Effect of acyl‐acceptor stepwise addition strategy using <i>alperujo</i> oil as a substrate in enzymatic biodiesel synthesis

Bonet-Ragel, Kírian; Canet, Albert; Benaiges, M. Dolors; Valero, Francisco

doi:10.1002/jctb.5399

Cited by 16 publications

(18 citation statements)

References 57 publications

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“…Also, consistent with previous results [37], both lipases were more strongly affected by ethanol than they were by methanol. However, this result conflicts with those for rROL in biodiesel synthesis reactions, where methanol proved more detrimental [15]. This contradiction can be ascribed to the alcohol interacting with the active site of the enzyme during biodiesel reactions, which is unlikely in a lipase-alcohol solution because the enzyme lid is closed.…”

Section: Stabilitymentioning

confidence: 77%

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Truncated Prosequence of Rhizopus oryzae Lipase: Key Factor for Production Improvement and Biocatalyst Stability

et al. 2019

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Recombinant Rhizopus oryzae lipase (mature sequence, rROL) was modified by adding to its N-terminal 28 additional amino acids from the C-terminal of the prosequence (proROL) to obtain a biocatalyst more suitable for the biodiesel industry. Both enzymes were expressed in Pichia pastoris and compared in terms of production bioprocess parameters, biochemical properties, and stability. Growth kinetics, production, and yields were better for proROL harboring strain than rROL one in batch cultures. When different fed-batch strategies were applied, lipase production and volumetric productivity of proROL-strain were always higher (5.4 and 4.4-fold, respectively) in the best case. rROL and proROL enzymatic activity was dependent on ionic strength and peaked in 200 mM Tris-HCl buffer. The optimum temperature and pH for rROL were influenced by ionic strength, but those for proROL were not. The presence of these amino acids altered lipase substrate specificity and increased proROL stability when different temperature, pH, and methanol/ethanol concentrations were employed. The 28 amino acids were found to be preferably removed by proteases, leading to the transformation of proROL into rROL. Nevertheless, the truncated prosequence enhanced Rhizopus oryzae lipase heterologous production and stability, making it more appropriate as industrial biocatalyst.

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Section: Stabilitymentioning

confidence: 77%

“…Lipases covalently linked to chemically modified supports [13] and cross-linked enzyme aggregates [14] have also been explored for biodiesel production. Furthermore, some authors have successfully prevented lipase inactivation by adding alcohol stepwise [15].…”

Section: Introductionmentioning

confidence: 99%

Truncated Prosequence of Rhizopus oryzae Lipase: Key Factor for Production Improvement and Biocatalyst Stability

et al. 2019

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“…Eversa-mCLEA was less active using short-chain primary alcohols (methanol and ethanol), which are the alcohols usually utilized in biodiesel production [ 67 , 68 , 69 ]. This lower activity can be explained by the fact of methanol and ethanol change the hydration layer of the enzyme [ 21 , 70 , 71 , 72 ], justifying the common practice of adding the alcohol in steps, mainly when methanol is used as an acyl acceptor ([ 24 ]). Similar results were obtained by Trivedi et al [ 66 ] using the formulation preceding Eversa (soluble lipase Callera Trans L, EP 258068) in the esterification reaction of short and long-chain primary alcohols (oleyl alcohol, octanol, and hexanol), for which the ester yield reduced as the length of the chain decreased.…”

Section: Resultsmentioning

confidence: 99%

Immobilization of Eversa® Transform via CLEA Technology Converts It in a Suitable Biocatalyst for Biolubricant Production Using Waste Cooking Oil

et al. 2021

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The performance of the previously optimized magnetic cross-linked enzyme aggregate of Eversa (Eversa-mCLEA) in the enzymatic synthesis of biolubricants by transesterification of waste cooking oil (WCO) with different alcohols has been evaluated. Eversa-mCLEA showed good activities using these alcohols, reaching a transesterification activity with isoamyl alcohol around 10-fold higher than with methanol. Yields of isoamyl fatty acid ester synthesis were similar using WCO or refined oil, confirming that this biocatalyst could be utilized to transform this residue into a valuable product. The effects of WCO/isoamyl alcohol molar ratio and enzyme load on the synthesis of biolubricant were also investigated. A maximum yield of around 90 wt.% was reached after 72 h of reaction using an enzyme load of 12 esterification units/g oil and a WCO/alcohol molar ratio of 1:6 in a solvent-free system. At the same conditions, the liquid Eversa yielded a maximum ester yield of only 34%. This study demonstrated the great changes in the enzyme properties that can be derived from a proper immobilization system. Moreover, it also shows the potential of WCO as a feedstock for the production of isoamyl fatty acid esters, which are potential candidates as biolubricants.

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“…The excess of ethanol (from 3 to 10 moles/mol of soybean oil) led to a reduction in the reaction yield up to 4 times. This result may be explained by the ability of the ethanol to alter the hydration layer of enzymes, leading to an activity reduction or even to an enzyme inactivation [65,66,67].…”

Section: Resultsmentioning

confidence: 99%

Improving the Yields and Reaction Rate in the Ethanolysis of Soybean Oil by Using Mixtures of Lipase CLEAs

et al. 2019

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Due to the heterogeneity of oils, the use of mixtures of lipases with different activity for a large number of glycerol-linked carboxylic acids that compose the substrate has been proposed as a better alternative than the use of one specific lipase preparation in the enzymatic synthesis of biodiesel. In this work, mixtures of lipases from different sources were evaluated in their soluble form in the ethanolysis of soybean oil. A mixture of lipases (50% of each lipase, in activity basis) from porcine pancreas (PPL) and Thermomyces lanuginosus lipase (TLL) gave the highest fatty acid ethyl ester (FAEE) yield (around 20 wt.%), while the individual lipases gave FAEE yields 100 and 5 times lower, respectively. These lipases were immobilized individually by the cross-linked enzyme aggregates (CLEAs) technique, yielding biocatalysts with 89 and 119% of expressed activity, respectively. A mixture of these CLEAs (also 50% of each lipase, in activity basis) gave 90.4 wt.% FAEE yield, while using separately CLEAs of PPL and TLL, the FAEE yields were 84.7 and 75.6 wt.%, respectively, under the same reaction conditions. The mixture of CLEAs could be reused (five cycles of 6 h) in the ethanolysis of soybean oil in a vortex flow-type reactor yielding an FAEE yield higher than 80% of that of the first batch.

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Effect of acyl‐acceptor stepwise addition strategy using alperujo oil as a substrate in enzymatic biodiesel synthesis

Cited by 16 publications

References 57 publications

Truncated Prosequence of Rhizopus oryzae Lipase: Key Factor for Production Improvement and Biocatalyst Stability

Truncated Prosequence of Rhizopus oryzae Lipase: Key Factor for Production Improvement and Biocatalyst Stability

Immobilization of Eversa® Transform via CLEA Technology Converts It in a Suitable Biocatalyst for Biolubricant Production Using Waste Cooking Oil

Improving the Yields and Reaction Rate in the Ethanolysis of Soybean Oil by Using Mixtures of Lipase CLEAs

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