scholarly journals Lipase from Rhizopus oryzae R1: in-depth characterization, immobilization, and evaluation in biodiesel production

Author(s):  
Shimaa E. Helal ◽  
Hemmat M. Abdelhady ◽  
Khadiga A. Abou-Taleb ◽  
Mervat G. Hassan ◽  
Mahmoud M. Amer

Abstract Background Rhizopus species is among the most well-known lipase producers, and its enzyme is suitable for use in many industrial applications. Our research focuses on the production of lipase utilizing waste besides evaluating its applications. Results An extracellular lipase was partially purified from the culture broth of Rhizopus oryzae R1 isolate to apparent homogeneity using ammonium sulfate precipitation followed by desalting via dialysis. The partially purified enzyme was non-specific lipase and the utmost activity was recorded at pH 6, 40 °C with high stability for 30 min. The constants Km and Vmax, calculated from the Lineweaver-Burk plot, are 0.3 mg/mL and 208.3 U/mL, respectively. Monovalent metal ions such as Na+ (1 and 5 mM) and K+ (5 mM) were promoters of the lipase to enhance its activity with 110, 105.5, and 106.5%, respectively. Chitosan was used as a perfect support for immobilization via both adsorption and cross-linking in which the latter method attained immobilization efficiency of 99.1% and reusability of 12 cycles. The partially purified enzyme proved its ability in forming methyl oleate (biodiesel) through the esterification of oleic acid and transesterification of olive oil. Conclusion The partially purified and immobilized lipase from Rhizopus oryzae R1 approved excellent efficiency, reusability, and a remarkable role in detergents and biodiesel production.

2021 ◽  
Vol 22 (21) ◽  
pp. 11956
Author(s):  
Saadiah A. Abdulmalek ◽  
Kai Li ◽  
Jianhua Wang ◽  
Michael Kidane Ghide ◽  
Yunjun Yan

This article describes the successful synthesis of a novel nanocomposite of superparamagnetic multi-walled nanotubes with a four-arm polyethylene glycol amine polymer (mMWCNTs@4-arm-PEG-NH2). This composite was then employed as a support for the covalent co-immobilization of Rhizopus oryzae and Candida rugosa lipases under appropriate conditions. The co-immobilized lipases (CIL-mMWCNTs@4-arm-PEG-NH2) exhibited maximum specific activity of 99.626U/mg protein, which was 34.5-fold superior to that of free ROL, and its thermal stability was greatly improved. Most significantly, CIL-mMWCNTs@4-arm-PEG-NH2 was used to prepare biodiesel from waste cooking oil under ultrasound conditions, and within 120 min, the biodiesel conversion rate reached 97.64%. This was due to the synergy effect between ROL and CRL and the ultrasound-assisted enzymatic process, resulting in an increased biodiesel yield in a short reaction time. Moreover, after ten reuse cycles, the co-immobilized lipases still retained a biodiesel yield of over 78.55%, exhibiting excellent operational stability that is attractive for practical applications. Consequently, the combined use of a novel designed carrier, the co-immobilized lipases with synergy effect, and the ultrasound-assisted enzymatic reaction exhibited potential prospects for future applications in biodiesel production and various industrial applications.


2016 ◽  
Vol 218 ◽  
pp. 1224-1229 ◽  
Author(s):  
Joana Rodrigues ◽  
Véronique Perrier ◽  
Jérôme Lecomte ◽  
Eric Dubreucq ◽  
Suzana Ferreira-Dias

Catalysts ◽  
2018 ◽  
Vol 8 (11) ◽  
pp. 527 ◽  
Author(s):  
Gaojian Ma ◽  
Lingmei Dai ◽  
Dehua Liu ◽  
Wei Du

Acidic oil, which is easily obtained and with lower cost, is a potential raw material for biodiesel production. Apart from containing large quantity of FFAs (free fatty acids), acidic oil usually contains some amount of inorganic acid, glycerides and some other complex components, leading to complicated effect on lipase’s catalytic performance. Exploring the efficient process of converting acidic oil for biodiesel production is of great significance to promote the use of acidic oil. A two-step conversion process for acidic soybean oil was proposed in this paper, where sulfuric acid-mediated hydrolysis was adopted first, then the hydrolyzed free fatty acid, collected from the upper oil layer was further subject to the second-step esterification catalyzed by immobilized lipase Novozym435. Through this novel process, the negative effect caused by harmful impurities and by-product glycerol on lipase was eliminated. A fatty acid methyl ester (FAME) yield of 95% could be obtained with the acid value decreased to 4 mgKOH/g from 188 mgKOH/g. There was no obvious loss in lipase’s activity and a FAME yield of 90% could be maintained with the lipase being repeatedly used for 10 batches. This process was found to have a good applicability to different acidic oils, indicating it has great prospect for converting low quality oil sources for biodiesel preparation.


2020 ◽  
Vol 12 (17) ◽  
pp. 7122
Author(s):  
Ludwika Tomaszewska-Hetman ◽  
Waldemar Rymowicz ◽  
Anita Rywińska

The study proposed the innovative low-cost strategy for erythritol production by Yarrowia lipolytica through developing a simple medium based on industrial waste by-products and a natural method for culture broth purification. Results obtained proved that corn steep liquor might successfully replace traditional sources of nitrogen and other nutrients without compromising activities of the enzymes responsible for erythritol production and its production level. As a consequence, a production process was performed where Y. lipolytica A-6 was able to produce 108.0 g/L of erythritol, with a production rate of 1.04 g/Lh and a yield of 0.45 g/g of the medium containing exclusively 220 g/L of crude glycerol derived from biodiesel production and 40 g/L of corn steep liquor. Moreover, a comparable concentration of erythritol (108.1 g/L) was obtained when a part of crude glycerol was exchanged for the crude fraction of fatty acids in the two-steps process. Next, the collected post-fermentation broths were used in the culture with Y. lipolytica Wratislavia K1 for natural purification. The process resulted in a high increase of erythritol selectivity from 72% to 97% and in the production of 22.0 g/L of biomass with 40.4% protein content, which enables its use as an attractive animal feedstuff.


2016 ◽  
Vol 19 (0) ◽  
Author(s):  
Ariela Veloso de Paula ◽  
Gisele Fátima Morais Nunes ◽  
Heizir Ferreira de Castro ◽  
Júlio César dos Santos

Summary A food grade lipase from Rhizopus oryzae immobilized on a hybrid polysiloxane-polyvinyl alcohol matrix (SiO2-PVA) was used as the biocatalyst to mediate the interesterification reactions of a blend containing 65% milkfat and 35% soybean oil. All the reactions occurred in an inert nitrogen atmosphere in cylindrical glass reactors (80 mL) with 40 g of the milkfat-soybean oil blend. The influence of the following variables was evaluated: biocatalyst loading (250-1500 activity units per gram of blend), biocatalyst moisture content (5-20%), temperature (45-60 °C) and incubation time (2-48 h). The reactions were monitored by determining the free fatty acid content, triacylglycerol (TAGs) composition in carbon species, and the consistency of the interesterified (IE) products. The reaction conditions were set based on the parameters that provided a high interesterification yield and good consistency of the final product within the ideal range (200 to 800 gf cm-2). Hence the best results were obtained using a biocatalyst loading of 500 U g-1 of blend with 10% moisture content at 45 °C for 4 h. Under these conditions the consistency of the interesterified product was 539.7 ± 38 gf cm-2. The results demonstrated the potential of the immobilized lipase to alter the TAGs profile of the milkfat-soybean oil blend, allowing for the production of structured lipids.


Catalysts ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 1052 ◽  
Author(s):  
Juvet Malonda Shabani ◽  
Omotola Babajide ◽  
Oluwaseun Oyekola ◽  
Leslie Petrik

Zeolites are aluminosilicate crystalline materials known for their unique characteristics, and have been prominent for nearly half a century due to their wide and important industrial applications. The production of zeolites, however, remains a challenge due to the high cost of commercial reagents conventionally used as feedstocks. In the current study, hydroxy sodalite (HS) zeolite samples were synthesised from coal fly ash feedstock by a direct hydrothermal synthesis method. The effects of hydrothermal crystallisation synthesis time on phase crystallinity, crystal size, and morphology of the formed HS were investigated. The prepared samples were characterised using XRD, SEM, EDS and FT-IR techniques. The XRD results of the samples prepared with varying synthesis times confirmed the formation of HS from low to high phase purity and crystallinity from 11 to over 98%. The SEM results reflected gradual variation in crystal morphology, of which highly crystalline HS samples were associated with hexagonal-cubic and cubic-platelet crystals. The FTIR, depicting zeolite characteristics of T–O and T–O–T stretching vibrations in the molecular framework, further confirmed the formation of HS zeolites for samples obtained above the 24-h synthesis time. These zeolite samples were then evaluated for their catalytic activities in the conversion of maggot oil to biodiesel. The application of the various hydroxy sodalite samples for the transesterification of maggot oil yielded up to 84.10% biodiesel (FAME) with physicochemical properties that were in compliance with the biodiesel specification standards. This study investigated the novel use of a coal fly ash-derived, heterogeneous HS catalyst in biodiesel production from maggot oil, and indicates its potential to enhance biodiesel yield and quality upon process optimisation tests.


2016 ◽  
Vol 27 (1) ◽  
pp. 13-21 ◽  
Author(s):  
Baskar Thangaraj ◽  
Zhaohua Jia ◽  
Lingmei Dai ◽  
Dehua Liu ◽  
Wei Du

Abstract Lipase-catalyzed biodiesel production is being the object of extensive research due to the demerits of chemical based catalytic system. Lipase immobilized on Fe3O4 magnetic nanoparticles has the integrated advantages of traditional immobilized lipase and free lipase for its rather fast reaction rate and easy separation. It has been demonstrated that free lipase NS81006 has potential in catalyzing the alcoholysis of renewable oils for biodiesel preparation. In this study, Fe3O4 magnetic nanoparticles functionalized with organosilane compounds like (3-aminopropyl)triethyloxysilane (APTES) and (3-mercaptopropyl)trimethoxysilane) MPTMS were used as carriers for lipase immobilization. Lipase NS81006 was covalently bound to the organosilane-functionalized magnetic nanoparticles by using glutaraldehyde cross-linking reagent. A biodiesel yield of 89% and 81% could be achieved by lipase immobilized on APTES-Fe3O4 and MPTMS-Fe3O4 magnetic nanoparticles respectively under optimized conditions of oil to methanol molar ratio 1:3 with three step addition of methanol, reaction temperature 45°C and reaction time duration 12 h. The lipases immobilized on magnetic nanoparticles could be recovered easily by external magnetic field for further use.


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