Catalysis Science & Technology
·
2018
·Vol. 8
·期 10
·2578-2587
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IF 5.365
Yanzi Wang,Jing Sun,Haohai Zhang,Zhiping Zhao,Wenfang Liu
Coenzyme regeneration is important for the industrial utilization of oxidoreductase. In this work, reduced nicotinamide adenine dinucleotide (NADH) was regenerated under visible light via a photosensitizer and an electron donor. First, several photosensitizers including 5,10,15,20-tetrakis(4-carboxyphenyl)porphyrin (TCPP), zinc porphyrin (ZnTCPP) and eosin Y (EY) with varied concentrations were compared. Next, using TCPP as the photosensitizer, the type and concentration of the electron donor, pH value and concentration of the buffer system for the reaction were optimized. Following that, TCPP was immobilized onto the surface of various supports for screening. Then, the immobilization conditions were optimized and the photocatalytic performance of silica-attached TCPP was studied. Lastly, the effect of a rhodium mediator on NADH regeneration was evaluated. The results show that the optimum concentration of TCPP, ZnTCPP and EY was 2, 1 and 4 mM, respectively, and the highest NADH yield of the TCPP system was 2.37 and 1.47 times those of the ZnTCPP and EY systems. With TCPP as the photosensitizer, triethanolamine was the most efficient electron donor and its appropriate concentration was 5 mM. The optimal pH value and concentration of the phosphate buffer was 7.0 and 0.05 M. Under these conditions, a high NADH yield of 81.5% was achieved after 60 min. For the supports examined, the thiol-containing silica microspheres coated with polydopamine and polyethyleneimine had the highest loading capacity of TCPP (9.15 mg per g-support). Via sensitization, the NADH yield was 9.64% after 90 min and was upgraded to 22.5% with the addition of the rhodium mediator.
研究领域:
生物化学
·
工业酶学
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产品关联
DOI
高影响因子
Research
·
2021
·
IF 0
Yuqing Cheng,Jiafu Shi,Yizhou Wu,Xueying Wang,Yiying Sun,Ziyi Cai,Yu Chen,Zhongyi Jiang
Solar-driven photocatalytic regeneration of cofactors, including reduced nicotinamide adenine dinucleotide (NADH), reduced nicotinamide adenine dinucleotide phosphate (NADPH), and reduced flavin adenine dinucleotide (FADH2), could ensure the sustainable energy supply of enzymatic reactions catalyzed by oxidoreductases for the efficient synthesis of chemicals. However, the elevation of cofactor regeneration efficiency is severely hindered by the inefficient utilization of electrons transferred on the surface of photocatalysts. Inspired by the phenomenon of ferredoxin-NADP+ reductase (FNR) anchoring on thylakoid membrane, herein, a homogeneous catalyst of rhodium (Rh) complex, [Cp∗Rh(bpy)H2O]2+, was anchored on polymeric carbon nitride (PCN) mediated by a tannic acid/polyethyleneimine (TA/PEI) adhesive layer, acquiring PCN@TA/PEI-Rh core@shell photocatalyst. Illuminated by visible light, electrons were excited from the PCN core, then transferred through the TA/PEI shell, and finally captured by the surface-anchored Rh for instant utilization during the regeneration of NADH. The TA/PEI-Rh shell could facilitate the electron transfer from the PCN core and, more importantly, achieved ~1.3-fold elevation of electron utilization efficiency compared with PCN. Accordingly, the PCN@TA/PEI-Rh afforded the NADH regeneration efficiency of 37.8% after 20 min reaction under LED light (405 nm) illumination, over 1.5 times higher than PCN with free Rh. Coupling of the NADH regeneration system with formate dehydrogenase achieved continuous production of formate from carbon dioxide (CO2). Our study may provide a generic and effective strategy to elevate the catalytic efficiency of a photocatalyst through intensifying the electron utilization.
研究领域:
生物化学
·
光催化材料科学
✓ 已验证
DOI
JOURNAL OF BIOTECHNOLOGY
·
2016
·
IF 2.667
Ping Wei,Jia-Xin Gao,Gao-Wei Zheng,Hong Wu,Min-Hua Zong,Wen-Yong Lou
The novel anti-Prelog stereospecific carbonyl reductase from Acetobacter sp. CCTCC M209061 was successfully expressed in E. coli combined with glucose dehydrogenase (GDH) to construct an efficient whole-cell biocatalyst with coenzyme NADH regeneration. The enzymatic activity of GAcCR (AcCR with a GST tag) reached 304.9 U/g-dcw, even 9 folds higher than that of wild strain, and the activity of GDH for NADH regeneration recorded 46.0 U/mg-protein in the recombinant E. coli . As a whole-cell biocatalyst, the recombinant E. coli BL21(DE3)pLysS (pETDuet- gaccr - gdh ) possessed a broad substrate spectrum for kinds of carbonyl compounds with encouraging yield and stereoselectivity. Besides, the asymmetric reduction of ethyl 4-chloroacetoacetate (COBE) to optically pure ethyl 4-chloro-3-hydroxybutyrate (CHBE) catalyzed by the whole-cell biocatalyst was systematically investigated. Under the optimal reaction conditions, the optical purity of CHBE was over 99% e.e. for ( S )-enantiomer, and the initial rate and product yield reached 8.04 μmol/min and 99.4%, respectively. Moreover, the space-time yield was almost 20 folds higher than that catalyzed by the wild strain. Therefore, a new, high efficiency biocatalyst for asymmetric reductions was constructed successfully, and the enantioselective reduction of prochiral compounds using the biocatalyst was a promising approach for obtaining enantiopure chiral alcohols.
研究领域:
生物工程
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合成生物学
✓ 已验证
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2017
·
IF 3.671
Mingan Yu,Duqiang Liu,Lili Sun,Jing Li,Qian Chen,Lijun Pan,Jingchuan Shang,Shurong Zhang,Wei Li
Ni 2+ -agarose bead-wrapped multi-enzyme/inorganic hybrid sphere composed of the immobilized enzymes as organic component and NaH 2 PO 4 and NaCl as inorganic component was developed by co-immobilizing extracellular His-tagged 3-quinuclidinone reductases and glucose dehydrogenase without pre-purification. The resulting biocatalysts has 3D porous architectures as confirmed by SEM and FESEM, and it enabled the continuous biotransformation of 3-quinuclidone to (R)-3-quinuclidinol with cofactor regeneration in situ. The 3D porous biocatalysts were formed via three steps: First, immobilization of the His-tagged enzymes directly from the cell lysates supernatant. Next, formation of enzyme aggregates, ribbons and gels. Finally, the enzymes, the formed aggregates/ribbons/gels and salt were incorporated to the foam and then covered the Ni 2+ -agarose bead. The technique made the immobilization of these enzymes effective such that specific enzyme loading of 60.8 mg/g support and enzyme loading efficiency of 92.3% were achieved. As a direct consequence, the biocatalyst catalyzed the conversion of 3-quinuclidinone (204 g/L) to (R)-3-quinuclidinol in 100% yield and 100% ee at 4.5 h, and the recyclability of the biocatalyst was excellent, retaining > 95% conversion yield and 100% ee even after the fifteenth runs. Overall, our strategy is demonstrated to be a promising method for developing efficient and robust biocatalyst for asymmetric synthesis.
研究领域:
生物化学
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酶工程
·
合成生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
RSC Advances
·
2019
·Vol. 9
·期 4
·2325-2331
·
IF 3.049
Yitong Chen,Baodi Ma,Songshuang Cao,Xiaomei Wu,Yi Xu
The production of (S)-N-boc-3-hydroxy piperidine (NBHP) via asymmetric bioreduction of 1-boc-3-piperidinone with reductase is impeded by the need for expensive coenzymes NAD(P)H. In order to regenerate the coenzyme in situ, the gene of alcohol dehydrogenase from Thermoanaerobacter brockii and glucose dehydrogenase from Bacillus subtilis were ligated into the multiple cloning sites of pRSFDuet-1 plasmid to construct the recombinant Escherichia BL21 (DE3) that co-expressing alcohol dehydrogenase and glucose dehydrogenase. Different culture conditions including the medium composition, inducer and pH etc were systematically investigated to improve the enzyme production. The enzyme activity was increased more than 11-fold under optimal culture condition, from 12.7 to 139.8 U L−1. In the further work, the asymmetric reduction of 1-boc-3-piperidinone by whole cells of recombinant E. coli was systematic optimized to increase the substrate concentration and reaction efficiency. At last, S-NBHP (>99% ee) was prepared at 500 mM substrate concentration without external addition of cofactors. The conversion of S-NBHP reached 96.2% within merely 3 h, corresponding a high space-time yield around 774 g L−1 d−1. All these results demonstrated the potential of recombinant E. coli BL21 (DE3) coupled expressing alcohol dehydrogenase and glucose dehydrogenase for efficient synthesis of S-NBHP.
研究领域:
生物技术
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药物化学
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酶学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
KOREAN JOURNAL OF CHEMICAL ENGINEERING
·
2021
·Vol. 38
·期 5
·1066-1077
·
IF 3.309
Lu Yuan,Dai Hongqian,Cheng Pengpeng,Shi Hanbing,Tang Lan,Sun Xingyuan,Ou Zhimin
A modular approach was applied for the synthesis of bienzyme-polymer nanoconjugates (nano-BECs) (50–70 nm) consisting of two enzymes (carbonyl reductase and glucose dehydrogenase) conjugated within a single universal polymer scaffold. The amount of the product ethyl (R)-2-hydroxy-4-phenylbutyrate (R-HPBE) with nano-BECs as the catalyst was 533 mM in a dibutyl phthalate-phosphate buffer (dibutyl phthalate-PB) biphasic system, while the amount of R-HPBE was 349 mM using carbonyl reductase-poly(acrylic acid) as the catalyst, indicating that the nano-BECs have an advantage for coenzyme regeneration. Compared with a single aqueous phase, the substrate treatment capacity was improved at the interface of the dibutyl phthalate-PB biphasic system. Under the optimal reaction conditions (35 °C, 40 h, dibutyl phthalate-PB 1: 1), nano-BECs can completely convert substrate into optically pure R-HPBE (enantiomeric excess (e.e.) >99.9%) in the organic-aqueous system.
研究领域:
生物催化
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酶工程
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纳米技术
·
有机合成
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）