Harnessing photosynthetic microorganisms for enhanced bioremediation of microplastics: A comprehensive review.

Mismanaged plastics, upon entering the environment, undergo degradation through physicochemical and/or biological processes. This process often results in the formation of microplastics (MPs), the most prevalent form of plastic debris (<1 mm). MPs pose severe threats to aquatic and terrestrial ecosystems, necessitating innovative strategies for effective remediation. Some photosynthetic microorganisms can degrade MPs but there lacks a comprehensive review. Here we examine the specific role of photoautotrophic microorganisms in water and soil environments for the biodegradation of plastics, focussing on their unique ability to grow persistently on diverse polymers under sunlight. Notably, these cells utilise light and CO2 to produce valuable compounds such as carbohydrates, lipids, and proteins, showcasing their multifaceted environmental benefits. We address key scientific questions surrounding the utilisation of photosynthetic microorganisms for MPs and nanoplastics (NPs) bioremediation, discussing potential engineering strategies for enhanced efficacy. Our review highlights the significance of alternative biomaterials and the exploration of strains expressing enzymes, such as polyethylene terephthalate (PET) hydrolases, in conjunction with microalgal and/or cyanobacterial metabolisms. Furthermore, we delve into the promising potential of photo-biocatalytic approaches, emphasising the coupling of plastic debris degradation with sunlight exposure. The integration of microalgal-bacterial consortia is explored for biotechnological applications against MPs and NPs pollution, showcasing the synergistic effects in wastewater treatment through the absorption of nitrogen, heavy metals, phosphorous, and carbon. In conclusion, this review provides a comprehensive overview of the current state of research on the use of photoautotrophic cells for plastic bioremediation. It underscores the need for continued investigation into the engineering of these microorganisms and the development of innovative approaches to tackle the global issue of plastic pollution in aquatic and terrestrial ecosystems.

A circular bioprocess application of algal-based substrate for Bacillus subtilis natto production of γ-PGA.

Poly-γ-glutamic acid (γ-PGA) is a bio-derived water-soluble, edible, hydrating, non-immunogenic polymer. Bacillus subtilis natto is a wild-type γ-PGA producer originally isolated from Japanese fermented natto beans whose activity has been shown to be enhanced through ion-specific activation of Extrachromosomal DNA maintenance mechanisms. Being a GRAS γ-PGA producer, this microorganism has attracted great interest in its use within an industrial context. Here we successfully synthesised amorphous, crystalline and semi-crystalline γ-PGA between 11-27 g/L. In line with circular economy principles, scalable macroalgal biomass has been evaluated as substrate for γ-PGA, displaying great potential in both yields and material composition. In this study whole cell, freeze dried seaweed -namely Laminaria digitata, Saccharina latissima and Alaria esculenta-were pre-treated by means of mechanical methods, sterilised and subsequently inoculated with B. subtilis natto. High shear mixing was found to be the most suitable pre-treatment technique. Supplemented L. digitata (9.1 g/L), S. latissima (10.2 g/L), A. esculenta (13 g/L) displayed γ-PGA yields comparable to those of standard GS media (14.4 g/L). Greatest yields of pure γ-PGA were obtained during the month of June for L. digitata (Avg. 4.76 g/L) comparable to those obtained with GS media (7.0 g/L). Further, pre-treated S. latissima and L. digitata complex media enabled for high molar mass (4,500 kDa) γ-PGA biosynthesis at 8.6 and 8.7 g/L respectively. Compared to standard GS media, algal derived γ-PGA displayed significantly higher molar masses. Further studies will be necessary to further evaluate the impact of varying ash contents upon the stereochemical properties and modify the properties of algal media based γ-PGA with the aid of key nutrients; however, the material synthesised to date can directly displace a number of fossil fuel derived chemicals in drug delivery applications, cosmetics, bioremediation, wastewater treatment, flocculation and as cryoprotectants.

The Enrichment of Whey Protein Isolate Hydrogels with Poly-γ-Glutamic Acid Promotes the Proliferation and Osteogenic Differentiation of Preosteoblasts.

Osseous disease accounts for over half of chronic pathologies, but there is a limited supply of autografts, the gold standard; hence, there is a demand for new synthetic biomaterials. Herein, we present the use of a promising, new dairy-derived biomaterial: whey protein isolate (WPI) in the form of hydrogels, modified with the addition of different concentrations of the biotechnologically produced protein-like polymeric substance poly-γ-glutamic acid (γ-PGA) as a potential scaffold for tissue regeneration. Raman spectroscopic analysis demonstrated the successful creation of WPI-γ-PGA hydrogels. A cytotoxicity assessment using preosteoblastic cells demonstrated that the hydrogels were noncytotoxic and supported cell proliferation from day 3 to 14. All γ-PGA-containing scaffold compositions strongly promoted cell attachment and the formation of dense interconnected cell layers. Cell viability was significantly increased on γ-PGA-containing scaffolds on day 14 compared to WPI control scaffolds. Significantly, the cells showed markers of osteogenic differentiation; they synthesised increasing amounts of collagen over time, and cells showed significantly enhanced alkaline phosphatase activity at day 7 and higher levels of calcium for matrix mineralization at days 14 and 21 on the γ-PGA-containing scaffolds. These results demonstrated the potential of WPI-γ-PGA hydrogels as scaffolds for bone regeneration.

Designing of Drug Delivery Systems to Improve the Antimicrobial Efficacy in the Periodontal Pocket Based on Biodegradable Polyesters.

Delivery systems for biologically active substances such as proanthocyanidins (PCANs), produced in the form of electrospun nonwoven through the electrospinning method, were designed using a polymeric blend of poly(L-lactide-co-glycolide) (PLGA)and poly[(R,S)-3-hydroxybutyrate] ((R,S)-PHB). The studies involved the structural and thermal characteristics of the developed electrospun three-dimensional fibre matrices unloaded and loaded with PCANs. In the next step, the hydrolytic degradation tests of these systems were performed. The release profile of PCANs from the electrospun nonwoven was determined with the aid of UV-VIS spectroscopy. Approximately 30% of the PCANs were released from the tested electrospun nonwoven during the initial 15-20 days of incubation. The chemical structure of water-soluble oligomers that were formed after the hydrolytic degradation of the developed delivery system was identified through electrospray ionization mass spectrometry. Oligomers of lactic acid and OLAGA oligocopolyester, as well as oligo-3-hydroxybutyrate terminated with hydroxyl and carboxyl end groups, were recognized as degradation products released into the water during the incubation time. It was also demonstrated that variations in the degradation rate of individual mat components influenced the degradation pattern and the number of formed oligomers. The obtained results suggest that the incorporation of proanthocyanidins into the system slowed down the hydrolytic degradation process of the poly(L-lactide-co-glycolide)/poly[(R,S)-3-hydroxybutyrate] three-dimensional fibre matrix. In addition, in vitro cytotoxicity and antimicrobial studies advocate the use of PCANs for biomedical applications with promising antimicrobial activity.

Polymers Use as Mulch Films in Agriculture-A Review of History, Problems and Current Trends.

The application of mulch films for preserving soil moisture and preventing weed growth has been a part of agricultural practice for decades. Different materials have been used as mulch films, but polyethylene plastic has been considered most effective due to its excellent mechanical strength, low cost and ability to act as a barrier for sunlight and water. However, its use carries a risk of plastic pollution and health hazards, hence new laws have been passed to replace it completely with other materials over the next few years. Research to find out about new biodegradable polymers for this purpose has gained impetus in the past few years, driven by regulations and the United Nations Organization's Sustainable Development Goals. The primary requisite for these polymers is biodegradability under natural climatic conditions without the production of any toxic residual compounds. Therefore, biodegradable polymers developed from fossil fuels, microorganisms, animals and plants are viable options for using as mulching material. However, the solution is not as simple since each polymer has different mechanical properties and a compromise has to be made in terms of strength, cost and biodegradability of the polymer for its use as mulch film. This review discusses the history of mulching materials, the gradual evolution in the choice of materials, the process of biodegradation of mulch films, the regulations passed regarding material to be used, types of polymers that can be explored as potential mulch films and the future prospects in the area.

Building a circular economy around poly(D/L-γ-glutamic acid)- a smart microbial biopolymer.

Bio-derived materials have long been harnessed for their potential as backbones of biodegradable constructs. With increasing understanding of organismal biochemistry and molecular genetics, scientists are now able to obtain biomaterials with properties comparable to those achieved by the petroleum industry. Poly-γ-glutamic acid (γ-PGA) is an anionic pseudopolypeptide produced and secreted by several microorganisms, especially Bacillus species. γ-PGA is polymerised via the pgs intermembrane enzymatic complex expressed by many bacteria (including GRAS member - Bacillus subtilis). γ-PGA can exist as a homopolymer of L- glutamic acid or D- glutamic acid units or it can be a co-polymer comprised of D and L enantiomers. This non-toxic polymer is highly viscous, soluble, biodegradable and biocompatible. γ-PGA is also an example of versatile chiral-polymer, a characteristic that draws great attention from the industry. Increased understanding in the correlation between microbial genetics, substrate compositions, fermentation conditions and polymeric chemical characteristics have led to bioprocess optimisation to provide cost competitive, non-petroleum-based, biodegradable solutions. This review presents detailed insights into microbial synthesis of γ-PGA and summaries current understanding of the correlation between genetic makeup of γ-PGA-producing bacteria, range of culture cultivation conditions, and physicochemical properties of this incredibly versatile biopolymer. Additionally, we hope that review provides an updated overview of findings relevant to sustainable and cost-effective biosynthesis of γ-PGA, with application in medicine, pharmacy, cosmetics, food, agriculture and for bioremediation.

Microbial Poly-γ-Glutamic Acid (γ-PGA) as an Effective Tooth Enamel Protectant.

Poly-γ-glutamic acid (γ-PGA) is a bio-derived water-soluble, edible, non-immunogenic nylon-like polymer with the biochemical characteristics of a polypeptide. This Bacillus-derived material has great potential for a wide range of applications, from bioremediation to tunable drug delivery systems. In the context of oral care, γ-PGA holds great promise in enamel demineralisation prevention. The salivary protein statherin has previously been shown to protect tooth enamel from acid dissolution and act as a reservoir for free calcium ions within oral cavities. Its superb enamel-binding capacity is attributed to the L-glutamic acid residues of this 5380 Da protein. In this study, γ-PGA was successfully synthesised from Bacillus subtilis natto cultivated on supplemented algae media and standard commercial media. The polymers obtained were tested for their potential to inhibit demineralisation of hydroxyapatite (HAp) when exposed to caries simulating acidic conditions. Formulations presenting 0.1, 0.25, 0.5, 0.75, 1, 2, 3 and 4% (w/v) γ-PGA concentration were assessed to determine the optimal conditions. Our data suggests that both the concentration and the molar mass of the γ-PGA were significant in enamel protection (p = 0.028 and p < 0.01 respectively). Ion Selective Electrode, combined with Fourier Transform Infra-Red studies, were employed to quantify enamel protection capacity of γ-PGA. All concentrations tested showed an inhibitory effect on the dissolution rate of calcium ions from hydroxyapatite, with 1% (wt) and 2% (wt) concentrations being the most effective. The impact of the average molar mass (M) on enamel dissolution was also investigated by employing commercial 66 kDa, 166 kDa, 440 kDa and 520 kDa γ-PGA fractions. All γ-PGA solutions adhered to the surface of HAp with evidence that this remained after 60 min of continuous acidic challenge. Inductively Coupled Plasma analysis showed a significant abundance of calcium ions associated with γ-PGA, which suggests that this material could also act as a responsive calcium delivery system. We have concluded that all γ-PGA samples tested (commercial and algae derived) display enamel protection capacity regardless of their concentration or average molar mass. However, we believe that γ-PGA D/L ratios might affect the binding more than its molar mass.