The hydrolases known as lipases (Triacyclglycerol acyl hydrolases, EC3.1.1.3) are specialised for the hydrolysis of fats into fatty acids and glycerol at the water-lipid interface. They are abounding in nature and have the ability to reverse the reaction in non-aqueous medium (van Schie et al. 2021). Clade Bernad was the first to identify lipase in pancreatic juice in 1856 as an enzyme that degraded insoluble oil droplets and transformed them into soluble compounds (Jamilu, Ibrahim, and Abdullahi 2022). There have been claims that various types of fungus, yeast, bacteria, animals, and plants are effective sources of lipase (Faryad, Ataa and Joyia, 2020). However, microbially produced lipases have become the focus of interest due to their multiple advantages, including ease of handling culture, ease of manufacturing scale-up, ease of genetic manipulation, seasonal changes, and safety and stability (Nema et al. 2019). According to Prem et al. (2020), most bacterial species are known to produce their highest levels of enzyme activity at neutral and alkaline pH levels. Excellent thermostability and other stress-tolerant traits are displayed by certain animals (Chandran et al., 2020). Some species exhibit great thermostability and other stress tolerant parameters.
Wide-temperature-window enzymes, particularly those that exhibit activity at lower temperatures, are necessary for several bioprocesses in the detergent, fermentation, and biomass conversion industries. These processes conserve energy conversion and prevent the growth of mesophilic bacteria (Atalah et al., 2019; Mrudula Vasudevan et al., 2019; Ramani et al., 2010; Kumar and Thakur, 2020). Enzymatic hydrolysis is a beneficial method, because it may be carried out at a lower temperature to conserve energy and because it has excellent selectivity, resulting in products with high purity and fewer byproducts (Ramani et al. 2010). It has been reported that lipase has thermostability and reacts to cold, which is useful for industrial application (Kumar and Thakur 2020). To produce commercial lipases, Bacillus species, Pseudomonas species, Staphylococcus species, and Burkholderia species are the most common bacterial sources (Bharathi and Rajalakshmi, 2019; Priyanka, Kinsella, et al., 2019)
The three-dimensional structures of lipases originating from diverse microbiological sources differ significantly in terms of sequence variety (Gupta et al. 2015). These characteristics make these enzymes distinct and particular to the kinds of bioconversion processes they catalyse, making them useful in a variety of industrial processes (Mehta, Bodh, and Gupta 2017). It has been discovered that they are helpful in catalysing a variety of reactions related to the food, pharmaceutical, dairy, fatty acid, leather, cosmetic, detergent, beverage, and paper sectors. (Sharma et al. 2017)
Eight families of bacterial lipases have been identified, and each family has unique characteristics that differ in terms of ideal pH and temperature, among other factors. (Rios et al. 2018). Except for a few acid ideal lipases made by cultures of Pseudomonas cepacia and Pseudomonas fluorescens, lipases from Pseudomonas sp usually have neutral or alkaline optimal pH (Liu, Li, and Yan 2017). The optimal pH activity spans from 4 to 9, the ideal temperature ranges from 25 to 70°C, and the enzyme molecular weight ranges from 16 to 96 kDa, depending on the species and genus of the microorganism from which lipases have been purified (Rios et al. 2018), however, very few exhibit catalytic properties over a wider temperature range. Lipases extract an acyl group from glycerides during hydrolysis to create a lipase–acyl complex, which subsequently transfers the acyl group to the O–H group in water. The hydrolysis of oils to produce fatty acids is the primary use of lipase in the oleochemical industry. Because free fatty acids are widely used in surfactants, soap production, the food sector, and biomedical applications, they are considered value-added goods. Traditionally, oil hydrolysis is accomplished by applying high pressure and temperature to a chemical catalyst.
The majority of organic chemists and pharmacologists prefer to use bacteria from the genera Pseudomonas and Burkholderia as catalysts due to their distinct characteristics, which include temperature stability, high enantio-selectivity, and activity in a wider pH range (Gurkok and Ozdal 2021). Because of this, a great deal of study has been done on the synthesis of lipase utilising Pseudomonas sp. from lipid substrates that are discarded as waste products from numerous industrial operations. Therefore, the synthesis of lipolytic enzymes from solid waste materials containing lipids can result in the creation of environmentally friendly technologies (Ramani et al. 2010).
Global production and improper handling of millions of tonnes of these waste lipids is currently occurring, which has a negative impact on the economy, the environment, and human health in addition to decreasing the materials' value as oleochemical resources (Lahiri et al. 2023). For instance, the incorrect disposal of used cooking oils, tallow waste, and grease traps in sewage causes accumulation and obstruction, which causes overflows, flooding during rainy seasons, the spread of viruses and rodents, pollution of water supplies, and damage to ecosystems.
This study focuses on the development of the lipase enzyme from bacterial species and produces feedstock for biodiesel production by hydrolysis of waste tallow using lipase to test the usefulness of the lipase activity in varied applications for beneficial results. The hydrolysed product may be effectively used as the feedstock for biodiesel production. The strains were identified by whole genome sequencing.