Substrate For Lipase Production Research Articles

Utilization of Moringa oleifera seed waste as substrate in lipase production under different pretreatments

Moringa industry in Sumenep Regency experienced growth in recent years along with the increase in export demand. Moringa seed waste, which is the coproduct of oil extraction contains a massive amount of lipid and biomass. As much as 30% of lipid residue from Moringa oleifera seeds waste are potentially usable as a substrate for lipase production. However, the phenolic compounds contained in the waste are difficult to degrade and have antimicrobial property which is needed to be removed. This study aims to determine the best pretreatment method in removing phenolic content in Moringa seed waste. The proposed pretreatment methods in this study including enzymatic pretreatment, Laccase Mediator System pretreatment, basic chemical pretreatment, acidic chemical pretreatment, and peroxide pretreatment. Lipase production carried out using Bacillus sp. SK II-5 isolates. Total protein content of lipases produced from various pretreatment then measured using the Bradford method. Lipase enzymatic activity determined qualitatively using the Quantofix formaldehyde test. Characterization of lipase based on isoelectric point. The result of this study is that laccase pretreatment is the best pretreatment method which capable of reducing total phenol concentration in Moringa seed waste by 70% to 3.87 mgGAE/g. The results were followed by a high total protein concentration of 0.43 mg/ml and lipase activity in the range of 100-200 ppm formaldehyde

Optimization of parameters for lipase production by Aspergillus niger NRRL-599 using response surface methodology

Background and objective Lipases are characterized to catalyze precise chemical transformation. That is they have widespread applications in detergents, cosmetics, food, organics synthesis, and pharmaceutical industries. In recent years, the study and development of lipase production in solid state fermentation, also using sequential statistical strategy as optimization for some vital factors are gaining more attention. This may be attributed to several advantages using techniques such as simplicity of fermentation media, no need for complex machinery. Equipment and control system, compactness of fermentation vessel due to lower water volume, high yields, less energy needed, and lower capital. Materials and methods Aspergillus niger NRRL-599 strain was tested for lipase production using different agroindustrial wastes including wheat bran, wheat germ cake oil, jojoba cake oil, almond cake oil, and olive oil as substrates under solid state fermentation. For further optimization, lipase activity was studied using two sequential optimization. Results and conclusion Olive oil waste was the most suitable substrate for lipase production (125 U/ml). Using one variable at a time, maximum lipase activity (200 U/ml) and specific enzyme activity (357.1 U/mg) were recorded in the presence of 5% w/v olive oil, 48 h inoculum age, 4% v/v inoculum size after 14 days fermentation at 30°C. The screening of the seven physiological factors using Plackett–Burman design showed that only three variables; that is tween 80, moisture content, and inoculum age affected significantly lipase production. Optimization by Box–Behnken design resulted in the highest lipase activity (420 U/ml) in which the most effective variables were inoculum size, inoculum age, moisture content, and tween 80 1%.

UTILIZATION OF WHEY AS A CHEAP SUBSTRATE FOR THE OPTIMIZATION OF LIPASE PRODUCTION BY Bacillus subtilis B10 ISOLATED FROM DAIRY INDUSTRY

Whey, a by-product of the dairy industry, acts as a prime source of environmental pollution due to its high organic load. Its utilization as a substrate for lipase production is an attractive option in facing the challenge associated with whey disposal. However, there are only very few studies that have reported the production of lipase using whey. In this study, 43 bacterial strains were isolated from dairy industry soil sample, of which, 26 were found to produce lipase. Since whey contains lactose as the sole carbon source, the ability of the isolates to utilize this sugar was tested and two of the isolates B3 and B10 were positive for growth in phenol red lactose broth. When they were cultured by submerged fermentation and their lipase activities quantified, strain B10 displayed 0.79U/ml of activity. In order to improve the production of lipase, one-factor-at-a-time method was used to study the impacts of oil inducers, nitrogen sources, mineral salts and whey concentration on the process. Statistical optimization was performed using the Box-Behnken design of Response Surface Methodology (RSM). From this design, 75% whey, 4% (v/v) sunflower oil, 0.625% (w/v) beef extract and 0.2% (w/v) CaCl2 were inferred to be the optimal conditions that resulted in a maximum lipase activity of 0.954U/ml. Analysis of variance indicated statistical significance of the model. Lipase being an industrially sought-after enzyme owing to its unique properties, this cost-effective route to its production from a waste substrate holds paramount environmental and economic significance.

The crude lecithin gum as a substrate for lipase production by Pseudomonas sp

The crude lecithin gum as a substrate for lipase production by Pseudomonas sp

An insight into microbial lipases and their environmental facet

Lipases are serine hydrolases that catalyze the hydrolysis and synthesis of esters formed from glycerol and long-chain fatty acids, by acting at the oil–water interface. Lipases from microbial sources have received heightened attention for an array of industrial applications, and these enzymes have been well exploited in the environmental sector as well. In this article, we present an overview of microbial lipase, including the microorganisms from which it could be produced; the application of recombinant DNA technology tools to produce lipase with enhanced properties, the effective use of waste materials as substrates for lipase production; the usage of statistical tools to efficiently optimize the production medium; lipase purification strategies; and the immobilization of the enzyme on a variety of support materials. The next section of the article provides a gist of its application in diversified spheres and focusses exclusively on the environmentally relevant ones. Lipase-catalyzed esterification, transesterification, and interesterification reactions, an emerging area of green chemistry; lipase-mediated in vitro biopolymer synthesis and degradation; and the application of lipase for remediating fat and oil constituents in wastewater are dealt with in-depth. When its full potential is harnessed, the enzyme could play a pivotal role in environmental management.

Lipase Production by Botryosphaeria ribis EC-01 on Soybean and Castorbean Meals: Optimization, Immobilization, and Application for Biodiesel Production

The effects of soybean and castorbean meals were evaluated separately, and in combinations at different ratios, as substrates for lipase production by Botryosphaeria ribis EC-01 in submerged fermentation using only distilled water. The addition of glycerol analytical grade (AG) and glycerol crude (CG) to soybean and castorbean meals separately and in combination, were also examined for lipase production. Glycerol-AG increased enzyme production, whereas glycerol-CG decreased it. A 2(4) factorial design was developed to determine the best concentrations of soybean meal, castorbean meal, glycerol-AG, and KH2PO4 to optimize lipase production by B. ribis EC-01. Soybean meal and glycerol-AG had a significant effect on lipase production, whereas castorbean meal did not. A second treatment (2(2) factorial design central composite) was developed, and optimal lipase production (4,820 U/g of dry solids content (ds)) was obtained when B. ribis EC-01 was grown on 0.5 % (w/v) soybean meal and 5.2 % (v/v) glycerol in distilled water, which was in agreement with the predicted value (4,892 U/g ds) calculated by the model. The unitary cost of lipase production determined under the optimized conditions developed ranged from US$0.42 to 0.44 based on nutrient costs. The fungal lipase was immobilized onto Celite and showed high thermal stability and was used for transesterification of soybean oil in methanol (1:3) resulting in 36 % of fatty acyl alkyl ester content. The apparent K m and V max were determined and were 1.86 mM and 14.29 μmol min(-1) mg(-1), respectively.

Production, characterization and applications of organic solvent tolerant lipase by Pseudomonas aeruginosa AAU2

Abstract Jatropha seedcake (JSC) represents an important environmental problem owing to its toxicity for the biodiesel producing sectors where they are generated in bulk as a major by-product. The aim of the present work was to study the feasibility of JSC as a substrate for lipase production under submerged condition. Pseudomonas aeruginosa AAU2 isolated from dumped rotting JSC was found to produce extracellular solvent tolerant lipase (0.432 U ml −1 ). A 11.4-fold increase in enzyme yield was obtained in an optimized medium containing 20 g JSC/l, 2.5 g potassium nitrate/l and 0.1 v/v% Tween 80 after 60 h of incubation at 30 °C under shake flask conditions (150 rpm). The pH and temperature optima of the partially purified AAU2 lipase were 7.5 and 40 °C, respectively. The AAU2 lipase was found to be a cysteine hydrolase and exhibited preference towards longer carbon chain fatty acid ester substrates over the shorter ones. The enzyme exhibited significant stability in the presence of commercial detergents namely, Ezee ® and Wheel ® . The AAU2 lipase efficiently catalyzed biodiesel production using Jatropha oil. In addition, the enzyme also hydrolyzed 86.49% of commercially available phorbol ester (phorbol 12-myristate 13-acetate) within 12 h of incubation at 37 °C, suggesting its application in detoxification of phorbol ester from JSC. These results demonstrate viable approach for utilization of JSC biomass and degradation of phorbol esters by P. aeruginosa AAU2.

Lipase from Penicillium camembertii KCCM 11268: Optimization of solid state fermentation and application to biodiesel production

Lipase was produced by Penicillium camembertii KCCM 11268 under solid state fermentation (SSF), and the production process was optimized by using statistical experimental designs. The initial moisture content, cultivation time, inoculum size and concentration of basal medium were considered as the factors of optimum conditions for SSF. P. Camembertii KCCM 11268 was cultivated in SSF using wheat bran as the substrate for lipase production. Under the optimized condition, lipase activity was reached around 7.8 U/ml after eight days fermentation. To partially purify the lipase, ammonium sulfate (80% saturation) was added to the crude lipase solution and concentrated using a diafiltration (VIVAFLOW 50). The concentrated lipase solution from P. Camembertii KCCM 11268 (PCL) was immobilized on silica gel by cross-linking method. Also, PCL was mixed with a commercial lipase solution from Candida rugosa (CRL), and this mixture was co-immobilized on silica gel. The immobilized and co-immobilized lipase activities were 1150.1 and 7924.8 U/g matrix, respectively. Palm oil and methanol were used as substrates and 1mmol of methanol was added every 1.5 h and 2 times during biodiesel production. The reaction was carried out at temperatures of 30, 40, 50, 60 and 70 °C. The maximum biodiesel conversion by co-immobilized lipase was 99% after 5 h at 50 °C.

Utilizing Agricultural Wastes as Substrates for Lipase Production by Candida rugosa NCIM 3462 in Solid-State Fermentation: Response Surface Optimization of Fermentation Parameters

Utilizing agricultural wastes for the production of industrially valuable enzymes like lipases in solid state fermentation holds a promising alternative for conventional processes due to the various obvious advantages, especially the cost-effectiveness of the process and sustainability. In this present work, the feasibility of exploiting various agricultural residues for lipase production was evaluated and optimization of the important parameters was done using statistical methods. Candida rugosa NCIM 3462 was used for the production of lipase in solid state fermentation. Agro industrial wastes such as sesame oil cake, groundnut oil cake and coconut oil cake were used for the solid state fermentation. The Energy dispersion spectrum of the solid substrates was used to study the elemental composition of oil cakes. The response surface methodology was employed to optimize the lipase production and to study the effect of temperature and substrate to moisture ratio. The maximum lipase activity of 22.40 U/g substrate was obtained using sesame oil cake which was 1.8 times the maximum activity obtained during the initial screening of variables by ‘one-factor-at-time’ approach. The optimized temperature and substrate to moisture ratio were found to be 32.3 °C and 1:3.23 g/ml respectively. A predictive model for the combined effects of the independent variables using response surface methodology and artificial neural network was proved to be excellent empirical model for lipase production by C. rugosa in solid state fermentation and aid in improving lipase fermentation utilizing agricultural residues as valuable substrates.

Optimization of process parameters for enhanced production of lipase by Penicillium notatum using agricultural wastes

In this study, cultural conditions were optimized for enhanced production of lipase byPenicillium notatum. Different agricultural wastes including canola oilseed cake, sesame oilseed cake, linseed oil cake, cotton oilseed cake, rice bran and wheat bran were used as substrates for their potential to be used for the production of lipase under solid state fermentation conditions. Among these, the canola oil seed cake proved to be the best substrate for production of lipase under solid state fermentation conditions. A maximum lipase activity of 5335 U/g dry substrate was observed after 96 h of incubation period using 60% moisture content, at pH 5, incubation temperature of 30°C and olive oil of 3%. Enrichment with maltose as carbon source enhanced the lipase production significantly, while nitrogen supplementation did not affect the lipase production significantly. The optimization of conditions led to two-fold enhancement of lipase activity as compared to the initial enzyme activity. Key words: Fermentation, Penicillium notatum, olive oil, lipase, culture conditions

Use of evolutionary operation (EVOP) factorial design technique to develop a bioprocess using grease waste as a substrate for lipase production

The aim of the present work was to develop a bioprocess using EVOP-factorial design technique employing grease waste as a substrate for the production of lipase. A newly isolated fungal strain of Penicillium chrysogenum was explored for the fermentation process. Solid-state fermentation (SSF) was carried out using grease waste and Czapek-dox medium, supplemented with wheat bran. The yield of lipase was 38U/ml when SSF was carried out at 32°C for 8days and grease:wheat bran:Czapek-dox media in 1:1:2 (w/w/v). Different physicochemical parameters affecting the production of lipase were optimized through evolutionary operation (EVOP) factorial design technique and after optimization yield was enhanced up to 46U/ml at 30°C, pH 7.0 with 1:1:2 (w/w/v) grease waste:wheat bran:Czapek-dox media. Industrial grease waste has never been reported before for the production of industrially important lipase enzyme.

Purification, characterization and application of acidic lipase from Pseudomonas gessardii using beef tallow as a substrate for fats and oil hydrolysis

Beef tallow, a slaughter house waste was used as a substrate for lipase production, employing Pseudomonas gessardii. The strain, P. gessardii was isolated from the beef tallow acclimatized soil. The crude lipase activity at 139 U/ml by volume was obtained at optimized conditions of pH 5.0 and temperature of 37 °C. After purification, a 7.59-fold purity of lipase with specific activity of 1120 U/mg protein and molecular mass of 92 kDa was obtained. The purified lipase showed maximum activity and stability at pH 5.0 and 30 °C. Ca 2+ had a stimulatory effect on the lipase activity compared to the other metal ions studied. The relative activity was enhanced with the addition of Triton X-100 with lower hydrophilic–lipophilic balance (HLB) value as 13.0 and DMSO with the lowest partition coefficient (log P) value, as 1.378. The amino acid composition and the functional groups of lipase were confirmed by HPLC and FT-IR spectroscopy. The purified lipase had the highest hydrolytic activity towards slaughterhouse wastes and vegetable oils. This work provides a potential biocatalyst for the wide applications in oleochemical and biotechnological industries.

Optimization of protease and lipase production by <italic>Bacillus pumilus</italic> SG 2 isolated from an industrial effluent

This investigation was aimed at isolating a potent bacterial strain which can produce both protease and lipase. Effluents of meat processing industry, dairy industry, food processing industry and oil industry were collected. Among the four positive strains screened, a bacterial strain identified as Bacillus pumilus SG2 isolated from food processing industry effluent was found to be a comparatively potent producer of both protease and lipase (52U/ml and 38U/ml respectively). The process parameters for the optimal production of both enzymes using this strain were studied. The optimum pH for protease production was 8.0 and that of lipase was 9.0. The optimum temperature was 37oC for production of both enzymes. The optimum incubation time was found to be 36 hours for protease and 63 hours for lipase. The best carbon source for enzyme production was glucose and the best nitrogen sources were yeast extract and casein. Addition of additives like SDS, Triton X-100 and Tween-20 influenced enzyme production. Whey was the best crude substrate that could be used for protease production and castor oil, gingelly oil and olive oil were the best substrates for lipase production. This study has indicated the possibility of producing both protease and lipase simultaneously on a common production medium by a single bacterial strain, Bacillus pumilus.

Candida cylindracea: A model organism to study the possibility of using olive mill wastewaters as a substrate for lipase production

Candida cylindracea: A model organism to study the possibility of using olive mill wastewaters as a substrate for lipase production