Engineered Bacteria-Based Living Materials for Biotherapeutic Applications.

Future advances in therapeutics demand the development of dynamic and intelligent living materials. The past static monofunctional materials shall be unable to meet the requirements of future medical development. Also, the demand for precision medicine has increased with the progressively developing human society. Therefore, engineered living materials (ELMs) are vitally important for biotherapeutic applications. These ELMs can be cells, microbes, biofilms, and spores, representing a new platform for treating intractable diseases. Synthetic biology plays a crucial role in the engineering of these living entities. Hence, in this review, the role of synthetic biology in designing and creating genetically engineered novel living materials, particularly bacteria, has been briefly summarized for diagnostic and targeted delivery. The main focus is to provide knowledge about the recent advances in engineered bacterial-based therapies, especially in the treatment of cancer, inflammatory bowel diseases, and infection. Microorganisms, particularly probiotics, have been engineered for synthetic living therapies. Furthermore, these programmable bacteria are designed to sense input signals and respond to disease-changing environments with multipronged therapeutic outputs. These ELMs will open a new path for the synthesis of regenerative medicines as they release therapeutics that provide in situ drug delivery with lower systemic effects. In last, the challenges being faced in this field and the future directions requiring breakthroughs have been discussed. Conclusively, the intent is to present the recent advances in research and biomedical applications of engineered bacteria-based therapies during the last 5 years, as a novel treatment for uncontrollable diseases.

Recent Advances in Synthetic, Industrial and Biological Applications of Violacein and Its Heterologous Production.

Violacein, a purple pigment first isolated from a gram-negative coccobacillus Chromobacterium violaceum, has gained extensive research interest in recent years due to its huge potential in the pharmaceutic area and industry. In this review, we summarize the latest research advances concerning this pigment, which include (1) fundamental studies of its biosynthetic pathway, (2) production of violacein by native producers, apart from C. violaceum, (3) metabolic engineering for improved production in heterologous hosts such as Escherichia coli, Citrobacter freundii, Corynebacterium glutamicum, and Yarrowia lipolytica, (4) biological/pharmaceutical and industrial properties, (5) and applications in synthetic biology. Due to the intrinsic properties of violacein and the intermediates during its biosynthesis, the prospective research has huge potential to move this pigment into real clinical and industrial applications.

Ovalbumin and Kappa-Carrageenan Mixture Suppresses the Oxidative and Structural Changes in the Myofibrillar Proteins of Grass Carp (Ctenopharyngodon idella) during Frozen Storage.

This study was done to analyze the cryoprotective influence of ovalbumin (OVA) with kappa-carrageenan (KC) in grass carp myofibrillar proteins during frozen storage. Ca2+-ATPase activity of MP was significantly reduced due to protein denaturation and showed a direct association with decreased sulphydryl (SH) contents and tertiary structural properties. Besides that, an increase in carbonyl, surface hydrophobicity, and dityrosine contents was observed. The addition of OVA-KC significantly restricted the decline in Ca2+-ATPase and SH groups, which were further confirmed by the retarded increase in carbonyls. Furthermore, the addition of OVA-KC increased the stability of α-helix contents. Moreover, MP treated with 6% OVA-KC also improved intermolecular interaction forces linked with gelling and water holding properties of MP. Therefore, it can be concluded that OVA-KC could be used as an effective cryoprotectant in fish and related products for preservation and commercialization.

Spray dried nanoemulsions loaded with curcumin, resveratrol, and borage seed oil: The role of two different modified starches as encapsulating materials.

Recently, food industries are directing on the promotion of innovative food matrices fortified with bioactive compounds in order to enhance the consumer's health. Octenyl succinic anhydride modified starches (OSA-MS) such as Hi-cap100 (HCP) and purity gum 2000 (PUG) were used to fabricate emulsions co-entrapped with borage seed oil (BSO), resveratrol (RES) and curcumin (CUR), which were further spray dried to obtain powders. The fabricated microcapsules loaded with BSO, RES, and CUR displayed excellent dissolution performance, high encapsulation efficiency (≈93.05%) as well as semi-spherical shape, revealed via scanning electron microscopy (SEM). We also evaluated the impact of storage time (4 weeks) and temperature (40 °C) on the physicochemical characterization of OSA-MS coated microcapsules. Microcapsules coated with HCP exhibited greater oxidative stability, lower water activity and moisture contents rather than PUG coated microcapsules during storage because of its good film-forming properties. Addition of CUR enhanced the oxidative stability and retention of bioactive compounds. HCP microcapsules loaded with BSO + RES + CUR presented supreme retention of RES (70.32%), CUR 81.6% and γ-linolenic acid (≈ 96%). Our findings showed that CUR acted as an antioxidant agent; also, lower molecular weight OSA-MS as wall material could be used for the entrapment of bioactive compounds and promotion of innovative food products.

Supercritical fluid extraction of four aromatic herbs and assessment of the volatile compositions, bioactive compounds, antibacterial, and anti-biofilm activity.

Artemisia arborescens, Artemisia abyssinica, Pulicaria jaubertii, and Pulicaria petiolaris are fragrant herbs traditionally used in medication and as a food seasoning. To date, there are no studies on the use of supercritical fluids extraction with carbon dioxide (SFE-CO2) on these plants. This study evaluates and compares total phenolic content (TPC), antioxidant activity by DPPH• and ABTS•+, antibacterial, and anti-biofilm activities of SFE-CO2 extracts. Extraction was done by SFE-CO2 with 10% ethanol as a co-solvent. A. abyssinica extract had the highest extraction yield (8.9% ± 0.41). The GC/MS analysis of volatile compounds identified 307, 265, 213, and 201compounds in A. abyssinica, A. arborescens, P. jaubertii, and P. petiolaris, respectively. The P. jaubertii extract had the highest TPC (662.46 ± 50.93 mg gallic acid equivalent/g dry extract), antioxidant activity (58.98% ± 0.20), and antioxidant capacity (71.78 ± 1.84 mg Trolox equivalent/g dry extract). The A. abyssinica and P. jaubertii extracts had significantly higher antimicrobial activity and were more effective against Gram-positive bacteria. B. subtilis was the most sensitive bacterium. P. aeruginosa was the most resistant bacterium. P. jaubertii extract had the optimum MIC and MBC (0.4 mg/ml) against B. subtilis. All SFE-CO2 extracts were effective as an anti-biofilm formation for all tested bacteria at 1/2 MIC. Meanwhile, P. jaubertii and P. petiolaris extracts were effective anti-biofilm for most tested bacteria at 1/16 MIC. Overall, the results indicated that the SFE-CO2 extracts of these plants are good sources of TPC, antioxidants, and antibacterial, and they have promising applications in the industrial fields.

Moringa Oleifera in Malnutrition: A Comprehensive Review.

Nutritional deficiency is a major concern in developing countries, resulting in serious health consequences like mental and physical growth retardation. Moringa oleifera (Moringa), a nutritious plant growing in tropical regions of developing countries, is a candidate for overcoming nutritional deficiency. Moringa leaves are rich in protein including sulphur containing amino acids. It contains high amounts of vitamin C than oranges, a higher concentration of vitamin A than carrots, higher calcium content than milk and more potassium than bananas. Moreover, there is 9 times more iron in moringa than spinach and 4 times more fiber than oats. This review enlightens and explores the nutritional diversification of Moringa oleifera and other benefits which make it a better choice to use in our daily diet to combat malnutrition.

Carotenoid-loaded nanocarriers: A comprehensive review.

Carotenoids retain plenty of health benefits and attracting much attention recently, but they have less resistance to processing stresses, easily oxidized and chemically unstable. Additionally, their application in food and pharmaceuticals are restricted due to some limitations such as poor bioavailability, less solubility and quick release. Nanoencapsulation techniques can be used to protect the carotenoids and to uphold their original characteristics during processing, storage and digestion, improve their physiochemical properties and enhance their health promoting effects. The importance of nanocarriers in foods and pharmaceuticals cannot be denied. This review comprehensively covers recent advances in nanoencapsulation of carotenoids with biopolymeric nanocarriers (polysaccharides and proteins), and lipid-based nanocarriers, their functionalities, aptness and innovative developments in preparation strategies. Furthermore, the present state of the art encapsulation of different carotenoids via biopolymeric and lipid-based nanocarriers have been enclosed and tabulated well. Nanoencapsulation has a vast range of applications for protection of carotenoids. Polysaccharides in combination with different proteins can offer a great avenue to achieve the desired formulation for encapsulation of carotenoids by using different nanoencapsulation strategies. In terms of lipid based nanocarriers, solid lipid nanoparticles and nanostructure lipid carriers are proving as the encouraging candidates for entrapment of carotenoids. Additionally, nanoliposomes and nanoemulsion are also promising and novel-vehicles for the protection of carotenoids against challenging aspects as well as offering an effectual controlled release on the targeted sites. In the future, further studies could be conducted for exploring the application of nanoencapsulated systems in food and gastrointestinal tract (GIT) for industrial applications.

Aptamer Induced Multicolored AuNCs-WS2 "Turn on" FRET Nano Platform for Dual-Color Simultaneous Detection of AflatoxinB1 and Zearalenone.

Mycotoxins posit serious threats to human and animal health, and numerous efforts have been performed to detect the multiple toxins by a single diagnostic approach. To best of our knowledge, for the first time, we synthesized an aptamer induced "turn on" fluorescence resonance energy transfer (FRET) biosensor using dual-color gold nanoclusters (AuNCs), l-proline, and BSA synthesized AuNCs (Lp-AuNCs and BSA-AuNCs), with WS2 nanosheet for simultaneous recognition of aflatoxinB1 (AFB1) and zearalenone (ZEN) by single excitation. Here, AFB1 aptamer stabilized blue-emitting AuNCs (AFB1-apt-Lp-AuNCs) (at 442 nm) and ZEN aptamer functionalized with red-colored AuNCs (ZEN-apt-BSA-AuNCs) (at 650 nm) were employed as an energy donor and WS2 nanosheet as a fluorescence quencher. With the addition of AFB1 and ZEN, the change in fluorescence intensity (F.I) was recorded at 442 and 650 nm and can be used for simultaneous recognition with a detection limit of 0.34 pg mL-1 (R2 = 0.9931) and 0.53 pg mL-1 (R2 = 0.9934), respectively. Most importantly, the semiquantitative determination of AFB1 and ZEN can also be realized through photovisualization. The current approach paves a new way to develop sensitive, selective, and convenient metal nanocluster-based fluorescent "switch-on" probes with potential applications in multipurpose biosensing.

A "turnon" aptasensor for simultaneous and time-resolved fluorometric determination of zearalenone, trichothecenes A and aflatoxin B1 using WS2 as a quencher.

A "turn on" time-resolved fluorometric aptasensor is described for the simultaneous detection of zearalenone (ZEN), trichothecenes A (T-2), and aflatoxin B1 (AFB1). Multicolor-emissive nanoparticles doped with lanthanide ions (Dy3+, Tb3+, Eu3+) were functionalized with respective aptamers and applied as a bioprobe, and tungsten disulfide (WS2) nanosheets are used as a quencher of time-resolved fluorescence. The assay exploits the quenching efficiency of WS2 and the interactions between WS2 and the respective DNA aptamers. The simultaneous recognition of the three mycotoxins can be performed in a single solution. In the absence of targets, WS2 is easily adsorbed by the mixed bioprobes via van der Waals forces between nucleobases and the WS2 basal plane. This brings the bioprobe and WS2 into close proximity and results in quenched fluorescence. In the presence of targets, the fluorescence of the bioprobes is restored because the analytes react with DNA probe and modify their molecular conformation to weaken the interaction between the DNAs and WS2. Under the optimum conditions and at an excitation wavelength of 273 nm, the time-resolved fluorescence intensities (peaking at 488, 544 and 618 nm and corresponding to emissions of Dy3+, Tb3+ and Eu3+) were used to quantify ZEN, T-2 and AFB1, respectively, with detection limits of 0.51, 0.33 and 0.40 pg mL-1 and a linear range from 0.001 to 100 ng mL-1. The three mycotoxins can be detected simultaneously without mutual interference. The assay was applied to the quantification of ZEN, T-2 and AFB1 in (spiked) maize samples. This homogeneous aptamer based assay can be performed within 1 h. Conceivably, it can become an alternative to other heterogeneous methods such as the respective enzyme-linked immunosorbent assays. Graphical abstract Schematic presentation of an aptasensor for simultaneous detection of zearalenone, trichothecenes A and aflatoxin B1 using aptamer modified time-resolved fluorescence nanoparticles as signalling probes and tungsten disulfide as the quencher. This assay shows lower detection limit and requires no washing steps.