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        <title>Frontiers in Catalysis | Biocatalysis section | New and Recent Articles</title>
        <link>https://www.frontiersin.org/journals/catalysis/sections/biocatalysis</link>
        <description>RSS Feed for Biocatalysis section in the Frontiers in Catalysis journal | New and Recent Articles</description>
        <language>en-us</language>
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        <pubDate>2026-05-14T17:10:33.209+00:00</pubDate>
        <ttl>60</ttl>
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        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2026.1778429</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2026.1778429</link>
        <title><![CDATA[Methyl-coenzyme M reductase in archaeal methanogenesis: evolution, mechanism, and biotechnological perspectives]]></title>
        <pubdate>2026-03-04T00:00:00Z</pubdate>
        <category>Review</category>
        <author>Alberto Vázquez-Salazar</author><author>Ricardo Hernández-Morales</author><author>Edgar Mixcoha</author><author>Ricardo Muñiz-Trejo</author><author>Israel Muñoz-Velasco</author>
        <description><![CDATA[Methanogenesis is an anaerobic, energy-conserving metabolism that converts CO2, acetate, and methylated compounds to methane, constituting a key terminal step in the mineralization of organic matter in many oxygen-limited ecosystems. In the modern biosphere, biological methane formation is overwhelmingly dominated by methanogenic archaea. Despite its phylogenetic constraint, this metabolism exerts disproportionate influence on global carbon cycling, radiative forcing, and methane-based energy systems. The defining biochemical signature of methanogenesis is methyl-coenzyme M reductase (MCR), a nickel tetrapyrrole enzyme that catalyzes the final methane-forming step through Ni-F430 chemistry. This cofactor chemistry enables selective C-H bond formation and cleavage under aqueous, physiologically compatible conditions. It can also function in the reverse direction in anaerobic methane-oxidizing archaea. This review examines archaeal methanogenesis as a case study in how biochemical constraint and evolutionary diversification jointly shape a single catalytic solution across multiple physiological contexts. We summarize the major routes of methane formation and the bioenergetic architectures that support them. We then evaluate competing scenarios for methanogenesis evolution in light of comparative genomics, geochemical constraints, and the rapidly expanding catalogue of methane-cycling archaea, emphasizing an evolutionary history marked by modular assembly, differential loss, and horizontal transfer rather than a simple pattern of vertical inheritance. Mechanistic sections focus on MCR and related alkyl-coenzyme M reductases, highlighting structural features, cofactor variation, post-translational modification repertoires, and recent advances that illuminate critical steps in Ni-F430 biosynthesis and ATP-dependent activation of the Ni(I) catalytic state. Finally, we discuss implications for biotechnology and catalysis, including selective methane abatement by mechanism-guided inhibition, strategies to enhance anaerobic digestion and biological biogas upgrading, and the use of MCR family enzymes as experimentally tractable platforms for selective anaerobic hydrocarbon transformations and bioinspired catalyst development.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2024.1470616</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2024.1470616</link>
        <title><![CDATA[Challenges and perspectives in using unspecific peroxygenases for organic synthesis]]></title>
        <pubdate>2024-10-15T00:00:00Z</pubdate>
        <category>Review</category>
        <author>Yawen Huang</author><author>Jiangtao Sha</author><author>Jie Zhang</author><author>Wuyuan Zhang</author>
        <description><![CDATA[In the past 20 years, unspecific peroxygenases (UPOs) have emerged as promising biocatalysts for various organic transformations. Particularly, we have witnessed great attention being paid to the screening of new enzymes and expansion of the substrates/products. However, challenges such as enzyme stability, low turnover numbers, and substrate specificity hinder their widespread utilization in practical organic synthesis. This review article provides a concrete and mini-overview of the challenges associated with using UPOs in organic synthesis and discusses strategies for enzyme engineering to overcome these limitations. The article highlights recent advancements in UPO research and presents potential solutions to enhance their catalytic efficiency, stability, substrate specificity, and regioselectivity. Additionally, the review outlines the current methodologies employed for directed evolution and protein engineering of UPOs, along with computational modeling approaches for rational enzyme design. By addressing the challenges and exploring avenues for enzyme engineering, this review aims to shed light on the prospects of UPOs in organic synthesis.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2024.1466176</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2024.1466176</link>
        <title><![CDATA[Editorial: Exploiting the power of biocatalysis: accessing optimized natural products analogues]]></title>
        <pubdate>2024-08-06T00:00:00Z</pubdate>
        <category>Editorial</category>
        <author>Mohamed Nagia</author><author>Mohammed N. A. Khalil</author><author>Islam El-Awaad</author>
        <description></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2024.1360129</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2024.1360129</link>
        <title><![CDATA[Adapting an acyl CoA ligase from Metallosphaera sedula for lactam formation by structure-guided protein engineering]]></title>
        <pubdate>2024-03-04T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Nikolas Capra</author><author>Chloé Lelièvre</author><author>Océane Touré</author><author>Aurélie Fossey-Jouenne</author><author>Carine Vergne-Vaxelaire</author><author>Dick B. Janssen</author><author>Andy-Mark W. H. Thunnissen</author><author>Anne Zaparucha</author>
        <description><![CDATA[The CoA ligase from Metallosphaera sedula (MsACL) can be used for the chemoenzymatic synthesis of amides from carboxylic acids. In this CoA-independent conversion, the enzyme catalyzes the adenylation of a carboxylic acid with the help of ATP, followed by the uncatalyzed cleavage of acyl-AMP by a nucleophilic amine to yield an amide. With ω-amino acids as substrates this reaction may result in formation of lactams, but unfortunately the substrate preference of the wild-type enzyme is rather limited. To allow structure-based protein engineering and expand the substrate scope of the enzyme, crystal structures of MsACL were solved in the thioesterification conformational state with AMP, CoA and with the reaction intermediate acetyl-AMP bound in the active site. Using substrate docking and by comparing the crystals structures and sequence of MsACL to those of related CoA ligases, mutations were predicted which increase the affinity in the carboxylic acid binding pocket for ω-amino acids. The resulting mutations transformed a non-active enzyme into an active enzyme for ε-caprolactam synthesis, highlighting the potential of the thermophilic CoA ligase for this synthetic and biotechnologically relevant reaction.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2024.1360702</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2024.1360702</link>
        <title><![CDATA[Novel concepts for the biocatalytic synthesis of second-generation biodiesel]]></title>
        <pubdate>2024-02-16T00:00:00Z</pubdate>
        <category>Review</category>
        <author>Androniki Spanou</author><author>Alexandra Moschona</author><author>Eleni Theodosiou</author><author>Sotiris I. Patsios</author><author>Ioannis V. Pavlidis</author>
        <description><![CDATA[Biodiesel is synthesized by the transesterification of triglycerides of oils with short-chain alcohols, such as methanol and ethanol. According to the Renewable Energy Directive guidelines (RED II 2018/2001/EU) the contribution of advanced biofuels, which do not include edible oils, towards the overall EU target, is at 1% in 2025 and at least 3.5% in 2030. Bioprocesses that valorize non-edible oils for the production of second-generation biodiesel could play a critical role in achieving this goal. Immobilized lipases, as well as other enzyme classes, such as cutinases and acyltransferases, are utilized as biocatalysts for this process. For the sustainability of the process, renewable materials can be used as immobilization matrices, or even enzymes anchored on the cells as whole-cell biocatalysts. Membrane reactors can also be employed to facilitate the enzymatic transesterification by conducting a continuous enzymatic reaction and simultaneously separate the products in a single operation. The advances on the aforementioned fast-pacing fields are presented in this work.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2023.1285074</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2023.1285074</link>
        <title><![CDATA[Biocatalytic route scouting and enzyme screening toward the synthesis of α-benzyl L-glutamate]]></title>
        <pubdate>2024-01-31T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Peter J. L. M. Quaedflieg</author><author>Lisanne M. H. Jente</author><author>Monika Müller</author><author>Linda Vermote</author><author>Victor Plesciuc</author><author>Jan-Metske van der Laan</author><author>Lone Nielsen</author><author>Martin Schürmann</author>
        <description><![CDATA[We here report four biocatalytic approaches for the synthesis of the protected amino acid building block α-benzyl L-glutamate. Screenings of these routes to identify active and selective enzymes were conducted, and major hits were confirmed in retest reactions. In the first approach, N-Boc L-glutamic acid is mono-benzylesterified by the protease Alcalase with 81% yield; and in the other three approaches, a biocatalytic γ-selective hydrolysis of α,γ-dibenzyl L-glutamate, a selective amide hydrolysis of α-benzyl L-glutamine, and a selective lactam hydrolysis of alpha-benzyl L-pyroglutamate is performed with up to 71% yield.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2023.1275281</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2023.1275281</link>
        <title><![CDATA[The chemoenzymatic synthesis of glycan-terminated oligo(Leu)x]]></title>
        <pubdate>2024-01-24T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Sarah P. Black</author><author>Melinda Liu</author><author>Cesar Castillo</author><author>Wynne Coradeli</author><author>Filbert Totsingan</author><author>Cody B. Edson</author><author>Sagar D. Khare</author><author>Richard A. Gross</author>
        <description><![CDATA[Introduction: Glycopeptides contain carbohydrate moieties (glycans) covalently attached to the side chain and/or terminal peptide units. Since glycans are present on cell surfaces, these constructs can potentially address a wide array of therapeutic functions. To overcome the deficiencies associated with current synthetic routes to glycopeptides, such as costly processes and toxic reagents, this work aimed to develop versatile environmentally friendly protease-catalyzed peptide synthesis routes to peptides decorated with a glycan at their N-terminus.Methods: “Grafters” were first synthesized that consist of a glycan conjugated directly, or through a spacer, to the amine group of L-Phe-ethyl ester (Phe-OEt). The role of Phe-OEt is to increase the conjugate’s recognition by the protease (papain) catalytic active site. A series of grafters were synthesized with variation of the glycan structure, linkage-chemistry, and presence of an oligo (ethylene glycol) “spacer” of varied length between the glycan and Phe-OEt moiety. High grafter efficiency will result by the successful acceptance of the grafter at the enzymes S1/S2 subsites, formation of an acyl enzyme complex and subsequent conversion to glycan-terminated oligo(Leu)x (x ≥ 1), as opposed to construction of non-glycan N-terminated oligo(Leu)x.Results and discussion: While glycan-Phe-OEt grafters without a spacer between the glycan and Phe-OEt resulted in low grafter efficiency (8.3% ± 2.0%), insertion of a short oligo (ethylene glycol) spacer between the glycan and Phe-OEt moieties (glycan-PEGn-Phe-OEt, n ≥ 3) increased the grafter efficiency by 3-fold–24.5% ± 1.8%. In addition, computational modeling was performed using Rosetta software provided insights on a molecular level of how grafter efficiency is influenced by the PEG spacer length.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2023.1231765</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2023.1231765</link>
        <title><![CDATA[Expression and characterization of PrnC—a flavin-dependent halogenase from the pyrrolnitrin biosynthetic pathway of Pseudomonas protegens Pf-5]]></title>
        <pubdate>2023-07-26T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Jan Gebauer</author><author>Jörg Pietruszka</author><author>Thomas Classen</author>
        <description><![CDATA[Introduction: The antimicrobial pyrrolnitrin from Pseudomonas strains is formed in four steps from tryptophan and comprises two flavin-dependent halogenases. Both PrnC and PrnA can carry out regioselective chlorination and bromination and are carrier protein-independent. Whilst the tryptophan halogenase PrnA has been studied in detail in the past, this study focuses on the pyrrole halogenating enzyme PrnC.Methods: The halogenating enzyme PrnC, as well as the essential electron suppliers, the flavin reductases, have been produced soluble in E. coli. Furthermore, a screening of a rational compound library revealed that the pyrrole is essential for substrate recognition; however, the substitution pattern of the benzene ring is not limiting the catalysis.Results and discussion: This renders PrnC to be a synthetically valuable enzyme for the synthesis of pyrrolnitrin congeners. For its natural substrate monodechloroaminopyrrolnitrin (MDA), the KM value was determined as 14.4 ± 1.2 µM and a kcat of 1.66 ± 0.02 min−1, which is comparable to other halogenases.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2023.1184959</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2023.1184959</link>
        <title><![CDATA[Exploring the selectivity and engineering potential of an NRPS condensation domain involved in the biosynthesis of the thermophilic siderophore fuscachelin]]></title>
        <pubdate>2023-06-16T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Y. T. Candace Ho</author><author>Thierry Izoré</author><author>Joe A. Kaczmarski</author><author>Edward Marschall</author><author>Minuri S. Ratnayake</author><author>Julien Tailhades</author><author>David L. Steer</author><author>Ralf B. Schittenhelm</author><author>Manuela Tosin</author><author>Colin J. Jackson</author><author>Max J. Cryle</author>
        <description><![CDATA[In nonribosomal peptide synthesis, condensation (C) domains are key catalytic domains that most commonly link carrier protein bound substrates to form peptides or depsipeptides. While adenylation domains have been well characterized due to their role in the selection of monomers and hence as gate keepers in nonribosomal peptide biosynthesis, C-domains have been the subject of debate as they do not have apparent “A-domain like” side chain selectivity for their acceptor substrates. To probe the selectivity and specificity of C-domains, here we report our biochemical and structural characterization of the C3-domain from the biosynthesis of the siderophore fusachelin. Our results show that this C-domain is not broadly flexible for monomers bearing significantly alternated side chains or backbones, which suggests there can be a need to consider C-domain specificity for acceptor substrates when undertaking NRPS engineering.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2023.1178345</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2023.1178345</link>
        <title><![CDATA[Structure and mutation of deoxypodophyllotoxin synthase (DPS) from Podophyllum hexandrum]]></title>
        <pubdate>2023-05-24T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Zoe Ingold</author><author>Gideon Grogan</author><author>Benjamin R. Lichman</author>
        <description><![CDATA[Deoxypodophyllotoxin synthase (DPS) is a 2-oxoglutarate (2-OG) dependent non-heme iron (II) dioxygenase that catalyzes the stereoselective ring-closing carbon-carbon bond formation of deoxypodophyllotoxin from the aryllignan (−)-yatein. Deoxypodophyllotoxin is a precursor of topoisomerase II inhibitors, which are on the World Health Organization’s list of essential medicines. Previous work has shown that DPS can accept a range of substrates, indicating it has potential in biocatalytic processes for the formation of diverse polycyclic aryllignans. Recent X-ray structures of the enzyme reveal possible roles for amino acid side chains in substrate recognition and mechanism, although a mutational analysis of DPS was not performed. Here, we present a structure of DPS at an improved resolution of 1.41 Å, in complex with the buffer molecule, Tris, coordinated to the active site iron atom. The structure has informed a mutational analysis of DPS, which suggests a role for a D224-K187 salt bridge in maintaining substrate interactions and a catalytic role for H165, perhaps as the base for the proton abstraction at the final rearomatization step. This work improves our understanding of specific residues’ contributions to the DPS mechanism and can inform future engineering of the enzyme mechanism and substrate scope for the development of a versatile biocatalyst.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2023.1147205</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2023.1147205</link>
        <title><![CDATA[Sustainable synthesis of L-phenylalanine derivatives in continuous flow by immobilized phenylalanine ammonia lyase]]></title>
        <pubdate>2023-05-04T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>David Roura Padrosa</author><author>Hansjoerg Lehmann</author><author>Radka Snajdrova</author><author>Francesca Paradisi</author>
        <description><![CDATA[The application of phenylalanine ammonia lyases (PALs) for the amination of a variety of cinnamic acids has been shown to be a cost-efficient method to produce a variety of phenylalanine analogues. Nonetheless, as many other biocatalytic tools, the process intensification, especially due to the high equivalents of ammonia needed, and the cost-efficiency of the catalyst production and use have been key points to further prove their usefulness. Here, we investigated the use of previously characterized PALs (AvPAL and PbPAL) for the amination of a series of substituted cinnamic acids. To enhance the process scalability and the reusability of the catalyst, we investigated the use of covalent immobilization onto commercially available supports, creating a heterogeneous catalyst with good recovered activity (50%) and excellent stability. The immobilized enzyme was also incorporated in continuous flow for the synthesis of 3-methoxy-phenyl alanine and 4-nitro-phenylalanine, which allowed for shorter reaction times (20 min of contact time) and excellent conversions (88% ± 4% and 89% ± 5%) respectively, which could be maintained over extended period of time, up to 24 h. This work exemplifies the advantages that the combination of enzyme catalysis with flow technologies can have not only in the reaction kinetics, but also in the productivity, catalyst reusability and downstream processing.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2023.1154452</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2023.1154452</link>
        <title><![CDATA[What’s new in flow biocatalysis? A snapshot of 2020–2022]]></title>
        <pubdate>2023-04-21T00:00:00Z</pubdate>
        <category>Mini Review</category>
        <author>Michele Crotti</author><author>Marina S. Robescu</author><author>Juan M. Bolivar</author><author>Daniela Ubiali</author><author>Lorena Wilson</author><author>Martina L. Contente</author>
        <description><![CDATA[Flow biocatalysis is a key enabling technology that is increasingly being applied to a wide array of reactions with the aim of achieving process intensification, better control of biotransformations, and minimization of waste stream. In this mini-review, selected applications of flow biocatalysis to the preparation of food ingredients, APIs and fat- and oil-derived commodity chemicals, covering the period 2020-2022, are described.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2023.1165079</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2023.1165079</link>
        <title><![CDATA[Impact of sunlight irradiation on CvFAP photodecarboxylation]]></title>
        <pubdate>2023-04-13T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Alexandre S. França</author><author>Gabriela C. Breda</author><author>Kleber T. De Oliveira</author><author>Rodrigo V. Almeida</author><author>Frank Hollmann</author><author>Rodrigo O. M. A. De Souza</author>
        <description><![CDATA[A visible-light-driven photocatalytic decarboxylation of palmitic acid and related fatty acids is described in this study. Remarkable decarboxylation rates have been observed with full conversion in less than 20 min. In this study, we have demonstrated that sunlight irradiation, even on cloudy days, can deliver similar results to traditional LED lamps while using much less energy and minimizing environmental impact. The findings indicate that the process of enzymatic decarboxylation could be useful for the production of different biofuels in the future.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2023.1114536</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2023.1114536</link>
        <title><![CDATA[Preparation of (3R)-quinuclidinol using heterogeneous biocatalytic hydrogenation in a dynamically-mixed continuous flow reactor]]></title>
        <pubdate>2023-03-08T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Sarah E. Cleary</author><author>Sofia Kazantzi</author><author>Joshua A. Trenchard</author><author>Martin Monedero</author><author>Jack W. Allman</author><author>Tara C. Lurshay</author><author>Xu Zhao</author><author>Michael B. C. Kenny</author><author>Holly A. Reeve</author>
        <description><![CDATA[Introduction: This work was carried out to understand if a heterogeneous biocatalytic hydrogenation system could be implemented in a slurry mode continuous flow reactor, as a “slot-in” alternative to a metal/carbon catalyst with minimal process or chemical engineering development.Method: Biocatalytic hydrogenation was compared to metal (Pd/C) catalysed hydrogenation for ketone to chiral alcohol conversion in both a continuous flow reactor (Coflore ACR, AM Technology) and analogous scaled down batch reactions.Results and discussion: Initial results demonstrated that batch reactions can achieve high conversions in 30 min, with relatively low biocatalyst loadings, meeting critical criteria for operation as continuous slurry mode process. Further results demonstrated full conversion of quinuclidinone to (3R)-quinuclidinol in continuous flow under mild conditions (35°C, 2 bar H2). On intensification of the process to higher substrate loading (50 mM), conversion was similar to with Pd/C, however the biocatalytic system achieved far higher turnover frequency and total turnover number (65 min−1 and 20,000, respectively) than the metal system (0.16 min−1, 37). Comparison to an analogous batch reaction highlights that the biocatalytic system has promise for further optimisation and intensification in the scalable Coflore ACR. Overall, biocatalytic hydrogenation is shown to offer a decarbonised biocatalytic route and a “slot-in” replacement to metal catalysts for hydrogenation reactions in continuous flow reactors.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2023.1112154</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2023.1112154</link>
        <title><![CDATA[Co-expression of thermophilic pectinases in a single host for cost-effective pectin bioconversion into D-galacturonic acid]]></title>
        <pubdate>2023-02-16T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Carol N. Flores-Fernández</author><author>Max Cárdenas-Fernández</author><author>Gary J. Lye</author><author>John M. Ward</author>
        <description><![CDATA[Co-expression of enzymes allow to produce multiple enzymes in a single host, representing a cost-effective alternative in biocatalytic processes which can be used for pectin bioconversion. Pectin-rich biomass is an abundant by-product from the fruit and sugar industries that is usually disposed in landfill or sold as a low value feedstock. The aim of this work was to co-express a thermophilic pectin methyl esterase (PME) and exo-polygalacturonases (exo-PGs) in a single host for pectin bioconversion into D-galacturonic acid (GalA) using different pectic substrates such as apple, citrus and sugar beet pectin. To achieve this, a PME from Bacillus licheniformis (BLI09) with either an exo-PG from Thermotoga maritima (TMA01) or from Bacillus licheniformis (BLI04) were cloned in pETDuet-1 and co-expressed in E. coli BL21 (DE3). Four co-expression plasmids containing both pectinases were constructed and factors such as the effect of the genes’ cloning order and their expression were evaluated. Co-expression constructs 3 and 4 (pETDuet-TMA01-BLI09 and pETDuet-BLI04-BLI09, respectively) showed better expression of both pectinases compared to co-expression constructs 1 and 2 (pETDuet-BLI09-TMA01 and pETDuet-BLI09-BLI04, respectively). Co-expression constructs 3 and 4 were the most efficient for pectin bioconversion into GalA reaching 3 and 2.5 mM GalA, respectively from apple and citrus pectin after 4 h reaction. In conclusion, this work demonstrates that the co-expression of pectinases can potentially contribute to reduce the cost associated to their production and purification as well as to increase their applicability for exploiting pectin-rich biomass to obtain bio-based chemicals.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2023.1105948</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2023.1105948</link>
        <title><![CDATA[Native amine dehydrogenases can catalyze the direct reduction of carbonyl compounds to alcohols in the absence of ammonia]]></title>
        <pubdate>2023-01-16T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Aurélie Fossey-Jouenne</author><author>Laurine Ducrot</author><author>Ewald P. J. Jongkind</author><author>Eddy Elisée</author><author>Anne Zaparucha</author><author>Gideon Grogan</author><author>Caroline E. Paul</author><author>Carine Vergne-Vaxelaire</author>
        <description><![CDATA[Native amine dehydrogenases (nat-AmDHs) catalyze the (S)-stereoselective reductive amination of various ketones and aldehydes in the presence of high concentrations of ammonia. Based on the structure of CfusAmDH from Cystobacter fuscus complexed with Nicotinamide adenine dinucleotide phosphate (NADP+) and cyclohexylamine, we previously hypothesized a mechanism involving the attack at the electrophilic carbon of the carbonyl by ammonia followed by delivery of the hydride from the reduced nicotinamide cofactor on the re-face of the prochiral ketone. The direct reduction of carbonyl substrates into the corresponding alcohols requires a similar active site architecture and was previously reported as a minor side reaction of some native amine dehydrogenases and variants. Here we describe the ketoreductase (KRED) activity of a set of native amine dehydrogenases and variants, which proved to be significant in the absence of ammonia in the reaction medium but negligible in its presence. Conducting this study on a large set of substrates revealed the heterogeneity of this secondary ketoreductase activity, which was dependent upon the enzyme/substrate pairs considered. In silico docking experiments permitted the identification of some relationships between ketoreductase activity and the structural features of the enzymes. Kinetic studies of MsmeAmDH highlighted the superior performance of this native amine dehydrogenases as a ketoreductase but also its very low activity towards the reverse reaction of alcohol oxidation.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2022.1089176</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2022.1089176</link>
        <title><![CDATA[Harnessing selenocysteine to enhance microbial cell factories for hydrogen production]]></title>
        <pubdate>2022-12-22T00:00:00Z</pubdate>
        <category>Perspective</category>
        <author>Armaan Patel</author><author>David W. Mulder</author><author>Dieter Söll</author><author>Natalie Krahn</author>
        <description><![CDATA[Hydrogen is a clean, renewable energy source, that when combined with oxygen, produces heat and electricity with only water vapor as a biproduct. Furthermore, it has the highest energy content by weight of all known fuels. As a result, various strategies have engineered methods to produce hydrogen efficiently and in quantities that are of interest to the economy. To approach the notion of producing hydrogen from a biological perspective, we take our attention to hydrogenases which are naturally produced in microbes. These organisms have the machinery to produce hydrogen, which when cleverly engineered, could be useful in cell factories resulting in large production of hydrogen. Not all hydrogenases are efficient at hydrogen production, and those that are, tend to be oxygen sensitive. Therefore, we provide a new perspective on introducing selenocysteine, a highly reactive proteinogenic amino acid, as a strategy towards engineering hydrogenases with enhanced hydrogen production, or increased oxygen tolerance.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2022.1021691</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2022.1021691</link>
        <title><![CDATA[Flavofun: Exploration of fungal flavoproteomes]]></title>
        <pubdate>2022-12-14T00:00:00Z</pubdate>
        <category>Review</category>
        <author>Bianca Kerschbaumer</author><author>Aleksandar Bijelic</author><author>Peter Macheroux</author>
        <description><![CDATA[Fungi produce a plethora of natural products exhibiting a fascinating diversity of chemical structures with an enormous potential for medical applications. Despite the importance of understanding the scope of natural products and their biosynthetic pathways, a systematic analysis of the involved enzymes has not been undertaken. In our previous studies, we examined the flavoprotein encoding gene pool in archaea, eubacteria, the yeast Saccharomyces cerevisiae, Arabidopsis thaliana, and Homo sapiens. In the present survey, we have selected the model fungus Neurospora crassa as a starting point to investigate the flavoproteomes in the fungal kingdom. Our analysis showed that N. crassa harbors 201 flavoprotein-encoding genes amounting to 2% of the total protein-encoding genome. The majority of these flavoproteins (133) could be assigned to primary metabolism, termed the “core flavoproteome”, with the remainder of flavoproteins (68) serving in, as yet unidentified, reactions. The latter group of “accessory flavoproteins” is dominated by monooxygenases, berberine bridge enzyme-like enzymes, and glucose-methanol-choline-oxidoreductases. Although the exact biochemical role of most of these enzymes remains undetermined, we propose that they are involved in activities closely associated with fungi, such as the degradation of lignocellulose, the biosynthesis of natural products, and the detoxification of harmful compounds in the environment. Based on this assumption, we have analyzed the accessory flavoproteomes in the fungal kingdom using the MycoCosm database. This revealed large differences among fungal divisions, with Ascomycota, Basidiomycota, and Mucoromycota featuring the highest average number of genes encoding accessory flavoproteins. Moreover, a more detailed analysis showed a massive accumulation of accessory flavoproteins in Sordariomycetes, Agaricomycetes, and Glomeromycotina. In our view, this indicates that these fungal classes are proliferative producers of natural products and also interesting sources for flavoproteins with potentially useful catalytic properties in biocatalytic applications.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2022.1049179</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2022.1049179</link>
        <title><![CDATA[Protein engineering of amine transaminases]]></title>
        <pubdate>2022-11-22T00:00:00Z</pubdate>
        <category>Review</category>
        <author>Qinglong Meng</author><author>Carlos Ramírez-Palacios</author><author>Hein J. Wijma</author><author>Dick B. Janssen</author>
        <description><![CDATA[Protein engineering is a powerful and widely applied tool for tailoring enzyme properties to meet application-specific requirements. An attractive group of biocatalysts are PLP-dependent amine transaminases which are capable of converting prochiral ketones to the corresponding chiral amines by asymmetric catalysis. The enzymes often display high enantioselectivity and accept various amine donors. Practical applications of these amine transaminases can be hampered by enzyme instability and by their limited substrate scope. Various strategies to improve robustness of amine transaminases and to redirect their substrate specificity have been explored, including directed evolution, rational design and computation-supported engineering. The approaches used and results obtained are reviewed in this paper, showing that different strategies can be used in a complementary manner and can expand the applicability of amine transaminases in biocatalysis.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/fctls.2022.1032088</guid>
        <link>https://www.frontiersin.org/articles/10.3389/fctls.2022.1032088</link>
        <title><![CDATA[Key technology to non-aqueous and multi-step biocatalysis: Pickering emulsions]]></title>
        <pubdate>2022-10-14T00:00:00Z</pubdate>
        <category>Perspective</category>
        <author>Marion B. Ansorge-Schumacher</author><author>Christoph Plikat</author>
        <description><![CDATA[Considering the importance of biocatalysis in chemical synthesis, technologies allowing full exploitation of its potential are urgently wanted. Eleven years ago, our team proposed Pickering emulsions as a concept to overcome the severe restrictions set by the general requirement for the presence of water. In this brief perspective, we demonstrate that the insights into bioactive Pickering emulsions gathered meanwhile strongly designate it a key technology to non-aqueous and multi-step biocatalysis. Mainly, this relates to the extensive compatibility of this system with different solvents, materials, biocatalysts, reactions and demands on productive use. We here give a brief overview of the most relevant details, including recent results from our own research.]]></description>
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