-w

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.