Microbial Lipases Research Articles

Comparative In-silico Analysis of Structural and Functional Attributes of Microbial Lipases

Lipase is the third most demanding enzyme group and is a major component of fat metabolism. It is omnipresent in nature. Microbial lipases are highly specific and economically efficient enzymes. They have applications in a variety of industries. In this study, the computational result elucidates structural and functional attributes of eight selected lipases from Uniprot. Primary, secondary, and tertiary structures obtained from Swiss and Profunc are analyzed. Further, they are validated with the help of Saves Server. COACH and COFACTOR is used for functional analysis. Lastly, phylogenetic tree is developed using MegaX software. Our in-silico analysis predicted lipases as alpha-beta-gamma protein with different ligand binding sites at different amino acid residue positions. Furthermore, they exhibited almost similar functional traits like catalytic activity and hydrolase activity. Overall, this study provides an insight into structural-functional characteristics of different lipases of microbial origin.

Enhancing Lipase Production of Bacillus salmalaya Strain 139SI Using Different Carbon Sources and Surfactants

Microbial lipase is one of the major sources of the enzyme that has been broadly exploited in the food, detergent, and pharmaceutical industries due to its high catalytic activity, high yield, and environmental friendliness and cost-effectiveness. Therefore, the aim of this study was to optimize the medium for the submerged fermentation for lipase production by a novel strain, Bacillus salmalaya strain 139SI. The media subjected to lipase production was Luria Bertani (LB) with different carbon sources and surfactants supplemented to determine which would give the highest lipase activity of Bacillus salmalaya. The Lipase activity of the supernatant containing lipase enzyme was ddetermined using the titrimetric method with hydrolysis reaction. Results showed that the olive oil that was used as a carbon source, induced the highest lipase activity (11.0 U/mL) compared to sunflower oil (9.6 U/mL) and cooking oil waste (7.8 U/mL). For surfactants, LB medium supplemented with tween 80 enhanced higher lipase activity (6.8 U/mL) compared to tween 20 (6.0 U/mL) and sodium dodecyl sulphate (SDS) (2.0 U/mL). Thus, it can be concluded that submerged fermentation allows optimization of the culture medium whereby, among carbon sources, olive oil induced the highest lipase production, whereas Tween 80 was the best lipase inducer compared to other surfactants.

Optimization of Enzymatic Degreasing of Sheep Leather for an Efficient Approach and Leather Quality Improvement Using Fractional Experimental Design.

Leather industry is making significant contributions to economic development. However, it is notably leading to a serious environmental pollution. Recently, the enzyme technology developments offer new opportunities for enzymatic application in leather making. In the present investigation, microbial lipases were studied and used in degreasing process of sheep leathers. In order to optimize degreasing efficiency, a fractional experimental design with four parameters (enzyme source, processing stage, lipase amount, and degreasing duration) was used. Lipases A from Aspergillus niger, F from Rhizopus oryzae, R from Penicillium roqueforti, and AY from Candida rugosa were selected for leather degreasing. Enzymatic treatment of sheep skin was carried out during two stages of beamhouse operations: deliming-bating and pickling. Obtained results showed that enzymatic degreasing efficiency is higher than those obtained with the conventional process. Lipase F from Rhizopus oryzae demonstrated the most interesting hydrolysis with yields of 58.3% and 37.2% for delimed and pickled skins, respectively. An enzymatic degreasing process on pickled leather using 0.125% (w/v) of lipase F during 3.5h is the most promising for an industrial application with a 76.03 of degreasing efficiency. Results of the physico-mechanical tests of leathers having undergone enzymatic treatment complied with industry requirement. The enzymatic treatment may be carried out in the same conditions as employed in leather manufacturing process. Results suggested that the enzymatic degreasing improves the leather quality and reduces the use of chemical compounds and surfactant.

Bioremediation of Vegetable Oil Contaminated Soil with Two Microbial Isolates

Vegetable oil spills are becoming frequent and are potentially more challenging than petroleum hydrocarbon spills. Microbial lipases occupy a place of prominence among biocatalysts and are often used for the remediation of vegetable oil spills. There is a need for extensive characterisation of lipase for the treatment of vegetable oil-polluted sites. This work was carried out to monitor the degradation pattern of vegetable oil. Two microbial isolates previously isolated from an oil mill in Ibadan, Nigeria were used for the bioremediation experiment. Soil samples (with some purposely contaminated with 2 different vegetable oils) collected from the Nursery section of the Microbiology department as well as soil samples from the oil mill were all subjected to varied treatment processes. Field bioremediation using the isolates was carried out for 12 weeks. The isolates were identified, and microbial load and residual oil weight were determined during the degradation period using standard methods. The two isolates were identified as Pseudomonas fluoresecens and Candida parapsilosis. The result of sterile soil samples with and without mixing option, from palm oil and palm kernel purposely contaminated soils for all the various treatments, showed a general increase in total viable counts from the 2nd week to the 12th week, however in the non-sterile counterpart there was a steady increase from the 2nd week to the 8th week and subsequently, a gradual decrease from the 10th to the 12th week. The residual oil weight in the sterile purposely palm oil-contaminated soil, treated with the consortium (POC) non-mixed gave a reduction of value from 0.335 g on day 0 to 0.13025 g by the 12th week. From the oil mill non-sterile, treated with (POC) non-mixed sample, the residual weight after 12 weeks of treatment was 0.0043 g from an initial weight of 0.01 g. The microorganisms Pseudomonas fluoresecens and Candida parapsilosis had the potential for the degradation of fatty waste. They could therefore be employed in the environmental cleanup of the vegetable oil spill sites.

Investigation of Lipolytic-Secreting Bacteria from an Artificially Polluted Soil Using a Modified Culture Method and Optimization of Their Lipase Production.

Compared to lipases from plants or animals, microbial lipases play a vital role in different industrial applications and biotechnological perspectives due to their high stability and cost-effectiveness. Therefore, numerous lipase producers have been investigated in a variety of environments in the presence of lipidic carbon and organic nitrogen sources. As a step in the development of cultivating the unculturable functional bacteria in this study, the forest soil collected from the surrounding plant roots was used to create an artificially contaminated environment for lipase-producing bacterial isolation. The ten strongest active bacterial strains were tested in an enzyme assay supplemented with metal ions such as Ca2+, Zn2+, Cu2+, Fe2+, Mg2+, K+, Co2+, Mn2+, and Sn2+ to determine bacterial tolerance and the effect of these metal ions on enzyme activity. Lipolytic bacteria in this study tended to grow and achieved a high lipase activity at temperatures of 35–40 °C and at pH 6–7, reaching a peak of 480 U/mL and 420 U/mL produced by Lysinibacillus PL33 and Lysinibacillus PL35, respectively. These potential lipase-producing bacteria are excellent candidates for large-scale applications in the future.

Preliminary Study of Actinomycetes as Potential Biocatalyst in Biodiesel Production Through Microbial Lipase Activity

Biocatalysts are biological substances that act as an accelerator of chemical reactions without causing influence on the living organism chemically. Biocatalysts based on renewable sources, biodegradable, tolerance to pH and temperature, and high selectivity to stereochemistry substrates and products. The utilization of biocatalyst is environmentally friendly and effective in production costs. This research aimed to determine the potential of actinomycetes as a biocatalyst in biodiesel production. This research method isolation and identification of actinomycetes isolates, conducting lipase activity test to determining lipase enzyme production of actinomycetes isolates. The data analysis in microbial identification was conducted by molecular identification by Gene bank through Basic Local Alignment Search Tool (BLAST). The lipase activity of actinomycetes analyzing by the value of lipase rate of test microbes. Based on chemical analysis of lipase activity, this research results show that actinomycetes T1A has the highest lipase activity by 14.4640 mU/g. Based on molecular identification analysis, actinomycetes T1A was identified as Streptomyces sp., 99% similar to Streptomyces phaeochromogenes.

A Valuable Product of Microbial Cell Factories: Microbial Lipase.

As a powerful factory, microbial cells produce a variety of enzymes, such as lipase. Lipase has a wide range of actions and participates in multiple reactions, and they can catalyze the hydrolysis of triacylglycerol into its component free fatty acids and glycerol backbone. Lipase exists widely in nature, most prominently in plants, animals and microorganisms, among which microorganisms are the most important source of lipase. Microbial lipases have been adapted for numerous industrial applications due to their substrate specificity, heterogeneous patterns of expression and versatility (i.e., capacity to catalyze reactions at the extremes of pH and temperature as well as in the presence of metal ions and organic solvents). Now they have been introduced into applications involving the production and processing of food, pharmaceutics, paper making, detergents, biodiesel fuels, and so on. In this mini-review, we will focus on the most up-to-date research on microbial lipases and their commercial and industrial applications. We will also discuss and predict future applications of these important technologies.

Study the Behavior of Activity and Express for Enzymes

Enzymes are be defined as soluble, colloidal, organic catalysts which are produced by living cells.Enzymes are biologic catalysts to accelerate the rate of biochemical reactions.in this study Quantification of the Enzyme Activity, Where: E = enzyme concentration; S = substrateconcentration; ES = enzyme-substrate complex concentration; k1, k3= 1st order reaction rate constants(t-1); P = product concentration; k2= 2nd reaction rate constant (M-1.t-1).At first, it must be stressed thatthe formation of ES (enzyme-substrate complex) is an obligatory step to any type of enzyme-catalyzedreaction. The ES, depending on the reaction conditions, can form the product (forward reaction) ornot (reward reaction). The immobilization technique allowed a more frequent use of biomaterials inindustry, therapeutics (confection of subcutaneous capsules for controlled delivering of hormones), andlab analysis (automatic dosing equipments, enzyme electrode, biosensors and enzyme immunoassays).In industry, the use of glucose isomerase for the conversion of glucose into fructose (high fructosecorn syrup), the aminoacylase for the separation of racemic mixture of amino acids, microbial lipase intriglyceride hydrolysis, and the lactase for removing lactose from milk and/or whey are stressed.

Polypropylene as a selective support for the immobilization of lipolytic enzymes: hyper‐activation, purification and biotechnological applications

AbstractLipases are the versatile biological catalysts used in a wide range of commercial synthetic applications. Microbial lipases have been widely used in the pharmaceutical, food, textile, oleochemical, paper, and bioenergy industries, in the composition of detergents, and in the production of polymers and enantiomerically pure chemical intermediates. Lipases have a unique lid opening mechanism that makes them interesting biocatalysts to catalyze reactions in hydrophobic environments. Water insoluble substrates have also been efficiently transformed by lipases in organic solvents. The use of lipases in industrial processes could be limited by the cost of their production, the difficulties in their recovery and disposal, or by the harsh conditions of some processes that could inactivate or denature the protein. The immobilization of enzymes helps overcome these physical and economical drawbacks, making the catalyst more active, stable, and reusable, resulting in the reduction of costs and of contaminating and harmful byproducts. Polypropylene has unique properties that aid in the activation of the lid opening mechanism and improve the catalytic efficiency of immobilized lipases. The aim of this review is to present an overview of the uses of polypropylene as a support for immobilization of lipolytic enzymes, the study of the immobilization procedure conditions, the characteristics of the immobilized enzymes, and the perspectives of their use in the future as a powerful tool in sustainable industry. © 2021 Society of Chemical Industry (SCI).

Lipolytic Activities of Bacteria and Fungi Isolated from Soil Samples

This study was carried out at the Department of Microbiology, Microbiology Laboratory, Ado-Ekiti State University, Ekiti State, Nigeria between July, 2018 to March, 2019. Due to the diverse biotechnological importance of lipases as a biocatalytic enzyme, extracellular production of microbial lipases has to gain lots of interest. This study, therefore, focused on the physicochemical parameters of lipase producing microorganisms from different soil samples. Microorganisms were isolated from four different soil samples using Nutrient Agar (NA) and Potato Dextrose Agar (PDA). The isolates were identified and characterized. Production, an assay for Lipase enzymes, purification, the effect of pH, Temperature and metal ion was investigated. The isolates were culturally, morphologically and biochemically characterized. Two of the bacteria strains (Bacillus sp. and Staphylococcus sp.) and four fungi (Fusarium sp., Aspergillus fumigatus, Aspergillus niger, and Trichophyton sp.) isolates were able to produce lipid using Sudan Black B Fat staining techniques. Fusarium sp. isolated from dumpsite soil had the highest specific lipase activity (21.16 µmol/min/ml) while Bacillus sp. isolated from red oil spill soil had the highest lipase activity (0.59 µmol/min/mg). The specific activity of partially purified lipase for Fusarium sp. was 2.39 µmol/min/mg while Bacillus sp. had a specific activity of 2.46 µmol/min/mg. 30oC - 50oC, pH 7.0 to 9.0 and KCl2 (139.672%) supported the highest production of lipase by the Bacillus sp. and Fusarium sp. This study demonstrated that the Bacillus sp. produced a high amount of lipase activity followed by Fusarium sp. Extensive and persistent screening for new microorganisms and their lipolytic activities will help to provide faster ways to solve most environmental soil pollution.

Industrial biotransformations catalyzed by microbial lipases: screening platform and commercial aspects.

The successfulness of a lipase-catalyzed industrial process depends on a proper lipase selection. In this work, an alternative screening platform for industrially important biotransformations catalyzed by microbial lipases was proposed. Thus, the reactivity of sixty lipase activities from spore-forming microorganisms towards hydrolytic and transesterification reactions by using p-nitrophenyl palmitate as a chromogenic acyl donor substrate was explored. Only three biocatalysts were capable of catalyzing all reactions tested. Fourteen biocatalysts did not show hydrolytic activity at all; however, they displayed transesterification activities using ethanol, starch, low-methoxyl (LM) pectin, high-methoxyl (HM) pectin, or vitamin C as acyl acceptors. Using heat-treated biocatalysts, hydrolytic activities were not highly correlated with the corresponding transesterification activities using ethanol (r = -0.058, p = 0.660), starch (r = 0.431, p = 0.001), LM pectin (r = -0.010, p = 0.938), HM pectin (r = 0.167, p = 0.202), and vitamin C (r = -0.048, p = 0.716) as acyl acceptor. In addition, to the best of our knowledge, several transesterification activities produced from microorganisms of the genus Bacillus, Brevibacillus, Lysinibacillus, Geobacillus, or Sporosarcina were reported for first time. Finally, the global lipase market was presented and segmented by date, application, geography and player highlighting the commercial contribution of microbial lipases.

Difference in Performance of EPI Pigs Fed Either Lipase-Predigested or Creon®-Supplemented Semielemental Diet.

Pancreatic enzyme replacement therapy (PERT) and fat predigestion are key in ensuring the optimal growth of patients with cystic fibrosis. Our study attempted to highlight differences between fat predigestion and conventional PERT on body composition of young pigs with exocrine pancreatic insufficiency (EPI). EPI and healthy pigs were fed with high-fat diet for six weeks. During the last two weeks of the study, all pigs received additional nocturnal alimentation with Peptamen AF (PAF) and were divided into three groups: H—healthy pigs receiving PAF; P—EPI pigs receiving PAF+PERT; and L—EPI pigs receiving PAF predigested with an immobilized microbial lipase. Additional nocturnal alimentation increased the body weight gain of EPI pigs with better efficacy in P pigs. Humerus length and area in pigs in groups L and P were lower than that observed in pigs in group H (p value 0.005-0.088). However, bone mineral density and strength were significantly higher in P and L as compared to that of H pigs (p value 0.0026-0.0739). The gut structure was improved in P pigs. The levels of neurospecific proteins measured in the brain were mainly affected in P and less in L pigs as compared to H pigs. The beneficial effects of the nocturnal feeding with the semielemental diet in the prevention of EPI pigs' growth/development retardation are differently modified by PERT or fat predigestion in terms of growth, bone properties, neurospecific protein distribution, and gut structure.

Supplementation of Bile Acids and Lipase in Broiler Diets for Better Nutrient Utilization and Performance: Potential Effects and Future Implications – A Review

Abstract Bile acids are used for better emulsification, digestion and absorption of dietary fat in chicken, especially in early life. Similarly, exogenous lipases have also been used for the improvement of physiological limitation of the chicken digestive system. Owing to potential of both bile acids and lipases, their use has been increased in recent years, for better emulsification of dietary fat and improvement of growth performance in broilers. In the past, pancreatic lipases were used for supplementation, but recently, microbial lipase is getting attention in poultry industry as a hydrolysis catalyst. Bile acids strengthen the defence mechanism of body against bacterial endotoxins and also play a key role in lipid regulation and sugar metabolism as signaling molecules. It has been demonstrated that bile acids and lipases may improve feed efficiency by enhancing digestive enzyme activity and ultimately leading to better fat digestion and absorption. Wide supplemental range of bile acids (0.004% to 0.25%) and lipases (0.01% to 0.1%) has been used in broiler diets for improvement of fat digestibility and their performance. Combinations of different bile acids have shown more potential to improve feed efficiency (by 7.14%) even at low (0.008%) levels as compared to any individual bile acid. Lipases at a lower level of 0.03% have exhibited more promising potential to improve fat digestibility and feed efficiency. However, contradicting results have been published in literature, which needs further investigations to elucidate various nutritional aspects of bile acids and lipase supplementation in broiler diet. This review focuses on providing insight on the mechanism of action and potential application of bile acids and lipases in broiler diets. Moreover, future implications of these additives in poultry nutrition for enhancing nutrient utilization and absorption are also discussed.

Various Perspectives on Microbial Lipase Production Using Agri-Food Waste and Renewable Products

Lipases are enzymes that catalyze various types of reactions and have versatile applications. Additionally, lipases are the most widely used class of enzymes in biotechnology and organic chemistry. Lipases can be produced by a wide range of organisms including animals, plants and microorganisms. Microbial lipases are more stable, they have substrate specificity and a lower production cost as compared to other sources of these enzymes. Although commercially available lipases are widely used as biocatalysts, there are still many challenges concerning the production of microbial lipases with the use of renewable sources as the main component of microbial growth medium such as straw, bran, oil cakes and industrial effluents. Submerged fermentation (SmF) and solid-state fermentation (SSF) are the two important technologies for the production of lipases by microorganisms. Therefore, this review focuses on microbial lipases, especially their function, specificity, types and technology production, including the use of renewable agro-industrial residues and waste materials.

Microbial halophilic lipases: A review.

Microbial lipases are commercially significant due to their versatile catalytic function of hydrolysis triacylglycerol. Among these, lipases from extremophiles are optimal for industrial application. Halophilic microorganisms living in a high salinity environment, such as the ocean, salt lakes, salt wells, and so on, produce halophilic lipases. In recent decades, many remarkable achievements have been made related to the properties and application of halophilic lipases. This review offers information collected over the last decades on halophilic lipase sources as well as advances in production, factors influencing activity, stability under various conditions, structural characteristics, progress in industrial applications such as food flavor modification, biodiesel production, and waste treatment, to provide theoretical and methodological references for the research in this direction.

Detection lipase gene of Pseudomonas aeruginosa from crude oil contaminated soil

The microbial lipases are industrially more substantial. The bacterial lipase enzymes can be extracellular and intracellular, and are extremely affected by bacterial nutrition and various physicochemical factors like temperature degree, pH, the sources of carbon and nitrogen, inorganic salts and agitation. The objective of current work is isolation and identify of P. aeruginosa from crude oil contaminated soil depending on PCR targeted lipA gene. 20g (15 samples) of crude oil contaminated soil were collected from north oil refineries/Kirkuk/Iraq. The isolates were identified according to microscopically diagnosis and colonial properties, biochemical tests, API-20E system with diagnosis by PCR technique based on gene lip A. PCR products by electrophoresis demonstrated only 8 isolates (53.33%) with positive results with lipA 558 from all isolates of P. aeruginosa.

Turning waste into valuable products: utilization of agroindustrial oily wastes as the low-cost media for microbial lipase production

The cost-effective production of lipase by microorganisms are currently needed. The bioconversion of agro-industrial oily wastes as an alternative oily substrate for lipase production in the form of solid-state fermentation (SSF) or submerged fermentation (SmF) would hold a potential role in future biotechnology. There are some studies revealed that yeast, fungi, and bacteria are the choice for lipase production such as from the genera Aspergillus, Penicillium, Rhizopus, Candida, Geotrichum, Stenotrophomonas, Lasiodiplodia, and Yarrowia, as well as Staphylococcus, Burkholderia, Enterococcus, Pediococcus, Pseudomonas, and Bacillus utilizing several residues such as oil cakes and oily wastewater. In this review, the microbial lipase production conducted by many scientists utilizing agro-industrial oily wastes as the substrates is summarized.

RETRACTED ARTICLE: Predigested high-fat meats based on Lactobacillus fermentum lipase enzyme immobilized on silver-alginate nanoparticle matrix

Lipase enzyme plays a part in the environmental and pharmaceutical industries, microbial lipase is the most potential source for industrial application. Lipases were extracted from endogenous isolate, sequentially purified by ammonium sulphate and Sephadex G-150 methods, and immobilized on silver-alginate nanoparticles to test the ability to used preparing predigested of high-fat meat to low fat meat. Lipase producing bacteria was identified as Lactobacillus fermentum, that extracted mesophilic lipases presented maximal activity at pH 8 and 40 °C. The purified enzyme was recovered up to 16% yield in 1.4 fold purification and specific activity 4 U/mg. Lipase activity exhibited the highest inhibition rate by adding 1% of non-ionic detergent of Triton X-100, Tween 80 and Tween 20 (60, 63 and 71%, respectively). Silver-alginate nanoparticles is a potential matrix for immobilization of biomolecules, especially enzymes. Immobilized lipase was preserved; the specific activity (2.3–2 U/mg) can even be reused more than three times, which attributed to hydrophobicity and flexibility of lipases. Predigested meat was obtained from treating high-fat meat with immobilized enzymes, as shown by losses of 40% of initial meat weight after 14 days of lipid digestion. Conjugation of silver-alginate matrix with lipase can controlled the digestion of meat fat and easy to get rid of in comparing to expressed enzyme. Concept of predigested meat could revolutionize the world of high-fat diets without interfering their dietary habits.

The influence of oriented external electric field on lipase catalyzed triglyceride hydrolysis

The use of microbial lipases for the hydrolysis of natural oils such as triglyceride esters is a green alternative to conventional processes to produce high value-added chemicals. Biocatalysis using lipases in aqueous solutions is limited often by instabilities, slow kinetics due to mass transfer limitations, and by challenges of economic lipase recycling. Prior work has shown that mass transfer rates and reaction rates can be intensified by increasing interfacial area using electrostatic spray reactors. However, the possible effect of oriented external electrical field on the lipase catalytic activity has hitherto not been considered. In this study, the performance of microbial lipase derived from Candida rugosa in aqueous solutions was analyzed in the presence of a steady DC externally applied electrical voltage. The reaction was conducted in three different batch type reactors, quiescent (fixed interface), stirred tank, and tubular flow system. In each of the three reactor systems it was concluded that the oriented external electrical field has a positive effect on reaction independently of interfacial area. Further studies conducted using reverse polarity, increased electrode distance and for an immobilized lipase system have shown that lipase undergoes conformational changes due to an oriented external electrical field is the main driving mechanism for this enhanced performance.

Production strategies and biotechnological relevance of microbial lipases: a review.

Lipases are enzymes that catalyze the breakdown of lipids into long-chain fatty acids and glycerol in oil-water interface. In addition, they catalyze broad spectrum of bioconversion reactions including esterification, inter-esterification, among others in non-aqueous and micro-aqueous milieu. Lipases are universally produced from plants, animals, and microorganisms. However, lipases from microbial origin are mostly preferred owing to their lower production costs, ease of genetic manipulation etc. The secretion of these biocatalysts by microorganisms is influenced by nutritional and physicochemical parameters. Optimization of the bioprocess parameters enhanced lipase production. In addition, microbial lipases have gained intensified attention for a wide range of applications in food, detergent, and cosmetics industries as well as in environmental bioremediation. This review provides insights into strategies for production of microbial lipases for potential biotechnological applications.