Generally Recognized As Safe Research Articles

The Frequently Used Industrial Food Process Additive, Microbial Transglutaminase: Boon or Bane.

Microbial transglutaminase (mTG) is a frequently consumed processed food additive, and use of its cross-linked complexes is expanding rapidly. It was designated as a processing aid and was granted the generally recognized as safe (GRAS) classification decades ago, thus avoiding thorough assessment according to current criteria of toxicity and public health safety. In contrast to the manufacturer's declarations and claims, mTG and/or its transamidated complexes are proinflammatory, immunogenic, allergenic, pathogenic, and potentially toxic, hence raising concerns for public health. Being a member of the transglutaminase family and functionally imitating the tissue transglutaminase, mTG was recently identified as a potential inducer of celiac disease. Microbial transglutaminase and its docked complexes have numerous detrimental effects. Those harmful aspects are denied by the manufacturers, who claim the enzyme is deactivated when heated or by gastric acidity, and that its covalently linked isopeptide bonds are safe. The present narrative review describes the potential side effects of mTG, highlighting its thermostability and activity over a broad pH range, thus, challenging the manufacturers' and distributers' safety claims. The national food regulatory authorities and the scientific community are urged to reevaluate mTG's GRAS status, prioritizing public health protection against the possible risks associated with this enzyme and its health-damaging consequences.

Screening, Synthesis, and Characterization of a More Rapidly Dissolving Celecoxib Crystal Form.

The prevalence of poor solubility in active pharmaceutical ingredients (APIs) such as celecoxib (CEL) is a major bottleneck in the pharmaceutical industry, leading to a low concentration gradient, poor passive diffusion, and in vivo failure. This study presents the synthesis and characterization of a new cocrystal of the API CEL. CEL is a nonsteroidal anti-inflammatory drug used for the treatment of osteoarthritis and rheumatoid arthritis. Computational screening was completed for CEL against a large library of generally recognized as safe (GRAS) coformers, based on molecular complementarity and hydrogen bond propensity (HBP). The generated list of 17 coformers with a likelihood for cocrystallization with CEL were experimentally screened using four techniques: liquid-assisted grinding (LAG), solvent evaporation (SE), gas antisolvent crystallization (GAS), and supercritical enhanced atomization (SEA). One new crystalline form was isolated, employing the liquid coformer N-ethylacetamide (NEA). This novel form, celecoxib-di-N-ethylacetamide (CEL·2NEA), was characterized by a variety of different techniques. The crystal structure was determined through single-crystal X-ray diffraction. Both NEA molecules are evolved from the crystal structure at a desolvation temperature of approximately 65 °C. The CEL·2NEA cocrystal exhibited a dissolution rate, with more than a twofold improvement in comparison to as-received CEL after only 15 min.

Workshop report: A study roadmap to evaluate the safety of recombinant human lactoferrin expressed in Komagataella phaffii intended as an ingredient in conventional foods – Recommendations of a scientific expert panel

Human milk lactoferrin (hmLF) is a glycoprotein with well-known effects on immune function. Helaina Inc. has used a glycoengineered yeast, Komatagaella phaffii, to produce recombinant human lactoferrin (Helaina rhLF, Effera™) that is structurally similar to hmLF with intended uses as a food ingredient. However, earlier FDA reviews of rhLF were withdrawn due to insufficient safety data and unanswered safety questions the experts and FDA raised about the immunogenicity/immunotoxicity risks of orally ingested rhLF. Helaina organized a panel of leading scientists to build and vet a safety study roadmap containing the studies and safety endpoints needed to address these questions. Panelists participated in a one-day virtual workshop in June 2023 and ensuing discussions through July 2023. Relevant workshop topics included physicochemical properties of LF, regulatory history of bovine LF and rhLF as food ingredients in the FDA's generally recognized as safe (GRAS) program, and synopses of publicly available studies on the immunogenicity/alloimmunization, immunotoxicology, iron homeostasis, and absorption, distribution, metabolism, and excretion of rhLF. Panelists concluded that the safety study roadmap addresses the unanswered safety questions and the intended safe use of rhLF as a food ingredient for adults and agreed on broad applications of the roadmap to assess the safety and support GRAS of other recombinant milk proteins with immunomodulatory functions.

The Use of Ultraviolet Irradiation to Improve the Efficacy of Acids That Are Generally Recognized as Safe for Disinfecting Fresh Produce in the Ready-to-Eat Stage.

Fresh-cut produce is usually produced under standardized disinfection processes, which are unavailable at the ready-to-eat stage. Currently, chemical sanitizers are used for washing, but their disinfection efficacy is limited. In this study, UV-C (1.03 kJ/m2) was combined with organic acids that are generally recognized as safe (GRAS), including citric, malic, acetic, and lactic acids (LAs), to wash lettuce and cherry tomatoes that are contaminated with Escherichia coli O157:H7 and Salmonella Typhimurium. The results showed that LA was the most effective treatment among the single treatments, with a pathogen reduction and cross-contamination incidence of 2.0-2.3 log CFU/g and 28-35%, respectively. After combining with UV-C, the disinfection efficacy and cross-contamination prevention capacity of the four GRAS acids significantly improved. Among the combination treatments, the highest pathogen reduction (2.5-2.7 log CFU/g) and the lowest cross-contamination incidence (11-15%) were achieved by LA-UV. The analyses of ascorbic acid, chlorophyll, lycopene, antioxidant capacity, and ΔE indicated that neither the single nor combination treatments negatively affected the quality properties. These results provide a potential hurdle technology for fresh produce safety improvement at the ready-to-eat stage.

Microenvironment-triggered cascade metal-polyphenolic nanozyme for ROS/NO synergistic hyperglycemic wound healing

Wound infection of hyperglycemic patient often has extended healing period and increased probability due to the high glucose level. However, achieving precise and safe therapy of the hyperglycemic wound with specific wound microenvironment (WME) remains a major challenge. Herein, a WME-activated smart L-Arg/GOx@TA-Fe (LGTF) nanozymatic system composed of generally recognized as safe (GRAS) compound is engineered. The nanozymatic system combining metal-polyphenol nanozyme (tannic acid-Fe3+, TA-Fe) and natural enzyme (glucose oxidase, GOx) can consume the high-concentration glucose, generating reactive oxygen species (ROS) and nitric oxide (NO) in situ to synergistically disinfect hyperglycemia wound. In addition, glucose consumption and gluconic acid generation can lower glucose level to promote wound healing and reduce the pH of WME to enhance the catalytic activities of the LGTF nanozymatic system. Thereby, low-dose LGTF can perform remarkable synergistic disinfection and healing effect towards hyperglycemic wound. The superior biosafety, high catalytic antibacterial and beneficial WME regulating capacity demonstrate this benign GRAS nanozymatic system is a promising therapeutic agent for hyperglycemic wound.

Characterizations on a GRAS Electrospun Lipid-Polymer Composite Loaded with Tetrahydrocurcumin.

Electrospun/sprayed fiber films and nanoparticles were broadly studied as encapsulation techniques for bioactive compounds. Nevertheless, many of them involved using non-volatile toxic solvents or non-biodegradable polymers that were not suitable for oral consumption, thus rather limiting their application. In this research, a novel electrospun lipid-polymer composite (ELPC) was fabricated with whole generally recognized as safe (GRAS) materials including gelatin, medium chain triglyceride (MCT) and lecithin. A water-insoluble bioactive compound, tetrahydrocurcumin (TC), was encapsulated in the ELPC to enhance its delivery. Confocal laser scanning microscopy (CLSM) was utilized to examine the morphology of this ELPC and found that it was in a status between electrospun fibers and electrosprayed particles. It was able to form self-assembled emulsions (droplets visualized by CLSM) to deliver active compounds. In addition, this gelatin-based ELPC self-assembled emulsion was able to form a special emulsion gel. CLSM observation of this gel displayed that the lipophilic contents of the ELPC were encapsulated within the cluster of the hydrophilic gelatin gel network. The FTIR spectrum of the TC-loaded ELPC did not show the fingerprint pattern of crystalline TC, while it displayed the aliphatic hydrocarbon stretches from MCT and lecithin. The dissolution experiment demonstrated a relatively linear release profile of TC from the ELPC. The lipid digestion assay displayed a rapid digestion of triglycerides in the first 3-6 min, with a high extent of lipolysis. A Caco-2 intestinal monolayer transport study was performed. The ELPC delivered more TC in the upward direction than downwards. MTT study results did not report cytotoxicity for both pure TC and the ELPC-encapsulated TC under 15 μg/mL. Caco-2 cellular uptake was visualized by CLSM and semi-quantified to estimate the accumulation rate of TC in the cells over time.

A Review of the Toxicity of Ingredients in e-Cigarettes, Including Those Ingredients Having the FDA's "Generally Recognized as Safe (GRAS)" Regulatory Status for Use in Food.

Some firms and marketers of electronic cigarettes (e-cigarettes; a type of electronic nicotine delivery system (ENDS)) and refill liquids (e-liquids) have made claims about the safety of ingredients used in their products based on the term "GRAS or Generally Recognized As Safe" (GRAS). However, GRAS is a provision within the definition of a food additive under section 201(s) (21 U.S.C. 321(s)) of the U.S. Federal Food Drug and Cosmetic Act (FD&C Act). Food additives and GRAS substances are by the FD&C Act definition intended for use in food, thus safety is based on oral consumption; the term GRAS cannot serve as an indicator of the toxicity of e-cigarette ingredients when aerosolized and inhaled (ie, vaped). There is no legal or scientific support for labeling e-cigarette product ingredients as "GRAS." This review discusses our concerns with the GRAS provision being applied to e-cigarette products and provides examples of chemical compounds that have been used as food ingredients but have been shown to lead to adverse health effects when inhaled. The review provides scientific insight into the toxicological evaluation of e-liquid ingredients and their aerosols to help determine the potential respiratory risks associated with their use in e-cigarettes.

Therapeutic cargo-free linalool-based nanoparticles attenuate inflammation by targeting NLRP3 inflammasome

Linalool is a commonly used monoterpene obtained from the essential oils of plants that impart multiple pharmacological effects. However, because of poor aqueous solubility, therapeutic efficacy is low, therefore effective methods are desired for improving the efficacy of linalool. In this study, instead of encapsulating linalool, we directly amalgamated linalool with glyceryl monostearate (GMS) and formulate linalool-based nanoparticles (Lin-NPs) using the hot melt method. Subsequently, we characterized Lin-NPs for targeting and suppressing the lipopolysaccharide (LPS)-induced inflammation in RAW264.7 cells. The hydrodynamic diameter of GMS-NPs was ∼175 nm but after amalgamating with linalool, the hydrodynamic diameter was found to be ∼214 nm. Furthermore, particle size and surface morphological characteristics were further confirmed with scanning electron microscopy and atomic force microscopy. The formulation of Lin-NPs was also demonstrated with Fourier-transform infrared spectroscopy (FTIR). These Lin-NPs were then assessed for their pharmacological efficacy in terms of their anti-inflammatory potential. These Lin-NPs effectively suppressed the ROS generation. Finally, investigated the effects of Lin-NPs on the NLRP3 inflammasome pathway as confirmed by the inhibition of NLRP-3, caspase-1 (inhibition of the inflammatory response initiator), and significant suppression of the other downstream biomarkers of the NLRP3 inflammasome pathway such as interleukin-1β and interleukin-18. In conclusion, amalgamation of linalool in GMS to develop Lin-NPs improved the efficacy of linalool, and plausible anti-inflammatory effects of Lin-NPs might be due to inhibition of NLRP3 inflammasome pathway. Moreover, the GMS and linalool both are listed under the FDA-approved Generally Recognized as Safe (GRAS) material and therefore Lin-NPs could supplement as potential NPs to combat inflammation.

Animal-free production of hen egg ovalbumin in engineered Saccharomyces cerevisiae via precision fermentation

To enable the sustainable production of ovalbumin (OVA) without relying on animal sources, the generally recognized as safe (GRAS) host Saccharomyces cerevisiae was used for the precision fermentation-based production of recombinant OVA. For this purpose, a signal peptide derived from EPX1, the most abundant extracellular protein produced by Pichia pastoris, was identified as a novel signal peptide for the efficient secretion of OVA in S. cerevisiae. To improve OVA secretion and cell growth, three helper proteins (PDI1, KAR2, and HAC1) present in the endoplasmic reticulum were expressed individually or in combination. Notably, the +P1/K2 strain coexpressing PDI1 and KAR2 with OVA produced 2 mg/L of OVA in the medium fraction; this value was 2.6-fold higher than the corresponding value for the control strain without helper proteins. Finally, a glucose-limited fed-batch fermentation process using the +P1/K2 strain yielded 132 mg/L of total OVA with 8 mg/L of extracellular OVA.

Characterization of a Food‐Safe Colorimetric Indicator Based on Black Rice Anthocyanin/PET Films for Visual Analysis of Fish Spoilage

ABSTRACTThe safety of food products is of prime importance for consumers and manufacturers. Many means can be used to validate a food product's safety before it is consumed. This study is about the preparation, characterization and evaluation of a generally recognized as safe (GRAS) sensitive colorimetric sensor that detects volatile gases (TVB‐N) resulting from fish spoilage, thus indicating the pH variation of packaged fish products. This is performed by coating a thin layer of ink sensors on the surface of the supporting matrix (corona‐treated PET). Various visual pH indicators were prepared based on black rice anthocyanin as an FDA‐approved dye. Black rice, which contains more than 80% cyanidin‐3‐glucoside, is the most prevalent anthocyanin component. Because of its low toxicity and high concentration, it can be utilized as a natural food colourant. pH indicators based on black rice can show distinct colours in various pH: from red (low pH) to violet (4–6) and deep purple/blue (6–7), blue (7–9) to yellowish/light brown (9–13) throughout the acid–base reaction by the analyte. The ink formulation was prepared by incorporating a binder system (PVOH‐PEG) for higher surface wettability, a crosslinking agent (citric acid) for higher adhesion, an antifoaming agent (natural vanillin) and acetic acid as a pH fixing agent. Corona treatments affected substrate surface chemistry in this study. The samples with thermal treatment passed the ASTM D3330 tape test, the 8000 passages for dry sponge and the 25 passages for wet sponge through the abrasion method. Anthocyanin concentration in formulated ink based on calculation by UV–vis spectra is 0.240 mg/100 g. Sensitivity tests towards TVB‐N gases were carried out at a temperature of 4°C to evaluate the performance of colorimetric films with formulated ink along with thermal treatment (temperature: 165°C, time: 5 min) using the volatile gases emitted by the fish sample inside the package.

Current status, challenges and prospects of antifouling materials for oncology applications.

Targeted therapy has become crucial to modern translational science, offering a remedy to conventional drug delivery challenges. Conventional drug delivery systems encountered challenges related to solubility, prolonged release, and inadequate drug penetration at the target region, such as a tumor. Several formulations, such as liposomes, polymers, and dendrimers, have been successful in advancing to clinical trials with the goal of improving the drug's pharmacokinetics and biodistribution. Various stealth coatings, including hydrophilic polymers such as PEG, chitosan, and polyacrylamides, can form a protective layer over nanoparticles, preventing aggregation, opsonization, and immune system detection. As a result, they are classified under the Generally Recognized as Safe (GRAS) category. Serum, a biological sample, has a complex composition. Non-specific adsorption of chemicals onto an electrode can lead to fouling, impacting the sensitivity and accuracy of focused diagnostics and therapies. Various anti-fouling materials and procedures have been developed to minimize the impact of fouling on specific diagnoses and therapies, leading to significant advancements in recent decades. This study provides a detailed analysis of current methodologies using surface modifications that leverage the antifouling properties of polymers, peptides, proteins, and cell membranes for advanced targeted diagnostics and therapy in cancer treatment. In conclusion, we examine the significant obstacles encountered by present technologies and the possible avenues for future study and development.

CLI: A new protocol for the isolation of Lactic Acid Bacteria from complex plant samples

Lactic Acid Bacteria (LAB) are predominantly probiotic microorganisms and the most are Generally Recognized As Safe (GRAS). LAB inhabit in the human gut ecosystem and are largely found in fermented foods and silage. In the last decades, LAB have also has been found in plant microbiota as a new class of microbes with probiotic activity to plants. For this reason, today the scientific interest in the study and isolation of LAB for agronomic application has increased. However, isolation protocols from complex samples such as plant tissues are scarce and inefficient.In this study, we developed a new protocol (CLI, Complex samples LAB Isolation) which yields purified LAB from plants. The sensitivity of CLI protocol was sufficient to isolate representative microorganisms of LAB genera (i.e. Leuconostoc, Lactococcus and Enterococcus).CLI protocol consists on five steps: i) sample preparation and pre-incubation in 1% sterile peptone at 30 °C for 24–48 h; ii) Sample homogenization in vortex by 10 min; iii) sample serial dilution in quarter-strength Ringer solution, iv) incubation in MRS agar plates with 0.2% of sorbic acid, with 1% of CaCO3, O2 < 15%, at pH 5.8 and 37 °C for 48 h.; v) Selection of single colonies with LAB morphology and CaCO3-solubilization halo. Our scientific contribution is that CLI protocol could be used for several complex samples and represents a useful method for further studies involving native LAB.

Classification of substances by health hazard using deep neural networks and molecular electron densities

In this paper we present a method that allows leveraging 3D electron density information to train a deep neural network pipeline to segment regions of high, medium and low electronegativity and classify substances as health hazardous or non-hazardous. We show that this can be used for use-cases such as cosmetics and food products. For this purpose, we first generate 3D electron density cubes using semiempirical molecular calculations for a custom European Chemicals Agency (ECHA) subset consisting of substances labelled as hazardous and non-hazardous for cosmetic usage. Together with their 3-class electronegativity maps we train a modified 3D-UNet with electron density cubes to segment reactive sites in molecules and classify substances with an accuracy of 78.1%. We perform the same process on a custom food dataset (CompFood) consisting of hazardous and non-hazardous substances compiled from European Food Safety Authority (EFSA) OpenFoodTox, Food and Drug Administration (FDA) Generally Recognized as Safe (GRAS) and FooDB datasets to achieve a classification accuracy of 64.1%. Our results show that 3D electron densities and particularly masked electron densities, calculated by taking a product of original electron densities and regions of high and low electronegativity can be used to classify molecules for different use-cases and thus serve not only to guide safe-by-design product development but also aid in regulatory decisions.Scientific contributionWe aim to contribute to the diverse 3D molecular representations used for training machine learning algorithms by showing that a deep learning network can be trained on 3D electron density representation of molecules. This approach has previously not been used to train machine learning models and it allows utilization of the true spatial domain of the molecule for prediction of properties such as their suitability for usage in cosmetics and food products and in future, to other molecular properties. The data and code used for training is accessible at https://github.com/s-singh-ivv/eDen-Substances.

Functional Food for Sarcopenia: Investigating the Impact of Myostatin Epitope Fusion Protein in Elderly Mice using Probiotic Bacillus subtilis

ABSTRACT Probiotics have a well-established history of positively impacting human health, particularly concerning immune function. Utilizing Generally Recognized as Safe (GRAS) strains to produce epitopes as probiotic foods exhibits significant potential in eliciting robust immune responses. The present study examines the effectiveness and safety of using a myostatin epitope fusion protein for increasing muscle mass in elderly mice. The fusion protein was developed with a targeting peptide for M-cells and was expressed in the food-grade host, Bacillus subtilis. An industrial potential formula was developed, which is composed of probiotic B. subtilis 3A16 and epitope expressing food-grade host B. subtilis WB800. This formula is safe and effective in increasing muscle mass in elderly mice, suggesting that it may be a potential treatment for sarcopenia and benefit human health as well.

Microstructural and physicochemical quality maintenance in green bell pepper infected with Botrytis cinerea and treated with thyme essential oil combined with carnauba wax.

Bell pepper presents rapid weight loss and is highly susceptible to gray mold caused by the fungus Botrytis cinerea. The most employed method to control this disease is the application of synthetic fungicides such as thiabendazole (TBZ); however, its continued use causes resistance in fungi as well as environmental problems. For these reasons, natural alternatives arise as a more striking option. Currently, bell pepper fruits are coated with carnauba wax (CW) to prevent weight loss and improve appearance. Moreover, CW can be used as a carrier to incorporate essential oils, and previous studies have shown that thyme essential oil (TEO) is highly effective against B. cinerea. Therefore, this study aimed to evaluate the effect of CW combined with TEO on the development of gray mold and maintenance of microestructural and postharvest quality in bell pepper stored at 13°C. The minimal inhibitory concentration of TEO was 0.5%. TEO and TBZ provoked the leakage of intracellular components. TEO and CW+TEO treatments were equally effective to inhibit the development of gray mold. On the quality parameters, firmness and weight loss were ameliorated with CW and CW+TEO treatments; whereas lightness increased in these treatments. The structural analysis showed that CW+TEO treatment maintained the cell structure reducing the apparition of deformities. The results suggest that CW+TEO treatment could be used as a natural and effective antifungal retarding the appearance of gray mold and maintaining the postharvest quality of bell pepper. PRACTICAL APPLICATION: CW and TEO are classified as generally recognized as safe (GRAS) by the US Food and Drug Administration (FDA). This combination can be employed on the bell pepper packaging system to extend shelf life and oppose gray mold developments. Bell pepper fruits are normally coated with lipid-base coatings such as CW before commercialization; therefore, TEO addition would represent a small investment without any changes on the packaging system infrastructure.

Abstract 3207: Chemical fabrication and characterization of a novel nanoparticle of the histone deacetylase (HDACi) romidepsin

Abstract Introduction: Histone Deacetylase inhibitors (HDACi) are important drugs for the treatment of patients with PTCL (peripheral T-cell lymphoma). Romidepsin (R), a potent Class I HDACi, is given intravenously, weekly for three to four consecutive weeks. R has a half-life of three hours and is extensively protein bound. The indication for R in PTCL was recently withdrawn due to a failed Phase 4 study. Given the need for new drugs in PTCL, and the importance of epigenetically targeted drugs for the disease, we developed a first-in-class nanoparticle of R (NR) deploying a proprietary nanochemistry platform. Methods: We modified the bulk nanoprecipitation method with FDA approved and Generally Recognized as Safe (GRAS) materials, specifically including PEGylated amphiphilic diblock copolymers, to develop NR combinatorially. By combining a drug and polymer solution with an anti-solvent, we produced core-shell structured NR particles with “stealth” properties. To achieve high R loading, stability, purity, scalability, and reproducibility, we devised a multi-pronged optimization system by modulating parameter sets in tandem. We determined the optimal parameters for the "lockdown" formulation. We purified NR by using centrifugal filters and determined stability by storing at 4°C. We used Cryo-electron microscopy (cryo-EM), Dynamic Light Scattering (DLS), and Liquid Chromatography-Mass Spectrometry (LC-MS) to determine the morphology, size, PDI, and drug concentration of NR. We developed fluorescent-NR with DiO dye to test bioavailability. We tested NR in T-cell lymphoma cell lines for histone acetylation and apoptosis induction. Furthermore, we investigated the in vivo pharmacokinetics and toxicity of NR in PTCL xenograft mouse model and quantified the drug by LC-MS. Results: The physiochemical analysis of NR demonstrated that the particles were spherical, with a hydrodynamic diameter of 46 ±5 nm and PDI of 0.15 - 0.2. NR had a negative zeta potential and an exceptional drug encapsulation efficiency of over 50% (&amp;gt;500 µg/mL). NR exhibited high stability with respect to size, PDI and drug concentration for over two years. Flow cytometry and western blot analyses indicate NR induces apoptosis and histone acetylation in a time and dose-dependent manner, equivalent to or better than that observed for R. NR’s AUC increased 25-fold when administered intravenously. NR had superior tolerability, and DiO-NR confirmed superior biodistribution in tumor in a PTCL xenograft mouse model. Conclusion: We developed a cutting-edge approach with minimum excipients, which enabled NR scale up with batch-to-batch reproducibility and high quality co-packaging of fluorescent dye and drug. NR improved R’s pharmacokinetic profile and efficacy in pre-clinical xenograft mice models. Our versatile technology enables Chemistry Materials and Controls products, an essential step in clinical product development. Citation Format: Samir Zuberi, Deepthi P. Damera, Ipsita Pal, Susan Walker, Ramesh Katla, Colin Haws, Akhil Gajjala, Andrea Joyner, Kallesh D. Jayappa, David J. Feith, Todd E. Fox, Thomas P. Loughran, Owen A. O'Connor, Anuradha Illendula. Chemical fabrication and characterization of a novel nanoparticle of the histone deacetylase (HDACi) romidepsin [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3207.

Evaluation of Probiotic Bacteria Isolated from Indigenous Honeybee Species of Pakistan

The honey bee gut is inhabited by many useful bacteria such as Lactic Acid Bacteria (LAB). These LAB possess active probiotic properties that are beneficial to other animals and even humans. The LAB is declared as generally recognized as safe (GRAS). The LAB is non-pathogenic and produces different metabolites having antimicrobial potential. In the current study LAB species isolated from four honey bee species (Apis melifera, Apis dorsata, Apis cerana, and Apis florae) prevalent in Pakistan were identified and investigated for their probiotic potential. The strains were identified using morphological, biochemical, and molecular techniques as genus Lactobacillus and Lactococcus. Current results confirmed that the honey bee gut has a rich source of lactic acid bacteria. Isolated strains were further evaluated for their ability to resist physiological stresses encountered in the human gastrointestinal tract. Safety testing of the isolates such as hemolytic assay revealed that these isolates have γ-hemolytic activity and were adjudged to be safe. Hence the present study concluded that honey bee gut is thesource of potential probiotic bacteria and can be used in pharmaceuticals, fermented foods, and nutraceuticals and may be used as a natural food preservative

Retrospective analysis of carcinogenicity assessments within FDA-notified GRAS determinations.

Frameworks have been developed to standardize the assessment of carcinogenic potential in the pharmaceutical and agrochemical industries, building upon decades of research. Carcinogenicity is also evaluated during the safety evaluation of food substances, using a comprehensive approach unique to each substance. To better understand these approaches, a retrospective assessment was conducted on the publicly available database of substances notified to the United States Food and Drug Administration (US FDA) as being Generally Recognized As Safe (GRAS). The data contained within these GRAS notifications (GRNs) were reviewed for the methods used to evaluate carcinogenic potential (genotoxicity studies, 2-year bioassays, other pre-clinical animal studies) to identify patterns that could provide an understanding of how this assessment has been conducted for different categories of food substances. While different approaches to the safety evaluation were required to adapt to the unique food substances, the data in all notifications supported the conclusion of safety. The evaluation of food substances for carcinogenic potential must consider all available data, including identifying the need for when more data must be generated to support an evaluation. Due to the complexity of substances used in food, ranging from defined chemical entities to minimally processed agricultural commodities to live microorganisms, the approach to conducting the safety evaluation of food substances must be able to adapt to the most relevant scientifically supported approach. This paper illustrates the data commonly used to support the safety of different types of food substances and proposes an approach familiar to other product sectors.

One-pot process for the biotransformation of vegetable oils into natural deca- and dodecalactones

Deca- and dodecalactones are highly desired natural compounds that are essential for creating flavor formulations with fruity, peachy, creamy, and floral notes. Although natural ingredients are preferred by consumers, these lactones cannot be extracted from natural sources. Therefore, the biotechnological processes that produce these compounds in their natural form are crucial for the flavor industry. Here, we report a study on the biotransformation of vegetable oils into natural deca- and dodecalactones. The proposed process is performed one-pot, through the sequential use of three different biotransformation steps, namely the lipase-mediated hydrolysis of the triglycerides, the use of probiotic bacteria for the hydration of the unsaturated fatty acids and the transformation of the obtained hydroxy-fatty acids into lactones derivatives employing Yarrowia lipolytica. By using a specific vegetable oil in combination with a selected bacterial strain, it is possible to obtain a preferred lactone derivative such as γ-dodecalactone, dairy lactone, tuberose lactone, or δ-decalactone in a concentration ranging from 0.9 to 1.5 g/L. Overall, our method is suitable for the industrial production of these lactones as it is easily scalable, it can be performed in only one bioreactor and it makes use of generally recognized as safe (GRAS) microorganisms.

Safety Regulations of Edible Ingredients Derived from Corynebacterium glutamicum

Corynebacterium glutamicum has been researched and developed as a strain that produces various edible ingredients, starting with glutamic acid. Due to environmental pollution and food security issues, studies on using C. glutamicum as a single-cell protein (SCP) are emerging. For the past 20 years, cases valid as safe among edible ingredients derived from C. glutamicum by the European Food Safety Authority (EFSA) or U.S. Food and Drug Administration (FDA) were L-glutamic acid, L-lysine, L-arginine, L-valine, L-isoleucine, L-histidine, L-threonine, L-methionine, L-tryptophan, L-glutamine, 2′-fucosyllactose, corn syrup fermentation product, and D-psicose 3-epimerase. According to validation, it is recommended that the final product should be free of viable cells and recombinant DNA of the production strain. Although there is a possibility that viable cells may be present in the final product, it can be considered safe if the strain qualifies for Qualified Presumption of Safety (QPS) or Generally Recognized as Safe (GRAS). Even if there is a possibility that recombinant DNA may be present in the final product, it can be considered safe as long as it is not a gene of concern for antibiotic resistance, toxicity, or pathogenicity. This review provides insights for future safety validation of edible ingredients derived from C. glutamicum, including SCPs.