Degumming Of Oil Research Articles

Enzymatic degumming of corn oil using phospholipase C from a selected strain of Pichia pastoris

Degumming is a necessary refining step for all crude vegetable oils. Purifine® PLC is a new enzyme used for industrial oil degumming. In the present study, enzymatic degumming trials were performed on crude corn oil using a commercial phospholipase C enzyme Purifine® PLC with the aim of determining its optimum process conditions. Enzymatic degumming applying 200 mg/kg of Purifine® PLC during 120 min at 60 °C, and a pH of 5.7 using chemical conditioning, resulted in a residual phosphorus content of 27 mg/kg and an absolute diacylglycerol increase of 0.54 wt%. Compared to water degumming, enzymatic degumming with Purifine® PLC provided a better degumming (67 mg/kg versus 27 mg/kg) and an increase in the diacylglycerol content. The pH adjustment of crude corn oil performed by means of a caustic pretreatment was not able to keep the pH at an optimal stable value due to the continuous release of acidic phosphate groups. Analysis of the composition of remaining phospholipids in the gums fraction showed that Purifine® PLC could effectively convert phosphatidylcholine and phosphatidylethanolamine into diacylglycerols, whereas it could not convert phosphatidylinositol and phosphatidic acid. These results confirm that Purifine® PLC degumming is a commercially feasible alternative to traditional degumming processes.

Recombinant Expression of Serratia marcescens Outer Membrane Phospholipase A (A1) in Pichia pastoris and Immobilization With Graphene Oxide-Based Fe3O4 Nanoparticles for Rapeseed Oil Degumming.

Enzymatic degumming is an effective approach to produce nutritional, safe, and healthy refined oil. However, the high cost and low efficiency of phospholipase limit the application of enzymatic degumming. In this study, an 879 bp outer membrane phospholipase A (A1) (OM-PLA1) gene encoding 292 amino acid residues was isolated from the genome of Serratia marcescens. The recombinant OM-PLA1 profile of appropriately 33 KDa was expressed by the engineered Pichia pastoris GS115. The OM-PLA1 activity was 21.2 U/mL with the induction of 1 mM methanol for 72 h. The expression efficiencies of OM-PLA1 were 0.29 U/mL/h and 1.06 U/mL/OD600. A complex of magnetic graphene oxide (MGO) and OM-PLA1 (MGO-OM-PLA1) was prepared by immobilizing OM-PLA1 with graphene oxide-based Fe3O4 nanoparticles by cross-linking with glutaraldehyde. The content of phosphorus decreased to 5.1 mg/kg rapeseed oil from 55.6 mg/kg rapeseed oil with 0.02% MGO-OM-PLA1 (w/w) at 50°C for 4 h. MGO-OM-PLA1 retained 51.7% of the initial activity after 13 times of continuous recycling for the enzymatic degumming of rapeseed oil. This study provided an effective approach for the enzymatic degumming of crude vegetable oil by developing a novel phospholipase and improving the degumming technology.

Degumming Role of Sunflower Oil (Helianthus annuus) on Biodiesel Quality

Biodiesel, known as fatty acid methyl ester, has become more attractive as an alternative fuel for diesel engine because of its environmental benefits and the fact that it is made from renewable resources such as vegetable oil and animal fats by reacting them with short chain alcohols. Biodiesel has become increasingly important due to the following advantages: it is renewable, biodegradable, non-toxic, and eco-friendly. Crude sunflower oil may be a cheap source for biodiesel production. However, it contains high amount of phosphorus, which can inhibit the action of the catalyst during transesterification. Phosphorus removal from vegetable oils applied for diesel fuel is an essential process, since the presence of phosphors in direct use to the diesel engine might cause plugging in the engine filters, lines and injectors. In the biodiesel production, the use of crude vegetable oils without degumming might decrease conversion rate and hard to separate glycerol from biodiesel, during and after trans-esterification reaction respectively.Degumming role of crude sunflower oil on physicochemical properties of biodiesel was studied with determination of following parameters: phosphorus content, acidity value, peroxide value, viscosity, density, iodine value and saponification value. Degumming of sunflower oil is realysed by water degumming and by electrolyte degumming followed by a fast decrease of residual phosphorus content. The compared results showed that degumming process of sunflower oil has a role on biodiesel quality.

Industrial uses of phospholipases: current state and future applications.

Phospholipids play a central role in all living organisms. Phospholipases, the enzymes aimed at modifying phospholipids, are consequently widespread in nature and play diverse roles, from lipid metabolism and cellular signaling in eukaryotes to virulence and nutrient acquisition in microbes. Phospholipases catalyze the hydrolysis of one or more ester or phosphodiester bonds of glycerophospholipids. The use of phospholipases with industrial purposes has constantly increased over the last 30years. This demand is rapidly growing given the ongoing improvements in protein engineering and the reduction of enzymes manufacturing costs, making them suitable for industrial use. Here, a general overview of phopholipases A, B, C, and D and their industrial application is presented along with potential new uses for these enzymes. We draw attention to commercial phospholipases used to improve the emulsifying properties of products in the baking, egg, and dairy industries. On the other hand, the improvement of oil degumming by phospholipases is thoroughly analyzed. Moreover, recent developments in enzymatic biodiesel production and the use of phospholipases for the synthesis of phospholipids with pharmaceutical or nutritional value are reviewed.

Soybean Oil Degumming by Immobilized Phospholipase A1

In the present study, we investigated the ability of an immobilized phospholipase A1 (PLA1) in degumming of soybean oil. The enzyme was immobilized by simple adsorption on bentonite without any further modification. The free and immobilized PLA1 were characterized by Fourier Transform InfraRed (FT-IR) spectroscopy and X-Ray Diffraction (XRD). The immobilization of PLA1 lowered the Energy of Activation (EA) from 155.64 to 27.13 kJ/mol, resulting in higher catalytic efficiency of PLA1. Thermal stability of the immobilized enzyme was found to be higher compared to free PLA1. Moreover, under the condition of pH 5.5 and T = 50 °C, the phosphorus content was reduced to less than 10 ppm after 4-5 h for free PLA1 and after 7 h for immobilized PLA1.

Degumming of crude vegetable oil by membrane separation: Probing structure-performance and stability of PVDF membranes

ABSTRACTStability of polymeric membrane in organic solvent during degumming process has been the main issue. In this work, various PVDF membranes were prepared under controlled conditions, characterized and tested for their degumming performance of crude peanut oil miscella. The optimally prepared PVDF membrane showed excellent phospholipid (gum) rejection of 95% with hexane and miscella permeate fluxes of 70 LMH and 46 LMH, respectively when operated at 10 bar and room temperature. At initial stage, the flux kept declining to about 10% for the best membrane but it remained stable afterwards.

The βγ-crystallin domain of Lysinibacillus sphaericus phosphatidylinositol phospholipase C plays a central role in protein stability.

βγ-crystallin has emerged as a superfamily of structurally homologous proteins with representatives across all domains of life. A major portion of this superfamily is constituted by microbial members. This superfamily has also been recognized as a novel group of Ca2+-binding proteins with a large diversity and variable properties in Ca2+ binding and stability. We have recently described a new phosphatidylinositol phospholipase C from Lysinibacillus sphaericus (LS-PIPLC) which was shown to efficiently remove phosphatidylinositol from crude vegetable oil. Here, the role of the C-terminal βγ-crystallin domain of LS-PIPLC was analyzed in the context of the whole protein. A truncated protein in which the C-terminal βγ-crystallin domain was deleted (LS-PIPLCΔCRY) is catalytically as efficient as the full-length protein (LS-PIPLC). However, the thermal and chemical stability of LS-PIPLCΔCRY are highly affected, demonstrating a stabilizing role for this domain. It is also shown that the presence of Ca2+ increases the thermal and chemical stability of the protein both in aqueous media and in oil, making LS-PIPLC an excellent candidate for use in industrial soybean oil degumming.

Intensification of degumming of crude soybean oil using ultrasound in combination with oxidizing agents

Intensified degumming of crude soybean oil was investigated using ultrasound assisted combination approaches based on longitudinal ultrasonic horn at a capacity of 5 L with comparison with the conventional processing based on only ultrasound. The combination of ultrasound and oxidizing agents such as hydrogen peroxide (H2O2), potassium persulphate (KPS), Peroxymono sulphate (PMS) and Fenton reagent have been investigated for establishing the efficacy for intensified degumming. Combination of ultrasound and H2O2 loading at 100 ppm resulted in 94.15% phospholipid reduction at 150 W power and ambient temperature. Enhanced extent of degumming was observed for combination of ultrasound with KPS and PMS as 95.10 and 95.55% respectively. Ultrasound combined with Fenton reagent resulted in significant increase in extent of degumming (98.28%). Best degumming protocol was established as the combination of ultrasound with UV and hydrogen peroxide giving 99.18% phospholipid removal in only 120 min of treatment. The degumming was also supported by positive changes in the oil characteristics in terms of reduction in acid value (0.74 as the final value) and better oxidative stability (TOTOX value of ˂ 4). The present work demonstrated intensified degumming protocol for crude soybean oil processing for obtaining more than 99% degumming with better characteristics of processed oil.

Cloning, expression and characterisation of phospholipase B from Saccharomyces cerevisiae and its application in the synthesis of l-alpha-glycerylphosphorylcholine and peanut oil degumming

ABSTRACTl-alpha-glycerylphosphorylcholine (GPC) has been shown to enhance cognitive performance. Meanwhile, vegetable oils must be refined to remove the impurities for them to be edible. Phospholipase B (PLB), having the ability of hydrolyzing both the sn-1 and sn-2 acyl ester bonds of phospholipids, can produce GPC using PC as substrate and transform the non-hydratable phospholipids into their hydratable forms. The Saccharomyces cerevisiae plb gene, which encodes PLB, was cloned and expressed in Pichia pastoris GS115 to produce recombinant PLB (rPLB). Fermentation optimisation yielded rPLB activity levels as high as 1723 U/mL. rPLB demonstrated maximum enzymatic activity at 40 °C and pH 5.5 and was stable at temperatures between 30 and 40 °C and pH values between 5.0 and 6.0. rPLB synthesised GPC with a conversion rate of 17% (w/w) and exhibited high degumming activity towards peanut oil, decreasing the phosphorus content from 91.8 to 3.7 mg/kg within 3 h. This study describes a candidate phospholipase for potential applications involving the modification of phospholipids and vegetable oil degumming.

Application of Computational Fluid Dynamics for Modeling of Hydrodynamics and Mass Transfer of Laboratory Scale Crude Palm Oil Degumming Process

In this research, computational fluid dynamics was applied to model a laboratory reactor for the degumming of crude palm oil (CPO) involving immiscible liquid-liquid mixing and being controlled by mass transfer. The fluid mixing of CPO and phosphoric acid in the reactor was simulated using multiphase mixture model and standard κ-ε turbulence model in steady state mode. The simulation predicts the distributions of the drop diameter, the dispersed-phase relative velocity, the drop Reynolds number and the interfacial area density. The mass transfer coefficient from experimental work is correlated using the model as Sherwood number Shd = 0.02576 Red0.673Sc0.431 with R2 being 0.91. Finally, the volumetric mass transfer coefficient was calculated and compared to the experiment result.

Ultrasound assisted enzymatic degumming of crude soybean oil

The present work deals with ultrasound assisted enzymatic degumming (UAED) of crude soybean oil quantifying the extent of degumming (EOD), cavitational yield and synergistic index (f) for the combination approaches. The effect of different operating parameters such as enzyme loading, pH, presence of water, temperature and ultrasonic power on the EOD has been investigated. Ultrasound combined with enzyme at loading of 2.0 ml/L resulted in EOD as 92.2% under ambient conditions. Addition of water (5%) in combination with ultrasound and enzyme at 2.0 ml/L loading and pH of 5 resulted in maximum EOD (98.4%) in 120 min of treatment. The extent of phospholipid separation was also observed to be dependent on the power dissipation and maximum phospholipids separation was obtained at 100 W. Scale-up studies were performed at 500 ml and 1 L operating volume under optimized conditions of 2.0 ml/L as the enzyme loading, pH of 5, 5% water addition and ultrasonic power of 100 W where 93.63% and 91.15% phospholipid separation respectively was obtained. The effects of ultrasonic treatment were also quantified in terms of the acid value reduction and oxidative stability for the processed oil. It was demonstrated that suitable reduction in acid value (final value less than 1) and oxidative stability (TOTOX less than 4) is effectively obtained using UAED. Overall the approach of UAED was established to show much higher efficacy for soybean oil processing as compared to only ultrasound or only enzymatic treatment.

Evaluation of Ultrafiltration Performance for Phospholipid Separation

Ultrafiltration membrane for degumming of crude palm oil has been applied as an alternative method since the membrane process required less procedure than the conventional degumming. This research focused on the examination of ultrafiltration performance for phospholipid separation from model crude palm oil degumming. Specifically, profile flux and rejection, as well as blocking mechanism, were investigated. Feed consisting of Refined Crude Palm Oil – Isopropanol – Lecithin mixtures were represented as crude palm oil degumming. Lecithin was denoted a phospholipid component, and the concentrations of lecithin in feed were varied to 0.1%, 0.2%, and 0.3%. The concentration of phospholipid was determined as phosphor content. At the concentration of lecithin in feed representing phospholipid concentration of 8,45 mg/kg, 8,45 mg/kg, 24,87 mg/kg and 57,58 mg/kg, respectively. Flux profiles confirmed that there was a flux decline during filtration. In addition, the lecithin concentrations do not significantly effect on further flux decline. Rejection characteristic and phospholipid concentration in the permeate showed that the phospholipid rejections by ultrafiltration were in the range of 23-79,5% representing permeate’s phospholipid concentration of 1,73 - 44,25 mg/kg. Evaluation of fouling mechanism by Hermia’s blocking model confirmed that the standard blocking is the dominant mechanism in the ultrafiltration of lecithin mixture.

Assessment of water degumming and acid degumming of mixed algal oil

Crude algal oil from Chlorella sp. contained high amount of phospholipids, causing lower conversion rate of transesterification and difficulty in glycerol separation. Since the crude algal oil was solid at ambient temperature that could not go to degumming process straightly, it was mixed with refined palm oil in the ratio of 30:70w/w to obtain suitable viscosity for further processing. The response surface methodology with Box-Behnken design was applied to investigate water and acid degumming processes. The studied parameters were temperature, time, and amount of degumming reagent (water or phosphoric acid). The greatest phospholipid removal reached 82.5% under the optimum condition of acid degumming with temperature 90°C, 60min and phosphoric acid 0.42wt.%. The water degumming at optimum condition (temperature 100°C, 30min and water 80wt.%) only gave 19.4% phospholipid removal. The major phospholipids in Chlorella sp. oil was considered to be the non-hydratable phospholipids.

Effect of Crambe abyssinica oil degumming in phosphorus concentration of refined oil and derived biodiesel

Abstract In the present study, the effect of different degumming processes on the phosphorus content of Crambe abyssinica oil and resultant biodiesel was evaluated. The non-edible oil was submitted to water degumming and chemical degumming with different concentrations of phosphoric acid and varying the acid to oil volume percentages. Phosphorus content of the products was measured by UV spectroscopy after sample calcination, according to NP 1994:2000. Biodiesel was produced by transesterification using a 6:1 methanol to oil molar ratio and 1 wt% sodium hydroxide as catalyst at 65 °C, during 1 h. The biodiesel produced directly from the crude oil presented high phosphorus concentration (>20 ppm); consequently, a degumming process was required to fulfil the quality standard (

Intensified degumming of crude soybean oil using cavitational reactors

A novel cavitation assisted degumming approach for the purification of crude soybean oil in terms of the removal of the total phospholipid content has been reported in the work with a detailed study into the effect of operating parameters such as temperature, hydrogen peroxide loading and introduction of ozone gas at varying flow rate. Ultrasonic horn, ultrasonic bath and hydrodynamic cavitation reactors have been used for comparing the efficacy of different types of cavitational reactors. Initially, extent of degumming was quantified at different temperatures and 60 °C was established as optimum to give significant extent of degumming in terms of phospholipids removal. Degumming in the presence of ozone gas gave unsatisfactory results because of the specific physical and chemical properties of crude soybean oil. Combination of ultrasound and hydrogen peroxide resulted in complete phospholipid degradation at capacity of 250 ml at optimized temperature in 100 min of treatment. Ultrasonic horn and bath were also used at 1 L operating volume and the extent of degumming was found to be 90.81% and 93.50% respectively. For large scale studies, hydrodynamic cavitation was operated at 4 bar pressure using slit venturi as the cavitating device which gave 93.65% as the extent of degumming. Overall, intensified degumming approach for processing of crude soybean oil was demonstrated to yield high quality oil in terms of the reduced phospholipids content. Cavitation based degumming process can be an excellent alternative approach to the conventional method with major benefits like reduction in time and energy requirements as demonstrated in the present work.

Production of biodiesel from three indigenous feedstock: Optimization of process parameters and assessment of various fuel properties

Biodiesel is a clean and sustainable fuel which has ability to replace diesel fuel used in power generation as well in transportation. This article explores Terminalia belerica Robx seed oil as an important bioresource for biodiesel synthesis. Extraction of terminalia oil from dried seeds and degumming of crude oil are also reported. Synthesis of biodiesel from waste vegetable oil and chicken fat oil as low‐cost biodiesel feedstocks has also been carried out. The oils are trans‐esterified using base homogeneous catalyst KOH and reaction variables were optimized for biodiesel synthesis. The biodiesel yields 89%, 89%, and 82% were obtained using feedstocks Terminalia oil, chicken fat oil, and waste vegetable oils, respectively, at individually optimized parameters such as molar ratio (methanol to oil), catalyst concentration, reaction temperature, reaction time, and agitation speed. The investigated fatty acid profile of feedstocks and different fuel properties of biodiesel were found to be as per ASTM D6751‐07b and EU 14214:2003 standards. This article deliberates the possibility of T. belerica seed oil, waste vegetable oil, and chicken fat oil as promising bioresources for biodiesel production considering the recycling and conversion of waste materials to value‐added products. © 2017 American Institute of Chemical Engineers Environ Prog, 36: 788–795, 2017

Development of a highly efficient oil degumming process using a novel phosphatidylinositol-specific phospholipase C enzyme.

Enzymatic degumming using phospholipase C (PLC) enzymes may be used in environmentally friendly processes with improved oil recovery yields. In this work, phosphatidylinositol-specific phospholipase C (PIPLC) candidates obtained from an in silico analysis were evaluated for oil degumming. A PIPLC from Lysinibacillus sphaericus was shown to efficiently remove phosphatidylinositol from crude oil, and when combined with a second phosphatidylcholine and phosphatidylethanolamine-specific phospholipase C, the three major phospholipids were completely hydrolyzed, providing an extra yield of oil greater than 2.1%, compared to standard methods. A remarkably efficient fed-batch Escherichia coli fermentation process producing ∼14g/L of the recombinant PIPLC enzyme was developed, which may facilitate the adoption of this cost-effective oil-refining process.

B. cereus phospholipase C engineering for efficient degumming of vegetable oil

Abstract Enzymatic phospholipid removal (degumming) is a fast-growing and environmentally friendly process for vegetable oil refining. Type C phospholipases (PLC) are the preferred enzymes since they provide an extra yield in the oil recovery. Bacillus cereus PLC can hydrolyze phosphatidylcholine (PC) but has a limited efficiency at removing phosphatidylethanolamine (PE), which together represent ∼70% of the phospholipids present in crude soybean oil. In the present work, we show that the B. cereus PLC mutant F66Y can remove up to 90% of PE while retaining its efficiency at hydrolyzing PC. Oil treatment with the engineered enzyme provides an extra yield of 1.84% making the B. cereus PLC F66Y mutant an attractive candidate for its industrial use.

Microbial Enzymatic Degumming of Crude Soybean Oil (Lecitase Novo Form Aspergillus Orizae)

Microbial Enzymatic Degumming of Crude Soybean Oil (Lecitase Novo Form Aspergillus Orizae)

Recombinant expression, characterization, and application of a phospholipase B from Fusarium oxysporum

In this study, a gene encoding a putative lipase from Fusarium oxysporum was optimized via codon optimization and expressed in Pichia pastoris KM71. The gene product was identified as a phospholipase B (PLB). The engineered P. pastoris was further cultured in a 3.6-L bioreactor. After optimization of the induction conditions, this system produced 6.6mgmL−1 protein and 6503.8UmL−1 PLB activity in the culture medium. Efficient expression of this PLB in P. pastoris should reduce the costs of production and application. The purified enzyme, with a specific activity of 1170Umg−1, was optimally active at pH 5.0 and 55°C. The results of a degumming experiment performed using the recombinant PLB showed that the phosphorus content of a test oil was decreased from 75.88ppm to 3.3ppm in 2h under optimal reaction conditions. This study provides a basis for the industrial use of F. oxysporum PLB in oil degumming applications.