The scientific community and industrial companies have discovered significant enzyme applications to plant material. This rise imparts to changing consumers’ demands while searching for ‘clean label’ food products, boosting the immune system, uprising resistance to bacterial and fungal diseases, and climate change challenges. First, enzymes were used for enhancing production yield with mild and not hazardous applications. However, enzyme specificity, activity, plant origin and characteristics, ratio, and extraction conditions differ depending on the goal. As a result, researchers have gained interest in enzymes’ ability to cleave specific bonds of macroelements and release bioactive compounds by enhancing value and creating novel derivatives in plant extracts. The extract is enriched with reducing sugars, phenolic content, and peptides by disrupting lignocellulose and releasing compounds from the cell wall and cytosolic. Nonetheless, depolymerizing carbohydrates and using specific enzymes form and release various saccharides lengths. The latest studies show that oligosaccharides released and formed by enzymes have a high potential to be slowly digestible starches (SDS) and possibly be labeled as prebiotics. Additionally, they excel in new technological, organoleptic, and physicochemical properties. Released novel derivatives and phenolic compounds have a significant role in human and animal health and gut-microbiota interactions, affecting many metabolic pathways. The latest studies have contributed to enzyme-modified extracts and products used for functional, fermented products development and sustainable processes: in particular, nanocellulose, nanocrystals, nanoparticles green synthesis with drug delivery, wound healing, and antimicrobial properties. Even so, enzymes’ incorporation into processes has limitations and is regulated by national and international levels.

The novelty of this study is twofold: AgNPs were obtained and characterized using Artemisia absinthium (A. absinthium), Humulus lupulus (H. lupulus), and Thymus vulgaris (T. vulgaris) plants extracts; moreover, a green and environmentally friendly approach for the synthesis of silver nanoparticles (AgNPs) using aqueous extracts was developed. This paper discusses new approaches about the synthesis of AgNPs. T. vulgaris, H. lupulus, and A. absinthium, which are renewable and common plants, perfect as reducing, stabilizing, and capping agent for green synthesis of silver nanoparticles (AgNPs). The extracts and synthesized AgNPs were characterized by various physico-chemical, phytochemical, morphological scanning electron microscope (SEM/EDS) and transmission electron microscope scanning (TEM), and antibacterial activity. The antioxidant activity of extracts and AgNPs were also assessed by 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS^•+), 2,2-diphenyl-1-picrylhydrazyl (DPPH^•), cupric reducing antioxidant capacity (CUPRAC), ferric reducing antioxidant power (FRAP), and trifluoperazine dihydrochloride (TFPH^•) scavenging assays. Extracts/AgNPs showed significant antioxidant activity in all cases. A. absinthium/AgNPs, H. lupulus/AgNPs, and T. vulgaris /AgNPs displayed activities against DPPH^• (0.14 ± 0.00; 0.11 ± 0.00 and 0.14 ± 0.00 mmol/g), ABTS^•+ (0.55 ± 0.05; 0.86 ± 0.05 and 0.55 ± 0.05 mmol/g), respectively. TEM analysis confirmed the average particle size, it estimated t A. absinthium/AgNPs–46 nm, H. lupulus/AgNPs size of synthesized particles was 42 nm and T. vulgaris/AgNPs–48 nm. SEM analysis revealed that T. vulgaris/AgNPs showed in solitary cases as snowflake-like, branched, but in a general spheric shape, H. lupulus/AgNPs were wedge-shaped, and A. absinthium/AgNPs were the spherical shape of the synthesized AgNPs. EDS analysis confirmed the purity of the synthesized AgNPs with a strong signal at 3.2 keV. A. absinthium/AgNPs, H. lupulus/AgNPs, and T. vulgaris/AgNPs exhibited higher antibacterial activity against all tested bacterial strains compared to their respective pure extracts. It is concluded that AgNPs synthesized in extracts have a broad range of biological applications, which can be used as an eco-friendly material without having negative effects on the environment.

Fungi, including filamentous organisms such as yeasts, play essential roles in various processes such as nutrient exchange in ecosystems, the cultivation of mushrooms, and solid-state fermentation (SSF). SSF involves microbial growth on solid substrates without free water, leading to the production of enzymes, bioactive compounds, and biofuels. This fermentation method offers advantages like lower production costs, reduced waste disposal issues, and the efficient utilization of agricultural residues and fruit and vegetable by-products. Filamentous fungi excel in SSF due to their enzyme secretion capacity and ability to produce valuable compounds. The process is influenced by biological, physico-chemical, and environmental factors, requiring careful optimization for optimal results. Fruit and vegetable by-products are increasingly recognized as valuable substrates for SSF, offering rich sources of bioactive compounds and high nutritional value. The optimization of SSF processes, compatibility with various substrates, and potential for producing diverse value-added products make SSF a promising method for sustainable resource utilization and enhanced product development. Future research should focus on improving process efficiency, expanding the substrate range, enhancing product quality and yield, and integrating SSF with other technologies for enhanced production capabilities.

Silver nanoparticles (AgNPs) biosynthesized using aqueous medical plant extracts as reducing and capping agents show multiple applicability for bacterial problems. The aim of this study was to expand the boundaries on AgNPs using a novel, low-toxicity, and cost-effective alternative and green approach to the biosynthesis of metallic NPs using Calendula officinalis (Calendula) and Hyssopus officinalis (Hyssopus) aqueous extracts. The formation of AgNPs was confirmed by transmission electron microscopy (TEM), scanning electron microscopy (SEM), and energy-dispersive spectroscopy (EDS) techniques. The effectiveness of biosynthesized AgNPs in quenching free radicals and inhibiting the growth of Gram-positive and Gram-negative microorganisms was supported by in vitro antioxidant activity assay methods and using the Kirby–Bauer disk diffusion susceptibility test, respectively. The elucidated antimicrobial and antioxidative activities of medical plant extracts were compared with data from the engineered biosynthetic AgNPs. The antimicrobial effect of engineered AgNPs against selected test cultures was found to be substantially stronger than for plant extracts used for their synthesis. The analysis of AgNPs by TEM revealed the presence of spherical-shaped nano-objects. The size distribution of AgNPs was found to be plant-type-dependent. The smaller AgNPs were obtained with Hyssopus extract (with a size range of 16.8 ± 5.8 nm compared to 35.7 ± 4.8 nm from Calendula AgNPs). The AgNPs’ presumably inherited biological functions of Hyssopus and Calendula medical plants can provide a platform to combat pathogenic bacteria in the era of multi-drug resistance.

Enzyme-assisted extraction is a valuable tool for mild and environmentally-friendly extraction conditions to release bioactive compounds and sugars, essential for silver nanoparticle (AgNP) green synthesis as capping and reducing agents. In this research, plant and fungal kingdoms were selected to obtain the enzyme-assisted extracts, using green synthesized AgNPs. For the synthesis, pseudo-cereal Fagopyrum esculentum (F. esculentum) and lichen Certaria islandica (C. islandica) extracts were used as environmentally-friendly agents under heating in an aqueous solution. Raw and enzyme-assisted extracts of AgNPs were characterized by physicochemical, phytochemical, and morphological characteristics through scanning and transmission electron microscopy (SEM and TEM), as well as Fourier transform infrared spectroscopy (FTIR). The synthesized nanoparticles were spherical in shape and well dispersed, with average sizes ranging from 10 to 50 nm. This study determined the total phenolic content (TPC) and in vitro antioxidant activity in both materials by applying standard methods. The results showed that TPC, ABTS^•+, FRAP, and DPPH^• radical scavenging activities varied greatly in samples. The AgNPs derived from enzymatic hydrolyzed aqueous extracts C. islandica and F. esculentum exhibited higher antibacterial activity against the tested bacterial pathogens than their respective crude extracts. Results indicate that the extracts’ biomolecules covering the AgNPs may enhance the biological activity of silver nanoparticles and enzyme assistance as a sustainable additive to technological processes to achieve higher yields and necessary media components.

Hippophae rhamnoides L. leaves possess a remarkable amount of polyphenols that could serve as a natural remedy in various applications. In comparison, numerous techniques, such as conventional and high-pressure techniques, are available for extracting the bioactive fractions from sea buckthorn leaves (SBL). However, enzyme-assisted extraction (EAE) of SBL has not been comprehensively studied. The aim of this study was to optimize critical EAE parameters of SBL using the cellulolytic enzyme complex, Viscozyme L, to obtain a high-yield extract with a high concentration of bioactive compounds. In order to determine the optimal conditions for EAE, the study employed a central composite design and response surface methodology to analyze the effects of four independent factors (pH, temperature, extraction time, and enzyme concentration) on two different responses. Our findings indicated that under optimal conditions (3:15 h extraction, temperature 45 °C, pH 4.9, and 1% Viscozyme L v/w of leaves DW), EAE yielded 28.90 g/100 g DW of the water-soluble fraction. Furthermore, the EAE-optimized liquid extract was continuously fermented using an ancient fermentation starter, Tibetan kefir grains, which possess lactic acid bacteria (LAB) and have significant potential for use in biopreservation. Interestingly, the results indicated various potential prebiotic characteristics of LAB. Additionally, alterations in the cell wall morphology of the SBL residue after EAE were examined using scanning electron microscopy (SEM). This study significantly optimized EAE parameters for sea buckthorn leaves, providing a promising natural source of bioactive compounds for various applications, such as nutraceuticals, functional foods, and high-value products.