Green manufacturing synthesis of nicotinamide mononucleotide: Pathways, catalysts, and prospects.

Bacillus subtilis a Generally Recognized As Safe (GRAS) microorganism, is an attractive chassis for producing high-value compounds in a safe and sustainable way. However, its potential for producing the C40 carotenoid lycopene has been limited by inefficient precursor supply and enzyme incompatibility. This study demonstrates that lycopene production in B. subtilis can be significantly enhanced through systematic metabolic engineering by rewiring the lycopene and methylerythritol phosphate (MEP) pathways. A synthetic lycopene biosynthesis pathway expressing the crtE gene from Pantoea agglomerans, which is commonly used for microbial lycopene production, failed to yield lycopene production in B. subtilis. However, replacing crtE with a multifunctional geranylgeranyl diphosphate synthase (GGPPS) from Archaeoglobus fulgidus successfully enabled lycopene synthesis. The optimization of the fermentation medium demonstrated that a combined carbon supply of glucose and glycerol markedly enhanced both cell growth and lycopene production in comparison with separate carbon sources. To further boost production, the methylerythritol phosphate (MEP) pathway was engineered by overexpressing the rate-limiting enzyme, 1-deoxy-D-xylulose-5-phosphate synthase (dxs), which resulted in a five-fold increase in lycopene titer after 72 h. Screening of various GGPPS enzymes revealed that idsA from Corynebacterium glutamicum was the most efficient, further increasing the yield. The final engineered strain achieved a lycopene titer of 55 mg/L in shake-flask cultivation, a significant improvement over the previously reported level in B. subtilis. These results demonstrate that targeted GGPPS selection and precursor pathway engineering are critical strategies for developing B. subtilis into a robust and sustainable platform for carotenoid production.

Solvent stress occurs during whole-cell biocatalysis of organic chemicals. Organic substrates and/or products may accumulate in the cellular membranes of whole cells, causing structural destabilization of the membranes, which leads to disturbances in cellular carbon and energy metabolism. Here, we investigate the effect of cyclohexanone on carbon metabolism in Escherichia coli BL21 and Corynebacterium glutamicum ATCC13032. Adding cyclohexanone to the culture medium (i.e., glucose mineral medium) resulted in a decreased specific growth rate and increased cellular maintenance energy in both strains of bacteria. Notably, carbon metabolism, which is mainly involved to increase cellular maintenance energy, was very different between the bacteria. Carbon flux into the acetic acid fermentation pathway was dominantly enhanced in E. coli, whereas the TCA cycle appeared to be activated in C. glutamicum. In fact, carbon flux into the TCA cycle in E. coli appeared to be reduced with increasing amounts of cyclohexanone in the culture medium. Metabolic engineering of E. coli cells to maintain or improve TCA cycle activity and, presumably, that of the electron transport chain, which are involved in regeneration of cofactors (e.g., NAD(P)H and ATP) and formation of toxic metabolites (e.g., acetic acid), may be useful in increasing solvent tolerance and biotransformation of organic chemicals (e.g., cyclohexanone).

The purpose of this study was to investigate whether intensive insulin treatment of dogs suffering from type 1 diabetes mellitus, resulting in tight glycemic control, could be reflected by changes in peripheral leukocyte metabolism. Specifically, plasma metabolites and enzyme activities were assessed. In addition, quantitative RT-PCR was used to determine changes in insulin signaling gene (insulin receptor substrate (IRS)-1, IRS-2 and phosphatidylinositol 3-kinase (PI3-K) P85alpha) mRNA levels in peripheral leukocytes. Lastly, leukocyte enzymes involved in cellular energy metabolism were examined for changes in glucose utilization. Our results indicated that intensive insulin treatment was successful in type 1 DM dogs, leading to tight glycemic control. The mean glucose concentration and glycated albumin percentage significantly decreased to 156 mg/dl and 15.6%, respectively, following treatment. In peripheral leukocytes, the IRS-2 and PI3-K p85alpha mRNA levels significantly increased, and a significant increase in pyruvate kinase and pyruvate carboxylase activity, two enzymes involved in cellular energy metabolism, was also observed post treatment. Therefore, the observed changes in insulin signaling pathway activity and cellular energy metabolism enzyme activity in peripheral leukocytes are considered to be characteristics of amelioration of glucose metabolism by insulin action. As such, peripheral leukocytes are sufficiently sensitive to monitor for improving glycemic control during intensive insulin treatment of type 1 DM dogs. Blood cells such as leukocytes are much more readily available than muscle or adipose tissue for studies in dogs.

Understanding and quantification of cosolvent influences on enzyme-catalyzed reactions are driven by a twofold interest. On the one hand, cosolvents can simulate the cellular environment for deeper understanding of in cellulo reaction conditions. On the other hand, cosolvents are applied in biotechnology to tune yield and kinetics of reactions. Further, cosolvents are even present inherently, for example, for reactions with cofactor regeneration or for enzymes that need cosolvents in a function of a stabilizer. As the experimental determination of yield and kinetics is costly and time consuming, this work aims at providing a thermodynamic predictive approach that might allow screening cosolvent influences on yield and Michaelis constants. Reactions investigated in this work are the reduction of butanone and 2-pentanone under the influence of 17 wt % of the cosolvent polyethylene glycol 6000, which is also often used as a crowder to simulate cellular environments. The considered reactions were catalyzed by a genetically modified alcohol dehydrogenase (ADH 270). Predictions of cosolvent influences are based on accounting for a cosolvent-induced change of molecular interactions among the reacting agents as well as between the reacting agents and the solvent. Such interactions were characterized by activity coefficients of the reacting agents that were predicted by means of electrolyte perturbed-chain statistical associating fluid theory. This allowed simultaneously predicting the cosolvent effects on yield and Michaelis constants for two-substrate reactions for the first time.

Cu/nucleotide coordination self-assembling to in situ regenerate NAD(P)+ and co-immobilize dehydrogenase with robust activity and stability

Combined cross-linked enzyme aggregates of glycerol dehydrogenase and NADH oxidase for high efficiency in situ NAD+ regeneration

eSports, as an active branch of the digital economy, has generated widespread influence globally and is exhibiting rapid development momentum in the Yangtze River Delta region. Based on an in-depth study of the eSports industry in this region, this paper analyzes the macro trends and micro mechanisms of the industry's development. Through empirical data and case studies, a systematic exploration of the development of the eSports industry in the Yangtze River Delta region is conducted. The research first clarifies the support policies for the eSports industry development by the governments of the three provinces and one city in the Yangtze River Delta and examines their effects. It reveals the positive role these policies play in promoting the formation and expansion of the regional game industry clusters. Secondly, it provides a detailed analysis of the evolution of internal enterprise cooperation and innovation models within the industry and uses models to predict main future trends of industry ecological changes. The paper also offers a rigorous assessment of the challenges, potential risks, and the impacts on the economic and cultural aspects faced by the industry's sustainable development, proposing strategies and recommendations to achieve ongoing advantages in global competition. Specifically, the research delves into how the eSports industry can leverage new information technologies, such as 5G networks, artificial intelligence, and big data analytics, as well as explores the potential for cross-integration with other industries, including education, tourism, and retail. The research also analyzes the region's global strategic layout of the eSports industry from an international perspective, focusing on the positive impact of international competitions on local economic development and brand image enhancement. In addition, through detailed studies of the eSports user group, it analyses how users' needs, consumer behavior, and market dynamics can promote the industry's healthy development. The paper also critically reflects on the social responsibility and ethical issues of quality development in the eSports industry, examining its potential effects on promoting social harmony and cultural dissemination, as well as its impact on the healthy growth of adolescents. By employing comprehensive research methods, this paper not only explores beneficial growth paths and models for the eSports industry in the Yangtze River Delta region but also provides strategic decision-making support for local businesses and government departments. It contributes to the mutual development and cooperation within and beyond the region's eSports industry chain. Towards the end of the study, the paper forecasts new opportunities and challenges for the Yangtze River Delta region in the global eSports industry due to the introduction of new technologies and changes in the international competitive landscape. It emphasizes the need to develop long-term plans and enhance the regional industrial support system. In conclusion, this research defines the position and role of the Yangtze River Delta region in the global eSports industry, analyzes key points for high-quality development strategies, and offers profound theoretical insights and practical guidance for promoting innovation, collaboration, and sustainable growth within the regional eSports industry.

This study aims to clarify the current status of development of China's sports industry, explore the fundamental meaning and basic elements of high-quality development of the sports industry, identify and analyze the main challenges faced by the sports industry, and provide theoretical inspiration and practical guidance for achieving sustainable and high-quality development of China's sports industry. This study adopted the literature method and logical analysis method. Through extensive reference to relevant literature and policy documents, the theoretical basis for the high-quality development of the sports industry was systematically sorted out;. Using the logical analysis method, its internal relationship and framework were explored, and the current status of the development of China's sports industry was comprehensively evaluated. The research results show that in the process of achieving high-quality development, China's sports industry is faced with problems such as insufficient technological innovation capabilities, unreasonable industrial structure, unbalanced regional development, and weak international competitiveness, which have become the main factors hindering the high-quality development of China's sports industry. In order to promote the high-quality development of China's sports industry, efforts should be made in the following aspects. Firstly, deepen the integration of science and technology in the sports industry, and enhance innovation capabilities and total factor productivity. Secondly, optimize the structure of the sports industry, and promote field collaboration and efficient allocation of resources. Thirdly, increase sports industry brand building and improve product quality and market influence. Fourthly, practice the green and low-carbon concept of sports, and balance environmental protection and economic growth. Last but not least, strengthen the collaboration between the government, enterprises and all sectors of society to form a joint force to promote the upgrading of China's sports industry. It is hoped that this study can provide a theoretical framework and practical guidance for the high-quality development of China's sports industry, provide inspiring value for relevant policy makers and practitioners in the sports industry, and enhance the international competitiveness of China's sports industry.

With the shortage of global energy resources and the increased awareness of environmental protection, in recent years, China's new energy vehicle industry has developed rapidly and become a major player in the global electric vehicle market. This paper will discuss the competitive situation of China's new energy vehicles in the global market and its impact on the activation of the global new energy vehicle market. By analyzing the policy support, technological innovation, market expansion and international competitiveness of China's new energy vehicle industry, this paper aims to reveal China's important position in the global new energy vehicle market and its far-reaching impact on the global industrial structure. In recent years, China's new energy vehicle industry has sprung up and developed rapidly, and has become a powerful force in the global electric vehicle market. In this ever-changing era of science and technology, the new energy vehicle industry is regarded as an important driving force for future global economic growth, and countries have increased investment and competed for market share. In terms of technological innovation, China's new energy automobile industry continues to increase investment in research and development, breakthroughs in key technologies, and has achieved remarkable results. From the perspective of policy support, the Chinese government attaches great importance to the development of the new energy automobile industry and has introduced a series of policy measures to encourage and promote the rapid development of the industry. From the perspective of market expansion, the market of new energy vehicles in our country shows a rapid growth trend. In terms of international competitiveness, China's new energy automobile industry has gradually risen, and has the strength to compete with developed countries such as Europe, America and Japan. With the continuous growth of China's new energy automobile industry, the export business has also achieved remarkable results, and the international market share has gradually increased. On a global scale, China's new energy automobile industry has formed a new pattern of competition with major economies. The competitive situation of China's new energy automobile industry in the global market has become increasingly prominent, which has a positive activation effect on the global new energy automobile market. Under the joint promotion of policy support, technological innovation, market expansion and international competitiveness, China's new energy vehicle industry will continue to play an important role in the global market and have a profound impact on the global industrial pattern. With the increasingly fierce competition in the field of new energy vehicles, the development of China's new energy vehicle industry will have a greater impact and influence on the global market.

Against the backdrop of accelerated global economic integration, the scale of international trade continues to expand. As a key node in the international logistics chain and an important hub of international trade, the logistics efficiency of ports plays a decisive role in regional economic development and even the stable operation of the global supply chain. Shanghai Port, as one of the top ports in terms of global cargo throughput and container throughput, is an important gateway for China's foreign trade and a key strategic resource for participating in international economic cooperation and competition. It not only undertakes the task of transferring and transporting a large amount of domestic goods, but also connects major economic regions around the world, occupying a core position in the global shipping network. However, with the profound adjustment of the global trade pattern, the rapid development of shipping technology, and the increasing demand for logistics services from customers, the logistics development of Shanghai Port is facing many challenges. On the one hand, competition among surrounding ports has intensified, with internationally renowned ports such as Singapore Port and Busan Port continuously improving their logistics service levels and competing for market share in the international shipping market, bringing enormous competitive pressure to Shanghai Port; On the other hand, there are still problems in the logistics operation of Shanghai Port, such as the need for further improvement of port infrastructure, insufficient integration and sharing of logistics information, and poor connection of multimodal transport, which make it difficult for logistics efficiency to meet the growing business needs and restrict the further improvement of Shanghai Port's international competitiveness. In this situation, in-depth research on strategies to improve the efficiency of Shanghai Port's port logistics will not only help Shanghai Port break through development bottlenecks, enhance its core competitiveness, consolidate its position in the global shipping market, but also provide useful references for other domestic ports, promote the overall high-quality development of China's port logistics industry, and better serve the country's opening-up strategy and economic development.

Against the backdrop of economic globalization and trade liberalization, financial services, as an emerging sector, have gradually become a focal point of attention for countries around the world. The development of financial services trade has accelerated the flow of capital between nations and effectively promoted global trade as well as the development of non-trade industries. This article, based on the background of the RCEP coming into effect, conducts a comparative analysis of the financial services trade competitiveness between China and RCEP member countries, and empirically studies the factors influencing China’s financial services trade competitiveness based on Porter’s Diamond Model.
 This study first starts with the current status of financial services trade among RCEP member countries, comparing the competitiveness of China’s financial services trade with other RCEP member countries using three commonly used indicators for evaluating international trade competitiveness: market share (MS), trade competitiveness index (TC), and revealed comparative advantage index (RCA). Subsequently, based on Porter’s competitive advantage theory, an evaluation system is constructed from six aspects: factor conditions, demand conditions, related and supporting industries, government, enterprise organization strategy and competition, and opportunities. The regression model is constructed using data from 2006 to 2021 combined with the principal component analysis method to empirically analyze the factors influencing China’s financial services trade competitiveness. The analysis results show that China’s financial services trade international competitiveness is relatively weak among RCEP member countries, but it has shown a strong growth trend in recent years. The impact of fiscal policy on the competitiveness of financial services trade is the largest, followed by technological and related industry factors, while human capital has the smallest contribution to the competitiveness of financial services trade.
 Based on the research results, in the future, China should strengthen the cultivation of talents in the financial services industry, promote consumer spending in the financial services sector, increase the capital and technological content of related financial industries, expand the scale of foreign capital utilization in the financial services industry, and improve the financial regulatory system to enhance China’s international competitiveness in financial services trade.

With the continuous development of benefits, China is slowly moving towards the era of service economy, and the service industry occupies an increasingly large position in our country's national economy, which also promotes the existence of weak industries in our country's international service trade competitiveness. As a kind of our country's service industry, financial service trade still has a big gap with other countries in terms of international competitiveness. Financial service trade is one of our country's comparative advantages, but with economic globalization, the connection between countries getting closer.In such an open era, factors such as the accelerated international capital flow, large capital flow, and fierce market competition have caused serious impacts to most countries in the world, and problems have arisen in the development of the domestic financial industry and the real economy industry. As a result, it will affect the overall competitiveness of the domestic financial industry, etc. These situations lead us to pay attention to some bad results brought about by financial services trade. It is necessary to analyze and study the reasons for these bad performances and give corresponding measures. This paper mainly conducts a relevant analysis from the research on the international competitiveness of China's financial services trade.

Competitiveness is a determining factor in the survival, stabilization and development of industries, as well as their individual industries, sub-industries and enterprises. In European and world markets, national and sectoral competitiveness focuses on the performance of enterprises and individual industries, on the one hand, and the competitiveness of enterprises and similar industries, on the other. Thus, in the modern conditions of international integration, innovative development, the international competitiveness of the enterprise acquires special significance. Today’s economy defines specific requirements for increasing the level of enterprises competitiveness, rapid response to changes in the economic situation in order to support sustainable development, strengthen financial stability, continuous improvement of internal business processes in accordance with changing market conditions. The wine industry is an important area of economic development in Ukraine and Odessa region. For the successful development of the viticulture and winemaking complex in Ukraine and in the region there are favorable soil and climatic conditions and sufficient provision of labor and other resources. However, today the stable and efficient operation of wineries is hampered by a number of problems, including the general crisis of the country’s economy, insecurity of domestic producers, lack of proper state support, military aggression and blockade of seaports in Odessa region. In this regard, the problem of increasing the international competitiveness of products and enterprises in general is of particular importance in the management system of enterprises in the wine industry. The tool for solving strategic tasks of increasing the international competitiveness of wineries is a comprehensive approach to developing and building a model of competitiveness of enterprises and products of the wine industry, the development of an effective mechanism for managing their competitiveness. The paper considers the theoretical and methodological foundations of the study of the winery’s international competitiveness. Factors influencing the winery’s international competitiveness are studied and the methods of assessing the winery’s international competitiveness are systematized.

The chemical engineering industry relies increasingly on enzyme-catalyzed reactions in the production of fine chemicals, value-added compounds, and commodity chemicals. However, many attractive transformations, such as the selective reduction of ketones and alkenes, depend on cofactors that are prohibitively expensive for application on an industrial scale. One such cofactor, nicotinamide adenine dinucleotide (NAD+/NADH), is utilized by >20% of all enzymes but is required in generally super-stoichiometric amounts and currently has no efficient means for recycling. Direct electrochemical reduction of NAD+ is an attractive approach for regeneration of the biological cofactor; however, electrolysis often results in relatively small amounts of biologically active NADH due to a parasitic dimerization. Recent work has demonstrated the ability of lower-cost NADH mimetics to exhibit enzymatic activity, sometimes exceeding that of NADH itself, although still required in at least stoichiometric amounts. Subsequently, we considered the possibility of designing NADH biomimetics to simultaneously maintain enzymatic activity and minimize deleterious side reactions, thereby allowing them to be electrochemically recycled. We will describe our recent progress in both measuring and modeling the electrochemical dimerization and regeneration rates of NADH mimetics. We have employed Electrochemical Impedance Spectroscopy to quantify rapid dimerization rates observed for electrochemically reduced NADH biomimetics. By combining this electroanalytical approach with density functional theory (DFT) modeling and machine learning algorithms, we are elucidating structure-function relationships to design more efficiently regenerable mimetics.

Guanidinoacetate methyltransferase (GAMT) catalyzes the last step of the biosynthetic pathway to creatine (Cr), the transfer of a methyl group from S-adenosylmethionine to guanidinoactate. Despite the importance of this methylation reaction in cellular energy metabolism and in methyl group metabolism, a systematic study of the expression and cellular localization of this enzyme is lacking. As a means for determining why the testis and seminal vesicles contain high levels of Cr, we analyzed GAMT protein and mRNA levels in mouse tissues by Western and Northern blot analyses. The results show that GAMT was regulated at tissue-specific levels; the enzyme was most highly expressed in testis, caput epididymis, ovary, and liver. Differences were also noted between sexes; the levels of GAMT mRNA and protein were higher in female liver than in male liver. Furthermore, as the male mouse developed from neonatal stages through sexual maturity, the GAMT protein level increased in testis but decreased in liver. Immunohistochemical labeling showed that GAMT was localized primarily in Sertoli cells of the testis and in microvilli of the epithelial cells lining the caput epididymis. GAMT expression in Sertoli cells was confirmed by biochemical and molecular analyses of purified testicular cells, testes from prepubertal mice of different ages, and germ cell-deficient mutant mouse testes. These results indicate that Cr is synthesized extensively in the epithelia of seminiferous tubules and caput epididymis, suggesting that GAMT or metabolic pathways related to Cr biosynthesis may be important for germ cell development or function.