Free Candida Rugosa Lipase–Catalyzed Synthesis Of Citronellyl Butyrate In n–Hexane By Direct Esterification: Effect Of Reaction Parameters

2012 ◽  
Author(s):  
Irvan Dahlan ◽  
Azlina Harun @ Kamaruddin ◽  
Ghasem D. Najafpour
Keyword(s):  
Butyric Acid ◽  
Active Sites ◽  
Immobilized Lipase ◽  
Candida Rugosa ◽  
Candida Rugosa Lipase ◽  
Molar Ratio ◽  
Esterification Reaction ◽  
Reaction Parameters ◽  
Direct Esterification ◽  
Competitive Nature

Sintesis sitronelil butirat melalui pengesteran langsung telah dikaji di dalam n–hexane sebagai pelarut organik yang dimangkinkan oleh lipase bebas dan lipase tersekatgerak daripada Candida rugosa. Kajian telah dijalankan untuk mengesahkan pengaruh pelbagai parameter pada pembentukan sitronelil butirat oleh lipase bebas daripada Candida rugosa, iaitu kesan kepekatan lipase, nisbah molar substrat, suhu, tiga jenis bahan sokongan untuk lipase tersekatgerak dan nisbah sekatgerak. Peningkatan kepekatan lipase bebas menyebabkan peningkatan kepada penukaran asid. Sifat peningkatan tidak berubah dari tindak balas pengesteran dapat diperhatikan pada kepekatan lipase tinggi yang memberikan kepekatan optimum lipase pada 3.33 g/l dengan penukaran asid sebanyak 92%. Kemungkinan ini disebabkan oleh tapak aktif lipase yang berlebihan yang berada di dalam zarah lipase pukal, yang tidak memberi sumbangan bererti kepada tindak balas. Aktiviti lipase didapati terencat dengan bertambahnya kepekatan asid butirik (pada kepekatan sitronelol tetap) dan sitronelol (pada kepekatan asid butirik tetap). Ini disebabkan adanya persaingan semulajadi pengikatan alkohol dan asid. Penukaran asid optimum diperolehi pada suhu 40°C selepas 24 jam pengeraman. Bagaimanapun, atas dari suhu ini, aktiviti pengesteran yang dimangkinkan oleh lipase mula menurun kerana penyahaslian protein. Daripada tiga jenis sokongan yang digunakan untuk lipase tersekatgerak, Amberlite MB–1 menunjukkan penukaran asid tertinggi berbanding dengan Amberlite XAD–1180 dan Celite 545. Penukaran asid optimum diperolehi pada nisbah sekatgerak 10 mg lipase/g penyokong. Pada nisbah sekatgerak ini, lipase mengoptimumkan sentuhan dengan permukaan penyokong dengan mengekalkan konformasi yang aktif pada tahap optimum. Kata kunci: Sitronelil butirat; lipase Candida rugosa; pengesteran langsung; lipase tersekatgerak; pelarut organik Free and immobilized Candida rugosa lipases were investigated for the synthesis of citronellyl butyrate by direct esterification reaction in n–hexane as organic solvent. A set of experiments was carried out to verify the influence of various parameters on the formation of citronellyl butyrate by free Candida rugosa lipase, such as lipase loading, substrate molar ratio, temperature, three kinds of support for immobilization, and ratio of immobilization. The conversion was increased with increasing lipase loading. The behavior of leveling–off in esterification was observed at higher lipase loading which gave the optimal amount of lipase loading at 3.33 g/l with 92% conversion. This might be due to the excess of lipase active sites, which remained inside the bulk of lipase particles, was not contributing significantly to the reaction. Increasing butyric acid and citronellol concentrations (at fixed citronellol and butyric acid concentrations, respectively) inhibited the lipase activity due to competitive nature of alcohol and acid binding. Optimal acid conversion was obtained at 40°C after 24–h incubation time. Above this temperature, however, the activity of lipase–catalyzed esterification begins to decrease due to denaturation of protein. From the three kinds of supports for immobilized lipase, Amberlite MB–1 showed the highest conversion compared to Amberlite XAD–1180 and Celite 545. The optimal acid conversion was obtained at lipase loading of 10 mg lipase/g support. At this loading, lipase attempts to optimize its contact with the surface of the support whereby optimum active conformation was retained. Key words: Citronellyl butyrate; Candida rugosa lipase; direct esterification; immobilized lipase; organic media

scholarly journals Biosynthesis of ethyl butyrate with immobilized Candida rugosa lipase onto modified Eupergit®C

Biocatalysis ◽  
2014 ◽  
Vol 1 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Daniele Spinelli ◽  
Simone Coppi ◽  
Riccardo Basosi ◽  
Rebecca Pogni
Keyword(s):  
Immobilized Lipase ◽  
Candida Rugosa ◽  
Product Yield ◽  
Candida Rugosa Lipase ◽  
Maximum Activity ◽  
Ethyl Butyrate ◽  
Molar Ratio ◽  
Practical Applications ◽  
Solvent Free Conditions ◽  
Immobilized Candida Rugosa Lipase

AbstractLipase from Candida rugosa was immobilized onto the modified Eupergit®C. The support was treated with ethylenediamine and subsequently activated with glutaraldehyde. Enzyme immobilization efficiency was 85%. The optimum pH was close to 6.5 for both the free and immobilized lipase. Immobilized lipase retained its maximum activity in a temperature range of 55 – 60°C. Subsequently, ethyl butyrate synthesis was investigated using immobilized enzyme by esterification of butyric acid with ethanol in solvent-free conditions (23% product yield) and using hexane as a solvent (65% product yield). The acid-alcohol molar ratio and different enzyme amounts were tested as efficient reaction parameters. The biocatalyst maintained 60% of its activity when reused in 8 successive batch reactions in organic solvent. Therefore, the immobilized lipase has demonstrated its potential in practical applications such as short-chain ester synthesis for the food industry.

scholarly journals Preparation of Immobilized Lipase Based on Hollow Mesoporous Silica Spheres and Its Application in Ester Synthesis

Molecules ◽  
2019 ◽  
Vol 24 (3) ◽  
pp. 395 ◽  
Author(s):  
Zhe Dong ◽  
Meng-Ying Jiang ◽  
Jie Shi ◽  
Ming-Ming Zheng ◽  
Feng-Hong Huang
Keyword(s):  
Reaction Time ◽  
Mesoporous Silica ◽  
Immobilized Lipase ◽  
Lipid Production ◽  
Candida Rugosa ◽  
Molar Ratio ◽  
Esterification Reaction ◽  
Enzymatic Esterification ◽  
Free System ◽  
Hollow Mesoporous Silica

In this study, Candida rugosa lipase (CRL) was immobilized into modified hollow mesoporous silica (HMSS) materials with different hydrophobicity. Among propyl-(C3), phenyl-(C6), octyl-(C8), and octadecyl-(C18) modified HMSS as well as native HMSS, taking advantage of more hydrophobic microenvironment, the HMSS-C18-CRL showed exceptional performance in enzymatic esterification reaction. Using the novel HMSS-C18 with immobilized CRL (HMSS-C18-CRL), we investigated the esterification of phytosterols with polyunsaturated fat acid (PUFA) in a solvent-free system for the production of phytosterols esters. Response surface methodology (RSM) was applied to model and optimize the reaction conditions, namely, the enzyme load (5–25%), reaction time (10–110 min), molar ratio of α-linolenic acid (ALA)/phytosterols (1:1–7:1) and represented by the letters E, T, and M respectively. Best-fitting models were successfully established by multiple regressions with backward elimination. The optimum production was achieved at 70 min for reaction time, 20% based on the weight of substrate for enzyme loading, and 5.6:1 for ALA/phytosterols molar ratio. Under optimized conditions, a conversion of about 90 ± 2% was achieved. These results indicated that HMSS-C18-CRL demonstrates to be a promising catalyst and can be potentially applied in the functional lipid production.

scholarly journals A STATISTICAL APPROACH FOR OPTIMIZING THE HIGH YIELD GREEN PRODUCTION OF THE FLAVOR ESTER BUTYL BUTYRATE

2017 ◽  
Vol 79 (7) ◽  
Author(s):  
Ida Nurhazwani Abd Rahman ◽  
Fatin Myra Abd Manan ◽  
Nur Haziqah Che Marzuki ◽  
Naji A. Mahat ◽  
Nursyafreena Attan ◽  
...  
Keyword(s):  
Molar Ratio ◽  
High Yield ◽  
Esterification Reaction ◽  
Reaction Parameters ◽  
Chemical Route ◽  
Butyl Butyrate ◽  
Novozyme 435 ◽  
Yield Production ◽  
Green Production ◽  
Percentage Conversion

Being the prevailing approach for producing esters such as butyl butyrate, the use of chemical route has been linked to numerous disadvantages. Hence, a green alternative method for higher yield production of butyl butyrate by esterification reaction utilizing Novozyme 435 as biocatalysts in a solvent-less system may prove useful. Such approach can be further improved by optimizing the relevant reaction parameters using the Response Surface Methodology by the Box-Benkhen Design attempted in this present study. The reaction parameters evaluated were: substrate molar ratio, time and temperature, and the response of each parameter was measured as percentage conversion yield. Using the Design Expert 7.1.6 optimization functions, the two sets of optimum conditions selected viz. [i] molar ratio butyric acid:butanol 1:3.93, 9.93 h at 56.09°C and [ii] molar ratio butyric acid:butanol 1:3.35, 9.79 h at 53.90°C had afforded the highest yield of butyl butyrate i.e. 99.62% and 99.55%, respectively. The ester product obtained from the reaction were confirmed as butyl butyrate by FTIR and GC. Therefore, the results substantiated the applicability of the RSM prediction technique as well as efficacy of Novozyme 435 as biocatalysts in the high yield solvent-less synthesis of butyl butyrate, adhering to the philosophy of Green Chemistry.

scholarly journals Improved Catalytic Performance of Carrier-Free Immobilized Lipase by Advanced Cross-Linked Enzyme Aggregates Technology

2021 ◽  
Author(s):  
Xia Jiaojiao ◽  
Yan Yan ◽  
Bin Zou ◽  
Adesanya Idowu Onyinye
Keyword(s):  
Low Cost ◽  
Immobilized Lipase ◽  
Candida Rugosa ◽  
Residual Activity ◽  
Candida Rugosa Lipase ◽  
Catalytic Performance ◽  
Functional Surface ◽  
Carrier Free ◽  
Effective Synthesis ◽  
Repeated Use

Abstract The cross-linked enzyme aggregates (CLEAs) are one of the technologies that quickly immobilize the enzyme without a carrier. This carrier-free immobilization method has the advantages of simple operation, high reusability and low cost. In this study, ionic liquid with amino group (1-aminopropyl-3-methylimidazole bromide，IL) was used as the novel functional surface molecule to modify industrialized lipase (Candida rugosa lipase, CRL). The enzymatic properties of the prepared CRL-FIL-CLEAs were investigated. The activity of CRL-FIL-CLEAs (5.51 U/mg protein) was 1.9 times higher than that of CRL-CLEAs without surface modification (2.86 U/mg protein). After incubation at 60℃ for 50 min, CRL-FIL-CLEAs still maintained 61% of its initial activity, while the value for CRL-CLEAs was only 22%. After repeated use for five times, compared with the 22% residual activity of CRL-CLEAs, the value of CRL-FIL-CLEAs was 51%. Further kinetic analysis indicated that the Km values for CRL-FIL-CLEAs and CRL-CLEAs were 4.80 mM and 8.06 mM, respectively, which was inferred that the affinity to substrate was increased after modification. Based on the above results, it was indicated that this method provided a new idea for the effective synthesis of immobilized enzyme.

scholarly journals Response surface methodology for optimizing the glycerolysis reaction of olive oil by Candida rugosa lipase

2014 ◽  
Vol 20 (1) ◽  
pp. 127-134 ◽  
Author(s):  
Kumar Singh ◽  
Mausumi Mukhopadhyay
Keyword(s):  
Response Surface Methodology ◽  
Reaction Time ◽  
Olive Oil ◽  
Water Content ◽  
Response Surface ◽  
Candida Rugosa ◽  
Candida Rugosa Lipase ◽  
Molar Ratio ◽  
Immobilized Candida Rugosa Lipase ◽  
Central Composite

In the present work, solvent free olive oil glycerolysis for the monoglycerides (MG) and diglycerides (DG) production with an immobilized Candida rugosa lipase was studied. MG and DG production were optimized using experiment design techniques and response surface methodology (RSM). RSM based on five-level, a five-variable central composite design (CCD) was used to optimize MG and DG production: reaction time, temperature, molar ratio of glycerol to oil, amount of lipase, and water content in glycerol. The reaction time, temperature, and amount of lipase were observed to be the most significant factors on the process response. The immobilized Candida rugosa lipase revealed optimum yield of MG and DG as 38.71 and 40.45 wt% respectively following a 5h reaction time with 0.025 g of lipase and 5% water content in glycerol at 40?C temperature. The yield of MG and DG production can be enhanced 1.5 fold by RSM.

scholarly journals Preparation and Candida rugosa Lipase Immobilization on Nylon-6 Grafted and Aminated (Polyvinyl Benzyl Chloride) Microfibers

2019 ◽  
Vol 14 (2) ◽  
pp. 369
Author(s):  
Nur Iilani Abd Halin ◽  
Maan Fahmi Rashid Al-Khatib ◽  
Hamzah Mohd. Salleh ◽  
Mohamed Mahmoud Nasef
Keyword(s):  
Benzyl Chloride ◽  
Immobilized Lipase ◽  
Candida Rugosa ◽  
Candida Rugosa Lipase ◽  
Nylon 6 ◽  
Covalent Attachment ◽  
Effect Of Temperature ◽  
P Value ◽  
Lipase Immobilization ◽  
The Stability

This paper demonstrates a simplified procedure for the preparation of a nylon-6 microfibers based support for the immobilization of Candida rugosa lipase via covalent attachment to enhance the stability and reusability of lipase. The preparation of the support was done by radiation induced graft copolymerization (RIGC) of vinyl benzyl chloride (VBC) onto nylon-6 microfibers followed by amination with ethanolamine to facilitate the immobilization of lipase. Fourier transfer infra red (FTIR) and scanning electron microscope (SEM) were used to study the chemical and physical changes following grafting, amination and immobilization. Response surface methodology (RSM) was applied for the optimization of lipase immobilization on the aminated microfibers. The optimization parameters were incubation time, pH, and lipase concentration. Moreover, this study investigated the effect of temperature, pH, and storage stability and reusability on the lipase in its immobilized and free forms. The developed model from RSM showed an R2 value of 0.9823 and P-value < 0.001 indicating that the model is significant. The optimum temperatures for both immobilized and free lipases were 45 °C, whereas the best pH values for lipase activity were at pH 8 and pH 7, respectively. This study also identifies values for KM and Vmax for both immobilized and free lipase accordingly. Based on the results, immobilized lipase had significantly improved the stability and reusability of lipase compared to that in free forms. Copyright © 2019 BCREC Group. All rights reserved 

scholarly journals Enzymatic synthesis optimization of isoamyl butyrate from fusel oil

2021 ◽  
Vol 43 ◽  
pp. e54966
Author(s):  
Andréia Anschau ◽  
Katira da Mota Huerta ◽  
Tatiane Vieira Rêgo ◽  
Janaina Mardioni Gonçalves de Oliveira ◽  
Carina Molins Borba ◽  
...  
Keyword(s):  
Butyric Acid ◽  
Immobilized Lipase ◽  
Scale Up ◽  
Maximum Yield ◽  
Fractional Factorial ◽  
Molar Ratio ◽  
Lipozyme Tl Im ◽  
Fusel Oil ◽  
Alcohol Production ◽  
Pharmaceutical Industries

Many food, cosmetic and pharmaceutical industries have increased their interest in short-chain esters due to their flavor properties. From the industrial standpoint, enzyme reactions are the most economical strategy to reach green products with neither toxicity nor damage to human health. Isoamyl butyrate (pear flavor) was synthesized by isoamyl alcohol (a byproduct of alcohol production) and butyric acid with the use of the immobilized lipase Lipozyme TL IM and hexane as solvents. Reaction variables (temperature, butyric acid concentration, isoamyl alcohol:butyric acid molar ratio and enzyme concentration) were investigated in ester conversion (%), concentration (mol L-1) and productivity (mmol ester g-1 mixture . h), by applying a sequential strategy of the Fractional Factorial Design (FFD) and the Central Composite Rotatable Design (CCRD). High isoamyl butyrate conversion of 95.8% was achieved at 24 hours. At 3 hours, the highest isoamyl butyrate concentration (1.64 mol L-1) and productivity (0.19 mmol ester g-1 mixture . h) were obtained under different reaction conditions. Due to high specificity and selectivity of lipases, process parameters of this study and their interaction with the Lipozyme TL IM are fundamental to understand and optimize the system so as to achieve maximum yield to scale up. Results show that fusel oil may be recycled by the green chemistry process proposed by this study.

scholarly journals Silica Coating of Metal-Loaded H-ZSM-22 to Form the Core-Shell Nanostructures: Characterization, Textural Properties, and Catalytic Potency in the Esterification of Oleic Acid

2021 ◽  
Vol 2021 ◽  
pp. 1-16
Author(s):  
Maryam Haghighi ◽  
Mehranoosh Fereidooni
Keyword(s):  
Oleic Acid ◽  
Electron Microscope ◽  
Active Sites ◽  
Maximum Yield ◽  
Operating Conditions ◽  
Molar Ratio ◽  
Emission Spectrometry ◽  
Esterification Reaction ◽  
Core Shell ◽  
The Core

In this study, ZSM-22 was synthesized using N,N-diethylaniline as a template through a hydrothermal method. The proton and various metals such as zirconium, strontium, and iron were immobilized on the surface of obtained zeolites through the ion exchange method. The catalysts were studied by Fourier-Transform Infrared Spectroscopy (FT-IR), X-Ray Diffraction (XRD), Brunauer–Emmett–Teller (BET) adsorption isotherms, Transmission Electron Microscope (TEM), Scanning Electron Microscope (SEM), Inductively Coupled Plasma-Optical Emission Spectrometry (ICP-OES) elemental analysis, and Temperature-Programmed Desorption of ammonia (TPD-NH3) technique for determining the number of acid sites. In the esterification reaction of oleic acid, the operating conditions such as catalyst dosage, temperature, molar ratio of methanol to oil, and reaction time were optimized and adjusted at 11 wt%, 70°C, 10 : 1, and 48 h subsequently. The maximum yield% of 48.07% was achieved in the presence of Zr-H-ZSM-22 at optimum conditions. In order to improve the efficiency of three zeolites Zr-H-ZSM-22, Fe-H-ZSM-22, and Sr-H-ZSM-22, the core-shell structures with SiO2 coating were prepared. Zr-H-ZSM-22@SiO2 was less active than Zr-H-ZSM-22 due to the SiO2 coverage of Lewis active sites.

scholarly journals The influence of composition of poly(n-isopropylacrylamide-co-itaconic acid) hydrogel on immobilized Candida rugosa lipase activity

2008 ◽  
Vol 62 (6) ◽  
pp. 339-344
Author(s):  
Nikola Milasinovic ◽  
Melina Kalagasidis-Krusic ◽  
Zorica Knezevic-Jugovic ◽  
Jovanka Filipovic
Keyword(s):  
Mechanical Properties ◽  
Itaconic Acid ◽  
Lipase Activity ◽  
Acid Content ◽  
Immobilized Enzyme ◽  
In Situ Polymerization ◽  
Immobilized Lipase ◽  
Candida Rugosa ◽  
Candida Rugosa Lipase

The application of lipases as catalysts in chemical reactions has been deterred by the high cost of isolation and purification of enzymes, the instability of their structure when they are isolated from their natural environment, contamination of products with residual protein, their sensitivity to process conditions, etc. These problems could be overcome using immobilized lipases. Immobilization is achieved by fixing enzymes to or within solid supports and as a result a heterogeneous system is obtained. The present paper reports on the immobilization of Candida rugosa lipase in hydrogels based on N-isopropylacrylamide and itaconic acid. Immobilization of lipase is carried out by two different methods. In the first method, lipase is added to the reaction mixture before polymerization and crosslinking (in situ polymerization), while in the second method the synthetized hydrogels are immersed in lipase solution and left to rich the equilibrium swelling. The specific activities of the immobilized lipase were determined in both cases and compared. The amount of the immobilized lipase is higher if the immobilization is carried out by immersing hydrogel in lipase solution. It was observed that in both cases lipase activity increases with an increase of the itaconic acid content up to 10 wt% and thereafter decreases. From the measurements of shear storage moduli (G') it was concluded that the increase of the itaconic acid content decreases the mechanical properties of the hydrogels. SEM analysis confirmed the highly porous structure of hydrogels. It was found that greater pores were achieved when the enzyme was immobilized by in situ polymerization. When the enzyme was immobilized by in situ polymerization the itaconic acid content had not great effect on the mass of the immobilized enzyme, except for the 100/0 sample. On the contrary, for the samples where the enzyme was immobilized by swelling, the increase of the itaconic acid content increases the mass of the immobilized enzyme. Concerning the activity of the immobilized lipase, the swelling degree and mechanical properties of the investigated hydrogels, the best results were performed by the 95/5 hydrogel sample.

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