DOI: 10.6084/m9.figshare.27016472.v1
title: Application of microbial enzymes in medicine and industry: current status and future perspectives

description: Microbes are a major source of enzymes due to their ability to be mass-cultivated and genetically modified. Compared with plant and animal enzymes, microbial enzymes are more stable and active. Enzymes are generally classified into six classes based on their reaction, substrate specificity and mechanism of action. In addition to their application in medicine for treating diseases, these compounds are used as anti-inflammatory, thrombolytic and digestive agents. However, challenges such as immunogenicity, tissue specificity and short <i>in vivo</i> half-life make clinical trials complex. Enzymes are metabolic catalysts in industry and their production and extraction must be optimized to preserve profitability due to rising demand. The present review highlights the increasing importance of bacterial enzymes in industry and medicine and explores methods for their production, extraction and purification. Enzymes are important substances made by the cells of plants and animals. They are catalysts, or substances that control how quickly chemical reactions occur. These reactions are the processes that keep all plants and animals functioning. They are present in almost every natural organism, from microorganisms to plants and mammals. But plants and animals produce small amounts of enzymes unsuitable for industrial applications. The use of microbial enzymes in industry offers many advantages over plant and animal enzymes. People use enzymes in industry and medicine. Enzymes help to heal cuts and to diagnose certain diseases. They are also an important part of the process called fermentation. In industries, they are applied in the textile, starch, bakery, and detergent industries. This helps turn milk into cheese and juice into wine, and it makes bread rise before it is baked. Enzymes are attractive in industry and medicine due to their high substrate affinity and specificity, low toxicity, efficient catalysis and minimal side effects. Bacteria exhibit rapid growth within a short time and enable relatively low-cost and straightforward processes such as purification, recovery and isolation, in comparison to animal and plant sources. Enzyme supplements are available in the form of capsules, tablets and powders, often comprising a mixture of several different enzymes. Enzymes are categorized based on their reaction mechanisms into six groups: oxidation-reduction (reductase), transition (transferase), hydrolysis (hydrolase), dissociation (lyase), isomerization (isomerase) and synthesis (ligase). Microbial enzymes are applied for diverse therapeutic purposes, including the treatment of genetic disorders, metabolic disorders, Gaucher's disease, Parkinson's disease, remove toxic substances, acting as anti-inflammatory agents, aiding digestion and their recent use in cancer treatment and combating infectious diseases. Important bacterial enzymes used in treatment include L-asparaginase, Serratiopeptidase, Collagenase, Nattokinase, EndoS, α-galactosidase, Prolyl endopeptidase, Cocaine esterase, IdeS, Glutaminase, Vibriolysin, Fibrinolytic enzyme, Methioninase, γ-glutamyl transpeptidase and Amine oxidase. The enzyme asparaginase possesses antilymphoma properties and is capable of converting extracellular asparagine into aspartic acid. Serratiopeptidase has demonstrated anti-inflammatory, analgesic, anti-biofilm, anti-edema and fibrinolytic activity. Collagenase is also used to treat diseases such as Peyronie's disease, burns, wounds and glaucoma. Nattokinase is a thrombolytic enzyme that degrades fibrin to dissolve blood clots. EndoS from Streptococcus pyogenes has been shown to be effective in reducing antibody-mediated red blood cell death. Streptokinase is an intravenous thrombolytic enzyme used for the treatment of cardiovascular diseases. Enzyme treatments, despite their potential applications, have limited FDA and EMA authorization due to issues like tissue specificity, short <i>in vivo</i> half-life and immunogenicity. A significant challenge with enzyme-based therapies is the immune response of patients following the introduction of exogenous recombinant enzymes, which can induce an immunogenic neo-antigenic response. Long treatments and repetitive exposure to enzymes after a prolonged treatment-free period can trigger more robust immune reactions compared with short-term treatments. Different hosts can yield varying levels of enzyme activity, stability and efficiency, all of which are crucial for therapeutic applications. The presence of proteolytic enzymes, as well as immune system rejection, can reduce their stability and lead to degradation via aggregation, hydrolysis, denaturation and oxidation. Scaling up from laboratory-scale to large-scale production has challenges and obstacles such as including maintaining enzyme activity and stability during the scale-up process, instability in pharmaceutical enzymes and an effective delivery system. The use of microbial enzymes in industry offers numerous advantages over plant and animal enzymes. Proteases, amylases and cellulases are essential microbial enzymes in the industry. Hydrolytic enzymes constitute the majority of industrial enzymes and are involved in decomposing various natural substances. Proteases, for instance, are crucial enzymes for the detergent and dairy industries, accounting for over 60% of the worldwide enzyme market. amylases and cellulases applied in the textile, starch, bakery and detergent industries. Submerged fermentation (SmF) and solid-state fermentation (SSF) are the primary methods for producing microbial enzymes. To utilize enzymes, they must first be extracted, typically employing two methods: (1) extraction of cells and (2) extraction of solid substrate cultures. Enzymes produced by microorganisms through culture or fermentation can be either extracellular or intracellular. The most common methods for bacterial enzyme purification in the industry include chromatography, electrophoresis and crystallization. Purification procedures utilize membrane systems such as microfiltration, ultrafiltration, nanofiltration and reverse osmosis. Mass spectrometric methods have increasingly been used to measure enzyme activity and kinetics.