scholarly journals Engineering of a Novel, Magnetic, Bi-Functional, Enzymatic Nanobiocatalyst for the Highly Efficient Synthesis of Enantiopure (R)-3-quinuclidinol

Catalysts ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 1126
Author(s):  
Qingman Li ◽  
Qihua Jiang ◽  
Pengcheng Gu ◽  
Lianju Ma ◽  
Yiwu Wang
Keyword(s):  
Glucose Dehydrogenase ◽  
Porous Silica ◽  
Temperature Tolerance ◽  
Cofactor Regeneration ◽  
Great Promise ◽  
Conversion Yield ◽  
Carbonyl Reduction ◽  
Multifunctional Enzymes ◽  
One Step ◽  
First Time

Ni2+-NTA-boosted magnetic porous silica nanoparticles (Ni@MSN) to serve as ideal support for bi-functional enzyme were fabricated for the first time. The versatility of this support was validated by one-step purification and immobilization of bi-functional enzyme MLG consisting of 3-Quinuclidinone reductase and glucose dehydrogenase, which can simultaneously catalyze both carbonyl reduction and cofactor regeneration, to fabricate an artificial bi-functional nanobiocatalyst (namely, MLG-Ni@MSN). The enzyme loading of 71.7 mg/g support and 92.7% immobilization efficiency were obtained. Moreover, the immobilized MLG showed wider pH and temperature tolerance and greater storage stability than free MLG under the same conditions. The nanosystem was employed as biocatalyst to accomplish the 3-quinuclidinone (70 g/L) to (R)-3-quinuclidinol biotransformation in 100% conversion yield with >99% selectivity within 6 h and simultaneous cofactor regeneration. Furthermore, the immobilized MLG retained up to 80.3% (carbonyl reduction) and 78.0% (cofactor regeneration) of the initial activity after being recycled eight times. In addition, the MLG-Ni@MSN system exhibited almost no enzyme leaching during biotransformation and recycling. Therefore, we have reason to believe that the Ni@MSN support gave great promise for constructing a new biocatalytic nanosystem with multifunctional enzymes to achieve some other complex bioconversions.

Efficient one-step biocatalytic multienzyme cascade strategy for direct conversion of phytosterol to C17-hydroxylated steroids

2021 ◽  
Author(s):  
Rui Tang ◽  
Xiaoxian Ren ◽  
Menglei Xia ◽  
Yanbing Shen ◽  
Linna Tu ◽  
...  
Keyword(s):  
Raw Materials ◽  
Hydroxysteroid Dehydrogenase ◽  
Direct Conversion ◽  
Cofactor Regeneration ◽  
Regeneration Strategy ◽  
Reduction Strategy ◽  
Mycobacterium Neoaurum ◽  
Carbonyl Reduction ◽  
One Step ◽  
Glucose Supplementation

Steroidal 17-carbonyl reduction is crucial to the production of natural bioactive steroid medicines, boldenone (BD) is one of the important C17-hydroxylated steroids. Although efforts have been made to produce BD through biotransformation, the challenge of complex transformation process, high substrates cost, and low catalytic efficiencies have yet to be mastered. Phytosterol (PS) is the most widely accepted substrate for the production of steroid medicines due to its similar foundational structure and ubiquitous sources. 17β-Hydroxysteroid dehydrogenase (17βHSD) and its native electron donor play significant roles in the 17β-carbonyl reduction reaction of steroids. In this study, we bridged 17βHSD with a cofactor regeneration strategy in Mycobacterium neoaurum to establish a one-step biocatalytic carbonyl reduction strategy for efficient biosynthesis of BD from PS for the first time. After investigating different intracellular electron transfer strategies, we rationally designed the engineered strain with co-expression of 17βhsd and glucose-6-phosphate dehydrogenase (G6PDH) gene in M. neoaurum . With establishment of an intracellular cofactor regeneration strategy, the ratio of [NADPH]/[NADP + ] was maintained at a relatively high level, the yield of BD increased from 17% (in MNR M3M- ayr1 S.c ) to 78% (in MNR M3M- ayr1 & g6p with glucose supplementation), and the productivity was increased by 6.5 times. Furthermore, under the optimal glucose supplementation condition, the yield of BD reached 82%, which is the highest yield reported by transformation from PS with one-step. This study demonstrated an excellent strategy for production of many other valuable carbonyl reduction steroidal products from natural cheap raw materials. Importance Steroid C17-carbonyl reduction is one of the important transformations for the production of valuable steroidal medicines or intermediates for further synthesis of steroidal medicines, but it remains a challenge through either chemical or biological synthesis. Phytosterol can be obtained from low-cost residue of waste natural materials, and it is preferred as the economical and applicable substrate for steroid medicines production by Mycobacterium . This study explored a green and efficient one-step biocatalytic carbonyl reduction strategy for direct conversion of phytosterol to C17-hydroxylated steroids by bridging 17β-Hydroxysteroid dehydrogenase with a cofactor regeneration strategy in Mycobacterium neoaurum . This work has practical value for the production of many valuable hydroxylated steroids from natural cheap raw materials.

Calorimetric Sensor for Ethanol Using Ni2+-nitrilotriacetic Acid (NTA) Resin Immobilized Alcohol Dehydrogenase (ADH)

2020 ◽  
Vol 16 (6) ◽  
pp. 795-799
Author(s):  
YongJin Li
Keyword(s):  
Cost Effective ◽  
Nitrilotriacetic Acid ◽  
Repeated Measurements ◽  
Great Promise ◽  
Calorimetric Sensor ◽  
Controlled Pore Glass ◽  
Pore Glass ◽  
One Step ◽  
Immobilization Matrix ◽  
Enzyme Thermistor

Background: A simple, fast and economic analytical method for the determination of ethanol is important for clinical, biological, forensic and physico-legal purposes. Methods: Ni2+-NTA resin was used as an immobilization matrix for the simple one-step purification/ immobilization of his6-tagged ADH. Different alcohols with a concentration range of 0.5-50% V/V, namely methanol, ethanol and propanol were measured using prepared ADH enzyme thermistor. The ethanol content of Tsingtao beer was tested as a real sample containing alcohol. Reproducibility and stability of prepared ADH enzyme thermistor were also investigated by repeated measurements. Results: In comparison to the controlled pore glass (a common used support for the immobilization of enzyme) used in thermal biosensor, the use of Ni2+-NTA resin not only led to simple one-step purification/ immobilization by his6-tagged ADH binding to Ni2+-NTA resin, but also made the immobilizing supports reusable. The prepared biosensor can be used to determine ethanol and methanol by the calorimetric measurement. A linear range of 1 -32% (V/V) and 2-20% (V/V) was observed for ethanol and methanol, respectively. The detection limits were 0.3% (V/V) and 1% (V/V) for ethanol and methanol, respectively. The tested ethanol concentration of Tsingtao beer was 4.5% V/V, which is comparable with the labeled alcohol by volume (ABV) 4.80%. Conclusion: Ni2+-NTA resin, as an immobilization matrix in ET sensor, provides a simple one-step purification/immobilization for His6-tagged recombinase and a reusable immobilization matrix. The prepared biosensor exhibits good repeatability and stability. Such a new biosensor shows great promise for rapid, simple, and cost-effective analysis of ethanol and methanol, both in qualitative and in quantitative tests.

scholarly journals Electrochromic Performance and Capacitor Performance of α-MoO3 Nanorods Fabricated by a One-Step Procedure

Coatings ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 783
Author(s):  
Ying Duan ◽  
Chen Wang ◽  
Jian Hao ◽  
Yang Jiao ◽  
Yanchao Xu ◽  
...  
Keyword(s):  
Growth Parameters ◽  
Average Diameter ◽  
Fast Response ◽  
Cycle Stability ◽  
Electrochromic Devices ◽  
Step Procedure ◽  
One Step ◽  
Supercapacitor Performance ◽  
First Time ◽  
Prepared Nanorods

In this paper, we propose for the first time the synthesis of α-MoO3 nanorods in a one-step procedure at mild temperatures. By changing the growth parameters, the microstructure and controllable morphology of the resulting products can be customized. The average diameter of the as-prepared nanorods is about 200 nm. The electrochromic and capacitance properties of the synthesized products were studied. The results show that the electrochromic properties of α-MoO3 nanorods at 550 nm have 67% high transmission contrast, good cycle stability and fast response time. The MoO3 nanorods also exhibit a stable supercapacitor performance with 98.5% capacitance retention after 10,000 cycles. Although current density varies sequentially, the nanostructure always exhibits a stable capacitor to maintain 100%. These results indicate the as-prepared MoO3 nanorods may be good candidates for applications in electrochromic devices and supercapacitors.

scholarly journals A Redox-Neutral, Two-Enzyme Cascade for the Production of Malate and Gluconate from Pyruvate and Glucose

2021 ◽  
Vol 11 (11) ◽  
pp. 4877
Author(s):  
Ravneet Mandair ◽  
Pinar Karagoz ◽  
Roslyn M. Bill
Keyword(s):  
Malate Dehydrogenase ◽  
Closed System ◽  
Glucose Dehydrogenase ◽  
Sulfolobus Solfataricus ◽  
Cofactor Regeneration ◽  
Triple Mutant ◽  
Thermococcus Kodakarensis ◽  
Platform Chemicals ◽  
Enzyme Cascade

A triple mutant of NADP(H)-dependent malate dehydrogenase from thermotolerant Thermococcus kodakarensis has an altered cofactor preference for NAD+, as well as improved malate production compared to wildtype malate dehydrogenase. By combining mutant malate dehydrogenase with glucose dehydrogenase from Sulfolobus solfataricus and NAD+/NADH in a closed reaction environment, gluconate and malate could be produced from pyruvate and glucose. After 3 h, the yield of malate was 15.96 mM. These data demonstrate the feasibility of a closed system capable of cofactor regeneration in the production of platform chemicals.

One-step synthesis of well-dispersed polypyrrole copolymers under gamma-ray irradiation

2021 ◽  
Vol 12 (5) ◽  
pp. 645-649
Author(s):  
Yu Gu ◽  
Yuqing Qiao ◽  
Yusen Meng ◽  
Ming Yu ◽  
Bowu Zhang ◽  
...  
Keyword(s):  
Room Temperature ◽  
Gamma Ray ◽  
Gamma Ray Irradiation ◽  
One Step ◽  
First Time

Herein, we report for the first time the synthesis of polypyrrole copolymers with good solvent-dispersibility under gamma-ray irradiation at room temperature in air.

scholarly journals Construction and characterization of a novel glucose dehydrogenase-leucine dehydrogenase fusion enzyme for the biosynthesis of l-tert-leucine

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Langxing Liao ◽  
Yonghui Zhang ◽  
Yali Wang ◽  
Yousi Fu ◽  
Aihui Zhang ◽  
...  
Keyword(s):  
Glucose Dehydrogenase ◽  
Catalytic Efficiency ◽  
Space Time ◽  
Cofactor Regeneration ◽  
Enzyme Complex ◽  
Regeneration System ◽  
Regeneration Rate ◽  
Leucine Dehydrogenase ◽  
Fusion Enzyme ◽  
Space Time Yield

Abstract Background Biosynthesis of l-tert-leucine (l-tle), a significant pharmaceutical intermediate, by a cofactor regeneration system friendly and efficiently is a worthful goal all the time. The cofactor regeneration system of leucine dehydrogenase (LeuDH) and glucose dehydrogenase (GDH) has showed great coupling catalytic efficiency in the synthesis of l-tle, however the multi-enzyme complex of GDH and LeuDH has never been constructed successfully. Results In this work, a novel fusion enzyme (GDH–R3–LeuDH) for the efficient biosynthesis of l-tle was constructed by the fusion of LeuDH and GDH mediated with a rigid peptide linker. Compared with the free enzymes, both the environmental tolerance and thermal stability of GDH–R3–LeuDH had a great improved since the fusion structure. The fusion structure also accelerated the cofactor regeneration rate and maintained the enzyme activity, so the productivity and yield of l-tle by GDH–R3–LeuDH was all enhanced by twofold. Finally, the space–time yield of l-tle catalyzing by GDH–R3–LeuDH whole cells could achieve 2136 g/L/day in a 200 mL scale system under the optimal catalysis conditions (pH 9.0, 30 °C, 0.4 mM of NAD+ and 500 mM of a substrate including trimethylpyruvic acid and glucose). Conclusions It is the first report about the fusion of GDH and LeuDH as the multi-enzyme complex to synthesize l-tle and reach the highest space–time yield up to now. These results demonstrated the great potential of the GDH–R3–LeuDH fusion enzyme for the efficient biosynthesis of l-tle.

The application of inexpensive and synthetic simple electrocatalyst CuFe-MoC@NG in immunosensor

The Analyst ◽  
2021 ◽  
Author(s):  
Lisha Mei ◽  
Wentang Zhao ◽  
Yiju Song ◽  
Li Zhang ◽  
Mengmeng Zhang ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Precious Metal ◽  
Doped Graphene ◽  
One Step ◽  
Metal Materials ◽  
First Time

In this study, we used inexpensive and synthetic simple electrocatalysts replacement conventional precious metal materials to reduce hydrogen peroxide (H2O2). We first time developed N-doped graphene-coated CuFe@MoC using one-step calcination...

Simultaneous Spectral Imaging of C2*/CH* at Low-to-High Pressure Combustion

2017 ◽  
Author(s):  
Jonathan Reyes ◽  
Kareem Ahmed
Keyword(s):  
Intensity Ratio ◽  
High Pressures ◽  
Diffusion Flames ◽  
Laminar Flames ◽  
Great Promise ◽  
Fuel Distribution ◽  
Premixed Laminar ◽  
The Cost ◽  
Indexing Technique ◽  
First Time

This paper presents the correlation of the intensity ratio of the C2* and CH* radicals to fuel-air measurements over a range of pressures using 93% octane gasoline as the fuel. The measurements are conducted for the first time at high pressures. The study utilizes beam splitting technology to simultaneously view C2* and CH* as a line of sight, global measurement at the cost of resolution. A heavily instrumented constant volume combustor, with optical access, was employed to acquire the data. The ratio of C2* and CH* has been proven to be a good index of the equivalence ratio of premixed laminar flames. This index is attained, quite simply, by filtering each at their respected emissive peaks and taking the ratio of C2* over CH*. This technique shows great promise for use in turbomachinery as it will allow for identification of rich and lean locations in a combustor. By knowing the fuel-air field, combustor inefficiencies can be addressed to allow for greater energy release in combustion. The issue lies with the application of the indexing technique. Presented data to date has been performed on laboratory based diffusion flames exhausting to atmosphere, or premixed, steady, combustor type flames at low pressure (1atm) conditions. These types of flames are not relevant for engine combustor conditions. Understanding the fuel distribution at relevant regimes will reveal where inefficiencies may lie in injector or combustor design. Propagating flame kernels pose a problem in that they do not produce as much light as a steady flame, this makes spectral data difficult to obtain. Steady flames also do not address the effects that pressure may have on the index of C2* and CH*. The authors of this work seek to address three main issues associated with the indexing technique: The feasibility of its application to combustors (hardware design), The ability to operate at low-light ignition events, and the effects pressure may have on the correlation of intensity ratio to the fuel-air measurement.

Facile one-step mechanochemical synthesis of [Cu(tu)]Cl·1/2H2O nanobelts for high-performance supercapacitor

RSC Advances ◽  
2015 ◽  
Vol 5 (48) ◽  
pp. 38527-38532 ◽  
Author(s):  
Di Guo ◽  
Hongxu Guo ◽  
Yingchang Ke ◽  
Dongfang Wang ◽  
Jianhua Chen ◽  
...  
Keyword(s):  
Mechanochemical Synthesis ◽  
Electrode Material ◽  
High Performance ◽  
Mechanochemical Process ◽  
One Step ◽  
First Time

A facile one-step mechanochemical process from CuCl2·2H2O and thiourea to fabricate novel [Cu(tu)]Cl·1/2H2O nanobelts has been observed for the first time, and the nanobelts were used as an electrode material for a supercapacitor.

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