Lipase For Biodiesel Production Research Articles

An encapsulated report on enzyme-assisted transesterification with an allusion to lipase.

Biodiesel is a renewable, sulfur-free, toxic-free, and low carbon fuel which possesses enhanced lubricity. Transesterification is the easiest method employed for the production of biodiesel, in which the oil is transformed into biodiesel. Biocatalyst-mediated transesterification is more advantageous than chemical process because of its non-toxic nature, the requirement of mild reaction conditions, absence of saponification, easy product recovery, and production of high-quality biodiesel. Lipases are found to be the primary enzymes in enzyme-mediated transesterification process. Currently, researchers are using lipases as biocatalyst for transesterification. Lipases are extracted from various sources such as plants, microbes, and animals. Biocatalyst-based biodiesel production is not yet commercialized due to high-cost of purified enzymes and higher reaction time for the production process. However, research works are growing in the area of various cost-effective techniques for immobilizing lipase to improve its reusability. And further reduction in the production cost of lipases can be achieved by genetic engineering techniques. The reduction in reaction time can be achieved through ultrasonic-assisted biocatalytic transesterification. Biodiesel production by enzymatic transesterification is affected by many factors. Various methods have been developed to control these factors and improve biodiesel production. This report summarizes the various sources of lipase, various production strategies for lipase and the lipase-mediated transesterification. It is fully focused on the lipase enzyme and its role in biodiesel production. It also covers the detailed explanation of various influencing factors, which affect the lipase-mediated transesterification along with the limitations and scope of lipase in biodiesel production.

Optimizing the catalytic activities of methanol and thermotolerant Kocuria flava lipases for biodiesel production from cooking oil wastes

In this study, two highly thermotolerant and methanol-tolerant lipase-producing bacteria were isolated from cooking oil and they exhibited a high number of catalytic lipase activities recording 18.65 ± 0.68 U/mL and 13.14 ± 0.03 U/mL, respectively. Bacterial isolates were identified according to phenotypic and genotypic 16S rRNA characterization as Kocuria flava ASU5 (MT919305) and Bacillus circulans ASU11 (MT919306). Lipases produced from Kocuria flava ASU5 showed the highest methanol tolerance, recording 98.4% relative activity as well as exhibited high thermostability and alkaline stability. Under the optimum conditions obtained from 3D plots of response surface methodology design, the Kocuria flava ASU5 biocatalyst exhibited an 83.08% yield of biodiesel at optimized reaction variables of, 60 ○C, pH value 8 and 1:2 oil/alcohol molar ratios in the reaction mixture. As well as, the obtained results showed the interactions of temperature/methanol were significant effects, whereas this was not noted in the case of temperature/pH and pH/methanol interactions. The obtained amount of biodiesel from cooking oil was 83.08%, which was analyzed by a GC/Ms profile. The produced biodiesel was confirmed by Fourier-transform infrared spectroscopy (FTIR) approaches showing an absorption band at 1743 cm−1, which is recognized for its absorption in the carbonyl group (C=O) which is characteristic of ester absorption. The energy content generated from biodiesel synthesized was estimated as 12,628.5 kJ/mol. Consequently, Kocuria flava MT919305 may provide promising thermostable, methanol-tolerant lipases, which may improve the economic feasibility and biotechnology of enzyme biocatalysis in the synthesis of value-added green chemicals.

Role of microbial lipases in transesterification process for biodiesel production

Bio-diesel is a fuel produced by transesterification reaction of oil or fat in the presence of a catalyst. It is an alternate renewable source for petrol and diesel, which helps in reducing greenhouse gases emissions. Chemical catalysts such as sodium hydroxide, potassium hydroxide, sulfuric acid are used in the conventional methods of biodiesel production. This review focusses on application of lipase enzyme in transesterification of different types of oils such as: soybean, sunflower, rapeseed, palm oil, and waste cooking oil. Worldwide production of biodiesel via lipase enzyme and its economic as well as future aspects are also discussed. Various researchers have used free lipase in biodiesel production however, some lipases are also utilized as immobilized form. There are number of fungal and bacterial species that have been used to synthesize lipases, some of the microbial species are as follows: Aspergillus niger, Burkholderia cepacia, Pseudomonas aeruginosa, Staphylococcus epidermidis etc. Solid-state fermentation and broth culture of fungus is suitable for crude lipase production. Immobilized lipase plays very significant role in transesterification process. Lipases enzymes act as a good catalyst therefore, its production and utilization may be a better alternative of chemical catalysts.

Hydrophobic pore space constituted in macroporous ZIF-8 for lipase immobilization greatly improving lipase catalytic performance in biodiesel preparation

BackgroundDuring lipase-mediated biodiesel production, by-product glycerol adsorbing on immobilized lipase is a common trouble that hinders enzymatic catalytic activity in biodiesel production process. In this work, we built a hydrophobic pore space in macroporous ZIF-8 (named as M-ZIF-8) to accommodate lipase so that the generated glycerol would be hard to be adsorbed in such hydrophobic environment. The performance of the immobilized lipase in biodiesel production as well as its characteristics for glycerol adsorption were systematically studied. The PDMS (polydimethylsiloxane) CVD (chemical vapor deposition) method was utilized to get hydrophobic M-ZIF-8-PDMS with hydrophobic macropore space and then ANL (Aspergillus niger lipase) was immobilized on M-ZIF-8 and M-ZIF-8-PDMS by diffusion into the macropores.ResultsANL@M-ZIF-8-PDMS presented higher enzymatic activity recovery and better biodiesel production catalytic performance compared to ANL@M-ZIF-8. Further study revealed that less glycerol adsorption was observed through the hydrophobic modification, which may attribute to the improved immobilized lipase performance during biodiesel production and ANL@M-ZIF-8-PDMS remained more than 96% activity after five cycles’ reuse. Through secondary structure and kinetic parameters’ analysis, we found that ANL@M-ZIF-8-PDMS had lower extent of protein aggregation and twice catalytic efficiency (Vmax/Km) than ANL@M-ZIF-8.ConclusionsHydrophobic pore space constituted in macroporous ZIF-8 for lipase immobilization greatly improved lipase catalytic performance in biodiesel preparation. The hydrophobic modification time showed negligible influence on the reusability of the immobilized lipase. This work broadened the prospect of immobilization of enzyme on MOFs with some inspiration.

Lipase Production by Solid-State Cultivation of Thermomyces Lanuginosus on By-Products from Cold-Pressing Oil Production

This study shows that by-products obtained after cold-pressing oil production (flex oil cake, hemp oil cake, hull-less pumpkin oil cake) could be used as substrates for the sustainable and cost-effective production of lipase when cultivating Thermomyces lanuginosus under solid-state conditions (T = 45 °C, t = 9 days). Lipase showed optimum activity at T = 40 °C. The produced lipase extract was purified 17.03-folds with a recovery of 1% after gel chromatography. Three different batch experiments were performed in order to test the possibility of using the lipase in biodiesel production. Experiments were performed with a commercial, unpurified enzyme, and partially purified lipase with sunflower oil and methanol as substrates in a batch reactor at 40 °C. During the experiments, the operational stability of the enzyme was studied. The obtained results clearly showed that produced crude and purified lipase can be used for biodiesel production, but the process needs some additional optimization. As for operation stability, it was noticed that the commercial enzyme was deactivated after 30 h, while produced crude enzyme remained 8.25% of its activity after 368 h.

Jet cutter as tool to immobilize lipase for biodiesel production

The use of lipases as a biocatalyst for industrial applications is an interesting route due to technical aspects but also to reduce environmental impacts caused by the use of chemical catalysts. Gel immobilization of the enzyme allows its reuse and avoids contamination of the product with residual portions of free enzyme. However, a typical technique available for enzyme immobilization is based on dripping driven by gravity which produces big particles and low rate of production. The reduction of size can improve the mass transfer by increasing the contact area. Thus, aiming to increase the rate of particles production and reduce the size of particles, the objective of this work was to encapsulate lipase, using a tool designed to cut the jet produced by pumping, called as Jet Cutter.

A robust process for lipase-mediated biodiesel production from microalgae lipid

Combination of two lipases for biodiesel production from microalgae lipid.

Recent developments in biocatalysis beyond the laboratory

Recent developments in biocatalysis, where implementation beyond the laboratory has been demonstrated, are explored: the use of transglutaminases to modify foods, reduce allergenicity and produce advanced materials, lipases for biodiesel production, and transaminases for biochemical production. The availability and application of enzymes at pilot and larger scale opens up possibilities for further improvements of biocatalyst-based processes and the development of new processes. Enzyme production, stability, activity, re-use, and product retrieval are common challenges for biocatalytic processes. We explore recent advances in biocatalysis within the process chain, such as protein engineering, enzyme expression, and biocatalyst immobilization, in the context of these challenges.

Efficient biodiesel production from phospholipids-containing oil: Synchronous catalysis with phospholipase and lipase

Recently, the use of low quality oils as potential alternative oil feedstock for biodiesel production has become more and more attractive, which provides a feasible solution to reduce the cost of biodiesel production. However, in the low quality oils, expect triglycerides, other lipids represented by phospholipids also exist. It was found that the presence of phospholipids showed adverse effect on enzyme catalyzed biodiesel production. Therefore, it is necessary to develop an effective method for the conversion of phospholipids-containing oils. In recent years, the employment of free lipase in catalyzing biodiesel from refined oils has received great attentions due to its advantages of lower cost and faster reaction rate. To further promote its application in efficient conversion of phospholipids-containing oils, a synchronous catalysis together with phospholipase and lipase for biodiesel production from phospholipids-containing oils was developed. A high FAME yield of 94.9% can be obtained even with 10% phospholipids contained in the oil feedstock. Further study showed that the biodiesel product could also meet the phosphorus demand of EN14214 standard. Moreover, the good reuse stability of the two enzymes further indicates that the combination catalysis of phospholipase and lipase is very promising for biodiesel production especially with high phospholipids-containing oils as feedstock.

New Lipase for Biodiesel Production: Partial Purification and Characterization of LipSB 25-4.

The lipolytic activities of 300 Streptomyces isolates were determined in Tributyrin and Rhodamine-B Agar. Lipase activities were also measured with p-nitrophenyl palmitate (p-NPP) as a substrate. The strain of Streptomyces bambergiensis OC 25-4 used in this study was selected among 300 strains of Streptomyces from MUCC as the best lipase producer. The incubation conditions were optimized and the inoculum amount, incubation period, effect of carbon and nitrogen sources, and rates of MgSO4 and CaCO3 were investigated. LipSB 25-4 (the lipase produced by S. bambergiensis OC 25-4 strain) was partially purified with ammonium sulphate precipitation, dialysis, and gel filtration chromatography 2.73-fold and with 92.12 U/mg specific activity. The optimal pH and temperature for LipSB 25-4 were determined as 8.0 and 50°C, respectively. The lipase has high stability in all pH and temperature values used in this study. While LipSB 25-4 was slightly activated in the presence of β-mercaptoethanol, it was slightly reduced by PMSF. The enzyme conserved approximately 75% of its activity at the end of 60 h, in the presence of methanol and ethanol. Since LipSB 25-4 displays high activity in the thermophilic conditions and stability in the presence of organic solvents, this lipase can catalyse the biodiesel production from olive oil by the transesterification reactions.

Synthesis of SBA-15 from low cost silica precursor obtained from sugarcane leaf ash and its application as a support matrix for lipase in biodiesel production

Ordered mesoporous silica material was synthesized from a low-cost precursor, sugarcane leaf ash, was used as a support matrix for lipase for the production of biodiesel. The mesoporous samples were characterized using Fourier transform infra red spectroscopy. The surface topography and morphology of the mesoporous materials were studied using scanning electron microscope. The pore diameter, pore volume, Brunauer Emmett and Teller surface area of the mesoporous material were determined by N2 gas adsorption technique. Different pore size Santa Barbara Acid-15 (SBA-15) samples were synthesized and their lipase immobilization capacity and specific enzyme activity of immobilization lipase were determined and compared. Lipase from Candida Antarctica immobilized on SBA-15 (C) had shown maximum percentage immobilization and specific enzyme activity. The immobilized lipase mesoporous matrix was used for biodiesel production from crude non-edible Calophyllum inophyllum oil. The percentage yield of fatty acid methyl ester, 97.6 % was obtained under optimized conditions: 100 mg of lipase immobilized on SBA-15, 6:1 methanol to oil molar ratio, the reaction of 2 g C. inophyllum oil with methanol.

Recent Strategy of Biodiesel Production from Waste Cooking Oil and Process Influencing Parameters: A Review

Cost of biodiesel produced from virgin vegetable oil through transesterification is higher than that of fossil fuel, because of high raw material cost. To minimize the biofuel cost, in recent days waste cooking oil was used as feedstock. Catalysts used in this process are usually acids, base, and lipase. Since lipase catalysts are much expensive, the usage of lipase in biodiesel production is limited. In most cases, NaOH is used as alkaline catalyst, because of its low cost and higher reaction rate. In the case of waste cooking oil containing high percentage of free fatty acid, alkaline catalyst reacts with free fatty acid and forms soap by saponification reaction. Also, it reduces the biodiesel conversions. In order to reduce the level of fatty acid content, waste cooking oil is pretreated with acid catalyst to undergo esterification reaction, which also requires high operating conditions. In this review paper, various parameters influencing the process of biofuel production such as reaction rate, catalyst concentration, temperature, stirrer speed, catalyst type, alcohol used, alcohol to oil ratio, free fatty acid content, and water content have been summarized.

A promising alternate lipase for biodiesel fuel production

By screening samples from different sources, a high-yield yet low-cost lipase of Enterobacter aerogenes was found and used in biodiesel production. The enzyme activity of Enterobacter aerogenes reached 34.16 U·mL−1 under the optimum conditions of initial pH 9.0, agitating at 30°C for 40 h, using NaHCO3 and glucose as carbon sources and peptone as nitrogen source, with 0.1% (w/v) MgSO4·7H2O. When adding 1.0% (w/v) olive oil to the culture, the enzyme activity reached 66.31 U·mL−1, which increased by 56.5% compared to that obtained by using 2.0% (w/v) olive oil as the sole carbon source. The lipase of Enterobacter aerogenes was immobilized by diatomite and used to catalyze biodiesel with rapeseed oil as raw materials, the transesterification rate was 92.91% under the optimum reaction conditions, which were using 1000 U immobilized lipase and 5 mL n-hexane, with ethanol as acyl acceptors, at the molar ratio of oil/ethanol of 1:4, and adding ethanol twice, at 35°C for 48 h. Therefore, the Enterobacter aerogenes will potentially serve as a promising alternative lipase for biodiesel production.

Development of recombinant Escherichia coli whole-cell biocatalyst expressing a novel alkaline lipase-coding gene from Proteus sp. for biodiesel production

A lipase-producing bacterium K107 was isolated from soil samples of China and identified to be a strain of Proteus sp. With genome-walking method, the open reading frame of lipase gene lipK107, encoding 287 amino acids, was cloned and expressed in a heterologous host, Escherichia coli BL21 (DE3). The recombinant lipase was purified and characterized, and the optimum pH of the purified LipK107 was 9, at 35 °C. The recombinant E. coli expressing lipK107 was applied in biodiesel production in the form of whole-cell biocatalyst. Activity of the biocatalyst increased significantly when cells were permeabilized with 0.3% (w/v) cetyl-trimethylammoniumbromide (CTAB). This transesterification was carried out efficiently in a mixture containing 5 M equivalents of methanol to the oil and 100% water by weight of the substrate. It was the first time to use E. coli whole-cell biocatalyst expressing lipase in biodiesel production, and the biodiesel reached a yield of nearly 100% after 12 h reaction at the optimal temperature of 15 °C, which was the lowest temperature among all the known catalyst in biodiesel production.

Desenvolvimento de um modelo da cinética enzimática da transesterificação de óleos vegetais para produção de biodiesel

Currently the technology of enzymatic production of biodiesel is more promising than that based on chemical methods, using acidic or basic catalysts. The enzyme lipase used as catalyst in this process can be isolated from microorganisms such as Pseudomonas fluorescens, P. cepacia, Candida antarctica among others. In this work, a mechanism for the transesterification of triglycerides catalyzed by lipase for production of biodiesel is proposed. The process was modeled in three consecutive reactions where diglycerides and monoglycerides were formed. Other three intermediate stages involving lipase were considered. The identification of parameters with the developed model was performed for reaction conditions where stoichiometric amount of reagents were used. The developed model was tested for different operational conditions with experimental data found in the literature. The simulation results showed good model flexibility.

Immobilization of Lipase on Macroporous Resin and Its Application in Synthesis of Biodiesel in Low Aqueous Media

Lipase from Candida sp. 99-125 was immobilized by physical adsorption onto macroporous resins. The results showed that the nonpolar resin NKA was the best carrier used in low aqueous media. 98.98 % of degree of immobilization could be achieved when the adsorption procedure was performed in the presence of heptane. The lipolytic activity and the apparent activity recovery of lipase adsorbed on resin in heptane were 4.07 and 3.43 times higher than that of lipase adsorbed in sodium phosphate buffer, respectively. The catalytic properties of immobilized lipase for production of biodiesel in low aqueous media were studied. Immobilized lipase displayed the highest activity when the crude enzyme/resin weight ratio was 1.92:1 and the water content (water/ oil weight ratio) was 15 % at 40 °C under pH 7.4. As lipase was adsorbed on NKA in heptane to produce biodiesel, the batch conversion rate reached 97.3 % when a three-step methanolysis protocol was used. After 19 consecutive batches, the conversion rate remained 70.2 %.