25333-42-0Relevant articles and documents
Structural basis for high substrate-binding affinity and enantioselectivity of 3-quinuclidinone reductase AtQR
Hou, Feng,Miyakawa, Takuya,Kataoka, Michihiko,Takeshita, Daijiro,Kumashiro, Shoko,Uzura, Atsuko,Urano, Nobuyuki,Nagata, Koji,Shimizu, Sakayu,Tanokura, Masaru
, p. 911 - 915 (2014)
(R)-3-Quinuclidinol, a useful compound for the synthesis of various pharmaceuticals, can be enantioselectively produced from 3-quinuclidinone by 3-quinuclidinone reductase. Recently, a novel NADH-dependent 3-quinuclidionone reductase (AtQR) was isolated from Agrobacterium tumefaciens, and showed much higher substrate-binding affinity (>100 fold) than the reported 3-quinuclidionone reductase (RrQR) from Rhodotorula rubra. Here, we report the crystal structure of AtQR at 1.72 A. Three NADH-bound protomers and one NADH-free protomer form a tetrameric structure in an asymmetric unit of crystals. NADH not only acts as a proton donor, but also contributes to the stability of the α7 helix. This helix is a unique and functionally significant part of AtQR and is related to form a deep catalytic cavity. AtQR has all three catalytic residues of the short-chain dehydrogenases/reductases family and the hydrophobic wall for the enantioselective reduction of 3-quinuclidinone as well as RrQR. An additional residue on the α7 helix, Glu197, exists near the active site of AtQR. This acidic residue is considered to form a direct interaction with the amine part of 3-quinuclidinone, which contributes to substrate orientation and enhancement of substrate-binding affinity. Mutational analyses also support that Glu197 is an indispensable residue for the activity.
Production of (R)-3-quinuclidinol by a whole-cell biocatalyst with high efficiency
Jia, Zhenhua,Ma, Hong,Huang, Yali,Huang, Yuanyuan,Ren, Pengju,Song, Shuishan,Hu, Meirong,Tao, Yong
, p. 316 - 323 (2018)
Optically pure (R)-3-quinuclidinol [(R)-3-Qui] is widely used as a chiral building block for producing various antimuscarinic agents. An asymmetric bioreduction approach using 3-quinuclidinone reductases is an effective way to produce (R)-3-Qui. In this study, a biocatalyst for producing (R)-3-Qui was developed by using Escherichia coli that coexpressed Kaistia granuli (KgQR) and mutant glucose dehydrogenase (GDH). KgQR catalyses the synthesis of (R)-3-Qui through the efficient reduction of 3-quinuclidinone. The specific activity of recombinant KgQR was 254 U/mg, and the Michaelis–Menten constant (Km) for 3-quinuclidinone was 0.51 mM. The thermal stability of KgQR was relatively high compared with ArQR. Approximately 73% of the residual activity remained after incubation in 0.2?M potassium phosphate buffer (KPB) (pH 7.0) for 8 h at 30 °C. In addition, 80% residual activity remained for the double-mutant GDH (Q252L and E170K) after incubation in a buffer (pH 7.0) for 8 h at 30 and 40 °C. 3-Quinuclidinone (242 g/L) can be reduced to (R)-3-Qui in 3 h by coexpressing KgQR and mutant GDH in E. coli. The conversion rate reached 80.6 g/L/h, which is the highest reported to date. The results demonstrates that this whole-cell biocatalyst will have a great potential in industrial manufacturing.
Highly efficient synthesis of (R)-3-quinuclidinol in a space-time yield of 916 g L-1 d-1 using a new bacterial reductase Ar QR
Zhang, Wen-Xia,Xu, Guo-Chao,Huang, Lei,Pan, Jiang,Yu, Hui-Lei,Xu, Jian-He
, p. 4917 - 4919 (2013)
A new keto reductase (ArQR), identified from Agrobacterium radiobacter ECU2556, can efficiently reduce 3-quinuclidinone in excellent enantioselectivity and high space-time yield for the synthesis of (R)-3-quinuclidinol, a chiral building block of many antimuscarinic agents. This is the first time that a high yield of (R)-3-quinuclidinol up to 916 g L-1 d-1 using a bioreduction approach is reported.
Borneol dehydrogenase from Pseudomonas sp. TCU-HL1 possesses novel quinuclidinone reductase activities
Chen, Hao-Ping,Ho, Tsung-Jung,Hung, Chien-Chi,Khine, Aye Aye,Lu, Pei-Chieh,Simaremare, Sailent Rizki Sari,Tung, Chi-Hua,Wu, Jia-Ru,Yiin, Lin-Ming
, (2021/08/30)
Borneol dehydrogenase (BDH) catalyses the last step of the camphor biosynthetic pathway in plants and the first reaction in the borneol degradation pathway in soil microorganisms. Native or engineered BDH can be used to produce optically pure borneol and camphor. The recently reported apo-form crystal structure of BDH (PDB ID: 6M5N) from Pseudomonas sp. TCU-HL1 superimposes well with that of 3-quinuclidinone reductase (QR) (PDB ID: 3AK4) from Agrobacterium tumefaciens. QR catalyses the conversion of 3-quinuclidinone into (R)-3-(?)-quinuclidinol, an important chiral synthone for several drugs. However, the kinetic parameter, kcat, of QR was not determined in the previous reports even though both BDH and QR have various potential industrial applications. Here, we aimed to further characterise their structural and functional relationship. Recombinant QR with the native sequence was cloned, expressed in E. coli, and purified. We found that 3-quinuclidinone can be used as an alternative substrate for BDH. Only (R)-3-(?)-quinuclidinol was detected in this BDH-catalysed reaction. The results of 3 D molecular docking simulation show that 3-quinuclidinone and (+)-/(-)- borneol were docked to two different parts of the QR active site. In contrast, all three compounds are docked uniformly to the alpha-1 helix of BDH. There results explain why BDH can turnover 3-quinuclidinone, while QR can not act on (+)-/(-)-borneol.
Rapid, Heterogeneous Biocatalytic Hydrogenation and Deuteration in a Continuous Flow Reactor
Thompson, Lisa A.,Rowbotham, Jack S.,Nicholson, Jake H.,Ramirez, Miguel A.,Zor, Ceren,Reeve, Holly A.,Grobert, Nicole,Vincent, Kylie A.
, p. 3913 - 3918 (2020/06/17)
The high selectivity of biocatalysis offers a valuable method for greener, more efficient production of enantiopure molecules. Operating immobilised enzymes in flow reactors can improve the productivity and handling of biocatalysts, and using H2 gas to drive redox enzymes bridges the gap to more traditional metal-catalysed hydrogenation chemistry. Herein, we describe examples of H2-driven heterogeneous biocatalysis in flow employing enzymes immobilised on a carbon nanotube column, achieving near-quantitative conversion in 2 gas as a clean reductant, in a completely atom-efficient process. The flow system is demonstrated for cofactor conversion, reductive amination and ketone reduction, and then extended to biocatalytic deuteration for the selective production of isotopically labelled chemicals.
