DOCSLIB.ORG

Immobilized Whole-Cell Transaminase Biocatalysts for Continuous-Flow Kinetic Resolution of Amines

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Immobilized Whole-Cell Transaminase Biocatalysts for Continuous-Flow Kinetic Resolution of Amines catalysts Article Immobilized Whole-Cell Transaminase Biocatalysts for Continuous-Flow Kinetic Resolution of Amines Zsófia Molnár 1,2,3, Emese Farkas 1, Ágnes Lakó 1 , Balázs Erdélyi 2, Wolfgang Kroutil 4 , Beáta G. Vértessy 3,5, Csaba Paizs 6 and László Poppe 1,6,7,* 1 Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, M˝uegyetemrkp. 3, 1111 Budapest, Hungary; molnar.zsofi[email protected] (Z.M.); [email protected] (E.F.); [email protected] (Á.L.) 2 Fermentia Microbiological Ltd., Berlini út 47–49, 1405 Budapest, Hungary; [email protected] 3 Institute of Enzymology, Research Center for Natural Sciences, Hungarian Academy of Science, Magyar tudósok krt. 2, 1117 Budapest, Hungary; [email protected] 4 Institute of Chemistry, University of Graz, NAWI Graz, BioTechMed Graz, BioHealth, Heinrichstraße 28, 8010 Graz, Austria; [email protected] 5 Department of Applied Biotechnology and Food Science, Budapest University of Technology and Economics, M˝uegyetemrkp. 3, 1111 Budapest, Hungary 6 Biocatalysis and Biotransformation Research Centre, Faculty of Chemistry and Chemical Engineering, Babe¸s-BolyaiUniversity of Cluj-Napoca, Arany János Str. 11, RO-400028 Cluj-Napoca, Romania; [email protected] 7 SynBiocat Ltd., Szilasliget u 3, H-1172 Budapest, Hungary * Correspondence: [email protected]; Tel.: +36(1)463-3299 Received: 29 March 2019; Accepted: 25 April 2019; Published: 10 May 2019 Abstract: Immobilization of transaminases creates promising biocatalysts for production of chiral amines in batch or continuous-flow mode reactions. E. coli cells containing overexpressed transaminases of various selectivities and hollow silica microspheres as supporting agent were immobilized by an improved sol-gel process to produce immobilized transaminase biocatalysts with suitable stability and mechanical properties for continuous-flow applications. The immobilized cell-based transaminase biocatalyst proved to be durable and easy-to-use in kinetic resolution of four racemic amines 1a–d. The batch and continuous-flow mode kinetic resolutions with transaminase biocatalyst of opposite stereopreference provided access to both enantiomers of the corresponding amines. By using the most suitable immobilized transaminase biocatalysts, this study describes the first transaminase-based approach for the production of both pure enantiomers of 1-(3,4-dimethoxyphenyl)ethan-1-amine 1d. Keywords: stereoselective biocatalysis; transaminase; kinetic resolution; flow chemistry; sol-gel; whole-cell immobilization 1. Introduction Enantiopure amines are essential chiral building blocks for the synthesis of a wide variety of active pharmaceutical ingredients. Chemical synthesis of these compounds usually employs transition metal catalysts of relatively high toxicity, and may require harsh reaction conditions. In recent years, there has been a growing interest in transaminases (TAs), which offer a sustainable alternative to these synthetic chemical processes [1–3]. Transaminases have been successfully used in the preparation of several pharmaceutically relevant compounds, like 3-aryl-γ-aminobutyric acid derivatives [4], sitagliptin [5] and valinol [6]. Transaminases can be applied for separation of the enantiomers from their racemates in kinetic resolution (KR) with a maximum of 50% yield, or for conversion of a prochiral carbonyl compound to a Catalysts 2019, 9, 438; doi:10.3390/catal9050438 www.mdpi.com/journal/catalysts Catalysts 2019, 9, 438 2 of 11 Catalysts 2019, 9, x FOR PEER REVIEW 2 of 11 tochiral a chiral amine amine in asymmetric in asymmetric synthesis synthesis with a with theoretical a theoretical yield of yield 100%. of Although 100%. Although asymmetric asymmetric synthesis synthesiscould provide could higher provide yields, higher it usually yields, su itff usuallyers from su disfavoredffers from reaction disfavored equilibrium reaction [7equilibrium]. Furthermore, [7]. Furthermore,the costs of enzyme the costs production of enzyme indicates production the need indi forcates biocatalyst the need recovery for biocatalyst in an economically recovery viablein an economicallyprocess [8]. These viable are process the most [8]. significant These are ditheffi cultiesmost significant hindering difficulties the widespread hindering industrial the widespread application industrialof transaminases. application Several of transaminases. strategies have beenSeveral developed strategies to overcomehave been the developed difficulties to arising overcome from the the difficultiesdisfavored arising reaction from equilibrium the disfavored in asymmetric reaction synthesis equilibrium with in TAs, asymmetric such as the synthesis removal with of products TAs, such by asextraction the removal [9], by of evaporation products by [10 extraction] or by coupled [9], by cascade evaporation reactions [10] [11 or,12 by]. Thecoupled overall cascade efficiency reactions of the [11,12].KR process The mayoverall be enhancedefficiency byof the recycling KR process the formed may be ketone enhanced to racemic by recycling amine bythe a formed proper ketone reductive to racemicamination amine method by a [proper13]. Although reductive these amination solutions method work [13]. well Although in batch reactions, these solutions for the work sustainable, well in batchindustrial reactions, production for the of sustainable, enantiopure industrial amines the pr intensificationoduction of enantiopure possibilities oaminesffered bythe immobilized intensification TA possibilitiesbiocatalysts andoffered by theby continuous-modeimmobilized TA biocatalysts operations areand needed by the [1continuous-mode]. This trend is indicated operations by are the neededsuccessful [1]. applicationsThis trend is ofindicated TAs in continuous-flowby the successful reactors applications which of haveTAs in been continuous-flow developed in reactors the past whichyears [14have–23 been]. developed in the past years [14–23]. A wide range of immobilization methods for transaminases have have been been reported over the last couple of of years. years. Transaminases Transaminases have have been been successfully successfully immobilized immobilized by byusing using different different kinds kinds of carriers,of carriers, such such as polymeric as polymeric resins [7,16,23], resins [7 ,functionalized16,23], functionalized cellulose cellulose[21], chitosan [21], [24–26], chitosan inorganic- [24–26], basedinorganic-based nanoflowers nanoflowers [27,28], macrocellular [27,28], macrocellular silica monoliths silica [19], monoliths MnO2 nanorods [19], MnO [29]2 nanorods or porous [29 glass] or metalporous affinity glass metalsupports affi nity[22]. supports In addition [22]. to Inimmobilization addition to immobilization of TAs in their of isolated TAs in theirand fully isolated or partiallyand fully purified or partially form, purified the entrapment form, the of entrapmentcell-free extracts, of cell-free or even extracts, whole cells or even with whole comprising cells with TA activitycomprising in different TA activity sol-gel in dimatricesfferent sol-gel[14,30,31] matrices proved [ 14to, 30be, 31an] provedexcellent to approach be an excellent to prepare approach high- performanceto prepare high-performance and stable immobilized and stable biocatalysts. immobilized In many biocatalysts. cases, utilization In many of cases, whole utilization cells over of enzymewhole cells solutions over enzymeis advantageous solutions due is advantageousto the lower production due to the costs, lower increased production stability costs, and increased easier handlingstability and[32]. easier Immobilize handlingd transaminases [32]. Immobilized have been transaminases successfully have employed been successfully in the preparation employed of severalin the preparationenantiopure ofcompounds, several enantiopure such as 1-phenox compounds,ypropane-2-amine such as 1-phenoxypropane-2-amine [33], 3-amino-1-Boc-piperidine [33], [34],3-amino-1-Boc-piperidine 1-methyl-3-phenylpropylamine [34], 1-methyl-3-phenylpropylamine [23,26] and mexiletine [16]. [23 ,26] and mexiletine [16]. Scheme 1. Continuous-flowContinuous-flow kinetickinetic resolutionresolution of of primary primary amines aminesrac rac-1a-1a–d–withd with immobilized immobilized whole-cell whole- celltransaminase transaminase biocatalysts. biocatalysts. This study aimed at the preparation of robust, stereoselective TA biocatalysts capable of operating This study aimed at the preparation of robust, stereoselective TA biocatalysts capable of under continuous-flow conditions to produce enantiopure amines. A promising sol-gel-based operating under continuous-flow conditions to produce enantiopure amines. A promising sol-gel- whole-cell immobilization method has already been applied in a previous work of our group for based whole-cell immobilization method has already been applied in a previous work of our group for the transaminase from Chromobacterium violaceum expressed in E. coli [14]. Hereby we report the Catalysts 2019, 9, 438 3 of 11 Catalysts 2019, 9, x FOR PEER REVIEW 3 of 11 immobilizationthe transaminase of fromfurtherChromobacterium transaminases with violaceum differentexpressed enantioselectivity in E. coli [14 and]. Hereby substrate we specificity. report the Theimmobilization immobilized of cells further proved transaminases to be easy-to with store, di ffcheaperent and enantioselectivity durable biocatalysts, and substrate and were specificity. applied successfullyThe immobilized in the cellscontinuous proved kinetic
Load more

Recommended publications
  • Improved Catalytic Performance of Carrier-Free Immobilized Lipase by Advanced Cross-Linked Enzyme Aggregates Technology
    Improved Catalytic Performance of Carrier-Free Immobilized Lipase by Advanced Cross-Linked Enzyme Aggregates Technology Xia Jiaojiao Jiangsu University of Science and Technology: Jiangsu University Yan Yan Jiangsu University Bin Zou ( [email protected] ) Jiangsu University https://orcid.org/0000-0003-3816-8776 Adesanya Idowu Onyinye Jiangsu University Research Article Keywords: lipase, carrier-free, immobilization, ionic liquid, stability Posted Date: May 24th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-505612/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/20 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 bromideIL) 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 modication (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 ve 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 anity to substrate was increased after modication.
    [Show full text]
  • Part I Principles of Enzyme Catalysis
    j1 Part I Principles of Enzyme Catalysis Enzyme Catalysis in Organic Synthesis, Third Edition. Edited by Karlheinz Drauz, Harald Groger,€ and Oliver May. Ó 2012 Wiley-VCH Verlag GmbH & Co. KGaA. Published 2012 by Wiley-VCH Verlag GmbH & Co. KGaA. j3 1 Introduction – Principles and Historical Landmarks of Enzyme Catalysis in Organic Synthesis Harald Gr€oger and Yasuhisa Asano 1.1 General Remarks Enzyme catalysis in organic synthesis – behind this term stands a technology that today is widely recognized as a first choice opportunity in the preparation of a wide range of chemical compounds. Notably, this is true not only for academic syntheses but also for industrial-scale applications [1]. For numerous molecules the synthetic routes based on enzyme catalysis have turned out to be competitive (and often superior!) compared with classic chemicalaswellaschemocatalyticsynthetic approaches. Thus, enzymatic catalysis is increasingly recognized by organic chemists in both academia and industry as an attractive synthetic tool besides the traditional organic disciplines such as classic synthesis, metal catalysis, and organocatalysis [2]. By means of enzymes a broad range of transformations relevant in organic chemistry can be catalyzed, including, for example, redox reactions, carbon–carbon bond forming reactions, and hydrolytic reactions. Nonetheless, for a long time enzyme catalysis was not realized as a first choice option in organic synthesis. Organic chemists did not use enzymes as catalysts for their envisioned syntheses because of observed (or assumed) disadvantages such as narrow substrate range, limited stability of enzymes under organic reaction conditions, low efficiency when using wild-type strains, and diluted substrate and product solutions, thus leading to non-satisfactory volumetric productivities.
    [Show full text]
  • L-Amino Acid Production by a Immobilized Double-Racemase Hydantoinase Process: Improvement and Comparison with a Free Protein System
    catalysts Article L-Amino Acid Production by a Immobilized Double-Racemase Hydantoinase Process: Improvement and Comparison with a Free Protein System María José Rodríguez-Alonso 1,2, Felipe Rodríguez-Vico 1,2, Francisco Javier Las Heras-Vázquez 1,2 and Josefa María Clemente-Jiménez 1,2,* 1 Department of Chemistry and Physic, University of Almeria, The Agrifood Campus of International Excellence, ceiA3, E-04120 Almería, Spain; [email protected] (M.J.R.-A.); [email protected] (F.R.-V.); [email protected] (F.J.L.H.-V.) 2 Research Centre for Agricultural and Food Biotechnology, BITAL, E-04120 Almería, Spain * Correspondence: [email protected]; Tel.: +34-950-015-055 Academic Editor: Manuel Ferrer Received: 4 May 2017; Accepted: 15 June 2017; Published: 20 June 2017 Abstract: Protein immobilization is proving to be an environmentally friendly strategy for manufacturing biochemicals at high yields and low production costs. This work describes the optimization of the so-called “double-racemase hydantoinase process,” a system of four enzymes used to produce optically pure L-amino acids from a racemic mixture of hydantoins. The four proteins were immobilized separately, and, based on their specific activity, the optimal whole relation was determined. The first enzyme, D,L-hydantoinase, preferably hydrolyzes D-hydantoins from D,L-hydantoins to N-carbamoyl-D-amino acids. The remaining L-hydantoins are racemized by the second enzyme, hydantoin racemase, and continue supplying substrate D-hydantoins to the first enzyme. N-carbamoyl-D-amino acid is racemized in turn to N-carbamoyl-L-amino acid by the third enzyme, carbamoyl racemase. Finally, the N-carbamoyl-L-amino acid is transformed to L-amino acid by the fourth enzyme, L-carbamoylase.
    [Show full text]
  • Screening of Macromolecular Cross-Linkers and Food-Grade Additives for Enhancement of Catalytic Performance of MNP-CLEA-Lipase of Hevea Brasiliensis
    IOP Conference Series: Materials Science and Engineering PAPER • OPEN ACCESS Screening of macromolecular cross-linkers and food-grade additives for enhancement of catalytic performance of MNP-CLEA-lipase of hevea brasiliensis To cite this article: Nur Amalin Ab Aziz Al Safi and Faridah Yusof 2020 IOP Conf. Ser.: Mater. Sci. Eng. 932 012019 View the article online for updates and enhancements. This content was downloaded from IP address 170.106.202.226 on 23/09/2021 at 18:45 1st International Conference on Science, Engineering and Technology (ICSET) 2020 IOP Publishing IOP Conf. Series: Materials Science and Engineering 932 (2020) 012019 doi:10.1088/1757-899X/932/1/012019 Screening of macromolecular cross-linkers and food-grade additives for enhancement of catalytic performance of MNP- CLEA-lipase of hevea brasiliensis. Nur Amalin Ab Aziz Al Safi1 and Faridah Yusof1 1 Department of Biotechnology Engineering, International Islamic University Malaysia. Abstract. Skim latex from Hevea brasiliensis (rubber tree) consist of many useful proteins and enzymes that can be utilized to produce value added products for industrial purposes. Lipase recovered from skim latex serum was immobilized via cross-linked enzyme aggregates (CLEA) technology, while supported by magnetic nanoparticles for properties enhancement, termed ‘Magnetic Nanoparticles CLEA-lipase’ (MNP-CLEA-lipase). MNP-CLEA-lipase was prepared by chemical cross-linking of enzyme aggregates with amino functionalized magnetic nanoparticles. Instead of using glutaraldehyde as cross-linking agent, green, non-toxic and renewable macromolecular cross-linkers (dextran, chitosan, gum Arabic and pectin) were screened and the best alternative based on highest residual activity was chosen for further analysis.
    [Show full text]
  • Chapter 2 Immobilization of Enzymes
    Chapter 2 Immobilization of Enzymes: A Literature Survey Beatriz Brena , Paula González-Pombo , and Francisco Batista-Viera Abstract The term immobilized enzymes refers to “enzymes physically confi ned or localized in a certain defi ned region of space with retention of their catalytic activities, and which can be used repeatedly and continuously.” Immobilized enzymes are currently the subject of considerable interest because of their advantages over soluble enzymes. In addition to their use in industrial processes, the immobilization techniques are the basis for making a number of biotechnology products with application in diagnostics, bioaffi nity chromatography, and biosensors. At the beginning, only immobilized single enzymes were used, after 1970s more complex systems including two-enzyme reactions with cofactor regeneration and living cells were developed. The enzymes can be attached to the support by interactions ranging from reversible physical adsorp- tion and ionic linkages to stable covalent bonds. Although the choice of the most appropriate immobilization technique depends on the nature of the enzyme and the carrier, in the last years the immobilization tech- nology has increasingly become a matter of rational design. As a consequence of enzyme immobilization, some properties such as catalytic activity or thermal stability become altered. These effects have been demonstrated and exploited. The concept of stabilization has been an important driving force for immobilizing enzymes. Moreover, true stabilization at the molecular level has been demonstrated, e.g., proteins immobilized through multipoint covalent binding. Key words Immobilized enzymes , Bioaffi nity chromatography , Biosensors , Enzyme stabilization , Immobilization methods 1 Background Enzymes are biological catalysts that promote the transformation of chemical species in living systems.
    [Show full text]
  • Recent Trends in Biomaterials for Immobilization of Lipases for Application in Non-Conventional Media
    catalysts Review Recent Trends in Biomaterials for Immobilization of Lipases for Application in Non-Conventional Media Robson Carlos Alnoch 1,2 , Leandro Alves dos Santos 3 , Janaina Marques de Almeida 2, Nadia Krieger 3 and Cesar Mateo 4,* 1 Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto 14040-900, São Paulo, Brazil; [email protected] 2 Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Cx. P. 19081, Centro Politécnico, Curitiba 81531-980, Paraná, Brazil; [email protected] 3 Departamento de Química, Universidade Federal do Paraná, Cx. P. 19061, Centro Politécnico, Curitiba 81531-980, Paraná, Brazil; [email protected] (L.A.d.S.); [email protected] (N.K.) 4 Departamento de Biocatálisis, Instituto de Catálisis y Petroleoquímica (CSIC), Marie Curie 2, Cantoblanco, Campus UAM, 28049 Madrid, Spain * Correspondence: [email protected]; Tel.: +34-915854768; Fax: +34-915854860 Received: 18 May 2020; Accepted: 18 June 2020; Published: 20 June 2020 Abstract: The utilization of biomaterials as novel carrier materials for lipase immobilization has been investigated by many research groups over recent years. Biomaterials such as agarose, starch, chitin, chitosan, cellulose, and their derivatives have been extensively studied since they are non-toxic materials, can be obtained from a wide range of sources and are easy to modify, due to the high variety of functional groups on their surfaces. However, although many lipases have been immobilized on biomaterials and have shown potential for application in biocatalysis, special features are required when the biocatalyst is used in non-conventional media, for example, in organic solvents, which are required for most reactions in organic synthesis.
    [Show full text]
  • Felodipine by Aspergillus Niger and Lipase AP6 Enzyme
    Advances in Microbiology, 2016, 6, 1062-1074 http://www.scirp.org/journal/aim ISSN Online: 2165-3410 ISSN Print: 2165-3402 Enantioselective Conversion of Racemic Felodipine to S(−)-Felodipine by Aspergillus niger and Lipase AP6 Enzyme Chandupatla Vijitha, Ettireddy Swetha, Ciddi Veeresham* University College of Pharmaceutical Sciences, Kakatiya University, Warangal, India How to cite this paper: Vijitha, C., Swetha, Abstract E. and Veeresham, C. (2016) Enantioselec- tive Conversion of Racemic Felodipine to The present study involves the enantioselective resolution of racemic Felodipine by S(−)-Felodipine by Aspergillus niger and using free and immobilized forms of microbial cultures as well as an enzyme (Lipase Lipase AP6 Enzyme. Advances in Microbi- AP6). Among the microbial cultures employed in the present study, Aspergillus ni- ology, 6, 1062-1074. http://dx.doi.org/10.4236/aim.2016.614099 ger, Sphingomonas paucimobilis, Cunninghamella elegans, Escherichia coli, Pseu- domonas putida and Cunninghamella blakesleeana were found to possess capability Received: November 22, 2016 of enantioselective resolution of racemic Felodipine. The enantiomeric excess (ee%) Accepted: December 24, 2016 Published: December 27, 2016 of Felodipine after reaction catalyzed by whole-cell A. niger and S. paucimobilis was found as 81.59 and 71.67%, respectively. Immobilization enhanced the enantioselec- Copyright © 2016 by authors and tivity (enantiomeric ratio (E)) of the biocatalysts and hence this led to enhanced en- Scientific Research Publishing Inc. antiomeric purity of the drug. The ee% values were found to be enhanced in reac- This work is licensed under the Creative Commons Attribution International tions catalyzed by A. niger and S. paucimobilis cultures after immobilization as 98.27 License (CC BY 4.0).
    [Show full text]
  • Immobilization of Cellulase for Industrial Production Gordana Hojnik Podrepšek, Mateja Primožiþ, Željko Knez, Maja Habulin*
    A publication of CHEMICAL ENGINEERING TRANSACTIONS The Italian Association VOL. 27, 2012 of Chemical Engineering Online at: www.aidic.it/cet Guest Editors: Enrico Bardone, Alberto Brucato, Tajalli Keshavarz Copyright © 2012, AIDIC Servizi S.r.l., ISBN 978-88-95608-18-1; ISSN 1974-9791 Immobilization of Cellulase for Industrial Production Gordana Hojnik Podrepšek, Mateja Primožiþ, Željko Knez, Maja Habulin* University of Maribor, Faculty of Chemistry and Chemical Engineering, Laboratory for Separation Processes and Product Design, Smetanova ul. 17, 2000 Maribor, Slovenia [email protected] Immobilized enzymes are used in analytical chemistry and as catalysts for the production of chemicals, pharmaceuticals and food. Because of their particular structure, immobilized enzymes require optimal conditions, different from those of soluble enzymes. Particle size, particle-size distribution, mechanical and chemical structure, stability and the catalytic activity, used for immobilization, must be considered. Generally, cellulases are used in various industries, including food, brewery and wine, agriculture, textile, detergent, animal feed, pulp and paper, and in research development. For the industrial application of cellulase, its immobilization, which allows the conditions of repeated use of the enzyme alongside retaining its activity, has been recently investigated. Celullase was immobilized with the use of glutaraldehyde, a covalent cross-linking agent in to cross-linked enzyme aggregates (CLEAs). The stability and activity of cross-linked cellulase, exposed to carbon dioxide under high pressure, were studied. Efficiency of enzyme immobilization was determined using Bradford method (Bradford, 1976). The activity of cross-linked cellulase was determined by spectrophotometric method. 1. Introduction Cellulase, a multicomponent enzyme, consisting of three different enzymes (endocellulase, cellobiohydrolase and ß-glucosidase) is responsible for bioconversion of cellulose into soluble sugar (Zhou et al., 2009).
    [Show full text]
  • Characterization of an Immobilized Amino Acid Racemase for Potential Application in Enantioselective Chromatographic Resolution Processes
    catalysts Article Characterization of an Immobilized Amino Acid Racemase for Potential Application in Enantioselective Chromatographic Resolution Processes Isabel Harriehausen 1,* , Jonas Bollmann 1, Thiane Carneiro 1, Katja Bettenbrock 1 and Andreas Seidel-Morgenstern 1,2,* 1 Department of Physical and Chemical Foundations of Process Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstraße 1, 39106 Magdeburg, Germany; [email protected] (J.B.); [email protected] (T.C.); [email protected] (K.B.) 2 Institute of Chemical Engineering, Otto von Guericke University, 39106 Magdeburg, Germany * Correspondence: [email protected] (I.H.); [email protected] (A.S.-M.); Tel.: +49-6110-401 (A.S.-M.) Abstract: Enantioselective resolution processes can be improved by integration of racemization. Applying environmentally friendly enzymatic racemization under mild conditions is in particular attractive. Owing to the variety of enzymes and the progress in enzyme engineering, suitable racemases can be found for many chiral systems. An amino acid racemase (AAR) from P. putida KT2440 is capable of processing a broad spectrum of amino acids at fast conversion rates. The focus Citation: Harriehausen, I.; Bollmann, of this study is the evaluation of the potential of integrating AAR immobilized on Purolite ECR 8309 J.; Carneiro, T.; Bettenbrock, K.; to racemize L- or D-methionine (Met) within an enantioselective chromatographic resolution process. Seidel-Morgenstern, A. Racemization rates were studied for different temperatures, pH values, and fractions of organic Characterization of an Immobilized co-solvents. The long-term stability of the immobilized enzyme at operating and storage conditions Amino Acid Racemase for Potential ◦ Application in Enantioselective was found to be excellent and recyclability using water with up to 5 vol% ethanol at 20 C could be Chromatographic Resolution demonstrated.
    [Show full text]
  • The Use of Immobilized Enzymes in the Food Industry: a Review
    C R C Critical Reviews in Food Science and Nutrition ISSN: 0099-0248 (Print) (Online) Journal homepage: http://www.tandfonline.com/loi/bfsn19 The use of immobilized enzymes in the food industry: A review Arun Kilara , Khem M. Shahani & Triveni P. Shukla To cite this article: Arun Kilara , Khem M. Shahani & Triveni P. Shukla (1979) The use of immobilized enzymes in the food industry: A review, C R C Critical Reviews in Food Science and Nutrition, 12:2, 161-198, DOI: 10.1080/10408397909527276 To link to this article: https://doi.org/10.1080/10408397909527276 Published online: 29 Sep 2009. Submit your article to this journal Article views: 73 View related articles Citing articles: 24 View citing articles Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=bfsn19 Download by: [Texas A&M University Libraries] Date: 09 January 2018, At: 11:05 December 1979 161 THE USE OF IMMOBILIZED ENZYMES IN THE FOOD INDUSTRY: A REVIEW* Authors: ArunKilara** Khem M. Shahani Department of Food Science and Technology University of Nebraska Lincoln, Nebraska Referee: Triveni P. Shukla Krause Milling Company Milwaukee, Wisconsin INTRODUCTION Enzymes are organic substances produced by living cells which catalyze physiologi- cally significant reactions. Enzymes often are defined as biocatalysts, and they possess greater catalytic activity than chemical catalysts. All known enzymes are protein in nature and are generally colloidal, thermolabile, have relatively high molecular weights, exhibit high degrees of stereochemical and substrate specificities, and can usu- ally be isolated from the living cell. There are a wide variety of enzymes which contrib- ute, in part, to the biological diversity observed in nature.
    [Show full text]
  • Improved Performance of Magnetic Cross-Linked Lipase Aggregates By
    molecules Article Improved Performance of Magnetic Cross-Linked Lipase Aggregates by Interfacial Activation: A Robust and Magnetically Recyclable Biocatalyst for Transesterification of Jatropha Oil Weiwei Zhang 1, Huixia Yang 1, Wanyi Liu 1, Na Wang 2,* and Xiaoqi Yu 2,* 1 School of Chemistry and Chemical Engineering, National Demonstration Center for Experimental Chemistry Education, Ningxia University, Yinchuan 750021, China; [email protected] (W.Z.); [email protected] (H.Y.); [email protected] (W.L.) 2 Key Laboratory of Green Chemistry Technology, Ministry of Education, College of Chemistry, Sichuan University, Chengdu 610064, China * Correspondence: [email protected] (N.W.); [email protected] (X.Y.); Tel.: +86-288-541-5886 (N.W. & X.Y.) Received: 18 November 2017; Accepted: 2 December 2017; Published: 7 December 2017 Abstract: Lipases are the most widely employed enzymes in commercial industries. The catalytic mechanism of most lipases involves a step called “interfacial activation”. As interfacial activation can lead to a significant increase in catalytic activity, it is of profound importance in developing lipase immobilization methods. To obtain a potential biocatalyst for industrial biodiesel production, an effective strategy for enhancement of catalytic activity and stability of immobilized lipase was developed. This was performed through the combination of interfacial activation with hybrid magnetic cross-linked lipase aggregates. This biocatalyst was investigated for the immobilization of lipase from Rhizomucor miehei (RML). Under the optimal conditions, the activity recovery of the surfactant-activated magnetic RML cross-linked enzyme aggregates (CLEAs) was as high as 2058%, with a 20-fold improvement over the free RML. Moreover, the immobilized RML showed excellent catalytic performance for the biodiesel reaction at a yield of 93%, and more importantly, could be easily separated from the reaction mixture by simple magnetic decantation, and retained more than 84% of its initial activities after five instances of reuse.
    [Show full text]
  • Development of Immobilized Biocatalysts for Continuous-Flow
    Budapest University of Technology and Economics Faculty of Chemical Technology and Biotechnology Department of Organic Chemistry and Technology George A. Olah Doctoral School DEVELOPMENT OF IMMOBILIZED BIOCATALYSTS FOR SELECTIVE BIOTRANSFORMATIONS PHD THESIS Emese Abaházi Supervisor: Prof. Dr. László Poppe 2018 2 DECLARATION 3 DECLARATION I, Emese Abaházi hereby declare, that this PhD thesis is my own work and I have used in this thesis only references listed in the list. Any parts that I have taken literally or in the same content but reworded from other sources are clearly indicated. Budapest, 02 September 2018. ……………………………… NYILATKOZAT Alulírott Abaházi Emese kijelentem, hogy ezt a doktori értekezést magam készítettem és abban csak a megadott forrásokat használtam fel. Minden olyan részt, amelyet szó szerint, vagy azonos tartalomban, de átfogalmazva más forrásból átvettem, egyértelműen, a forrás megadásával megjelöltem. Budapest, 2018. szeptember 2. ……………………………… 4 TABLE OF CONTENTS DEVELOPMENT OF IMMOBILIZED BIOCATALYSTS FOR SELECTIVE BIOTRANSFORMATIONS ............ 1 DECLARATION ..................................................................................................................................... 3 NYILATKOZAT ..................................................................................................................................... 3 TABLE OF CONTENTS ........................................................................................................................... 4 ACKNOWLEDGEMENT / KÖSZÖNETNYILVÁNÍTÁS..............................................................................
    [Show full text]