Enhanced Lipase Production Research Articles

Surfactant-facilitated metabolic induction enhances lipase production from an optimally formulated waste-derived substrate mix using Aspergillus niger: A case of machine learning modeling and metaheuristic optimization

This study introduces a novel method to enhance lipase production by integrating machine learning (ML) models and optimization algorithms. Six ML models, including support vector regression (SVR), kernel ridge regression (KRR), extreme gradient boosting (XGB), extreme learning machine (ELM), random forest (RF), and artificial neural networks (ANN), were employed to predict lipase activity in a multi-substrate system using avocado seed, coconut pulp, and palm oil mill effluent (POME) with Aspergillus niger. SVR proved the most effective (R2 = 0.9738; RMSE = 7.0089). Further optimization using manta ray foraging optimization (MFRO), particle swarm optimization (PSO), and genetic algorithm (GA) identified optimal substrate loadings, achieving a maximum lipase activity of 194.38 U/gds. The addition of a mixture of surfactants (Tween 80, Tween 20, Triton X-100) further increased lipase production to 520.95 U/gds (168.3 % increase). Global sensitivity analysis (GSA) confirmed the important roles of avocado seed, POME, and surfactants in enhancing lipase production. This approach represents a significant advancement in bioprocess scalability.

Enhanced lipase production and characterization from Aeromonas media VBC8: Applications in biodegradation of lubricating oil waste

This study aimed to explore the potential of a novel indigenous strain for the improved production of lipase from castor oil-contaminated soil. Among the various isolates, Aeromonas media VBC8 was found to be the most effective for lipase production. The effect of different inducer oils (olive, peanut, soybean, rice bran, sunflower, coconut, sesame, and fish liver oil) on the biomass of A. media VBC8 and its lipase activity was determined. Among the various oils assessed, fish liver oil exhibited highest lipase activity, with 89 U/mL with 9.1 g/L of biomass. Furthermore, Box-Behnken Design was used to optimize the cultural conditions resulting in an enhanced lipase activity of 1156 U/mL. The lipase was purified through ammonium salt (60 w/v %) precipitation, desalting and ion exchange column, achieving a yield of 16 % and specific activity of 98.4 U/mL. The purified lipase remained active over a wide range of pH 4.0–11.0 and temperature of 10–80 °C with maximum activity at pH 8.0 and 40 °C. SDS-PAGE analysis revealed the lipase's molecular weight to be 94 kDa. The study also evaluated the role of crude and purified lipase in the biodegradability of lubricating oil waste, achieving a maximum fatty acid conversion of 39 and 76 %, respectively, after 7 h incubation at room temperature.

Optimization of Submerged Fermentation Parameters for Enhanced Lipase Production from Newly Isolated Lipolytic Yeasts using Agro Waste Substrates

Fungi are currently recognized as the most preferred sources of lipase due to their ability to produce copious amount of extracellular lipase with lower production’s and extraction’s costs. Lipase production was carried out in agro wastes formulated-liquid fermentation media at 300C, at 130rpm, at initial pH of 7 for 96 hours using newly isolated LipA gene identified yeasts [Pichia kudriavzevii ENS1 [Accession No: OL546800], Candida orthopsilosis PMS3 [Accession No: OL546803] and Saccharomyces cerevisiae PMS8 [Accession No: OL546805] at inoculum size of 2%. Agrowastes such as Mango kernel and orange peel were used as alternative carbon sources; beans hull and groundnut shell as nitrogen sources while lipidic sources such as olive oil, soyabeans oil, palm oil, groundnut oil and coconut oil were used as inducers. Single factor optimization process was carried out to determine the effects of carbon sources, nitrogen sources, inducers, incubation time, inoculum size, pH and carbon: nitrogen ratio on lipase production and cell growth. Extraction by filtration, centrifugation and acetone precipitation; Quantitative lipase assay using titrimetric method and Cell growth by measuring optic density at 600nm were done. Proximate analysis of the agro-waste substrates showed higher percentage of carbohydrate and crude fat in mango kernel powder at 68.05±0.56% and 19.54±0.62% while groundnut shell recorded higher crude protein at 11.63±0.05%. Single factor optimization study showed optimum lipase production and cell growth at 50.83±1.18u/ml/min and 0.69±0.00[OD] with Candida orthopsilosis PMS3[Accession No: OL546803] using hot water treated mango kernel hydrolysate as carbon source. The results from this study showed that mango kernel would be an economical alternative carbon source for industrial-scale lipase production.

Correction to: Isolation, screening, and optimization of culture parameters for enhanced lipase production from Aspergillus eucalypticola strain CBS 122,712 and its application

Correction to: Isolation, screening, and optimization of culture parameters for enhanced lipase production from Aspergillus eucalypticola strain CBS 122,712 and its application

Isolation, screening, and optimization of culture parameters for enhanced lipase production from Aspergillus eucalypticola strain CBS 122,712 and its application

Isolation, screening, and optimization of culture parameters for enhanced lipase production from Aspergillus eucalypticola strain CBS 122,712 and its application

Enhancement of Lipase Production based on improved Bacillus Licheniformis for Catabolizing of Edible Oil Wastes

Enhancement of Lipase Production based on improved Bacillus Licheniformis for Catabolizing of Edible Oil Wastes

Enhanced lipase production from a ternary substrate mix of agricultural residues: A case of optimization of microbial inducers and global sensitivity analysis

In this study, a ternary substrate mix made up of coconut pulp waste, banana peels and pineapple peels was optimally formulated to produce lipase. The optimum mix of coconut pulp waste (7.46 g), banana peels (0.01 g) and pineapple peels (7.53 g) gave a maximum lipase activity of 339.4 U/gds. The effect of inducers (castor oil, jatropha oil and olive oil) on lipase production from the substrate mix was investigated using response surface methodology (RSM) and artificial neural networks (ANN). ANN predictions were marginally superior to those of RSM. Optimized levels of castor oil (1.99%w/w), jatropha oil (0.02%w/w) and olive oil (1.99%w/w) yielded a maximum lipase activity of 488.7 U/gds which was a 30.6% increase compared to the un-induced system. Global sensitivity analysis revealed that olive oil was the most sensitive inducer. Thus, biological induction can enhance the production of lipase from the ternary substrate mix.

Isolation, Optimization and Molecular Characterization of Novel Lipase Producing Lysinibacillus macroides FS1 from Oil Rich Sample for Biodiesel Production

Lipase is one of the promising biocatalyst in field of Enzymatic Biodiesel production. Screening of lipase producing bacteria and its media optimization facilitates enhanced lipase production. Considering the advantages of enzymatic biodiesel production using lipases the objective of present work was to isolate lipase producing bacteria and its media optimization. Bacterial strain were isolated from oil rich sample showing maximum zone of hydrolysis on TBA Media was selected and identified as Lysinibacillus macroides FS1 by 16SrDNA gene sequence analysis. Lipase production was carried out in production media and lipase activity of 3.1U/ ml was assayed by titrimetric method. Various media parameters were optimized and maximal lipase activity of 16.75U/ml were observed at 48 hr of incubation in optimized media with Honge oil as inducer, Galactose as carbon source , Ammonium Chloride and Beef extract as source of nitrogen, pH 7 and 37ºC temperature with agitation speed of 120 rpm. Under optimized condition 5.4 fold increases in the lipase activity than innate activity was observed. 50.74% yield was observed with specific activity of 21.25U/mg after purification with 30% ammonium sulphate. Purified lipase was stable in presence of methanol at 37°C. Results obtained shows that Lipase from Lysinibacillus macroides FS1 was promising biocatalyst for Biodiesel production.

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.

DETECTION OF SOME TYPES OF OILS THAT INDUCE LIPASE PRODUCTION IN SOME BACTERIAL ISOLATES

Bacterial lipases are a highly versatile class of hydrolytic enzymes which catalyze the hydrolysis of triglycerol to glycerol and fatty acids. They play an important role in biotechnological and industrial processes, including oleo chemical, food, pharmaceutical detergent formulation, cosmetic, leather, textile, and paper industries applications. Tremendous interest among scientists and industrialists has been arisen in the last decade in microbial lipases due to their versatility and ease of mass production.In the current work, natural oils such as Triglyceride, Tween 20, Tween 80, Olive oil, Sweet almond, Celery oil, Sesame, Rosemary oils were implemented to enhance lipase production by soil and pathogenic bacterial isolates. Results revealed that triglyceride and tween 80 had the best enhancing activity for lipase production in most isolates. It was also shown that Pseudomonas and Bacillus were the most potential lipase producing bacteria.A novel modified tributary agar was prepared by the addition of Sudan blue dye and used for the detection of lipase producers. It was shown to give a very clear zone even in the case of weak producing strains as compared to the original medium.Specific lipase activity had been determined Staphylococcus aureus (in the present research. The highest lipase activity was demonstrated by Pseudomonas (4.5u/ml), while Staphylococcus aureus did not show any lipase activity.

Enhanced lipase production from organic solvent tolerant Pseudomonas aeruginosa UKHL1 and its application in oily waste-water treatment

Abstract Lipase, as a handy hydrolytic enzyme, is gaining importance in environmental management like oil and fat-containing wastewater treatment. In this study, we have focused on cost-effective production of lipase by Pseudomonas aeruginosa UKHL1 and its applicability in oily wastewater treatment. Plackett-Burman randomized factorial design with 20 experiments recognized ammonium oxalate, peptone, yeast extract, coconut oil, NaCl, and CaCl2 as significant factors among the screened 13 factors by one factor at a time approach (OFAT). Further fine-tuning of three most significant media components (ammonium oxalate (0.01% w/v), peptone (0.246% w/v) and coconut oil (0.1% v/v)) was done by Central Composite Design experiment of Response Surface Method with 20 runs. Lipase production by P. aeruginosa UKHL1 was improved 9.4 and 3.91-fold after 24 h and 36 h, respectively under optimized conditions. Strain in this study also showed good tolerance to iso-octane and n-hexadecane solvents having higher log_P value. P. aeruginosa UKHL1 strain degraded 37% oil after 72 h in an experimental setup under laboratory condition.

Molecular, physiological, and biochemical characterization of extracellular lipase production by Aspergillus niger using submerged fermentation.

BackgroundExtracellular production of fungal lipases especially the lipases obtained from the Aspergilli has gained immense interest in recent years due to its diverse biotechnological applications. In this study, we focused on determining the fermentation parameters required for the optimal lipase production.MethodsA total of 256 fungal isolates were obtained from oil seeds. From each genus, one isolate was selected to evaluate lipase production using phenol red and tributyrin plate assays. Lipase activity was estimated using the spectrophotometric pNPP hydrolysis assay. The highest lipase producer isolates were identified using 18S ribosomal RNA gene sequencing. The genetic variability was determined by random amplified polymorphic DNA (RAPD) analysis and the dendrogram was constructed using the unweighted pair group method with arithmetic averages method. The isolates were examined in a submerged fermentation culture (Smf) to measure the effect of temperature, pH, incubation time, carbon source, nitrogen source, inoculum volume, and lipid source on lipase production.ResultsEleven isolates belonging to the genus Aspergillus were analyzed for lipase production where they were found to be the highest lipase producers among various fungal genera. All the tested isolates were identified as A. niger using 18s rRNA sequencing. Genetic diversity was evaluated among all of the studied A. niger isolates using RAPD primers. The RAPD primers were used to amplify 285 loci, of which five were polymorphic (1.75%) and seven were monomorphic (2.45%). Thus, a high level of genetic diversity was observed among all isolates. The tributyrin test and the lipase activity assay identified five strains of A. niger as high lipase producers, and their optimal enzyme activities were 709.74, 532.54, 735.64, 794.62, and 787.69 U/ml. The optimal conditions for lipase production were as follows: 40 °C, pH 7.5, 1% fructose as the carbon source, 1% yeast extract as the nitrogen source, 2% palm oil, 2.5 × 107 spores/ml suspension, and 3 days of incubation.ConclusionsThe current study provides a comprehensive characterization of the optimal conditions, which are essential to enhance lipase production in five A. niger isolates.

Potential biodiesel production from Brazilian plant oils and spent coffee grounds by Beauveria bassiana lipase 1 expressed in Aspergillus nidulans A773 using different agroindustry inputs

This paper describes the obtaining of fatty acids ethyl esters from Açaí (Euterpe oleracea Martius), Buriti (Mauritia flexuosa) and spent coffee grounds oils using a Beauveria bassiana lipase expressed in Aspergillus nidulans, which gene was cloned in pExpyr vector. Carbon sources were tested to enhance recombinant lipase secretion. Cassava peels, corn meal, corn syrup, sorghum seed and wheat bran induced higher lipase activity. Monosaccharides, when combined to previously cited carbon sources, interfered at lipase production: fructose and glucose enhanced lipase production, especially on sorghum seed and wheat bran cultures; while xylose diminished its production. Lipase was immobilized on hydrophobic resins, and derivatives were characterized using pNPP transesterification with ethanol as co-substrate. Sepabead-C18 derivative demonstrated as better pNPP transesterification catalyzer, being 2-fold higher than butyl derivative. The Sepabead-C18 derivative was biochemically characterized using different temperatures and solvents. The transesterification of Açaí, Buriti and coffee spent grounds oils were performed with cyclohexane, and the product content was quantified using gas-chromatography as 17.1%, 5.0%, and 3.4%, respectively. The results demonstrated the potential of immobilized heterologous lipases to produce ethyl ester from Brazilian natural oils and a common food waste. This work will improve the enzymatic methodologies for biodiesel industry.

Enhancement of lipase production by ethyl methane sulfonate mutagenesis of soil fungal isolate

Strain improvement through random mutagenesis is an extremely developed practice and it plays an important role in the economic growth of microbial agitation processes. The present study comprises of genetic improvement of fungus isolated from petrol pump soil by ethyl methane sulfonate (EMS) mutagenesis for increased production of extracellular lipase. Random mutagenesis was performed by incubating the spore suspension of fungus with EMS at a concentration of 5% (v/v) and 8% (v/v) for 30, 60 and 90 min, respectively. Control set was prepared by incubating the spore suspension with sterile distilled water. Control plate showed maximum number of fungal colonies whereas number of colonies was decreased as we increased exposure time of EMS from 30 to 90 min. The lipase activity of six mutagenic strains and wild strain was determined under submerged fermentation and solid state fermentation. Treated culture named as EMS5%-60min (obtained after 60 min exposure with 5% EMS) exhibited maximum activity (32.09 ± 1.84 IU/ml/min) in SmF as compared to wild strain (8.77 ± 3.52 IU/ml/min) and another treated strain named as EMS8%-90min (obtained after 90 min exposure with 8% EMS) exhibited maximum activity (7.99 ± 0.12 IU/g/min) in SSF as compared to wild strain (1.77 ± 0.71 IU/g/min). The activity of mutagenic strain i.e. EMS5%-60min was increased to 365.90% as compared to 100% activity of wild strain in SmF whereas activity of another mutagenic strain i.e. EMS8%-90min was increased to 451.41% as compared to 100% activity of wild strain in SSF.

Statistical Optimization by Response Surface Methodology to Enhance Lipase Production by Aspergillus fumigatus

Background:Lipases have various commercial applications and microorganisms serve as a potential source of production.Objective:The aim of this paper was to study the effect of interactions among different production parameters on lipase yield ofAspergillus fumigatus.Method:Plackett Burman and Central Composite Design (CCD) were established by using Design Expert software 10.0.Results:In the present study, interactions were studied for six different variables such as inoculum size, pH, temperature, galactose concentration, peptone concentration and incubation time. In Plackett-Burman design, galactose concentration, peptone concentration, pH and incubation time were found to be important factors. Using the statistical approach, the optimum factors were found to be as: galactose concentration (1.5%), peptone concentration (1.8%), pH (10.0) and incubation time (72 h) at 45°C under response surface curves. Upon statistical analysis, the coefficient of determination (R2) obtained was 0.9318 which showed that the model was significant.Conclusion:The statistical tools used predicted the optimal conditions for the production of the lipase. The optimized parameters were galactose concentration 1.5%, peptone concentration 1.4%, temperature 45°C, pH 10.0 and incubation time of 72 h for obtaining a maximum lipase activity of 6.22 U/ml.

Optimization of culture conditions for enhanced lipase production by an indigenous Bacillus aryabhattai SE3-PB using response surface methodology

Lipases are enzymes that hydrolyze fats into fatty acids and glycerol at the water–lipid interface and are also involved in a variety of bioconversion reactions in non-aqueous and micro-aqueous environments. In this study, we optimized the culture conditions for extracellular lipase production by an indigenous lipase-producing bacterial strain isolated from lipid-rich wastewater, using response surface methodology. The studied isolate was identified as Bacillus aryabhattai SE3-PB by polymerase chain reaction and analysis of 16S rDNA. Sunflower oil was found to induce maximum lipase production. Face centered central composite design revealed that temperature (40 °C), pH (7.6), inoculum volume (2.8%, v/v), agitation (193 rpm) and inducer oil concentration (2%, v/v) significantly influenced lipase production at the respective optimum conditions. The coincidence of observed lipase production (264.02 ± 1.94 U/mL) with predicted lipase yield (265.82 U/mL) coupled with a high correlation coefficient (R2 = 0.9919, P < 0.01) confirmed the validity of the model. A 7.2-fold increase in lipase production was obtained in the optimized medium compared to the basal medium. These findings provide the first report on lipase production and optimization by B. aryabhattai SE3-PB and suggest a rational choice of optimum processing conditions for commercial lipase production by B. aryabhattai SE3-PB.

Effect of Environmental and Nutritional Parameters on the Extracellular Lipase Production by &lt;i&gt;Aspergillus niger&lt;/i&gt;

Abstract- The present investigation was carried out to evaluate the effect of different growth conditions on lipase production byAspegillus niger. The extracellular lipase producing fungus was isolated from spent bleaching earths. Optimization of physical and chemical parameters was done for maximum lipase production using this isolate. Growth of the organism and lipase production were measured usig varying pH (4 – 9), incubation temperature (20 – 30 °C), incubation time (8 – 80 hrs.), carbon sources, nitrogen sources, and shaking speed. Enhanced lipase production was observed at 24 °C, pH 7 and after 72hrs of incubation. Olive oil 5 % was observed as the most effective carbon source and Yeast extract 1.0 % as the most effective nitrogen source for lipase production. The optimum shaking value to get maximum lipase activity byAspergillusnigerwas 200 rpm.

Effect of Environmental and Nutritional Parameters on the Extracellular Lipase Production by &lt;i&gt;Aspergillus niger&lt;/i&gt;

Abstract- The present investigation was carried out to evaluate the effect of different growth conditions on lipase production byAspegillus niger. The extracellular lipase producing fungus was isolated from spent bleaching earths. Optimization of physical and chemical parameters was done for maximum lipase production using this isolate. Growth of the organism and lipase production were measured usig varying pH (4 – 9), incubation temperature (20 – 30 °C), incubation time (8 – 80 hrs.), carbon sources, nitrogen sources, and shaking speed. Enhanced lipase production was observed at 24 °C, pH 7 and after 72hrs of incubation. Olive oil 5 % was observed as the most effective carbon source and Yeast extract 1.0 % as the most effective nitrogen source for lipase production. The optimum shaking value to get maximum lipase activity byAspergillusnigerwas 200 rpm.

Coconut oil induced production of a surfactant-compatible lipase from Aspergillus tamarii under submerged fermentation.

Filamentous fungi are efficient producers of lipases. The present study focuses on identification of a potent lipolytic fungus and enhancement of lipase production through optimization of nutritional and cultural conditions under submerged fermentation. Molecular characterization of the fungus by 18S rDNA sequencing revealed its identity as Aspergillus tamarii with 98% homology. Maximum lipase production was noted in mineral salts medium supplemented with coconut oil (2.5%, v/v). A combination of ammonium chloride (2%, w/v) and tryptone (2%, w/v) facilitated maximum lipase production at pH 5 of the production medium. A carbon: nitrogen ratio of 1:4 led to significant (p < 0.00008) increase in the enzyme production in the presence of surfactant cetyltrimethylammonium bromide (0.5%, w/v). Maximum lipase activity (2,32,500 ± 192 U/ml/min) was recorded after 7 days of incubation at 25 °C on a rotary shaker at 120 rpm. A 9.8-fold increase in lipase activity was recorded after optimization of the process parameters. Addition of crude lipase enhanced the oil stain removal activity of a commercially available detergent by 2.2-fold. The current findings suggest the potentiality of this fungal lipase to be used in detergent formulation.

Purification and Characterization of Extracellular Lipase from Serratia marcescens VITSD2

Lipases represent a group of extracellular enzymes and hold a prominent place among the enzymes. In the present research work Serratia marcescens isolated from soil was selected for lipase production. Effects of various parameters such as substrate, pH, temperature, incubation time and inoculum size were analysed for the enhanced lipase production. The optimum substrate was found to be olive oil with the concentration of 0.7 mL in 100 mL of production medium. Serratia marcescens produced maximum lipase after 48 h of incubation at 30 °C with the optimum pH as 7.0. The inoculum concentration of 4 % was more favourable for the production of lipase. The extracted lipase was purified by two steps: ultra-filtration and Carboxymethyl(CM)-Cellulose chromatography. The specific activity of the purified enzyme was determined to be 65.0 U/mg with 2.55 purification fold and yield of 67.5 %. Polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed the presence of a single band with a molecular weight of around 35 kDa. Based on Fourier transformation-infrared spectroscopy analysis the functional groups of lipase were determined and high performance liquid chromatography analysis with the retention time of 1.995 min reveals the presence of lipase in the purified extract.