共查询到20条相似文献,搜索用时 15 毫秒
1.
Emily Radley John Davidson Jake Foster Richard Obexer Elizabeth L. Bell Anthony P. Green 《Angewandte Chemie (International ed. in English)》2023,62(52):e202309305
The development and implementation of sustainable catalytic technologies is key to delivering our net-zero targets. Here we review how engineered enzymes, with a focus on those developed using directed evolution, can be deployed to improve the sustainability of numerous processes and help to conserve our environment. Efficient and robust biocatalysts have been engineered to capture carbon dioxide (CO2) and have been embedded into new efficient metabolic CO2 fixation pathways. Enzymes have been refined for bioremediation, enhancing their ability to degrade toxic and harmful pollutants. Biocatalytic recycling is gaining momentum, with engineered cutinases and PETases developed for the depolymerization of the abundant plastic, polyethylene terephthalate (PET). Finally, biocatalytic approaches for accessing petroleum-based feedstocks and chemicals are expanding, using optimized enzymes to convert plant biomass into biofuels or other high value products. Through these examples, we hope to illustrate how enzyme engineering and biocatalysis can contribute to the development of cleaner and more efficient chemical industry. 相似文献
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Jie Qiao Yijie Sheng Minghui Wang Anni Li Dr. Xiujuan Li Prof. Dr. He Huang 《Angewandte Chemie (International ed. in English)》2023,62(12):e202300320
Obtaining a robust and applicable enzyme for bioethanol production is a dream for biorefinery engineers. Herein, we describe a general method to evolve an all-round and interpretable enzyme that can be directly employed in the bioethanol industry. By integrating the transferable protein evolution strategy InSiReP 2.0 (In Silico guided Recombination Process), enzymatic characterization for actual production, and computational molecular understanding, the model cellulase PvCel5A (endoglucanase II Cel5A from Penicillium verruculosum) was successfully evolved to overcome the remaining challenges of low ethanol and temperature tolerance, which primarily limited biomass transformation and bioethanol yield. Remarkably, application of the PvCel5A variants in both first- and second-generation bioethanol production processes (i. Conventional corn ethanol fermentation combined with the in situ pretreatment process; ii. cellulosic ethanol fermentation process) resulted in a 5.7–10.1 % increase in the ethanol yield, which was unlikely to be achieved by other optimization techniques. 相似文献
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Carboxysome-Inspired Electrocatalysis using Enzymes for the Reduction of CO2 at Low Concentrations**
Dr. Samuel J. Cobb Azim M. Dharani Dr. Ana Rita Oliveira Prof. Inês A. C. Pereira Prof. Erwin Reisner 《Angewandte Chemie (International ed. in English)》2023,62(26):e202218782
The electrolysis of dilute CO2 streams suffers from low concentrations of dissolved substrate and its rapid depletion at the electrolyte-electrocatalyst interface. These limitations require first energy-intensive CO2 capture and concentration, before electrolyzers can achieve acceptable performances. For direct electrocatalytic CO2 reduction from low-concentration sources, we introduce a strategy that mimics the carboxysome in cyanobacteria by utilizing microcompartments with nanoconfined enzymes in a porous electrode. A carbonic anhydrase accelerates CO2 hydration kinetics and minimizes substrate depletion by making all dissolved carbon available for utilization, while a highly efficient formate dehydrogenase reduces CO2 cleanly to formate; down to even atmospheric concentrations of CO2. This bio-inspired concept demonstrates that the carboxysome provides a viable blueprint for the reduction of low-concentration CO2 streams to chemicals by using all forms of dissolved carbon. 相似文献
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Dr. Fabio Pirro Dr. Salvatore La Gatta Dr. Federica Arrigoni Dr. Antonino Famulari Dr. Ornella Maglio Prof. Pompea Del Vecchio Prof. Mario Chiesa Prof. Luca De Gioia Prof. Luca Bertini Dr. Marco Chino Prof. Flavia Nastri Prof. Angela Lombardi 《Angewandte Chemie (Weinheim an der Bergstrasse, Germany)》2023,135(1):e202211552
De novo metalloprotein design is a remarkable approach to shape protein scaffolds toward specific functions. Here, we report the design and characterization of Due Rame 1 (DR1), a de novo designed protein housing a di-copper site and mimicking the Type 3 (T3) copper-containing polyphenol oxidases (PPOs). To achieve this goal, we hierarchically designed the first and the second di-metal coordination spheres to engineer the di-copper site into a simple four-helix bundle scaffold. Spectroscopic, thermodynamic, and functional characterization revealed that DR1 recapitulates the T3 copper site, supporting different copper redox states, and being active in the O2-dependent oxidation of catechols to o-quinones. Careful design of the residues lining the substrate access site endows DR1 with substrate recognition, as revealed by Hammet analysis and computational studies on substituted catechols. This study represents a premier example in the construction of a functional T3 copper site into a designed four-helix bundle protein. 相似文献
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酶立体选择性的定向进化及其高通量筛选方法 总被引:3,自引:0,他引:3
定向进化技术已成为开发新型生物催化剂的有力工具,特别是在对酶结构或催化机理信息缺乏的情况下。酶的立体选择性是个比较难处理的参数,其在定向进化过程中的技术瓶颈是建立快速有效的高通量筛选方法。本文概述了在酶立体选择性的定向进化方面所取得的进展,着重论述了酶立体选择性的高通量筛选方法。 相似文献
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Efficiency and evolution of enzyme catalysis. 总被引:2,自引:0,他引:2
A new function derived from kinetic data, the efficiency function, can be used to quantify the efficiency of a catalyst. For freely diffusing species the maximum efficiency is unity. The enzyme triose phosphate isomerase has an efficiency of 0.6 and is thus almost a perfect catalyst. The efficiency of the acetate ion as catalyst for the same reaction is 2.5 × 10?11. This increase in catalytic efficiency is discussed in terms of three types of evolutionary improvement: the uniform binding of the substrate to the enzyme, changes in the internal thermodynamics of the bound states, and more effective catalysis of elementary steps. These concepts are illustrated for triose phosphate isomerase. 相似文献
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Uwe T. Bornscheuer 《Angewandte Chemie (International ed. in English)》1998,37(22):3105-3108
On the way to a combinatorial biotechnology? The directed evolution of enzymes promises a rapid access to effective biocatalysts. New molecular biology techniques for random mutagenesis in combination with high-throughput screening might revolutionize the creation of enzymes with new and improved properties. 相似文献
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Norbert Strter William N. Lipscomb Thomas Klabunde Bernt Krebs 《Angewandte Chemie (International ed. in English)》1996,35(18):2024-2055
Numerous studies, both in enzymatic and nonenzymatic catalysis, have been undertaken to understand the way by which metal ions, especially zinc ions, promote the hydrolysis of phosphate ester and amide bonds. Hydrolases containing one metal ion in the active site, termed mononuclear metallohydrolases, such as carboxypeptidase. A and thermolysin were among the first enzymes to have their structures unraveled by X-ray crystallography. In recent years an increasing number of metalloenzymes have been identified that use two or more adjacent metal ions in the catalysis of phosphoryl-transfer reactions (R-OPO3 + R′-OH → R′-OPO3 + R-OH; in the case of the phosphatase reaction R′-OH is a water molecule) and carbonyl-transfer reactions, for example, in peptidases or other amidases. These dinuclear metalloenzymes catalyze a great variety of these reactions, including hydrolytic cleavage of phosphomono-, -di- and -triester bonds, phosphoanhydride bonds as well as of peptide bonds or urea. In addition, the formation of the phosphodiester bond of RNA and DNA by polymerases is catalyzed by a two-metal ion mechanism. A remarkable diversity is also seen in the structures of the active sites of these di- and trinuclear metalloenzymes, even for enzymes that catalyze very similar reactions. The determination of the structure of a substrate, product, stable intermediate, or a reaction coordinate analogue compound bound to an active or inactivated enzyme is a powerful approach to investigate mechanistic details of enzyme action. Such studies have been applied to several of the metalloenzymes reviewed in this article; together with many other biochemical studies they provide a growing body of information on how the two (or more) metal ions cooperate to achieve efficient catalysis. 相似文献
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Emil Thomas Kaiser 《Angewandte Chemie (International ed. in English)》1988,27(7):913-922
Protein engineering has as its goals the design and construction of new peptides and proteins with novel binding and catalytic properties. In one approach to protein engineering, new active sites have been introduced into naturally occurring proteins either by site-directed mutagenesis or by chemical modification. Providing that important changes in the tertiary structures do not result from such alterations, at least a portion of the binding site of the original protein should be available for the formation of complexes between the altered enzyme and its substrates. Many examples of active-site mutations have been described, including the generation by us of a cysteine mutant of alkaline phosphatase. A fundamental limitation of the site-directed mutagenesis methodology is that replacements of residues are restricted to the twenty naturally occurring amino acids. The alternative, chemical modification, is difficult to carry out for the specific replacement of one amino acid by another. However, we have shown that through such modification coenzyme analogues can be introduced covalently into appropriate positions in proteins, allowing us to produce semisynthetic enzymes with catalytic activities radically altered from those of their precursor proteins. In another approach to protein engineering efforts have focused on the construction of systems where, as a first approximation, folding can be neglected and the preparation of secondary structural units is the target. Examples of the successful design of biologically active peptides and proteins along such lines, taken from our own work, include molecules mimicking apolipoproteins, toxins, and many hormones. In recent studies we have progressed to the stage where we are starting to combine the two general approaches to protein engineering we have described and are able to construct small enzymes like ribonuclease T1 and its structural analogues. 相似文献
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Dr. Yunling Deng Dr. Sudharsan Dwaraknath Wenhao O. Ouyang Cory J. Matsumoto Stephanie Ouchida Prof. Dr. Yi Lu 《Angewandte Chemie (International ed. in English)》2023,62(20):e202215719
While native CO2-reducing enzymes display remarkable catalytic efficiency and product selectivity, few artificial biocatalysts have been engineered to allow understanding how the native enzymes work. To address this issue, we report cobalt porphyrin substituted myoglobin (CoMb) as a homogeneous catalyst for photo-driven CO2 to CO conversion in water. The activity and product selectivity were optimized by varying pH and concentrations of the enzyme and the photosensitizer. Up to 2000 TON(CO) was attained at low enzyme concentrations with low product selectivity (15 %), while a product selectivity of 74 % was reached by increasing the enzyme loading but with a compromised TON(CO). The efficiency of CO generation and overall TON(CO) were further improved by introducing positively charged residues (Lys or Arg) near the active stie of CoMb, which demonstrates the value of tuning the enzyme secondary coordination sphere to enhance the CO2-reducing performance of a protein-based photocatalytic system. 相似文献
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Alexander D. Ryabov 《Angewandte Chemie (International ed. in English)》1991,30(8):931-941
Most specialists doing organometallic chemistry have little understanding of what modern biochemistry is. On the other hand, most biochemists believe that organometallic chemistry stands much apart from the problems they study. But the real distance, if any, between these magnificent pyramids of modern science is progressively decreasing. Their interaction has given birth to a new branch of science, organometallic biochemistry, the general aspects of which are discussed here. 相似文献
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Matúš Gajdoš Jendrik Wagner Felipe Ospina Antonia Köhler Dr. Martin K. M. Engqvist Prof. Dr. Stephan C. Hammer 《Angewandte Chemie (International ed. in English)》2023,62(7):e202215093
Enantioselective synthesis of chiral alcohols through asymmetric addition of water across an unactivated alkene is a highly sought-after transformation and a big challenge in catalysis. Herein we report the identification and directed evolution of a fatty acid hydratase from Marinitoga hydrogenitolerans for the highly enantioselective hydration of styrenes to yield chiral 1-arylethanols. While directed evolution for styrene hydration was performed in the presence of heptanoic acid to mimic fatty acid binding, the engineered enzyme displayed remarkable asymmetric styrene hydration activity in the absence of the small molecule activator. The evolved styrene hydratase provided access to chiral alcohols from simple alkenes and water with high enantioselectivity (>99 : 1 e.r.) and could be applied on a preparative scale. 相似文献
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Dr. Binbin Gu Prof. Dr. Bernd Goldfuss Prof. Dr. Jeroen S. Dickschat 《Angewandte Chemie (International ed. in English)》2023,62(1):e202215688
The sesterviolene synthase from Streptomyces violens was identified and represents the second known sesterterpene synthase from bacteria. Isotopic labelling experiments in conjunction with DFT calculations were performed that provided detailed insight into its complex cyclisation mechanism. Enzyme engineering through site-directed mutagenesis gave access to a high-yielding enzyme variant that provided six additional minor products and the main product in sufficient quantities to study its chemistry. 相似文献
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Christian A. Gomez Dibyendu Mondal Qian Du Natalie Chan Jared C. Lewis 《Angewandte Chemie (International ed. in English)》2023,62(15):e202301370
FeII- and α-ketoglutarate-dependent halogenases and oxygenases can catalyze site-selective functionalization of C−H bonds via a variety of C−X bond forming reactions, but achieving high chemoselectivity for functionalization using non-native functional groups remains rare. The current study shows that directed evolution can be used to engineer variants of the dioxygenase SadX that address this challenge. Site-selective azidation of succinylated amino acids and a succinylated amine was achieved as a result of mutations throughout the SadX structure. The installed azide group was reduced to a primary amine, and the succinyl group required for azidation was enzymatically cleaved to provide the corresponding amine. These results provide a promising starting point for evolving additional SadX variants with activity on structurally distinct substrates and for enabling enzymatic C−H functionalization with other non-native functional groups. 相似文献
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Lemeng Chao Seino Jongkees 《Angewandte Chemie (Weinheim an der Bergstrasse, Germany)》2019,131(37):12880-12890
Carbohydrates are attached and removed in living systems through the action of carbohydrate‐active enzymes such as glycosyl transferases and glycoside hydrolases. The molecules resulting from these enzymes have many important roles in organisms, such as cellular communication, structural support, and energy metabolism. In general, each carbohydrate transformation requires a separate catalyst, and so these enzyme families are extremely diverse. To make this diversity manageable, high‐throughput approaches look at many enzymes at once. Similarly, high‐throughput approaches can be a powerful way of finding inhibitors that can be used to tune the reactivity of these enzymes, either in an industrial, a laboratory, or a medicinal setting. In this review, we provide an overview of how these enzymes and inhibitors can be sought using techniques such as high‐throughput natural product and combinatorial library screening, phage and mRNA display of (glyco)peptides, fluorescence‐activated cell sorting, and metagenomics. 相似文献