Corner Engineering: Tailoring Enzymes for Enhanced Resistance and Thermostability in Deep Eutectic Solvents.

Deep eutectic solvents (DESs), heralded for their synthesis simplicity, economic viability, and reduced volatility and flammability, have found increasing application in biocatalysis. However, challenges persist due to a frequent diminution in enzyme activity and stability. Herein, we developed a general protein engineering strategy, termed corner engineering, to acquire DES-resistant and thermostable enzymes via precise tailoring of the transition region in enzyme structure. Employing Bacillus subtilis lipase A (BSLA) as a model, we delineated the engineering process, yielding five multi-DESs resistant variants with highly improved thermostability, such as K88E/N89 K exhibited up to a 10.0-fold catalytic efficiency (kcat /KM ) increase in 30 % (v/v) choline chloride (ChCl): acetamide and 4.1-fold in 95 % (v/v) ChCl: ethylene glycol accompanying 6.7-fold thermal resistance improvement than wild type at ≈50 °C. The generality of the optimized approach was validated by two extra industrial enzymes, endo-ß-1,4-glucanase PvCel5A (used for biofuel production) and esterase Bs2Est (used for plastics degradation). The molecular investigations revealed that increased water molecules at substrate binding cleft and finetuned helix formation at the corner region are two dominant determinants governing elevated resistance and thermostability. This study, coupling corner engineering with obtained molecular insights, illuminates enzyme-DES interaction patterns and fosters the rational design of more DES-resistant and thermostable enzymes in biocatalysis and biotransformation.

Revealing the biotoxicity of phosphorene oxide nanosheets based on the villin headpiece.

Phosphorene, a novel member of the two-dimensional nanomaterial family, has demonstrated great potential in biomedical applications, such as photothermal therapy, drug delivery and antibacterial. However, phosphorene is unstable and easily oxidized in an aerobic environment. In this paper, using larger-scale molecular dynamics simulations, we investigated the disruption of phosphorene oxide (PO) to the structure of a model protein, villin headpiece subdomain (HP35). It shows that the disruption of PO nanosheets to the protein structure is enhanced with increasing oxidation concentration of PO, while PO's oxidation mode has very little effect on the PO-HP35 interaction. PO with a low oxidation concentration has certain biocompatibility to HP35. Oxygen atoms filling into the groove region in the puckered surface of phosphorene enhance the dispersion interaction between phosphorene and HP35, which enhances the disruption of phosphorene to the structure of HP35. Compared with the dispersion interaction, the electrostatic interaction between PO and the protein has a negligible effect on the structural damage of HP35. These findings might shed light on the biological toxicity of PO nanosheets and would be helpful for future potential biomedical applications of PO nanosheets, such as nanodrugs and antibacterial agents.

Evolving Robust and Interpretable Enzymes for the Bioethanol Industry.

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.

Global plastic upcycling during and after the COVID-19 pandemic: The status and perspective.

Plastic pollution has become one of the most pressing environmental issues worldwide since the vast majority of post-consumer plastics are hard to degrade in the environment. The coronavirus disease (COVID-19) pandemic had disrupted the previous effort of plastic pollution mitigation to a great extent due to the overflow of plastic-based medical waste. In the post-pandemic era, the remaining challenge is how to motivate global action towards a plastic circular economy. The need for one package of sustainable and systematic plastic upcycling approaches has never been greater to address such a challenge. In this review, we summarized the threat of plastic pollution during COVID-19 to public health and ecosystem. In order to solve the aforementioned challenges, we present a shifting concept, regeneration value from plastic waste, that provides four promising pathways to achieve a sustainable circular economy: 1) Increasing reusability and biodegradability of plastics; 2) Transforming plastic waste into high-value products by chemical approaches; 3) The closed-loop recycling can be promoted by biodegradation; 4) Involving renewable energy into plastic upcycling. Additionally, the joint efforts from different social perspectives are also encouraged to create the necessary economic and environmental impetus for a circular economy.

Bioconversion of corn fiber to bioethanol: Status and perspectives.

Due to the rising demand for green energy, bioethanol has attracted increasing attention from academia and industry. Limited by the bottleneck of bioethanol yield in traditional corn starch dry milling processes, an increasing number of studies focus on fully utilizing all corn ingredients, especially kernel fiber, to further improve the bioethanol yield. This mini-review addresses the technological challenges and opportunities on the way to achieving the efficient conversion of corn fiber. Significant advances during the review period include the detailed characterization of different forms of corn kernel fiber and the development of off-line and in-situ conversion strategies. Lessons from cellulosic ethanol technologies offer new ways to utilize corn fiber in traditional processes. However, the commercialization of corn kernel fiber conversion may be hampered by enzyme cost, conversion efficiency, and overall process economics. Thus, future studies should address these technical limitations.

Discovery and mechanism-guided engineering of BHET hydrolases for improved PET recycling and upcycling.

Although considerable research achievements have been made to address the plastic crisis using enzymes, their applications are limited due to incomplete degradation and low efficiency. Herein, we report the identification and subsequent engineering of BHETases, which have the potential to improve the efficiency of PET recycling and upcycling. Two BHETases (ChryBHETase and BsEst) are identified from the environment via enzyme mining. Subsequently, mechanism-guided barrier engineering is employed to yield two robust and thermostable ΔBHETases with up to 3.5-fold enhanced kcat/KM than wild-type, followed by atomic resolution understanding. Coupling ΔBHETase into a two-enzyme system overcomes the challenge of heterogeneous product formation and results in up to 7.0-fold improved TPA production than seven state-of-the-art PET hydrolases, under the conditions used here. Finally, we employ a ΔBHETase-joined tandem chemical-enzymatic approach to valorize 21 commercial post-consumed plastics into virgin PET and an example chemical (p-phthaloyl chloride) for achieving the closed-loop PET recycling and open-loop PET upcycling.

Influence of carbon tariffs on China's export trade.

The database of the Global Trade Analysis Project (GTAP) and its energy-environmental model, known as GTAP-E, are used in this study to simulate the effect of the simultaneous and separate carbon tariff impositions of EU, the USA, and Japan on the export and export structure in China. Simulation results show that the carbon tariff impositions of developed countries on China will decrease the export to EU, the USA, and Japan but increase the export of China to other countries associated with the trade diversion in China. The USA and EU impose carbon tariffs on China, which will have a serious impact on China's export trade, especially for the export trade of energy-intensive industries. When Japan imposes carbon tariff on the exports of China, the positive influence on the export trade is weaker compared to the situation that the USA and EU imposed on China. Furthermore, imposing carbon tariffs on China will improve its trade structure; promote its agriculture, petroleum, and natural gas exploration and electricity industries; reduce its export trade volume of coal mining, petroleum products, and energy-intensive and other industries; decrease the export trade share of energy-intensive industries; and increase the export trade share and services of other industries. In this regard, China should reduce the carbon content of export products initiatively. On the one hand, China can solve this problem by levying carbon tax, developing emerging industries, and strengthening the research and development of low-carbon technologies. On the other hand, China should actively participate in the formulation of international standards of carbon tariff and become a participant in the international emission reduction rules in the field of climate change.