Search

Polyhydroxyalkanoate (PHA), specifically poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(3HB-co-3HHx), PHBHHx) with physical properties governed by the 3-hydroxyhexanoate (3HHx) mole fraction, is a promising bioplastic. Although engineered strains used to produce P(3HB-co-3HHx) with various 3HHx mole contents and fermentation techniques have been studied, mass production with specific 3HHx fractions and monomers depends on the batch, supply of substrates, and strains, resulting in the time-consuming development of strains and complex culture conditions for P(3HB-co-3HHx). To overcome these limitations, we blended poly(3-hydroxybutyrate) [(P(3HB), produced from C. necator H16] and P(3HB-co-20 mol%3HHx) [from C. necator 2668/pCB81] to prepare films with various 3HHx contents. We evaluated the molecular weight and physical, thermal, and mechanical properties of these films and confirmed the influence of the 3HHx monomer content on the mechanical and thermal properties as well as degradability of the blended P(3HB-co-3HHx) films containing various 3HHx mole fractions, similar to that of original microbial-based P(3HB-co-3HHx). Moreover, the degradation rate analyzed via Microbulbifer sp. was >76% at all blending ratios within 2 days, whereas a weaker effect of the 3HHx mole fraction of the blended polymer on degradation was observed. P(3HB-co-3HHx) could be produced via simple blending using abundantly produced P(3HB) and P(3HB-co-20 mol%HHx), and the resulting copolymer is applicable as a biodegradable plastic. [ABSTRACT FROM AUTHOR]

Although efforts have been undertaken to produce polyhydroxyalkanoates (PHA) with various monomers, the low yield of PHAs because of complex metabolic pathways and inhibitory substrates remains a major hurdle in their analyses and applications. Therefore, we investigated the feasibility of mass production of PHAs containing 5-hydroxyvalerate (5HV) using δ-valerolactone (DVL) without any pretreatment along with the addition of plant oil to achieve enough biomass. We identified that PhaCBP-M-CPF4, a PHA synthase, was capable of incorporating 5HV monomers and that C. necator PHB−4 harboring phaCBP-M-CPF4 synthesized poly(3HB-co-3HHx-co-5HV) in the presence of bean oil and DVL. In fed-batch fermentation, the supply of bean oil resulted in the synthesis of 49 g/L of poly(3HB-co-3.7 mol% 3HHx-co-5.3 mol%5HV) from 66 g/L of biomass. Thermophysical studies showed that 3HHx was effective in increasing the elongation, whereas 5HV was effective in decreasing the melting point. The contact angles of poly(3HB-co-3HHx-co-5HV) and poly(3HB-co-3HHx) were 109 and 98°, respectively. In addition, the analysis of microbial degradation confirmed that poly(3HB-co-3HHx-co-5HV) degraded more slowly (82% over 7 days) compared to poly(3HB-co-3HHx) (100% over 5 days). Overall, the oil-based fermentation strategy helped produce more PHA, and the mass production of novel PHAs could provide more opportunities to study polymer properties. [ABSTRACT FROM AUTHOR]

Indigo is a unique blue dye that has been used in the textile industry for centuries and is currently mass-produced commercially through chemical synthesis. However, the use of toxic substrates and reducing agents for chemical synthesis is associated with environmental concerns, necessitating the development of eco-friendly alternatives based on microbial production. In this study, a robust industrial strategy for indigo production was developed using Pseudomonas putida KT2440 as the host strain, which is characterized by its excellent ability to degrade aromatic compounds and high resistance to environmental stress. By introducing the genes tryptophanase (tnaA) and Flavin-containing monooxygenase (FMO), a P. putida HI201 strain was constructed to produce indigo from tryptophan. To enhance the indigo yield, culture conditions, including medium composition, temperature, tryptophan concentration, and shaking speed, were optimized. Under optimal conditions such as TB medium, 15 mM tryptophan, 30°C, 200 rpm, P. putida HI201 biosynthesized 1.31 g/L indigo from tryptophan in a fed-batch fermentation system. The introduction of tnaA and FMO genes also enabled the production of indigo in various P. putida species, and the indigo-producing strain had a blue color, which served as a visual indicator. This study presents a strategy for using P. putida as a host for robust and sustainable microbial production of indigo, highlighting the strain's applicability and efficiency in environment friendly dye synthesis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

With an increase in the commercialization of bioplastics, the importance of screening for plastic-degrading strains and microbes has emerged. Conventional methods for screening such strains are time-consuming and labor-intensive. Therefore, we suggest a method for quickly and effectively screening plastic-degrading microbial strains through dual esterase assays for soil and isolated strains, using p -nitrophenyl alkanoates as substrates. To select microbe-abundant soil, the total amount of phospholipid fatty acids (PLFAs) included in each soil sample was analyzed, and esterase assays were performed for each soil sample to compare the esterase activity of each soil. In addition, by analyzing the correlation coefficients and sensitivity between the amount of PLFAs and the degree of esterase activity according to the substrate, it was confirmed that substrate p NP-C2 is the most useful index for soil containing several microbes having esterase activity. In addition, esterase assays of the isolated strains allowed us to select the most active strain as the degrading strain, and 16S rRNA results confirmed that it was Bacillus sp. N04 showed the highest degradation activity for polybutylene succinate (PBS) as measured in liquid culture for 7 days, with a degradation yield of 99%. Furthermore, Bacillus sp. N04 showed degradation activity against various bioplastics. We propose the dual application of p -nitrophenyl alkanoates as an efficient method to first select the appropriate soil and then to screen for plastic-degrading strains in it, and conclude that p NP-C2 in particular, is a useful indicator.