scholarly journals Protein Engineering Approaches to Enhance Fungal Laccase Production in S. cerevisiae

2021 ◽  
Vol 22 (3) ◽  
pp. 1157
Author(s):  
Pablo Aza ◽  
Felipe de Salas ◽  
Gonzalo Molpeceres ◽  
David Rodríguez-Escribano ◽  
Iñigo de la Fuente ◽  
...  
Keyword(s):  
Protein Engineering ◽  
Directed Evolution ◽  
Signal Peptide ◽  
Laccase Activity ◽  
Laccase Production ◽  
Wide Range ◽  
Mating Factor ◽  
Conserved Amino Acids ◽  
Basidiomycete Fungi

Laccases secreted by saprotrophic basidiomycete fungi are versatile biocatalysts able to oxidize a wide range of aromatic compounds using oxygen as the sole requirement. Saccharomyces cerevisiae is a preferred host for engineering fungal laccases. To assist the difficult secretion of active enzymes by yeast, the native signal peptide is usually replaced by the preproleader of S. cerevisiae alfa mating factor (MFα1). However, in most cases, only basal enzyme levels are obtained. During directed evolution in S. cerevisiae of laccases fused to the α-factor preproleader, we demonstrated that mutations accumulated in the signal peptide notably raised enzyme secretion. Here we describe different protein engineering approaches carried out to enhance the laccase activity detected in the liquid extracts of S. cerevisiae cultures. We demonstrate the improved secretion of native and engineered laccases by using the fittest mutated α-factor preproleader obtained through successive laccase evolution campaigns in our lab. Special attention is also paid to the role of protein N-glycosylation in laccase production and properties, and to the introduction of conserved amino acids through consensus design enabling the expression of certain laccases otherwise not produced by the yeast. Finally, we revise the contribution of mutations accumulated in laccase coding sequence (CDS) during previous directed evolution campaigns that facilitate enzyme production.

A comparative study on the laccase activity of four basidiomycete fungi with different lignocellulosic residues via solid-state fermentation

BioResources ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. 3017-3031
Author(s):  
Mei-Ling Han ◽  
Qi An ◽  
Kai-Yue Ma ◽  
Wen-Ning An ◽  
Wen-Yao Hao ◽  
...  
Keyword(s):  
Oryza Sativa ◽  
Solid State ◽  
Comparative Study ◽  
Sorghum Bicolor ◽  
Solid State Fermentation ◽  
Laccase Activity ◽  
Industrial Applications ◽  
Laccase Production ◽  
Lignocellulosic Residues ◽  
Basidiomycete Fungi

The laccase producing abilities of four Basidiomycete fungi species were compared using solid-state fermentation using four different lignocellulosic residues. The biosynthetic potential of the Basidiomycetes was highly dependent on the type of fungi. In general, the laccase secreting ability of Cerrena unicolor Han 849 was greater than Lenzites betulinus Han 851, Stropharia rugosoannulata Han 1321, and Auricularia heimuer Han 1333. The maximum laccase production of C. unicolor Han 849 was approximately 11.25, 122.26, and 15.27 times higher than L. betulinus Han 851, S. rugosoannulata Han 1321 and A. heimuer Han 1333, respectively. Different species of fungi had a preference in lignocellulosic residues. The presence of Firmiana platanifolia was conducive to secreting laccase via C. unicolor Han 849 during solid-state fermentation. A continuous and stable laccase production via C. unicolor Han 849 was an obvious advantage of solid-state fermentation with any of the four lignocellulosic residues used. The maximum laccase production of C. unicolor Han 849 using Firmiana platanifolia was approximately 2.12, 1.68, and 6.13 times higher than Populus beijingensis, Sorghum bicolor, and Oryza sativa, respectively. These findings will be helpful for developing new productivity strains in industrial applications and selecting suitable lignocellulosic residues for laccase production.

scholarly journals Droplet Microfluidics-Enabled High-Throughput Screening for Protein Engineering

Micromachines ◽  
2019 ◽  
Vol 10 (11) ◽  
pp. 734 ◽  
Author(s):  
Lindong Weng ◽  
James E. Spoonamore
Keyword(s):  
Protein Engineering ◽  
Directed Evolution ◽  
High Throughput ◽  
Protein Design ◽  
High Throughput Screening ◽  
Fluid Phase ◽  
Droplet Microfluidics ◽  
New Paradigm ◽  
Wide Range ◽  
Challenges And Opportunities

Protein engineering—the process of developing useful or valuable proteins—has successfully created a wide range of proteins tailored to specific agricultural, industrial, and biomedical applications. Protein engineering may rely on rational techniques informed by structural models, phylogenic information, or computational methods or it may rely upon random techniques such as chemical mutation, DNA shuffling, error prone polymerase chain reaction (PCR), etc. The increasing capabilities of rational protein design coupled to the rapid production of large variant libraries have seriously challenged the capacity of traditional screening and selection techniques. Similarly, random approaches based on directed evolution, which relies on the Darwinian principles of mutation and selection to steer proteins toward desired traits, also requires the screening of very large libraries of mutants to be truly effective. For either rational or random approaches, the highest possible screening throughput facilitates efficient protein engineering strategies. In the last decade, high-throughput screening (HTS) for protein engineering has been leveraging the emerging technologies of droplet microfluidics. Droplet microfluidics, featuring controlled formation and manipulation of nano- to femtoliter droplets of one fluid phase in another, has presented a new paradigm for screening, providing increased throughput, reduced reagent volume, and scalability. We review here the recent droplet microfluidics-based HTS systems developed for protein engineering, particularly directed evolution. The current review can also serve as a tutorial guide for protein engineers and molecular biologists who need a droplet microfluidics-based HTS system for their specific applications but may not have prior knowledge about microfluidics. In the end, several challenges and opportunities are identified to motivate the continued innovation of microfluidics with implications for protein engineering.

The Problem of Identifying and Modeling the Relationship between Monetary Factor and Inflation in the Russian Economy

2008 ◽  
pp. 61-76
Author(s):  
A. Porshakov ◽  
A. Ponomarenko
Keyword(s):  
Money Supply ◽  
Money Demand ◽  
Russian Economy ◽  
Long Run ◽  
Complex Approach ◽  
Wide Range ◽  
Long Time ◽  
The Relationship ◽  
Supply Side Factors

The role of monetary factor in generating inflationary processes in Russia has stimulated various debates in social and scientific circles for a relatively long time. The authors show that identification of the specificity of relationship between money and inflation requires a complex approach based on statistical modeling and involving a wide range of indicators relevant for the price changes in the economy. As a result a model of inflation for Russia implying the decomposition of inflation dynamics into demand-side and supply-side factors is suggested. The main conclusion drawn is that during the recent years the volume of inflationary pressures in the Russian economy has been determined by the deviation of money supply from money demand, rather than by money supply alone. At the same time, monetary factor has a long-run spread over time impact on inflation.

Introduction: Emotion, History and the Arts

2018 ◽  
Vol 7 (2) ◽  
pp. 117-128 ◽  
Author(s):  
Erin Sullivan ◽  
Marie Louise Herzfeld-Schild
Keyword(s):  
Historical Change ◽  
Carson Mccullers ◽  
Emotional Life ◽  
Review Section ◽  
The Arts ◽  
Wide Range ◽  
War Correspondents ◽  
History Of ◽  
Johan Huizinga

This introduction surveys the rise of the history of emotions as a field and the role of the arts in such developments. Reflecting on the foundational role of the arts in the early emotion-oriented histories of Johan Huizinga and Jacob Burkhardt, as well as the concerns about methodological impressionism that have sometimes arisen in response to such studies, the introduction considers how intensive engagements with the arts can open up new insights into past emotions while still being historically and theoretically rigorous. Drawing on a wide range of emotionally charged art works from different times and places—including the novels of Carson McCullers and Harriet Beecher-Stowe, the private poetry of neo-Confucian Chinese civil servants, the photojournalism of twentieth-century war correspondents, and music from Igor Stravinsky to the Beatles—the introduction proposes five ways in which art in all its forms contributes to emotional life and consequently to emotional histories: first, by incubating deep emotional experiences that contribute to formations of identity; second, by acting as a place for the expression of private or deviant emotions; third, by functioning as a barometer of wider cultural and attitudinal change; fourth, by serving as an engine of momentous historical change; and fifth, by working as a tool for emotional connection across communities, both within specific time periods but also across them. The introduction finishes by outlining how the special issue's five articles and review section address each of these categories, while also illustrating new methodological possibilities for the field.

Short Films from a Small Nation

2018 ◽  
Author(s):  
C. Claire Thomson
Keyword(s):  
Public Information ◽  
Industrial Process ◽  
Actor Network Theory ◽  
Documentary Films ◽  
Wide Range ◽  
Short Films ◽  
Post War ◽  
The 1960S ◽  
Art And Architecture

The first book-length study in English of a national corpus of state-sponsored informational film, this book traces how Danish shorts on topics including social welfare, industry, art and architecture were commissioned, funded, produced and reviewed from the inter-war period to the 1960s. For three decades, state-sponsored short filmmaking educated Danish citizens, promoted Denmark to the world, and shaped the careers of renowned directors like Carl Th. Dreyer. Examining the life cycle of a representative selection of films, and discussing their preservation and mediation in the digital age, this book presents a detailed case study of how informational cinema is shaped by, and indeed shapes, its cultural, political and technological contexts.The book combines close textual analysis of a broad range of films with detailed accounts of their commissioning, production, distribution and reception in Denmark and abroad, drawing on Actor-Network Theory to emphasise the role of a wide range of entities in these processes. It considers a broad range of genres and sub-genres, including industrial process films, public information films, art films, the city symphony, the essay film, and many more. It also maps international networks of informational and documentary films in the post-war period, and explores the role of informational film in Danish cultural and political history.

Quantitative and Qualitative Methods of Studying the Processes of Socialization of a Modern Teenager

2020 ◽  
pp. 102-109
Author(s):  
Svetlana Alekseevna Raschetina ◽  
Keyword(s):  
Research Methods ◽  
Interpersonal Relations ◽  
Internet Communication ◽  
Socialization Process ◽  
Wide Range ◽  
The Family ◽  
Modern Family ◽  
Making Friends ◽  
Main Support

Relevance and problem statement. Modern unstable society is characterized by narrowing the boundaries of controlled socialization and expanding the boundaries of spontaneous socialization of a teenager based on his immersion in the question arises about the importance of the family in the process of socialization of a teenager in the conditions of expanding the space of socialization. There is a need to study the role of the family in this process, to search, develop and test research methods that allow us to reveal the phenomenon of socialization from the side of its value characteristics. The purpose and methodology of the study: to identify the possibilities of a systematic and anthropological methodology for studying the role of the family in the process of socialization of adolescents in modern conditions, testing research methods: photo research on the topic “Ego – I” (author of the German sociologist H. Abels), profile update reflexive processes (by S. A. Raschetina). Materials and results of the study. The study showed that for all the problems that exist in the family of the perestroika era and in the modern family, it acts for a teenager as a value and the first (main) support in the processes of socialization. The positions well known in psychology about the importance of interpersonal relations in adolescence for the formation of attitudes towards oneself as the basis of socialization are confirmed. Today, the frontiers of making friends have expanded enormously on the basis of Internet communication. The types of activities of interest to a teenager (traditional and new ones related to digitalization) are the third pillar of socialization. Conclusion. The “Ego – I” method of photo research has a wide range of possibilities for quantitative and qualitative analysis of the socialization process to identify the value Pillars of this process.

Victorian Culture and Classical Antiquity

2011 ◽  
Author(s):  
Simon Goldhill
Keyword(s):  
Nineteenth Century ◽  
Roman Empire ◽  
Classical Antiquity ◽  
Victorian Culture ◽  
Wide Range ◽  
Victorian Art ◽  
Victorian Science ◽  
Role Of Religion ◽  
Nineteenth Century Novels

How did the Victorians engage with the ancient world? This book is an exploration of how ancient Greece and Rome influenced Victorian culture. Through Victorian art, opera, and novels, the book examines how sexuality and desire, the politics of culture, and the role of religion in society were considered and debated through the Victorian obsession with antiquity. Looking at Victorian art, it demonstrates how desire and sexuality, particularly anxieties about male desire, were represented and communicated through classical imagery. Probing into operas of the period, the book addresses ideas of citizenship, nationalism, and cultural politics. And through fiction—specifically nineteenth-century novels about the Roman Empire—it discusses religion and the fierce battles over the church as Christianity began to lose dominance over the progressive stance of Victorian science and investigation. Rediscovering some great forgotten works and reframing some more familiar ones, the book offers extraordinary insights into how the Victorian sense of antiquity and our sense of the Victorians came into being. With a wide range of examples and stories, it demonstrates how interest in the classical past shaped nineteenth-century self-expression, giving antiquity a unique place in Victorian culture.

Role of Cardiac MRI in Detecting Familial Hypertrophic Cardiomyopathy: Review

2016 ◽  
Vol 1 (1) ◽  
pp. 4
Author(s):  
Marymol Koshy ◽  
Bushra Johari ◽  
Mohd Farhan Hamdan ◽  
Mohammad Hanafiah
Keyword(s):  
Hypertrophic Cardiomyopathy ◽  
Magnetic Resonance ◽  
Genetic Disorder ◽  
Familial Hypertrophic Cardiomyopathy ◽  
Non Invasive ◽  
Wide Range ◽  
Proper Diagnosis ◽  
Magnetic Resonance Imaging Mri ◽  
Potential Use

Hypertrophic cardiomyopathy (HCM) is a global disease affecting people of various ethnic origins and both genders. HCM is a genetic disorder with a wide range of symptoms, including the catastrophic presentation of sudden cardiac death. Proper diagnosis and treatment of this disorder can relieve symptoms and prolong life. Non-invasive imaging is essential in diagnosing HCM. We present a review to deliberate the potential use of cardiac magnetic resonance (CMR) imaging in HCM assessment and also identify the risk factors entailed with risk stratification of HCM based on Magnetic Resonance Imaging (MRI).

Synthetic and Mechanistic Studies of a Versatile Heteroaryl Thioether Directing Group for Pd(II) Catalysis

2019 ◽  
Author(s):  
Andrew Romine ◽  
Kin Yang ◽  
Malkanthi Karunananda ◽  
Jason Chen ◽  
Keary Engle
Keyword(s):  
Mechanistic Studies ◽  
Salvianolic Acid ◽  
Wide Range ◽  
Computational Data ◽  
Synthetic Utility ◽  
Directing Group ◽  
High Yields ◽  
Julia Olefination ◽  
Reaction Progress Kinetic Analysis

A weakly coordinating monodentate heteroaryl thioether directing group has been developed for use in Pd(II) catalysis to orchestrate key elementary steps in the catalytic cycle that require conformational flexibility in a manner that is difficult to accomplish with traditional strongly coordinating directing groups. This benzothiazole thioether, (BT)S, directing group can be used to promote oxidative Heck reactivity of internal alkenes providing a wide range of products in moderate to high yields. To demonstrate the broad applicability of this directing group, arene C–H olefination was also successfully developed. Reaction progress kinetic analysis provides insights into the role of the directing group in each reaction, which is supplemented with computational data for the oxidative Heck reaction. Furthermore, this (BT)S directing group can be transformed into a number of synthetically useful functional groups, including a sulfone for Julia olefination, allowing it to serve as a “masked olefin” directing group in synthetic planning. In order to demonstrate this synthetic utility, natural products (+)-salvianolic acid A and salvianolic acid F are formally synthesized using the (BT)S directed C–H olefination as the key step.

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