Biocatalysis with Undissolved Solid Substrates and Products

Halling, Peter J.; Carrea, Giacomo; Riva, Sergio

doi:10.1002/9783527621729.ch12

Cited by 4 publications

(2 citation statements)

References 78 publications

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“…Although it has been described that Ca 2+ is a requirement apparently no significant for PLDs from bacteria , previous studies in our group have showed better results in the presence of Ca 2+ ions. Besides, in precipitation‐driven catalysis systems, such as the present study, the addition of specific counter‐ions that form poorly soluble salts with an ionic form of the product of the reaction (and not with the substrates) leads to product salts precipitation from the reaction mixture . Table shows the influence of CaCl 2 at different concentrations ranging from 33 to 201 mmol/L.…”

Section: Resultsmentioning

confidence: 84%

Novel glyceryl ethers phospholipids produced by solid to solid transphosphatidylation in the presence of a food grade phospholipase D

Arranz-Martínez

Casado

Reglero

et al. 2017

Euro J Lipid Sci & Tech

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Enzymatic transphosphatidylation between batyl alcohol and phospatidylcholine has been investigated. In situ product removal methodology and the utilization of a green biphasic medium consisting of a buffer and an organic phase comprised of generally recognized as safe flavoring additives, have been established as milestones to increase the applicability of the process. Different organic phases (limonene, pinene, ethyl isovalerate, ethyl lactate, ethyl butyrate, hexanal, and cyclopentanone) and one aqueous phase (0.1 M acetate buffer at pH = 5.5) were studied. The relative proportion of organic and aqueous phases was also investigated. Different molar ratios alkylglycerol/phosphatidylcholine (10:1, 5:1, 2:1, 1:1, and 1:2) were also evaluated. Addition of calcium chloride and the reaction temperature have been adjusted in the enzymatic transphosphatidylation. Finally, the percentage of phospholipase D and the molar concentration of the reactants were also optimized. The ratio transphosphatidylation to hydrolysis was evaluated for the different biphasic reaction mixtures tested. The best results (ca. 70% w/w of glyceryl ether phospholipid) were achieved utilizing 5% (w/w) of phospholipase D, equimolar ratio of reactants and limonene/acetate buffer at a volumetric ratio 1:3. The volumetric productivity of the process was then optimized up to 126 mM and scaled‐up to 50 g of reaction mixture with similar results in terms of composition and yield. This study reports a procedure for the production of tailor‐made phospholipids from commercial sources with high volumetric yield. These highly valuable ingredients are intended to be used as both delivery systems and bioactive lipids. Practical applications: Solid to solid transphosphatidylation based on in situ product removal has been developed for the synthesis of a novel glyceryl ether phospholipid, incorporating batyl alcohol into the polar head of the phosphatidylcholine in the presence of a food grade PLD. Enzymatic transphosphatidylation usually reported in the literature utilize very low concentration of phospholipid and a large excess of the other reactant. In this sense, the present study evaluated different molar ratios alkylglycerol/phosphatidylcholine. Besides, regarding novel phospholipid as food for human use, the utilization of toxic organic solvents was avoided in its production, replacing them by GRAS organic solvents. Hence, this procedure readily scalable is intended to be used for obtaining this novel glyceryl ether phospholipid with capabilities to perform as both novel delivery systems and bioactive lipids. In addition, a very simple downstream process for product recovery has been also described. Schematic representation of a solid to solid transphosphatidylation reaction between phosphatidylcholine and batyl alcohol in a biphasic reaction medium comprised of an acetate buffer and an organic solvent generally recognized as safe (limonene).

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Section: Resultsmentioning

confidence: 84%

Novel glyceryl ethers phospholipids produced by solid to solid transphosphatidylation in the presence of a food grade phospholipase D

Arranz-Martínez

Casado

Reglero

et al. 2017

Euro J Lipid Sci & Tech

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“…This might be explained by the presence of the IR particles as suspended substrate, representing a condition of strong mass transfer limitation, especially in the beginning of the incubation. The presence of solid substrates has important consequences for the reaction kinetics and thermodynamics, affecting directly the reactions yields …”

Section: Discussionmentioning

confidence: 99%

In vitro studies on the degradation of poly(cis‐1,4‐isoprene)

Andler

Altenhoff

Mäsing

et al. 2018

Biotechnology Progress

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Cleavage of the backbone of poly(cis-1,4-isoprene) (IR) in solid rubber material was accomplished by the addition of partially purified latex clearing protein (Lcp1 ) using a 200-mL enzyme reactor. Two strategies for the addition of Lcp1 were studied revealing that the daily addition of 50 µg mL of Lcp1 for 5 days was clearly a more efficient regime in comparison to a one-time addition of 250 µg of Lcp1 at the beginning. Soluble oligo(cis-1,4-isoprene) molecules occurred as degradation products and were identified by ESI-MS and GPC. Oxygenase activity of Lcp1 with solid IR particles as substrate was shown for the first time by measuring the oxygen consumption in the reaction medium. A strong decrease of the dissolved oxygen concentration was detected at the end of the assay, which indicates an increase in the number of cleavage reactions. The oligo(cis-1,4-isoprene) molecules comprised 1 to 11 isoprene units and exhibited an average molecular weight (M ) of 885 g mol . Isolation of the oligo(cis-1,4-isoprene) molecules was achieved by using silica gel column chromatography. The relative quantification of the isolated products was performed by HPLC-MS after derivatization with 2,4-dinitrophenilhydrazyne yielding a concentration of total degradation products of 1.62 g L . Analysis of the polymer surface in samples incubated for 3 days with Lcp1 via ATR-FTIR indicated the presence of carbonyl groups, which occurred upon the cleavage reaction. This study presents a cell-free bioprocess as an alternative rubber treatment that can be applied for the partial degradation of the polymer. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:890-899, 2018.

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Process Considerations for the Application of Enzymes

Kara

Liese

2019

Industrial Enzyme Applications

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Biocatalysis with Undissolved Solid Substrates and Products

Cited by 4 publications

References 78 publications

Novel glyceryl ethers phospholipids produced by solid to solid transphosphatidylation in the presence of a food grade phospholipase D

Novel glyceryl ethers phospholipids produced by solid to solid transphosphatidylation in the presence of a food grade phospholipase D

In vitro studies on the degradation of poly(cis‐1,4‐isoprene)

Process Considerations for the Application of Enzymes

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