The rapid rise of antimicrobial resistance demands therapeutic strategies that extend beyond conventional antibiotics. However, most existing reviews describe emerging alternatives without systematically linking their mechanistic advances to translational readiness and clinical implementation barriers. This review addresses this gap by integrating evidence across multiple beyond-antibiotic approaches, including antimicrobial peptides, bacteriophage therapy, CRISPR-based antimicrobials, nanotechnology-enabled delivery systems, anti-virulence agents, host-directed immunotherapies, microbiome modulation (engineered probiotics and fecal microbiota transplantation), and drug-repurposing or combination therapies. The principal contribution of this synthesis is a comparative framework that maps mechanisms of action, engineering innovations, and translational evidence across these diverse strategies. Advances such as peptidomimetics, engineered phages, and nanoparticle carriers that enhance stability, targeting, and therapeutic efficacy are highlighted, along with synergistic strategies including phage-antibiotic and CRISPR-nanocarrier combinations. The review further identifies major barriers limiting clinical translation, including delivery efficiency, toxicity and ecological concerns, large-scale production challenges, cost, inconsistent clinical outcomes, and regulatory fragmentation for biologics and live therapeutics. To facilitate clinical implementation, the study proposes a translational roadmap emphasizing standardized evaluation assays, physiologically relevant infection models, integrated rapid diagnostics, and regulatory frameworks tailored for emerging antimicrobial platforms, thereby supporting the development of sustainable therapies for the post-antibiotic era.

Amidst the escalating crisis of antimicrobial resistance globally, genome mining has emerged as a promising field for the discovery of newer antibiotics from microbial sources. Secondary metabolites like non-ribosomal peptides, polyketides, and ribosomally synthesized post-translationally modified peptides are synthesized by biosynthetic gene clusters and exhibit diverse pharmacological activities. Advanced sequencing technologies and informatics studies have made high-precision identification and prediction of cluster function possible. Computational tools like antiSMASH, BAGEL, PRISM, and RiPPMiner are the core of BGC classification and characterization of derived metabolites from microbial genomes. Heterologous expression, microbial co-culture, elicitor induction, and genetic regulation have been used in various strategies to induce cryptic or silent gene clusters, leading to improved production of novel compounds. The combination of bioinformatics and synthetic biology has yielded higher precision in prediction and understanding of biosynthesis. Therefore, genome mining is an economical and productive approach for the discovery of next-generation antimicrobials, offering a potential solution to the global healthcare catastrophe caused by multidrug-resistant pathogens.

Urate oxidase (uricase; EC 1.7.3.3) catalyzes the oxidation of uric acid to allantoin, a highly soluble metabolite that is readily excreted. Lack of functional urate oxidase in human’s leads to hyperuricemia, which is associated with gout, tumor lysis syndrome and renal complications. The present study reports statistical optimization, purification, and comprehensive characterization of a novel extracellular urate oxidase secreted by Arthrobacter creatinolyticus SA1. Plackett–Burman Design screening followed by Central Composite Design identified inoculum volume, initial pH, peptone, and yeast extract as key determinants for enzyme production. Under optimized conditions, the enzyme yield 36.18 IU/mL was obtained. The enzyme was purified by ammonium sulfate fractionation and DEAE–Sepharose chromatography with an overall purification of 5.8-fold. SDS–PAGE revealed a 30 kDa subunit, while native-PAGE indicated a homo-tetrameric enzyme with an estimated molecular weight of 120 kDa. The purified enzyme displayed optimal activity at pH 9.0 and 30 °C. The Km and Vmax of the enzyme were 14.57 mM and 142.85 µmol/min/mg, respectively. Urate oxidase SA1 showed moderate thermostability, exhibiting half-life of about 2 h at 50 °C. MALDI–TOF/TOF peptide mapping confirmed identity of enzyme with urate oxidase (P78609.1). In vitro assays using hyperuricemic human serum demonstrated dose-dependent uric acid degradation, reducing levels from 7.50 to 2.73 mg/dL within 12 h at 1.3 IU/mg, the observed results were found comparable to commercial uricase. These findings suggest that extracellular urate oxidase SA1 may serve as a promising candidate for further development for the management of hyperuricemia and gout.

Brewer’s spent grains (BSG), a by-product in beer brewing, have historically been relegated to animal fodder. This study delves into the untapped potential of BSG as a valuable source of fermentable sugars, specifically xylose. Proximate analysis confirmed the presence of xylan-rich hemicellulose (21.5 ± 0.32

The study employed response surface methodology (RSM) to optimize physicochemical variables for extracellular collagenase production by gram negative bacterial strain Chryseobacterium contaminans KU665299 under submerged fermentation. It is also revealing the ability of collagenase to degrade collagen, main structural protein in human blood. The study successfully enhanced collagenase activity by 1.2 folds through Response Surface Methodology (RSM) and 5.33 folds through purification of enzyme using ammonium sulfate precipitation and DEAE-Sepharose chromatography (specific activity with 538.0 U/mg). SDS-PAGE analysis identified its molecular weight as 32 kDa. Optimal conditions for the enzyme’s activity were pH 7.5 and 40 °C. Kinetic studies of collagenase KU665299 revealed specificity for collagen, with Km and Vmax values of 0.059 mg/l and 588.24 µmol/min/mg, respectively. Zinc and calcium ions enhanced activity, while EDTA and DTT strongly inhibited it. The purified collagenase demonstrated remarkable efficiency in digesting blood clots, fully dissolving 1 ml clots within 40 min at 37 °C, showcasing significant thrombolytic potential. The study successfully optimized and characterized a novel collagenase from C. contaminans KU665299, revealing its high specificity, stability, and efficiency in degrading collagen and its promising ability to rapidly digest blood clots for potential thrombolytic properties.

Alginate is a major extra polymeric substance in the biofilm formed by mucoid Pseudomonas aeruginosa. It is the main proven perpetrator of lung infections in patients suffering from cystic fibrosis. Alginate lyases are very important in the treatment of cystic fibrosis. This study evaluated the role of standalone and in conjugation, effect of alginate lyase of SG4 + isolated from Paenibacillus lautus in enhancing in vitro bactericidal activity of gentamicin and amikacin on mucoid P. aeruginosa. Using Response Surface Methodology (RSM) alginate lyase SG4 + production was optimized in shake flask and there 8.49-fold enhancement in enzyme production. In fermenter, maximum growth (10.15 mg/ml) and alginate lyase (1.46 International Units) production, 1.71-fold was increased using Central Composite Design (CCD). Further, fermentation time was reduced from 48 to 20 h. To the best of our knowledge this is the first report in which CCD was used for fermenter studies to optimize alginate lyase production. The Km and Vmax of purified enzyme were found to be 2.7 mg/ml and 0.84 mol/ml-min, respectively. The half-life (t 1/2) of purified alginate lyase SG4 + at 37 °C was 180 min. Alginate lyase SG4 + in combination with gentamicin and amikacin eradiated 48.4- 52.3

Alginate Oligosaccharides (AOs) are bioactive compounds prepared through the enzymatic degradation of alginate polysaccharides by alginate lyase. These Oligosaccharides have become a focus of growing interest in scientific community, largely because of their array of bioactive attributes, which include immunomodulatory, antioxidant, anti-inflammatory and prebiotic effects. This review critically examines the current scientific literature on Alginate Oligosaccharides (AOs), encompassing in vitro studies, animal models and preliminary human clinical trials. It also addresses the safety profiles of AOs, focusing on toxicological assessments, to provide a comprehensive view of their risk-to-benefit ratio. Additionally, the review assesses the existing information regarding the impact of Alginate Oligosaccharides (AOs) on human health. It aims to identify the current research gaps and outline potential future directions for their therapeutic application.

Fungal infections, that are becoming more common by the day, can mean a life-threatening menace to people who are immunological or medically compromised. Fungi are eukaryotic creatures that have a similarity with humans at molecular level and have many biochemical targets which are also found in eukaryotic cells. Due to this similarity, it is difficult to design drugs that target only fungal cells without affecting human cells. As a result, many antifungals have side effect. Therefore, such antifungals are required which target fungal cells but are nontoxic to human cells. To date, only a few numbers of antifungal medications, such as polyenes, azoles, echinocandins, and flucytosine, are available for the treatment of invasive fungal infections. Furthermore, toxicity, drug interaction and the emergence of fungal resistance to different fungicides limits the use of current antifungal drugs. This results in significant morbidity and mortality rates. Natural compounds with fungistatic and fungicidal activity to treat fungal infections can be found in plants. Secondary bioactive metabolites such as terpenoids, phenolics, flavonoids, saponins, and alkaloids, which have antifungal properties, are abundant in plants. Traditional medicine systems cite several medicinal plants for the treatment of both animal and human mycoses, thus offering them a promising future as a potential source for anti-fungal medicines.

Aim In the present study, malic acid in combination with sodium hypochlorite is evaluated for the eradication of biofilms formed by Cronobacter sakazakii strains individually and in a cocktail on different abiotic surfaces.Methods and results The biofilm formation by five strains of C. sakazakii and their cocktail culture on different substrates was studied in Tryptone Soy Broth (TSB) and reconstituted Powdered Infant Formula (PIF). Further, the effect of temperature (4, 27, 37, and 50 degrees C) and contact time (10, 20, 30, 40, 50, and 60 min) on the antibiofilm potential of the test solution (0.0625 mol l-1 malic acid and 0.00004 mol l-1 sodium hypochlorite) against the biofilm formed by C. sakazakii cocktail culture was investigated on these surfaces. The effect was evaluated in terms of viable cell count and biofilm texture using scanning electron microscopy (SEM). Principal Component Analysis (PCA) revealed that the maximum biofilm reduction was observed for stainless steel at 4 degrees C after 60 min of contact, whereas at 25, 37, and 50 degrees C, maximum biofilm reduction was observed for polycarbonate. For glass and polyurethane, maximum log reductions were observed at 50 degrees C. The SEM images revealed cell surface deformation and disruption in biofilms after treatment with the test solution.Conclusions The antibiofilm potential was observed to be greatly affected by contact time and temperature. These results indicated that the combination of malic acid and NaOCl can effectively kill and remove C. sakazakii biofilms from food contact surfaces and enteral feeding tubes.

Alginate, a complex polysaccharide predominantly found in seaweeds, is degraded into alginate oligosaccharides by the enzyme alginate lyase, an enzyme of high interest for its diverse applications in medicine and industry. In this study, our primary focus was to isolate microorganisms with robust alginate lyase-producing capabilities from marine algae. Through systematic screening methodologies and 16S rRNA gene sequencing, the isolate was taxonomically classified as belonging to the Paenibacillus genus and has been archived in gene bank with the accession number OM980630. A comprehensive optimization process was executed, testing 13 different growth media to ascertain the most conducive environment for both bacterial proliferation and enzyme activity. It was determined that Medium M-7 yielded the most promising results. Furthermore, the presence of alginate in the culture medium not only acted as a carbon source but also as an inducer for alginate lyase production. Recognizing the crucial role of alginate lyase in medical treatments, especially in managing conditions like cystic fibrosis, and its importance in industrial processes, specifically for generating bioactive oligosaccharides with applications including antitumor, antidiabetic, antihypertensive, anti-inflammatory, antimicrobial, antioxidant, anticancer, immunomodulatory, and anti-radiation activities, the identification of a new microbial source for alginate lyase production could represent a significant advancement in both healthcare and industry.

Deterioration of food during storage is a major cause of concern for food industries especially, in tropical countries. Food is rich in nutrients and apposite for the growth and reproduction of pathogenic microorganisms. As per the UNEP Food Waste Index Report (2021), approximately 17% of the total global food available to consumers ended up in the trash barrels of households, restaurants, retailers and other food services and its major cause is microbial contamination. Microbial contamination can cause significant changes in the characteristics of food such as nutritional value, texture, smell and flavour. Some microbial agents produce toxins and chemicals that can cause serious damage to human health. Because of this, various physical and chemical methods are used by the food industry to produce food products that are free from contamination but these methods can also alter the sensorial and nutritional properties of food. Considering the interest of consumers towards the food that is nutritionally balanced, good in taste, and free from any chemical treatment, lactic acid bacteria (LAB) can play a great role as an antimicrobial alternative. Various fermented foods naturally contain LAB which are Gram-positive, cocci, or rod in shape and are known to restrict the growth of food spoilage and foodborne pathogens because of their antimicrobial activities. These antimicrobial activities are due to the production of several antimicrobial metabolites, particularly bacteriocins. By emphasizing the significance of LAB, this review offers a thorough overview of emerging approaches to the control of foodborne pathogens.

Candida albicans infections are a substantial cause of illness burden in immunocompromised patients. The opportunistic pathogen, C. albicans is polymorphic in nature which shows commensalism in humans, and colonizes mostly mucosal surfaces of the human body, including the respiratory, urinary, genital, gastrointestinal tracts and oropharyngeal cavity. It can cause infections which range from superficial infections such as oral thrush to serious infections like candidemia and disseminated candidiasis. These infections occur when Candida spp. undergo a reversible morphological transition from yeast to filamentous form. Significant efforts are focused on the mechanisms that control this transition. Antifungal drugs such as azoles, polyenes, allylamines and echinocandins are most commonly used in the treatment of Candida infected patients. The toxicity of antifungal medications on human cells, coupled with the increasing resistance of Candida infections to these drugs, highlights the necessity for the development of novel strategies to enhance the well-being of patients affected by such infections. Lactobacillus spp. found in the human microbiome are the natural competitors of Candida spp. and have the potential to control fungal growth. Lactic acid bacteria are cocci or rod-shaped, Gram-positive, non-spore forming bacteria. LAB produces lactic acid and other metabolites which suppress filamentation, a key virulence feature of C. albicans. This review provides a comprehensive overview of novel strategies in the prevention of Candida infections by highlighting the significance of LAB.

The current study determined the antibacterial and antibiofilm activity of six organic acids including acetic acid, citric acid, formic acid, tartaric acid, malic acid and maleic acid along with different chemicals (disinfectants, surfactants and chelating agents) either alone or in a combination of organic acid with chemicals against C. sakazakii. All the organic acids and only three chemicals viz., EDTA, NaCl, NaOCl were found to possess antiCronobacter activity and were evaluated for antibiofilm efficacy. All the agents tested for antibiofilm activity were quite effective against formation of biofilm as well as prefomed mature biofilms. Subtractive screening based on antibiofilm activity of different combinations revealed malic acid (0.0625 mol l- 1) with NaOCl (0.00004 mol l- 1) as the most effective combination with fractional inhibitory concentration index (FICI) of 0.38. Biofilm microstructures were rarely observed after treatment as revealed by light microscopy and SEM analysis and a reduction of 7.06 +/- 0.40 log CFU ml-1 in viable cell counts as compared to control groups. The findings indicate that combination of malic acid with NaOCl has synergistic effect and may have implications in its usage in eradication of C. sakazakii biofilms from abiotic surfaces in production plant and clinical settings.

Cronobacter sakazakii is an opportunistic foodborne pathogen of concern for foods having low water activity such as powdered infant formula (PIF). Its survival under desiccated stress can be attributed to its ability to adapt effectively to many different environmental stresses. Due to the high risk to neonates and its sporadic outbreaks in PIF, C. sakazakii received great attention among the scientific community, food industry and health care providers. There are many extrinsic and intrinsic factors that affect C. sakazakii survival in low-moisture foods. Moreover, short- or long-term pre-exposure to sub-lethal physiological stresses which are commonly encountered in food processing environments are reported to affect the thermal resistance of C. sakazakii. Additionally, acclimation to these stresses may render C. sakazakii resistance to antibiotics and other antimicrobial agents. This article reviews the factors and the strategies responsible for the survival and persistence of C. sakazakii in PIF. Particularly, studies focused on the influence of various factors on thermal resistance, antibiotic or antimicrobial resistance, virulence potential and stress-associated gene expression are reviewed.

Collagenase is an endopeptidase responsible for the hydrolysis of native collagen into peptide fragments.Microbial collagenases are truly promising enzymes in relation to their extensive biological and industrial applications.Collagen being the most abundant constituent of extracellular matrix (ECM) in vertebrates opens a wide array of biotechnological and therapeutic applications for microbial collagenases.In the present study, a cost-effective and eco-friendly production of extracellular collagenase from novel, nonpathogenic, Chryseobacterium contaminans KU665299 was carried out utilizing slaughter house waste material as sole source.The complete digestion of waste material resulted in good yield of collagenase utilizing goat skin and auricular cartilage.

Candida species are considered to be an opportunistic pathogen that leaves asymptomatically in warm blooded animals like human, birds etc. It can cause serious and lethal disease in healthy individuals. It generally causes infection of skin, oral cavity esophagus, gastrointestinal tract, vagina and vascular system of humans. It is a common cause of nosocomial infection in hospitals and most of the Candida infections occur in patients who are immunocompromised. Virulence factor like adhesion, morphogenesis, phospholipase are responsible for pathogenesis. Genus Candida include about 200 species but normally C. albicans, C. glabrata, C. tropicalis, C. parapsilosis, and C. krusei are five species of Candida involved in 90% of invasive infections. Candida infections can be superficial or invasive. Superficial infections often affect the skin or mucous membrane and can be treated successfully with antifungal drugs whereas invasive fungal infections are often life-threatening. Different chemical drugs such as azoles, polyenes, echinocandins and nucleoside analogues are in use to treat the infection caused by C. albicans, but due to their regular use, C. albicans can show resistance to these drugs and mortality rate increases. Resistance to various drugs is due to multiple factors, viz., reduced accumulations of drug in the cells, decreased target affinity and counteraction of the drug effect. Biofilms production is the another major cause of resistance to different drug groups and to overcome this emergence, different strategies are improvise which include the new triazoles, peptides, use of natural compounds, vaccinations, antibodies, cytokines therapy, and low level laser therapy.

Antibiotics are used to treat and prevent infections, but the overuse of drugs makes the microorganisms drug resistant, and with the time they become resistant to more than one drug. These bacteria are known as multidrug-resistant bacteria. Resistivity of bacteria against the antibiotics makes them more hazardous for human beings and animals too and those bacterial infections become untreatable and lead to death of the person. The resistivity to bacteria is provided by the bacterial plasmids, which contains antibiotic resistant genes. This problem can be overcome by removing bacterial plasmids from the bacterial cells. This can be done by treatment with plasmid curing agents, but various plasmid curing agents such as acridine orange or SDS are toxic and mutagenic in nature. Various investigations have been undertaken to determine the antibacterial effect and plasmid curing ability of extracellular and intracellular extracts of lactic acid bacteria (LAB) to cure plasmids of antibiotic resistant bacterial isolates. The microtiter plate method was used to determine the minimum inhibitory concentration of LAB extracts. Plasmid curing mediated by LAB extracts resulted in the loss of antibiotics resistance encoded in plasmids, as well as the antibiotics resistance profile of cured strains. The extracellular extracts of LAB were considered a natural and harmless source to combat plasmid borne multidrug-resistant bacteria.