Amine Donor Research Articles

Electrocatalytic Nitrite Reduction in Neutral Water with Ni(II) and Co(II) Macrocycle Complexes: Catalytic Evaluation and Mechanistic Elucidation

AbstractNitrite anions, of anthropogenic origin, in the environment disrupt the nitrogen cycle making the electrocatalytic reduction of nitrite a significant objective. Homogeneous Ni(II) and Co(II) complexes bearing a macrocyclic supporting ligand consisting of a tridentate redox active bis(imino)pyridine moiety coupled with a tertiary amine donor site are effective and selective for the electrocatalytic reduction of nitrite to ammonium ion and hydroxylamine in buffered (pH 7) aqueous solutions. Controlled potential coulometry at potentials from −0.98 to −1.05 V vs Ag/AgCl in 4‐morpholinepropanesulfonic acid (MOPS) buffer yielded ammonium as the major product with Faradaic efficiencies ranging from 76–90 %. A foot‐of‐the‐wave analysis yielded calculated turn‐over frequencies of 61 s−1and 28 s−1 for the Ni and Co complexes, respectively. The Ni complex displayed a higher selectivity for NH4+/NH2OH than the Co analog. A computational examination of the catalytic mechanism of the Ni complex was used to support and elucidate the proposed chemical steps, provide some energetic details of the electron and proton transfers, and present a rationale for the selectivity of this reduction.

Biotransamination with racemic amines as amine donors: kill three birds with one stone through a dual-enzyme cascade

A novel dual-enzyme cascade process utilizing carbonyl reductase and ω-transaminase enzyme was developed to remove by-products via the asymmetric reduction and increase product formation, making it ideal for both industrial and academic applications.

Biocatalytic reductive aminations with NAD(P)H-dependent enzymes: enzyme discovery, engineering and synthetic applications.

Chiral amines are pivotal building blocks for the pharmaceutical industry. Asymmetric reductive amination is one of the most efficient and atom economic methodologies for the synthesis of optically active amines. Among the various strategies available, NAD(P)H-dependent amine dehydrogenases (AmDHs) and imine reductases (IREDs) are robust enzymes that are available from various sources and capable of utilizing a broad range of substrates with high activities and stereoselectivities. AmDHs and IREDs operate via similar mechanisms, both involving a carbinolamine intermediate followed by hydride transfer from the co-factor. In addition, both groups catalyze the formation of primary and secondary amines utilizing both organic and inorganic amine donors. In this review, we discuss advances in developing AmDHs and IREDs as biocatalysts and focus on evolutionary history, substrate scope and applications of the enzymes to provide an outlook on emerging industrial biotechnologies of chiral amine production.

Antibacterial properties of enzymatically treated PET fibers functionalized by nitric oxide

At present, microbes have enormous potential to become a major global public health issue. For example, Escherichia coli is the prominent cause of cholecystitis, urinary tract infections, and other infections. Due to its outstanding antibacterial properties, nitric oxide (NO) is essential for biological processes. Additionally, enzymatic hydrolysis using polyethylene terephthalate hydrolase (PETase) is one of the promising methods for PET upcycling. First, recombinant PETase was used to enzymatically treat waste PET fibers, and polyethylenimine (PEI) was added as a secondary amine donor. Subsequently, the aminated PET fiber was inserted into a reactor charged with NO gas (10 atm, 3 days) to obtain N-diazeniumdiolate (NONOate) products that can inhibit bacteria growth. In this study, the first strategy for antibacterial applications by NO-releasing PETase-hydrolyzed PET fibers was demonstrated. NO-conjugated PET fibers were successfully prepared which exhibits a continuous NO release profile over 12 h. The surface properties of functionalized PET fibers were successfully confirmed by fourier-transform infrared spectroscopy (FTIR), scanning electron microscope (SEM), and Griess assay. The antibacterial test indicated a reduction of Escherichia coli by 90.2% and Staphylococcus aureus by 71.1% after exposure to the functionalized material. Therefore, this novel antibacterial agent may offer great potential applications in the medical field.

NIR-II emissive donor-acceptor-donor fluorophores for dual fluorescence bioimaging and photothermal therapy applications.

Fluorescence bioimaging with near-infrared II (NIR-II) emissive organic fluorophores has proven to be a viable noninvasive diagnostic technique. However, there is still the need for the development of fluorophores that possess increased stability as well as functionalities that impart stimuli responsiveness. Through strategic design, we can synthesize fluorophores that possess not only NIR-II optical profiles but also pH-sensitivity and the ability to generate heat upon irradiation. In this work, we employ a donor-acceptor-donor (D-A-D) design to synthesize a series of NIR-II fluorophores. Here we use thienothiadiazole (TTD) as the acceptor, 3-hexylthiophene (HexT) as the π-spacer and vary the alkyl amine donor units: N,N-dimethylaniline (DMA), phenylpiperidine (Pip), and phenylmorpholine (Morp). Spectroscopic analysis shows that all three derivatives exhibit emission in the NIR-II region with λemimax ranging from 1030 to 1075 nm. Upon irradiation, the fluorophores exhibited noticeable heat generation through non-radiative processes. The ability to generate heat indicates that these fluorophores will act as theranostic (combination therapeutic and diagnostic) agents in which simultaneous visualization and treatment can be performed. Additionally, biosensing capabilities were supported by changes in the absorbance properties while under acidic conditions as a result of protonation of the alkyl amine donor units. The fluorophores also show minimal toxicity in a human mammary cell line and with murine red blood cells. Overall, initial results indicate viable NIR-II materials for multiple biomedical applications.

Conversion of amino-terephthalonitriles to multi-substituted single benzene fluorophores with utility in bioimaging.

Substitution of two fluorine atoms of the tetrafluoroterephthalonitrile (TFTN) ring (ortho to each other) by amine nucleophiles through SNAr chemistry is achievable. However, tri- and tetra-substitution towards multi-substituted single benzene fluorophores (SBFs) is harder due to increased electron richness of the TFTN moiety. Tertiary amine donors promote the molecule towards such multi-substitution guided by the steric obstruction to intramolecular charge transfer to the TFTN ring. Contrarily, secondary amine substituents with better lone pair donation to the TFTN ring cannot induce the SNAr pathway and instead promote hydrolysis of the nitrile groups of the TFTN moiety. Theoretical investigations have helped unearth the reasons for this observed difference in chemical reactivities and also explain the differences in the emission spectra. Finally, the success of the synthetic method towards multi-substitution is showcased through creation of a highly lipophilic SBF bearing an octyl unit and demonstrating its utility in in vitro cellular imaging.

The influence of hindered rotation on electron transfer and exchange interaction in triarylamine-triptycene-perylene diimide triads.

Stretched electron-donor-bridge-acceptor triads that exhibit intramolecular twisting degrees of freedom are capable of modulating exchange interaction (J) as well as electronic couplings through variable π-overlap at the linear bond links, affecting the rate constants of photoinduced charge separation and recombination. Here we present an in-depth investigation of such effects induced by methyl substituents leading to controlled steric hindrance of intramolecular twisting around biaryl axes. Starting from the parent structure, consisting of a triphenyl amine donor, a triptycene (TTC) bridge and a phenylene-perylene diimide acceptor (Me0), one of the two phenylene linkers attached to the TTC was ortho-substituted by two methyl groups (Me2, Me3), or both such phenylene linkers by two pairs of methyl groups (Me23). Photoinduced charge separation (kCS) leading to a charge-separated (CS) state was studied by fs-laser spectroscopy, charge recombination to either singlet ground state (kS) or to the first excited local triplet state of the acceptor (kT) by ns-laser spectroscopy, whereby kinetic magnetic field effects in an external magnetic field were recorded and analysed using quantum dynamic simulations of the spin dependent kinetics of the CS state. Kinetic spectra of the initial first order rate constants of charge recombination (k(B)) exhibited characteristic J-resonances progressing to lower fields in the series Me0, Me2, Me3, Me23. From the quantum simulations, the values of the parameters J, kS, kT and kSTD, the singlet/triplet dephasing constant, were obtained. They were analysed in terms of molecular dynamics simulations of the intramolecular twisting dynamics based on potentials calculated by density functional theory. Apart from kT, all of the parameters exhibit a clear correlation with the averaged cosine square products of the biaryl angles.

Synthetic Advances for Mechanistic Insights: Metal-Oxygen Intermediates with a Macrocyclic Pyridinophane System.

ConspectusMetalloenzymes, which are proteins containing earth-abundant transition-metal ions as cofactors in the active site, generate various metal-oxygen intermediates via activating a dioxygen molecule (O2) to mediate vital metabolic functions, such as the oxidative metabolism of xenobiotics and the biotransformation of naturally occurring molecules. By replicating the active sites of metalloenzymes, many bioinorganic chemists have studied the geometric and electronic properties and reactivities of model complexes to understand the nature of enzymatic intermediates and develop bioinspired metal catalysts. Among the reported model complexes, nonporphyrinic macrocyclic ligands are the predominant coordination system widely used in stabilizing and isolating diverse metal-oxygen intermediates, which allows us to extensively investigate the physicochemical characteristics of the analogs of reactive intermediates of metalloenzymes. In particular, it has been reported that the ring size of the macrocyclic ligands, defined by the number of atoms in the macrocyclic ring, drastically affects the identity of the metal-oxygen intermediate. Thus, systematic modification of the macrocyclic ligands has been a great subject being examined in various inorganic fields.In this Account, we describe synthetic advances of a macrocyclic ligand system by introducing pyridine donors into a 12-membered tetraazamacrocyclic ligand (12-TMC) that initially has 4 amine donors. Interestingly, the backbone of the pyridinophane ligand with 2 pyridine and 2 amine donors in a 12-membered ring is shown to be much more folded than in other macrocyclic ligands, thereby allowing the axial and equatorial donors to separately control the electronic structure of metal complexes. Then, we looked over independent electronic and steric effects on metal-oxygen species with thorough physicochemical analysis. The NiIII-peroxo complexes exhibit nucleophilic reactivity dependent on the steric hindrance of the second coordination sphere. Furthermore, the C-H bond strength of the second coordination sphere has also been an important factor in determining the stability of MnIV-bis(hydroxo) intermediates. Electronic tuning on CoIII-hydroperoxo intermediates results in a trend between the electron-donating abilities of para-substituents on pyridine in the pyridinophane ligand and electrophilic reactivities, from which mechanistic insights into the metal-hydroperoxo species have been gained. Importantly, the metal-oxygen intermediates supported by the pyridinophane ligand system have revealed quite challenging chemical reactions, including dioxygenase-like nitrile activation by CoIII-peroxo intermediates and the oxidation of aldehyde and aromatic compounds by manganese-oxygen intermediates. Based on the fine substitution of donors, we have addressed that those novel reactions originated from the unique framework of the pyridinophane system incorporating spin-crossover behavior and high redox potentials of the metal-oxygen intermediates. These results will be valuable for the structure-activity relationship of metal-oxygen intermediates, giving a better understanding on the enzymatic coordination system where amino acid ligands vary for specific chemical reactions.

Aluminum complexes of a hemilabile pyrrolylaldiminate ligand

AbstractOverviewThis study describes the synthesis, characterization, and reactivity of aluminum complexes containing the amine‐functionalized pyrrolylaldiminate ligand N‐(2‐dimethylaminoethyl)‐5‐tert‐butyl‐pyrrol‐2‐ylaldiminate ([PyImN]−).SynthesisThe condensation reaction of N,N‐dimethylethylenediamine with 5‐tert‐butylpyrrole‐2‐carbaldehyde in methanol at room temperature produces H[PyImN]. Alkane elimination reactions of MeAlCl2 with H[PyImN] in toluene at 110°C and AlR3 (R = Me, Et) with H[PyImN] in toluene at −35°C afford [PyImN]AlCl2 and [PyImN]AlR2, respectively.CharacterizationSolution NMR studies reveal the coordination preferences of [PyImN]—whose hemilabile amine donor is bound to the aluminum center in [PyImN]AlCl2 but unbound in [PyImN]AlR2 (R = Me, Et).CatalysisThe catalytic activities of [PyImN]AlMe2 in controlled ring‐opening polymerization of ε‐caprolactone are discussed.

Mechanistic Insights into Nitrile Activation by Cobalt(III)-Hydroperoxo Intermediates: The Influence of Ligand Basicity.

The versatile applications of nitrile have led to the widespread use of nitrile activation in the synthesis of pharmacologically and industrially valuable compounds. We reported the activation of nitriles using mononuclear cobalt(III)-hydroperoxo complexes, [CoIII(Me3-TPADP)(O2H)(RCN)]2+ [R = Me (2) and Ph (2Ph)], to form cobalt(III)-peroxyimidato complexes, [CoIII(Me3-TPADP)(R-C(=NH)O2)]2+ [R = Me (3) and Ph (3Ph)]. The independence of the rate on the nitrile concentration and the positive Hammett value of 3.2(2) indicated that the reactions occur via an intramolecular nucleophilic attack of the hydroperoxide ligand to the coordinated nitrile carbon atom. In contrast, the previously reported cobalt(III)-hydroperoxo complex, [CoIII(TBDAP)(O2H)(CH3CN)]2+ (2TBDAP), exhibited the deficiency of reactivity toward nitrile. The comparison of pKa values and redox potentials of 2 and 2TBDAP showed that Me3-TPADP had a stronger ligand field strength than that of TBDAP. The density functional theory calculations for 2 and 2TBDAP support that the strengthened ligand field in 2 is mainly due to the replacement of two tert-butyl amine donors in TBDAP with methyl groups in Me3-TPADP, resulting in the compression of the Co-Nax bond lengths. These results provide mechanistic evidence of nitrile activation by the cobalt(III)-hydroperoxo complex and indicate that the basicity dependent on the ligand framework contributes to the ability of nitrile activation.

Synthesis and Bioluminescence of 'V'-Shaped Firefly Luciferin Analogues Based on A Novel Benzobisthiazole Core.

The design of π-extended conjugation 'V'-shaped red shifted bioluminescent D-luciferin analogues based on a novel benzobisthiazole core is described. The divergent synthetic route allowed access to a range of amine donor substituents through an SN Ar reaction. In spectroscopic studies, the 'V'-shaped luciferins exhibited narrower optical band gaps, more red-shifted absorption and emission spectra than D-luciferin. Their bioluminescence characteristics were recorded against four different luciferases (PpyLuc, FlucRed, CBR2 and PLR3). With native luciferase PpyLuc, the 'V'-shaped luciferins demonstrated more red-shifted emissions than D-luciferin (λbl =561 nm) by 60 to 80 nm. In addition, the benzobisthiazole luciferins showed a wide range of bioluminescence spectra from the visible light region (λbl =500 nm) to the nIR window (>650 nm). The computational results validate the design concept which can be used as a guide for further novel D-luciferin analogues based upon other 'V'-shaped heterocyclic cores.

Biotransamination of Furan-Based Aldehydes with Isopropylamine: Enzyme Screening and pH Influence.

Furan-based amines are highly valuable compounds which can be directly obtained via reductive amination from easily accessible furfural, 5-(hydroxymethyl)furfural (HMF) and 2,5-diformylfuran (DFF). Herein the biocatalytic amination of these carbonyl derivatives is disclosed using amine transaminases (ATAs) and isopropylamine (IPA) as amine donors. Among the different biocatalysts tested, the ones from Chromobacterium violaceum (Cv-TA), Arthrobacter citreus (ArS-TA), and variants from Arthrobacter sp. (ArRmut11-TA) and Vibrio fluvialis (Vf-mut-TA), afforded high levels of product formation (>80 %) at 100-200 mM aldehyde concentration. The transformations were studied in terms of enzyme and IPA loading. The pH influence was found as a key factor and attributed to the imine/aldehyde equilibrium that can arise from the high reactivity of the carbonyl substrates with a nucleophilic amine such as IPA.

Cr(II) and Cr(III) NCN pincer complexes: synthesis, structure, and catalytic reactivity

The synthesis, characterization, and reactivity of several new Cr(II) and Cr(III) complexes featuring an NCN pincer ligand with an arene backbone connected to amine donors NEt2 and NiPr2 via CH2-linkers is described. Reacting the in situ lithiated ligand precursor N(C–Br)NCH2-Et with [CrCl3(THF)3] resulted in the formation of the Cr(III) complex trans-[Cr(κ3NCN-NCNCH2-Et)(Cl)2(THF)]. Upon reaction of lithiated N(C–Br)NCH2-iPr with a suspension of anhydrous CrCl2, the Cr(II) complex [Cr(κ2NC-NCNCH2-iPr)2] is formed featuring two NCN ligands bound in κ2NC-fashion. In contrast, when lithiated N(C–Br)NCH2-iPr is reacted with a homogeneous solution of anhydrous CrX2 (X = Cl, Br), complexes [Cr(κ3NCN-NCNCH2-iPr)X] are obtained. Treatment of [Cr(κ3NCN-NCNCH2-iPr)Cl] with 1 equiv of PhCH2MgCl and LiCH2SiMe3 afforded the alkyl complexes [Cr(κ3NCN-NCNCH2-iPr)(CH2Ph)] and [Cr(κ3NCN-NCNCH2-iPr)(CH2SiMe3)]. All Cr(II) complexes exhibit effective magnetic moments in the range of 4.7–4.9 µB which is indicative for d4 high spin systems. If a solution of lithiated N(C–Br)NCH2-iPr is treated with CrCl2, followed by addition of an excess of Na[HB(Et)3], the dimeric complex [Cr(κ2NC-NCNCH2-iPr)(μ2-H)]2 is obtained bearing two bridging hydride ligands. [Cr(κ3NCN-NCNCH2-iPr)(CH2SiMe3)] turned out to be catalytically active for the hydrosilylation of ketones at room temperature with a catalyst loading of 1 mol%. X-ray structures of all complexes are presented.Graphical abstract

Amine Synthesis Using the Amine Donor N-Phenyl Putrescine and the Johnson Matthey Transaminase Biocatalyst Library

Chiral amines are important building blocks in the pharmaceutical, agrochemical and chemical industries. There is a drive to augment traditional transition metal catalysts with ‘green’ alternatives such as biocatalysts. Transaminase (TA) biocatalysts can be used in combination with ‘smart’ sacrificial amine donors to synthesise a variety of aliphatic and aromatic amines from the corresponding aldehydes and ketones. Despite their enormous potential, the unfavourable reaction equilibrium often limits the widespread application of TAs for industrial synthesis. Recently we disclosed a new biomimetic amine donor N-phenyl putrescine (NPP), which was inspired by the biosynthesis of the dipyrroloquinoline alkaloids. NPP was demonstrated to have good activity with a library of commercial and wild-type TAs (total 25 TAs). This work focused on exploring the use of NPP with the Johnson Matthey TA kit (17 biocatalysts; eight S-selective and nine R-selective) and three different amine acceptors (vanillin, benzaldehyde and acetophenone). NPP worked well with all 17 TAs and gave the corresponding amine products vanillylamine, benzylamine and methylbenzylamine (MBA) in up to 85% high-performance liquid chromatography (HPLC) yield. From the screen, STA-14 was identified as a good biocatalyst for further analysis and used in a comparative screen of NPP versus the commonly used donor iPrNH2. It was found that NPP was the best amine donor and used to prepare S-methylbenzylamine in >99.5% enantiomeric excess (e.e.). This work, combined with our previous study, highlights the potential of NPP in the biocatalytic synthesis of amines.

Efficient bioamination of lignin-derived vanillin to vanillylamine in dibutyl phthalate-water system

Lignin is one of the earth's most abundant renewable resources. Hence, achieving sustainable industry development in the future relies on obtaining a range of high-value chemicals from abundant and renewable biological resources. Vanillylamine is a basic upgrading product of vanillin in organic synthesis. It is also an important raw material for synthetic drugs, precursors, and additives. In this study, a two-phase reaction medium composed of organic solvent-water was established, and Escherichia coli CV was applied as ω-transaminase biocatalyst to the biosynthesis of vanillylamine from lignin-derived vanillin. Under optimized reaction conditions (pH 6.5, 35 °C) in a dibutyl phthalate (DBP)-water medium (1:9, vol/vol), vanillin (50 mM) was effectively transformed into vanillylamine by CV whole-cell biocatalyst containing ω-transaminase with amine donor L-Alanine (L-Ala:vanillin=12:1, mol/mol). Also, benzaldehyde, anisaldehyde, and veratraldehyde (50 mM) could be fully converted into the corresponding amines. This work provides an efficient and environmentally friendly method for the preparation of vanillylamine and its derivatives from lignin-based feedstocks.

Structure of the imine reductase from Ajellomyces dermatitidis in three crystal forms.

The NADPH-dependent imine reductase from Ajellomyces dermatitidis (AdRedAm) catalyzes the reductive amination of certain ketones with amine donors supplied in an equimolar ratio. The structure of AdRedAm has been determined in three forms. The first form, which belongs to space group P3121 and was refined to 2.01 Å resolution, features two molecules (one dimer) in the asymmetric unit in complex with the redox-inactive cofactor NADPH4. The second form, which belongs to space group C21 and was refined to 1.73 Å resolution, has nine molecules (four and a half dimers) in the asymmetric unit, each complexed with NADP+. The third form, which belongs to space group P3121 and was refined to 1.52 Å resolution, has one molecule (one half-dimer) in the asymmetric unit. This structure was again complexed with NADP+ and also with the substrate 2,2-difluoroacetophenone. The different data sets permit the analysis of AdRedAm in different conformational states and also reveal the molecular basis of stereoselectivity in the transformation of fluorinated acetophenone substrates by the enzyme.

Chemobiocatalytic transfromation of biomass into furfurylamine with mixed amine donor in an eco-friendly medium

Biobased furfurylamine (FAM) is a versatile platform molecule for producing additives, pharmaceuticals, and pesticides. Recombinant E. coli HNND-AlaDH was created by co-expressing L-alanine dehydrogenase (AlaDH) and mutated Aspergillus terreus ω-transaminase (HNND), aiming to convert furfural (FUR) into FAM using inexpensive L-alanine and isopropylamine as mixed amine donors. In ChCl:FA:OA (10 wt%), pineapple peel, bagasse, barley shell, peanut shell, and corn stalk could be efficiently transformed into FUR under 170 °C for 10 min. Pineapple peel produced a high titer of FUR (183.3 mM). Additionally, the viscosity, surface tension and polarity of ChCl:FA:OA were explored. The biomass-derived FUR was fully transformed to FAM by HNND-AlaDH with amine donor (1:1:1 of L-Ala/isopropylamine/FUR mol/mol/mol) within 300 min. Accordingly, the FAM productivity was 0.58 g/(g xylan in pineapple peel). This chemobiocatalytic strategy established through the combination of chemocatalysis and biocatalysis could be applied to convert renewable biomass into valuable organic amines.

Fused Double Donor Design with a Cross-Conjugated Dibenzosilin for Dye-Sensitized Solar Cells.

Dye-sensitized solar cells (DSCs) can provide a clean energy solution to growing energy demands. In order to have devices of high performance, sensitizers that are able to absorb in the near-infrared region (NIR) are needed. Stronger electron donors are needed for intramolecular charge-transfer sensitizers to access longer wavelength photons. Thus, two novel organic dyes with a cross-conjugated dibenzosilin double donor design are studied herein. The double donor delocalizes multiple filled orbitals across both amine donors due to the fused design that planarizes the donor as observed computationally, which improves intramolecular charge-transfer strength. The dyes are studied via density functional theory (DFT), optical spectroscopy, electrochemistry, and in DSC devices. The studies indicate that the dye design can reduce recombination losses, allowing for improved DSC device performances relative to a single arylamine donor. The reduction in recombination losses is attributed to the six alkyl chains that are incorporated into the donor, which offer good surface protection.

Mapping roles of active site residues in the acceptor site of the PA3944 Gcn5-related N-acetyltransferase enzyme.

An increased understanding of how the acceptor site in Gcn5-related N-acetyltransferase (GNAT) enzymes recognizes various substrates provides important clues for GNAT functional annotation and their use as chemical tools. In this study, we explored how the PA3944 enzyme from Pseudomonas aeruginosa recognizes three different acceptor substrates, including aspartame, NANMO, and polymyxin B, and identified acceptor residues that are critical for substrate specificity. To achieve this, we performed a series of molecular docking simulations and tested methods to identify acceptor substrate binding modes that are catalytically relevant. We found that traditional selection of best docking poses by lowest S scores did not reveal acceptor substrate binding modes that were generally close enough to the donor for productive acetylation. Instead, sorting poses based on distance between the acceptor amine nitrogen atom and donor carbonyl carbon atom placed these acceptor substrates near residues that contribute to substrate specificity and catalysis. To assess whether these residues are indeed contributors to substrate specificity, we mutated seven amino acid residues to alanine and determined their kinetic parameters. We identified several residues that improved the apparent affinity and catalytic efficiency of PA3944, especially for NANMO and/or polymyxin B. Additionally, one mutant (R106A) exhibited substrate inhibition toward NANMO, and we propose scenarios for the cause of this inhibition based on additional substrate docking studies with R106A. Ultimately, we propose that this residue is a key gatekeeper between the acceptor and donor sites by restricting and orienting the acceptor substrate within the acceptor site.

Highly efficient synthesis of chiral lactams by using a ω-transaminase from Bacillus megaterium and its mutant enzymes

In this study, a series of chiral lactams were successfully prepared by one-pot two-step reactions. It included a transamination reaction catalyzed by a ω-transaminase from Bacillus megaterium (ω-TABm), and subsequent spontaneous cyclisation. The different amine donors were firstly screened with ethyl levulinate as an amine acceptor, and both the excellent conversion and enantiomeric excess (e.e.p) of >99% could be achieved with 4-bromophenylethylamine as an amine donor, however, the long reaction time of 24 h was unfavorable for its application. After the amino acid residue Thr120 situated in the small pocket was mutated to Ser, the T120S variant could exhibit 10.6-fold higher activity comparing to that of wild type, and achieve the reaction with both conversion and e.e.p of >99% within 3 h. The T120S variant could also be applied to prepare the other chiral γ- and δ-lactams with excellent conversion and e.e.p. Furthermore, the inclusion bodies of ω-TABm and its variants could be aggregated as “native-like structures” to continuously prepare the chiral lactam for 18 h. The aim of the study is to provide a simply and efficiently biocatalytic route for preparing chiral lactams.