Enzymatic Degumming Research Articles

Research on Alkaline Pectinase for Hemp Degumming

Alkaline pectinase has a broad application prospect in textile field. In hemp degumming, enzymatic degumming is preferred because of the better fiber quality and less environmental pollution. In this thesis, we used the recombinant strain Bacillus subtilis TCCC11485 constructed in our laboratory to express high-yield alkaline pectinase. The optimal conditions of the alkaline pectinase for hemp degumming were determined by single factor experiments as: bath ratio 1:36, pH 8, 45°C, enzyme activity 600 U/mL, degumming for 6 h. Under the above conditions, the residual gum content of fiber is 32.76%. Further, the scanning electron microscopy (SEM) was also used to compare quality of the hemp fibers produced by enzymatic-, chemical-degumming. The results showes that the fibers are in higher quality by alkaline pectinase degumming, suggesting the prospect of commercial applications in domestic textile industry.

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 (10–50mg/kg), pH (4.5–6), temperature (45–65°C), water amount (1–3%), ultrasonic power (0.06–0.09W/cm3) and reaction time were investigated subsequently. A suitable ultrasonic power of 0.07W/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.

Optimization of magnetic immobilized phospholipase A1 degumming process for soybean oil using response surface methodology

This study focused on optimization of processing conditions of enzymatic degumming process for soybean oil using phospholipase A1 immobilized onto magnetic nanoparticles. A response surface methodology was developed and used to obtain optimum processing conditions. Four variables (temperature, reaction time, enzyme dosage, and added water) were investigated based on two response functions (phosphorus and free fatty acids (FFA) contents in degummed soy oil). For each response, second-order polynomial models were developed using multiple linear regression analysis. The optimum operating parameters of enzymatic degumming process were as follows: temperature of 56 °C, reaction time of 6.3 h, enzyme dosage of 0.10 g/kg, and added water of 2.13 ml/100 g. According to these optimum conditions, final residual phosphorus and FFA contents of degummed soy oil were reduced, respectively, to 10.38 mg/kg and 1.09/100 g using magnetic immobilized phospholipase A1. This finding is applicable for the physical refining of soybean oil or refining crude oils from field- and frost-damaged beans which have high content of non-hydratable phosphatides.

Comparison of acid degumming and enzymatic degumming process for Silybum marianum seed oil

In this study the effects of processing conditions of acid degumming and enzymatic degumming on the removal of phospholipids from Silybum marianum seed oil were investigated and the degumming efficiency was compared based on the phospholipid content. An orthogonal array experimental design was performed to optimise the process of citric acid degumming. Based on range analysis and analysis of variance, the optimal processing conditions were determined to be a citric acid dosage of 3 g kg(-1) , a degumming temperature of 70 °C, a water addition of 40 mL kg(-1) and a degumming time of 30 min. Under these conditions the phospholipid content of degummed S. marianum seed oil was reduced from 273.0 to 128.1 mg kg(-1) . In the case of enzymatic degumming, the effects of enzyme reaction time and enzyme dosage were investigated using single-factor experiments. The optimal processing conditions were found to be an enzyme reaction time of 6 h and an enzyme dosage of 100 mg kg(-1) oil. Under these conditions the phospholipid content of degummed S. marianum seed oil was reduced to 17.95 mg kg(-1) . The results indicated that enzymatic degumming is more effective than acid degumming.

Emulsifier and antioxidant properties of by‐products obtained by enzymatic degumming of soybean oil

AbstractThe enzymes used in degumming, called phospholipases, specifically act on phospholipids without degrading the oil itself. Degumming using a phospholipase C enzyme allows to meet all market specifications while it increases the oil yield. The aim of this study was to evaluate antioxidant and emulsifier properties of the recovered gum (RG) obtained by enzymatic oil degumming process of crude soybean oil subjected to modifications as deoiling (RG deoiled) or ethanol fractionation (RG soluble and insoluble). RG soluble allowed obtaining more stable oil‐in‐water emulsions (30:70 w/w) in comparison with those by‐products assayed at different concentrations (0.1–1.0%). Also, deoiled soybean lecithin (DSL) and RG deoiled had a similar behavior in relation to the kinetic destabilization (% backscattering profiles), despite the different degumming processes used to obtain these samples. The study on induction times (Metrohm Rancimat) showed a significant antioxidant effect (p<0.05) against a refined sunflower oil associated with all the by‐products analyzed. However, RG soluble and DSL showed a strong effect on the oil stability at high concentrations (1000–2000 ppm). These results showed that the deoiled recovered gum and its derivates obtained by ethanol fractionation are a potential alternative for industrial application as additive.Practical applications: The economic benefits of enzymatic degumming process have also been quantified by several oilseed processors. This process allows obtaining a by‐product with a high concentration of different phospholipids. This study intends to increase the commercial value of this recovered gum contributing to the food industry with useful information about their functional properties.

An Effective Degumming Enzyme fromBacillussp. Y1 and Synergistic Action of Hydrogen Peroxide and Protease on Enzymatic Degumming of Ramie Fibers

Enzymatic degumming, as an alternative to chemical processing, has attracted wide attention. However, to date, little information about other enzyme components with effective degumming except pectinase has been reported, and there is no report about the effect of bleaching agent (H2O2) on enzymatic degumming and combining enzymatic degumming and H2O2 bleaching process. In this study, we found that the crude enzyme of wild-type Bacillus sp. Y1 had a powerful and fast degumming ability. Its PGL activity was the highest at pH 9.6–10.0 and 60°C and stable at pH 7–10.5 and 30–50°C, having a wide scope of pH and temperature. Its PGL also had a high H2O2 tolerance, and the gum loss and brightness of fibers could be significantly improved when H2O2 was added into it for degumming. The synergistic action was also found between it and H2O2 on the degumming and bleaching of ramie fibers. All showed that it was very suitable for a joint process of enzymatic degumming and H2O2 bleaching. It also contained more proteins compared with a control pectinase, and its high protease content was further substantiated as a factor for effective degumming. Protease and pectinase also had a synergistic action on degumming.

Enzymatic Degumming of Silk with Microbial Proteases

Degumming of Chinese bivoltine silk with alkaline proteases from various microbial sources was investigated and compared with commercial enzymes. Among the proteases tested, two fungal and two actinomycete proteases were promising, which showed weight loss similar to conventional method (19.58% to 21.78%). Conidiobolus brefeldianus and BOA-2 proteases were best enzymes, which showed weight loss similar to conventional method with low enzyme concentrations and in shorter time. No significant differences were found in tensile strength or elongation at break by enzymatic degumming indicating that there was no damage to the fiber. Scanning electron micrographs showed the sericin deposits were removed and the fibers were separated.

DEGUMMING OF RAW SILK FABRIC WITH HELP OF MARINE EXTRACELLULAR PROTEASE

Protease secreting microbe was isolated and charact erized on the basis of their morphological, biochem ical, physiological and 16S rDNA based molecular properties. The extracellular protease was quantified and characterized. Protease was used for different time (4, 8, 12 and 24 h) at different temperature (RT a nd 37°C) for optimization of the degumming process for raw silk fabric with enzyme dosage (0.2-1 unit/cm 2 of fabric). Post-enzymatic treatment, the fabric quali ty and texture was compared with conventionally tre ated as well as untreated fabric in terms of degumming l oss, tensile strength and yarn count and colour fas tness to light/water. The isolate SM1 ( Bacillus thuringensis ) was able to grow in Carbon Minimal Salt Medium (CMSM) with jaggery or tamarind as the carbon source (0.3% w/v). Energy Dispersive X-Ray Fluorescense (EDXRF) data showed intracellular accumulation of heavy metal by the isolate. Extracellular protease w as able to degum silk fabric within 4 h at RT with enz yme concentration of 0.8unit/cm 2 and the maximum degumming loss was 21.72%. Post enzymatic degumming, a shiny texture was observed under Environmental Scanning Electron Microscope (ESEM) and the yarn volume also increased. Utilization of CMSM made the process cost effective during large scale application. Intracellular metal accumulation and growth in a wide range of temperature and pH made the isolate a potential candidate for bioremediation . Extracellular protease with significant degumming p roperty could be used as an eco friendly approach a s compared to the conventional chemical treatment.

Current and future technologies for the sustainable and cost‐efficient production of high quality food oils

AbstractThe production of high quality food oils in a cost‐efficient and also sustainable way requires a new refining approach and “next generation” oil refining processes are developed to meet the new refining standards. Nano‐neutralization was recently successfully introduced in chemical refining as first industrial application of the nano‐reactor® technology. Proven advantages include significant lower chemicals consumption and a higher refined oil yield. A better understanding of the working principle will definitively result in more applications in edible oil processing. Enzymatic degumming was (re‐)discovered as efficient degumming technique making not only degummed oils suitable for physical refining giving but also giving a higher yield. Bleaching is gradually converted from a single stage “bleaching” into a multi‐stage adsorptive purification process in which a wide range of unwanted components (soaps, phospholipids, oxidation products, trace metals, contaminants, etc.) are removed prior to deodorization. This is achieved by a more systematic use of more efficient adsorbents (bleaching earths, silica hydrogels, and activated carbon). Recent trends and developments in deodorization include more efficient heat recovery, improved stripping tray design and integration of packed column strippers, reducing heat load by application of dual temperature deodorizers and low pressure (ice condensing) vacuum systems and reducing oil losses and increasing the value of the deodorizer distillate by improved (dual) vapor scrubbers. Other technologies like short path distillation and supercritical CO2 technology have also been investigated but are not (yet) broadly applied because of too high operating cost. Although there is the growing importance of oil quality and sustainable processing, potential cost saving and/or oil yield increase remain the prime parameters for the implementation of a new process.

Biodiesel production from crude canola oil by two-step enzymatic processes

Crude canola oil (CCO) contains about 100–300 ppm of phospholipids, which have shown negative effects on biodiesel/buffer solution phase separation, resulting in low biodiesel production yield. Therefore, phospholipids should be removed before transesterification by a degumming process for efficient production of biodiesel. In this study, two-step enzymatic processes (degumming and transesterification) were carried out for the production of biodiesel from CCO. Degumming of CCO was performed using phospholipase A2 as a degumming reagent. The initial phospholipid content was reduced to less than 5 ppm by enzymatic degumming. The effects of three formulations of enzyme catalyst on the efficiency of transesterification were investigated. As a result, conversion rates of degummed CCO to fatty acid methyl esters (FAME) were 68.56%, 70.15%, and 84.25%, respectively. Lipase formulation composed of a 1:1 (vol:vol) enzyme mixture of Rhizopus oryzae and Candida rugosa showed the best performance among those tested. In order to recover and reuse the lipase catalyst efficiently, a 1:1 enzyme mixture of R. oryzae and C. rugosa was immobilized on silica gel. The immobilized lipase was used in subsequent transesterification optimization experiments. Optimization of transesterification was performed by response surface methodology (RSM). A total of 20 experiments based on RSM were carried out, and the optimal reaction conditions appeared to be 24.4% (w/w) immobilized catalyst, 13.5% (w/w) buffer solution, and 15.8% (w/w) methanol based on oil mass. Conversion rate of degummed CCO to FAME was determined to be 88.9% under optimal conditions.

Crude Canola Oil의 효소 탈검과 수용성 탈검에 관한 연구

In this study, degumming process was carried out for reducing to less than 10 ppm of phosphorus contents and primary properties of crude canola oil including 0.64 mgKOH/g of acid value, 0.09% of water contents, 0.13% of insoluble impurities, and 40 ppm of phosphorus contents. Efficiency of water degumming and enzymatic degumming was compared for the selection of suitable process obtaining feedstock of biodiesel. Degumming method was deter- mined for preparation of raw material of biodiesel, and reaction conditions were also established. The most effective conditions for water degumming were 2% distilled water (w/w oil), 30 o C of reaction temperature, 900 rpm of agitation speed, and 30 min of reaction time, respectively. In case of enzymatic degumming, optimal conditions were found to be 90 ppm of phospholipase A2 (w/w oil), 50 o C of reaction temperature at pH 5, respectively. When comparing water degumming with enzymatic degumming, efficiency of enzymatic degumming was better than water degumming. How- ever, water degumming method was much more suitable for the production of biodiesel feedstock considering reaction time and process feasibility.

Optimization of degumming process for soybean oil by phospholipase B

AbstractBACKGROUND: Enzymatic degumming, the ‘EnzyMax® process’, in which a phospholipase (type A1, A2 or B) was used to convert nonhydratable phospholipids into their hydratable forms. Compared with conventional methods, enzymatic degumming offers a safe biological route and eco‐friendly solution to industrial processes. To date, only phospholipases A1 and A2 have been used for enzymatic oil‐degumming. In this study, phospholipase B from Pseudomonas fluorescens BIT‐18 was applied for the first time in soybean oil degumming.RESULTS: Three major factors (temperature, pH and PLB dosage) were screened out through Plackett–Burman design. Then, response surface modeling combined with central composite design and regression analysis were employed for optimization of the final degumming process. The optimum conditions for the minimum residual phosphorus content in the oil were achieved at 40 °C, pH 4.7 and with PLB dosage of 500 U kg−1. Under optimal conditions, the residual phosphorus content decreased to 4.9 mg kg−1, which was comparable with predicted response values.CONCLUSION: These results suggested that Plackett‐Burman design combined with response surface modeling were proved effective in determining the optimum soybean oil degumming conditions. The results also revealed that phospholipase B from Pseudomonas fluorescens BIT‐18 was a good candidate for degumming various vegetable oils. Copyright © 2011 Society of Chemical Industry

Enzymatic degumming

AbstractThe oldest enzymatic degumming process (the Lurgi EnzyMax® process) was launched in 1992. It used porcine phospholipase A2, which has the disadvantages of limited availability and not being kosher/halal. To overcome these disadvantages, various microbial enzymes have been developed; they have different specificities and therefore offer different advantages. Phospholipase C for instance has the advantage that it leads to the formation of diacylglycerols that remain in the oil being degummed. This constitutes a significant yield improvement which also results from the formation of lysophospholipids that retain less oil than their precursors. In the laboratory, a fine dispersion of the aqueous enzyme solution in the oil can be maintained so that the phospholipase enzymes can be made to interact with non‐hydratable phosphatides (NHP) in the oil phase and catalyse their hydrolysis. On an industrial scale, dispersions coalesce before the enzymatic NHP hydrolysis is complete. Accordingly, enzymatic degumming processes that claim NHP‐removal and a low residual phosphorus content in the enzymatically degummed oil are invariably preceded by an acid treatment in which a degumming acid (citric acid) is finely dispersed into the oil to be degummed and made to react with the NHP present in the oil before the enzyme is added. This enzyme then only interacts with the phospholipids present in the water phase. This raises the question whether the yield increase resulting from the use of enzymes should be realised by treating the oil to be degummed or the gums that have already been isolated from the oil during a degumming treatment. Lack of experimental evidence prevents a firm answer to this question but the arguments in favour of treating the gums look more impressive than what can be said in favour of treating the oil. In short: Enzymes do not degum the oil but can be used to de‐oil the gums.

Optimize the Condition for Enzymatic Degumming of Crude Soybean Oil

Tropical Agricultural Research and Extension is a peer-reviewed international scientific journal covering a wide range of subject areas in tropical and subtropical agriculture published quarterly by the Faculty of Agriculture, University of Ruhuna, Sri Lanka. The journal is also available on the Faculty of Agriculture, University of Ruhuna website https://www.agri.ruh.ac.lk/tare/index.htm .

Degumming rice bran oil using phospholipase‐A 1

The efficacy of enzymatic degumming was assessed using the third generation phospholipase-A1, Lecitase®-Ultra (EC 3.1.1.3) from Thermomyces lanuginosa/Fusarium oxysporum with different qualities of crude rice bran oil. The phosphorus content in the oil reduced to ∼10 mg/kg from an initial level of 390 mg/kg after 2 h of incubation period at 50°C. However, in the solvent-phase degumming, there was practically no phospholipid reduction at lower water content (2%) due to the poor contact between the highly nonpolar solvent and the aqueous phase (citric acid, NaOH, and enzyme solutions). Increasing the water content to 20% reduced the phosphorus level in the degummed-oil to 71 mg/kg but did not match the performance of oil-phase degumming. The degumming efficiency of Lecitase®-Ultra was effective in oil-phase and suitable for practical application. Solvent-phase enzymatic degumming offers more benefits but needs greater efforts to overcome the challenges.

Enzymatic degumming

AbstractThe first enzymatic degumming process to be used industrially was the EnzyMax® process that was launched in 1992; it used porcine phospholipase A2 (PLA2). Subsequently, various microbial phospholipases (PLases) with different specificities have been developed. They have the advantages of being kosher/halal and of having a non‐limited availability and lower cost.The first of these microbial enzymes were the phospholipases A1 (Lecitase® Novo and Ultra) and more recently, a phospholipase C (Purifine®) and a lipid acyl transferase (LysoMax®) with PLA2 acitivity have also become available in commercial quantities. These enzymes have different specificities. The Lecitases® and the LysoMax® enzymes catalyse the hydrolysis of all common phosphatides and differ in this respect from the Purifine® enzyme, which is specific for phosphatidyl choline and phosphatidyl ethanolamine. These phosphatides are hydrolysed to oil‐soluble diacylglycerol and water‐soluble phosphate esters. Since these diacylglycerols remain in the oil during refining, they contribute to the oil yield. That also holds for the sterol and stanol fatty esters formed as a consequence of the phosphatide hydrolysis catalysed by the LysoMax® enzyme. In addition, all enzymes cause less oil to be retained by the gums by decreasing the amount of gums and/or their oil retention, which also contributes to an improved oil yield.On the other hand and contrary to common belief, the enzymes are incapable of catalysing the hydrolysis of non‐hydratable phosphatides (alkaline earth salts of phosphatidic acid) under industrial conditions. Consequently, the industrial enzymatic degumming step has to be preceded by a chemical degumming step to arrive at a degummed oil with a sufficiently low residual phosphorus content that can be physically refined. Accordingly, it might well be preferable to limit the oil treatment to said chemical degumming and produce oil with a low residual phosphorus content and gums, and then treat the gums separately to recover their fatty matter, whereby this recovery can be enzymatic or chemical.

Purification of soybean oil with phospholipase Al

An enzymatic degumming step in the production of edible soybean oil is carried out. The phosphatides are present in free hydratable form (HP) or in nonhydratable form (NHP). The main characteristic of the discovery is the use of phospholipid A1 (Lecithin Ultra) enzyme, which catalyzes reactions at specific temperatures. The mechanism includes the conversion of nonhydratable phospholipids into water-soluble lysophospholipids, which are then removed by centrifuge, yielding degumming oil low in phosphorus. The effects of important determining factors affecting oil degumming such as enzyme dosage, temperature and pH are investigated.

Enzymes in lipid production and modification – Current knowledge and future perspectives

AbstractMicrobial enzymes are today used in a range of lipid processing from refining through to fat modification. The range of options is increasing following new research into enzyme types and functionalities. Within the near future, improved methods of enzymatic degumming will be developed as well as alternative enzymes for interesterification and synthesis of nutritionally interesting fats. An enzymatic route to Biod‐diesel is also a distinct possibility focussing on the waste oils and fats which are today difficult to process through chemical esterification.

Insight into the Enzymatic Degumming Process of Soybean Oil

AbstractAn enzymatic degumming trial of soybean oil was carried out at a capacity of 400 tons/day by applying microbial phospholipase A1 from Thermomyces lanuginosus/Fusarium oxysporum. When the pH was kept in the range of 4.8–5.1, less than 10 mg/kg of phosphorous content of The oil was obtained. The gum and oil were easily separated after centrifugation and the oil loss was minimal under the process conditions. Through analysis of phospholipids compounds in the gum by Electrospray Ionization‐Mass Spectrometer and phosphorous content, it could be seen that both glycerophospholipids and lysophospholipids existed with contents of 45.7 and 54.3%, respectively. The performance of enzymatic degumming was found to be related to the production of glycerophospholipids.

Optimization of enzymatic degumming process for rice bran oil using response surface methodology

Response surface methodology was used to determine the optimum processing conditions for enzymatic degumming of rice bran oil. Reaction time, enzyme dosage, level of water added and temperature were the factors investigated with respect to phosphorus and free fatty acids contents. A D-optimal design, with four variables and two response functions, was employed to study the effect of the individual variables on the response functions. For each response, second-order polynomial models were developed using multiple linear regression analysis. Applying desirability function method, optimum operating conditions were found to be reaction time of 4.07 h, enzyme dosage of 50 mg/kg, added water of 1.5 ml/100 g and temperature of 49.2 °C. At this optimum point, phosphorous and free fatty acids contents of degummed oil were found to be 8.86 mg/kg and 2.01 g/100 g as oleic acid, respectively.