The effect of ultrasound on enzymatic degumming process of rapeseed oil by the use of phospholipase A1

Comparative studies of enzymatic degumming process of rapeseed oil were carried out in mechanical-stirring and ultrasonic-assisted mechanical-stirring systems. The influences of enzyme dosage (10–50 mg/kg), pH (4.5–6), temperature (45–65 °C), water amount (1–3%), ultrasonic power (0.06–0.09 W/cm³) and reaction time were investigated subsequently. A suitable ultrasonic power of 0.07 W/cm³ was determined to guarantee satisfactory degumming efficiency and enzyme activity. Compared to the mechanical-stirring system, optimum temperature of phospholipase A (PLA) in the ultrasonic-assisted mechanical-stirring system was about 5 °C higher, while the effects of pH on both of the two systems were quite similar. Less time and water were used in the ultrasonic-assisted mechanical-stirring system for enzymatic degumming. The study on the quality changes of degummed oils showed that ultrasound could accelerate the oxidation of edible oils due to the effect of cavitation, thus more attention should be paid on the oxidative stability in the further application.     

Study on ultrasound-assisted oxidative desulfurization for crude oil

The existence of sulfur compounds in crude oil will bring many problems such as corrosion, catalyst poisoning and pollution to the petroleum processing process. Therefore, how to reduce the sulfur content as much as possible in the process of crude oil processing has become an important research topic in the petroleum processing industry. In this paper, ultrasonic-oxidative desulfurization is studied. The effects of reaction temperature, reaction time, amount of oxidant and demulsifier on desulfurization rate are investigated. And the effect of oxidative desulfurization and single oxidative desulfurization under ultrasonic treatment are compared. It is found that the addition of ultrasonic treatment can enhance the desulfurization effect of desulfurizer, the desulfurization efficiency can be increased by about 10% under ultrasonic treatment (100 W, 70 kHz); ultrasonic wave plays an auxiliary role in the system, it can promote heterogeneous reactions, improve the activity of oxidants, and promote the degradation of macromolecular compounds. Finally, physical desulfurization, chemical desulfurization and biological desulfurization technologies are compared.     

Ultrasound-assisted desalination of crude oil: The influence of mixing extent,crude oil species,chemical demulsifier and operation variables

Coalescence of water droplets in crude oil has been effectively promoted by chemical demulsifiers integrated with ultrasound. Temporary images of water droplets in W/O emulsions were directly monitored using a metallurgical microscope. Water droplets achieved expansion of 118% at 40 min ultrasonic irradiation time under well mixing conditions. However, water droplets in heavy crude oil undergo less aggregation than those in light crude oil, due to resistance of mobility in highly viscous fluid. In the absence of chemical demulsifiers, water droplets enveloped by native surfactants appeared to aggregate arduously because of occurrence of interfacial tension gradients. Influential significance analyses have been executed by a factorial design method on operation variables, including acoustic power intensity, operation temperature, ultrasonic irradiation time and chemical demulsifier dosages. In this work, the outcomes indicate that the optimal operating conditions for desalination of crude oil assisted by ultrasound were as follows: acoustic power intensity = 300 W, operation temperature = 90℃, ultrasonic irradiation time = 75 min and chemical demulsifier dosages = 54 mg/L. Besides, it was found that the most influential importance of operation parameter was temperature, followed with acoustic power intensity, ultrasonic irradiation time and chemical demulsifier dosages.     

Ultrasound assisted enzymatic conversion of non edible oil to methyl esters

Conventional and ultrasound-assisted hydrolysis and subsequent esterification of Nagchampa oil under mild operating conditions have been investigated with an objective of intensification of methyl esters production using a sustainable approach. The effect of ratio of reactants, temperature, enzyme loading, pretreatment of enzyme (using ultrasonic irradiations) on the hydrolysis and esterification reaction has been studied. Optimum conditions for hydrolysis were observed to be 1:1 weight ratio of oil: water for Lip Z and 1:3 for Lip 2 enzymes, enzyme loading of 400 units for Lip Z and 800 mg for Lip 2 enzymes and reaction time of 6 h. In the case of esterification reaction, optimum conditions obtained were oil to methanol molar ratio of 1:2, enzyme loading of 1000 mg and reaction time of 20 h. Use of pretreated enzyme (using ultrasonic irradiations) was found to increase the extent of esterification reaction from 75% to 92.5%. It was observed that use of ultrasound in the reaction significantly intensified the esterification reaction with time requirement reducing from 20 h for conventional stirring based approach to only about 7.5 h in the presence of ultrasound. The extent of esterification obtained with sonicated enzyme also increased to 96% from 75% with unsonicated enzyme.     

Ultrasound assisted enzymatic depolymerization of aqueous guar gum solution

The present work investigates the effectiveness of application of low intensity ultrasonic irradiation for the intensification of enzymatic depolymerization of aqueous guar gum solution. The extent of depolymerization of guar gum has been analyzed in terms of intrinsic viscosity reduction. The effect of ultrasonic irradiation on the kinetic and thermodynamic parameters related to the enzyme activity as well as the intrinsic viscosity reduction of guar gum using enzymatic approach has been evaluated. The kinetic rate constant has been found to increase with an increase in the temperature and cellulase loading. It has been observed that application of ultrasound not only enhances the extent of depolymerization but also reduces the time of depolymerization as compared to conventional enzymatic degradation technique. In the presence of cellulase enzyme, the maximum extent of depolymerization of guar gum has been observed at 60 W of ultrasonic rated power and ultrasonic treatment time of 30 min. The effect of ultrasound on the kinetic and thermodynamic parameters as well as the molecular structure of cellulase enzyme was evaluated with the help of the chemical reaction kinetics model and fluorescence spectroscopy. Application of ultrasound resulted in a reduction in the thermodynamic parameters of activation energy (E_a), enthalpy (ΔH), entropy (ΔS) and free energy (ΔG) by 47%, 50%, 65% and 1.97%, respectively. The changes in the chemical structure of guar gum treated using ultrasound assisted enzymatic approach in comparison to the native guar gum were also characterized by FTIR. The results revealed that enzymatic depolymerization of guar gum resulted in a polysaccharide with low degree of polymerization, viscosity and consistency index without any change in the core chemical structure which could make it useful for incorporation in food products.     

Ultrasound assisted intensification of biodiesel production using enzymatic interesterification

Ultrasound assisted intensification of synthesis of biodiesel from waste cooking oil using methyl acetate and immobilized lipase obtained from Thermomyces lanuginosus (Lipozyme TLIM) as a catalyst has been investigated in the present work. The reaction has also been investigated using the conventional approach based on stirring so as to establish the beneficial effects obtained due to the use of ultrasound. Effect of operating conditions such as reactant molar ratio (oil and methyl acetate), temperature and enzyme loading on the yield of biodiesel has been investigated. Optimum conditions for the conventional approach (without ultrasound) were established as reactant molar ratio of 1:12 (oil:methyl acetate), enzyme loading of 6% (w/v), temperature of 40 °C and reaction time of 24 h and under these conditions, 90.1% biodiesel yield was obtained. The optimum conditions for the ultrasound assisted approach were oil to methyl acetate molar ratio of 1:9, enzyme loading of 3% (w/v), and reaction time of 3 h and the biodiesel yield obtained under these conditions was 96.1%. Use of ultrasound resulted in significant reduction in the reaction time with higher yields and lower requirement of the enzyme loading. The obtained results have clearly established that ultrasound assisted interesterification was a fast and efficient approach for biodiesel production giving significant benefits, which can help in reducing the costs of production. Reusability studies for the enzyme were also performed but it was observed that reuse of the catalyst under the optimum experimental condition resulted in reduced enzyme activity and biodiesel yield.     

Techno-economic comparative assessment of the ultrasound,electrostatic and microwave supported coalescence of binary water droplets in crude oil

In this study, comparative assessment of the technical performance, energy usage and economic impact of ultrasound, electrostatics and microwave on the coalescence of binary water droplets in crude oil was conducted. The effect of different oil properties such as crude oil viscosity (10.6–106 mPa s) and interfacial tension (IFT) (20–250 mN/m) on the coalescence time and energy consumption was examined. In addition, operation conditions such as inlet emulsion flow velocity (10–100 mm/s), electric field type, ultrasound frequency and applied voltage amplitude (0–30 kV) were evaluated. The numerical models showed good agreement with experimental findings in the literature. Moreover, the process time of the dewatering process increased with rising inlet flow velocities. The elevation of the coalescence time with velocity can be attributed to the increasing effect of flow disturbance, and the reduction of the emulsion residence time. As regards the IFT, the coalescence time reduced as the IFT was increased. This can be associated with the improved stability of emulsions formed at lowered IFT. As the maximum droplet size is directly proportional to the IFT, lowering the IFT reduces the peak diameter of the droplets that are present in the emulsion. Moreover, the coalescence time followed the order: ultrasound < microwave < electrostatics approaches under varying IFT. The coalescence energy increased from ∼15 J, ∼90 J and ∼25 mJ to ∼61 J, ∼235 J and ∼26 mJ for microwave, electrostatics and ultrasound techniques, respectively, as the viscosity was raised from 10.6 to 106 mPa s. Ultrasound coalescence showed significant energy and economic savings in comparison to microwave and electro-coalescence. Hence, ultrasound coalescence would be a potential method for standalone or integrated demulsification over the two other techniques. However, there are indications that beyond a viscosity of 300 mPa s, the effect of ultrasound becomes weak with significant hindrance to droplet movement and accumulation. This analysis provides fundamental insights on the comparative behavior of the three emulsion separation techniques.     

Ultrasound assisted enzyme catalyzed hydrolysis of waste cooking oil under solvent free condition

The present work demonstrates the hydrolysis of waste cooking oil (WCO) under solvent free condition using commercial available immobilized lipase (Novozyme 435) under the influence of ultrasound irradiation. The process parameters were optimized using a sequence of experimental protocol to evaluate the effects of temperature, molar ratios of substrates, enzyme loading, duty cycle and ultrasound intensity. It has been observed that ultrasound-assisted lipase-catalyzed hydrolysis of WCO would be a promising alternative for conventional methods. A maximum conversion of 75.19% was obtained at mild operating parameters: molar ratio of oil to water (buffer pH 7) 3:1, catalyst loading of 1.25% (w/w), lower ultrasound power 100 W (ultrasound intensity – 7356.68 W m⁻²), duty cycle 50% and temperature (50 °C) in a relatively short reaction time (2 h). The activation energy and thermodynamic study shows that the hydrolysis reaction is more feasible when ultrasound is combined with mechanical agitation as compared with the ultrasound alone and simple conventional stirring technique. Application of ultrasound considerably reduced the reaction time as compared to conventional reaction. The successive use of the catalyst for repetitive cycles under the optimum experimental conditions resulted in a loss of enzymatic activity and also minimized the product conversion.     

Spectrophotometric determination of vanadium in crude oil

Spectrophotometric method has been developed for the determination of vanadium after derivatization with 2-pyrrolealdehyde phenylsemicarbazone (PPS). The linear calibration curve was obtained with 2.5- vanadium. Copper(II), cobalt(II), iron(II) and palladium(II) could also be determined separately using PPS with linear calibration curves within 2.5-12.5, 5-15, 2.5-15 and 1- at 362, 355, 355 and 365 nm, respectively. The vanadium in crude oil was determined with relative deviation (RSD) of 2.5-5.0%. The methods have been applied for the analysis of copper from copper wires, cobalt from pharmaceutical preparation and palladium from palladium on barium sulphate with RSD within 2.6-4.5%.     

Ultrasound microwave-assisted enzymatic production and characterisation of antioxidant peptides from sweet potato protein

Herein, we investigated the effects of ultrasound microwave (UM)-assisted hydrolysis using Alcalase (ALC), Flavourzyme (FLA), and their combination (ALC + FLA), on the production of sweet potato protein hydrolysates (SPPH). UM-assisted enzymatic hydrolysis significantly increased the degree of hydrolysis of SPPH compared with untreated (UN) samples. Fractions with differences in molecular weight (MW) of >10, 3–10, and < 3 kDa in SPPH from UM-assisted ALC, FLA, and ALC + FLA hydrolysis displayed higher antioxidant activities than those from UN samples. MW < 3 kDa fractions of SPPH from UM-assisted ALC and ALC + FLA hydrolysis treatments presented much stronger Fe²⁺-chelating activity (98.48% and 98.59%), ·OH scavenging activity (67.11% and 60.06%), and higher ORAC values (110.32 and 106.32 µg TE/mL), from which diverse peptides with potential antioxidant activities were obtained by semi-preparative HPLC and LC-MS/MS. All identified peptide sequences exhibited at least three potential antioxidant amino acids. Additionally, changes in peptide conformational structure and antioxidant amino acid composition were revealed by structure–activity relationship analysis. Thus, ultrasound microwave treatment has great potential in antioxidant peptides production.     

Ultrasound irradiation promoted enzymatic alcoholysis for synthesis of monoglyceryl phenolic acids in a solvent-free system

Monoglyceryl phenolic acids (MPAs) were known as the natural hydrophilic antioxidants which could be used in different fields such as food, pharmaceutical, cosmetic etc. A novel enzymatic route of MPAs synthesis by the alcoholysis of phenolic acid ethyl esters with glycerol under ultrasound irradiation in solvent free system was developed. Optimization of reaction parameters shows that a high conversion of above 97.4% can be obtained under the following conditions: phenolic acid ethyl esters to glycerol molar ratio of 1:10, with 6% catalyst (Novozym 435), at 60 °C and 200 rpm, with ultrasound input of 250 W, at 20 kHz frequency. Compared to the conventional stirring method, the activation energy for phenolic acid ethyl esters conversion was decreased from 65.0 kJ/mol to 32.1 kJ/mol under ultrasound promotion; the apparent kinetic constant (V_m/K_m) increased above 1.2-folds; the lipase amount decreased to 50%; the time required for the maximum conversion reduced up to 3-folds without damaging the lipase activity, which is the fastest report for enzymatic synthesis of MPAs.     

Process optimization and characterization of arachidonic acid oil degumming using ultrasound-assisted enzymatic method

Ultrasound assisted enzymatic method was applied to the degumming of arachidonic acid (ARA) oil produced by Mortierella alpina. The conditions of degumming process were optimized by response surface methodology with Box- Behnken design. A dephosphorization rate of 98.82% was achieved under optimum conditions of a 500 U/kg of Phospholipase A₁ (PLA₁) dosage, 2.8 mL/100 g of water volume, 120 min of ultrasonic time, and 135 W of ultrasonic power. The phosphorus content of ultrasonic assisted enzymatic degumming oil (UAEDO) was 4.79 mg/kg, which was significantly lower than that of enzymatic degumming oil (EDO, 17.98 mg/kg). Crude Oil (CO), EDO and UAEDO revealed the similar fatty acid compositions, and ARA was dominated (50.97 ~ 52.40%). The oxidation stability of UAEDO was equivalent to EDO and weaker than CO, while UAEDO presented the strongest thermal stability, followed by EDO and CO. Furthermore, aldehydes, acids and alcohols were identified the main volatile flavor components for the three oils. The proportions of major contributing components such as hexanal, nonanal, (E)-2-nonanal, (E, E)-2,4-decadienal, (E)-2-nonenal and aldehydes in UAEDO and EDO were all lower than CO. Overall, Ultrasound assisted enzymatic degumming proved to be an efficient and superior method for degumming of ARA oil.     

Applying ultrasonic fields to separate water contained in medium-gravity crude oil emulsions and determining crude oil adhesion coefficients

Separating produced water is a key part of production processing for most crude oils. It is required for quality reasons, and to avoid unnecessary transportation costs and prevent pipework corrosion rates caused by soluble salts present in the water. A complicating factor is that water is often present in crude oil in the form of emulsions. Experiments were performed to evaluate the performance of ultrasonic fields in demulsifying crude oil emulsions using novel pipe-form equipment. A horn-type piezoelectric ultrasonic transducer with a frequency of 20 kHz and power ranging from 80 W to 1000 W was used for experimental purposes. The influences of the intensity of ultrasonic fields, ultrasonic irradiation time, and the initial water content of crude oils were evaluated to establish the rate of water segregation from oil. The experiments applied ultrasonic-field intensities of $0.25$ W/cm³, $0.5$ W/cm³, $0.75$ W/cm³ and $1$ W/cm³ to synthetic emulsions with 10%, 15%, 20%, and 25% of the water in crude oil. Crude oil demulsification occurred for each ultrasonic field intensity tested for all the samples tested. Function β involving adhesion coefficients was expressed in terms of wave-field intensity and initial concentration of water in each of the three crude oil samples tested. The experiments demonstrated that despite the absence of any chemical demulsifier involved, water separation caused by applying ultrasonic fields was effective and occurred rapidly. As the intensity of the ultrasonic field applied increased, the amount of water segregated from crude oil also increased. Subjected to constant field intensity, higher initial water cuts (up to 15% or so) in the crude oil samples and higher ultrasonic irradiation times, resulted in greater segregation of water from crude oil in percentage terms. However, in samples with initial water cuts of 20+% long irradiation times (~5 min), resulted in a decline in water separation compared to 2-min tests. Ultrasonic field treatments offer commercially-viable and environmentally-friendly alternatives to treatments using chemical demulsifiers as they reduce desalination requirements of wastewater.     

Ultrasound-assisted lipase-catalyzed transesterification of soybean oil in organic solvent system

This work reports the transesterification of soybean oil with ethanol using two commercial immobilized lipases under the influence of ultrasound irradiation. The experiments were performed in an ultrasonic water bath, following a sequence of experimental designs to assess the effects of temperature, enzyme and water concentrations, oil to ethanol molar ratio and output irradiation power on the reaction yield. Results show that ultrasound-assisted lipase-catalyzed transesterification of soybean oil with ethanol might be a potential alternative route to conventional alkali-catalyzed method, as high reaction yields (∼90 wt.%) were obtained at mild irradiation power supply (∼100 W), and temperature (60 °C) in a relatively short reaction time, 4 h, using Lipozyme RM IM as catalyst. The repeated use of the catalyst under the optimum experimental condition resulted in a decay in both enzyme activity and product conversion after two cycles. The use of Novozym 435 led to lower conversions (about 57%) but the enzyme activity was stable after eight cycles of use, showing, however, a reduction in product conversion after the forth cycle.     

Detrended fluctuation analysis on spot and futures markets of West Texas Intermediate crude oil

In this paper, we study the auto-correlations and cross-correlations of West Texas Intermediate (WTI) crude oil spot and futures return series employing detrended fluctuation analysis (DFA) and detrended cross-correlation analysis (DCCA). Scaling analysis shows that, for time scales smaller than a month, the auto-correlations and cross-correlations are persistent. For time scales larger than a month but smaller than a year, the correlations are anti-persistent, while, for time scales larger than a year, the series are neither auto-correlated nor cross-correlated, indicating the efficient operation of the crude oil markets. Moreover, for small time scales, the degree of short-term cross-correlations is higher than that of auto-correlations. Using the multifractal extension of DFA and DCCA, we find that, for small time scales, the correlations are strongly multifractal, while, for large time scales, the correlations are nearly monofractal. Analyzing the multifractality of shuffled and surrogated series, we find that both long-range correlations and fat-tail distributions make important contributions to the multifractality. Our results have important implications for market efficiency and asset pricing models.     

Intensification of ultrasonic-assisted crude oil demulsification based on acoustic field distribution data

Water removal is an essential step during crude oil production due to several problems such as increased transportation costs and high corrosion rate due to dissolved salts. Indirect low frequency ultrasonic energy (US), using baths, has been recently proposed as an effective alternative for crude oil demulsification. However, the reactor position during sonication and its influence on the demulsification efficiency for crude oil has not been evaluated. In this sense, the aim of this study was to develop an automated system based on an open source hardware for mapping the acoustic field distribution in an US bath operating at 35 kHz using a hydrophone. Data acquired with this system provided information to evaluate the demulsification efficiency in the different positions of the US bath and correlate it with the acoustic intensity distribution. The automated 3D-mapping system revealed a higher acoustic intensity in the regions immediately above the transducers (ca. 0.6 W cm⁻²), while the other regions presented a relatively lower intensity (ca. 0.1 W cm⁻²). Experimental data demonstrated that reactors positioned in the most intense acoustic regions provided a much higher efficiency of demulsification in comparison with the ones positioned in the less intense acoustic field regions. Demulsification efficiency up to 93% was obtained with 15 min of sonication (100% amplitude) using few amount of chemical demulsifier. Hence, this work demonstrated that the information acquired with the developed mapping system could be used for inducing a higher efficiency of demulsification only by finding the more suitable position of reactor in the US bath, which certainly will help development of appropriate reactors design when looking for such approach.     

Multifractal analysis on international crude oil markets based on the multifractal detrended fluctuation analysis

The multifractal nature of WTI and Brent crude oil markets is studied employing the multifractal detrended fluctuation analysis. We find that two crude oil markets become more and more efficient for long-term and two Gulf Wars cannot change time scale behavior of crude oil return series. Considering long-term influence caused by Gulf Wars, we find such “turning windows” in generalized Hurst exponents obtained from three periods divided by two Gulf Wars so that WTI and Brent crude oil returns possess different properties above and below the windows respectively. Comparing with the results obtained from three periods we conclude that, before the First Gulf War, international crude oil markets possessed the highest multifractality degree, small-scope fluctuations presented the strongest persistence and large-scope fluctuations presented the strongest anti-persistence. We find that, for two Gulf Wars, the first one made a greater impact on international oil markets; for two markets, Brent was more influenced by Gulf Wars. In addition, we also verified that the multifractal structures of two markets’ indices are not only mainly attributed to the broad fat-tail distributions and persistence, but also affected by some other factors.     

Ultrasound enhanced butyric acid-lauric acid designer lipid synthesis: Based on artificial neural network and changes in enzymatic structure

Ultrasound is a green technology for intensifying enzymatic reactions. In this study, an ultrasonic water bath with equipment parameters of 28 kHz, 1750.1 W/m², 60% duty cycle was used to assist the synthesis of butyric acid-lauric acid designer lipid (BLDL), which was catalyzed by Lipozyme 435. A convincing three-layer feed-forward artificial neural network (ANN) model was established (R² = 0.949, RMSE = 4.759, ADD = 7.329) to accurately predict the optimal parameters combination, which was described as 13.72 mL reaction volume, 15.49% enzyme loading, 0.253 substrate molar ratio (tributyrin/lauric acid), 56.58 °C reaction temperature and 120 min reaction time. The ultrasonic assistance increased actual butyric acid conversion rate by 11.38%, and also enhanced the consumption rate of tributyrin and lauric acid during the reaction. Meanwhile, the esterification activity of Lipozyme 435 was enhanced and its effectiveness up to 6 cycles. Structurally, ultrasound assistance significantly disrupted the secondary structure of the Lipozyme 435: reduced the content of α-helices, increased the content of β-sheet and β-turn. In addition, sonication caused an increase in crevice and micro-damage on the surface of the immobilized enzyme. In conclusion, low-intensity ultrasound at 28 kHz improved the synthesis efficiency of BLDL, which was scientifically predicted by ANN model, and the change of enzyme structure may be the vital reason for ultrasound enhanced reaction. However, the effect of ultrasound on immobilized enzymes’ activity needs to be further explored.     

Ultrasound assisted modulation of yeast growth and inactivation kinetics

The yeast Saccharomyces cerevisiae is well known for its application in the food industry for the purpose of developing fermented food. The ultrasound (US) technology offer a wide range of applications for the food industry, including the enhancement of fermentation rates and inactivation of microbial cells. However, a better understanding and standardization of this technology is still required to ensure the scaling-up process. This study investigated the effect of the US technology on the growth of S. cerevisiae using frequencies of 20, 25, 45 and 130 kHz, treatment periods from 2 to 30 min. Furthermore, yeast kinetics subjected to US treatments were evaluated using modelling tools and scanning electron microscopy (SEM) analysis to explore the impact of sonication on yeast cells. Yeast growth was monitored after different US treatments plotting optical density (OD) at 660 nm for 24 h at 30 ⁰C. Growth curves were fitted using models of modified Gompertz and Scale-Free which showed good parameters of the fit. In particular, US frequencies of 45 and 130 kHz did not have a disruptive effect in lag phase and growth rate of the yeast populations, unlike the frequency of 20 kHz. Moreover, inactivation curves of yeast cells obtained after exposure to 20 and 25 kHz also observed the best fit using the Weibull model. US frequency of 20 kHz achieved significant reductions of 1.3 log cfu/mL in yeast concentration and also induced important cell damage on the external structures of S. cerevisiae. In conclusion, the present study demonstrated the significant effect of applying different US frequencies on the yeast growth for potential application in the food industry.     

Ultrasound assisted enzymatic pre-treatment of high fat content dairy wastewater

This paper illustrates the application of ultrasound in a dairy waste water treatment for the removal of fat using enzyme as a catalyst. Lipase Z was used to perform the enzymatic pre-hydrolysis of a synthetic dairy wastewater containing around 2000 mg/L of fat content coupled with ultrasound irradiation. Different process parameters like effect of enzyme loading, temperature, ultrasound power, frequency, duty cycle and speed of agitation are optimized. The maximum hydrolysis of 78% is achieved at 0.2% enzyme loading (w/v), 30 °C temperature, 165 W of ultrasonication power at 25 kHz and 66% duty cycle. It was observed that the enzymatic pre-hydrolysis under the influence of ultrasound drastically reduces the reaction time from 24 h to 40 min as compared to conventional stirring with improved yield.