This Volume provides protocols for the biochemical analysis of hydrocarbon- and lipid-relevant products, cell components and activities of microbes that interact with hydrophobic compounds. They include methods for the extraction, purification and characterisation of surface tension-reducing bioemulsifiers and biosurfactants that increase the surface area and hence bioavailability of hydrophobic substrates. Protocols for the isolation and biochemical analysis of lipids and polyhydroxyalkanoates, food storage products made during nutrient abundance that represent important biotechnological products, are presented. The extraction of membrane lipid rafts, sub-organelles that fulfil important functional roles for the cell membrane, and the isolation and characterisation of membrane phospholipid biomarkers, are also described. The purification and characterisation of integral membrane hydrocarbon-oxidising enzymes are addressed. Lastly, two generic methods for the genetic analysis of catabolic pathways and analysis of ligand binding are presented. Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.

This Volume presents protocols for systems and synthetic biology applications in the field of hydrocarbon and lipid microbiology. It complements another Volume that describes generic protocols for wet experimental and computer-based systems and synthetic biology studies. The protocols in this Volume demonstrate how to employ systems and synthetic biology approaches in the design of microbes for the production of esters, isoprenoids, hydrophobic polymers, rhamnolipid biosurfactant, and peptide antimicrobial and thioether-stabilised molecules. Also presented is a protocol for the engineering of transcription factor-based biosensors for intracellular products, and another for the creation of a synthetic hydroxylase with novel activity for the selective oxyfunctionalisation of linear alkanes. Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.

This book covers the latest developments in enzyme immobilization with its wide applications, such as for industry, agriculture, medicine, and the environment. Topics covered include basics of enzyme immobilization, its implication in therapeutics and disease diagnostics, and its significance in solving environmental problems. This is an ideal book for researchers, graduate and postgraduate students, as well as scientists in industry, agriculture and health sectors. This book is a complete summary of enzyme immobilization and also thoroughly covers all the latest research. This book covers: The last one-hundred years of innovative research done in enzyme immobilization Recent developments in immobilization techniques, such as types of matrices, immobilization methods, and linking agents, as well as enzyme immobilization without any matrices and its properties The physiological and industrial significance of enzymes from plants and the implementation of immobilized enzymes in the treatment of waste water and polluted air Biomedical and bioanalytical applications of immobilized enzymes

This book delves into the recent developments in the microscale and microfluidic technologies that allow manipulation at the single and cell aggregate level. Expert authors review the dominant mechanisms that manipulate and sort biological structures, making this a state-of-the-art overview of conventional cell sorting techniques, the principles of microfluidics, and of microfluidic devices. All chapters highlight the benefits and drawbacks of each technique they discuss, which include magnetic, electrical, optical, acoustic, gravity/sedimentation, inertial, deformability, and aqueous two-phase systems as the dominant mechanisms utilized by microfluidic devices to handle biological samples. Each chapter explains the physics of the mechanism at work, and reviews common geometries and devices to help readers decide the type of style of device required for various applications. This book is appropriate for graduate-level biomedical engineering and analytical chemistry students, as well as engineers and scientists working in the biotechnology industry.

This book describes the latest research advances, innovations, and applications in the field of water management and environmental engineering as presented by leading researchers, engineers, life scientists and practitioners from around the world at the Frontiers International Conference on Wastewater Treatment (FICWTM), held in Palermo, Italy in May 2017. The topics covered are highly diverse and include the physical processes of mixing and dispersion, biological developments and mathematical modeling, such as computational fluid dynamics in wastewater, MBBR and hybrid systems, membrane bioreactors, anaerobic digestion, reduction of greenhouse gases from wastewater treatment plants, and energy optimization. The contributions amply demonstrate that the application of cost-effective technologies for waste treatment and control is urgently needed so as to implement appropriate regulatory measures that ensure pollution prevention and remediation, safeguard public health, and preserve the environment. The contributions were selected by means of a rigorous peer-review process and highlight many exciting ideas that will spur novel research directions and foster multidisciplinary collaboration among different water specialists.

This Volume presents generic protocols for wet experimental and computer-based systems and synthetic biology approaches relevant to the field of hydrocarbon and lipid microbiology. It complements a second Volume that describes protocols for systems and synthetic biology applications. The wet experimental tools presented in this Volume include protocols for the standardisation of transcriptional measurements, application of uracil excision-based DNA editing for, inter alia, multi-gene assembly, the use of fluxomics to optimise "reducing power availability", and the incorporation of non-canonical amino acids into proteins for optimisation of activities. Phenome-ing microbes, using a combination of RNA-seq and bioinformatic algorithms, is presented, as is an illustration, using methylotrophs as an example, of how the different key omics approaches constitute a pipeline for functional analysis, acquisition of a systems overview, and metabolic optimisation. Complementary computational tools that are presented include protocols for probing the genome architecture of regulatory networks, genome-scale metabolic reconstruction, and bioinformatic approaches to guide metabolic engineering. The Volume also includes an overview of how synthetic biology approaches can be used to improve biocontainment. Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.

This Volume describes methods for investigating microbes in their natural environment and how to obtain representative samples and preserve them for subsequent analyses. Chapters are arranged according to the environments under investigation, which include: oil reservoirs, fracking fluids, aquifers, coal beds, oil sands and their tailing ponds, lakes, rivers, leaves, polar seas and ice, the sea-surface microlayer, mud flats, microbialites, and deep-sea fauna. A variety of downstream analytical procedures are described, including: nucleic-acid extraction and preparation for high-throughput sequencing, fluorescence in-situ hybridisation, and cultivation of aerobic and anaerobic hydrocarbon-degrading microbes. Though most chapters focus on hydrocarbon-rich environments, many of the approaches used are generic, and as such will be of value to researchers embarking on studies of microbes and their processes in the field. Hydrocarbon and Lipid Microbiology Protocols There are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.

A unique feature of this book is its focus on nanotechnological solutions for the production of bioenergy and biofuels. Coverage includes topics such as nanobiotechnology, microalgae, biofuel cells, biomass pretreatment, and biomass conversion. An international team of experts also addresses the need to precisely characterize nanoparticles and the role of catalysts. The range of topics addressed, together with a chapter on risk management, make this book a highly useful resource for a broad readership including physicists, chemists, microbiologists, biotechnologists, food technologists, agricultural engineers, and nanotechnologists.

This book connects a retrosynthetic or disconnection approach with synthetic methods in the preparation of target molecules from simple, achiral ones to complex, chiral structures in the optically pure form. Retrosynthetic considerations and asymmetric syntheses are presented as closely related topics, often in the same chapter, underlining the importance of retrosynthetic consideration of target molecules neglecting stereochemistry and equipping readers to overcome the difficulties they may encounter in the planning and experimental implementation of asymmetric syntheses. This approach prepares students in advanced organic chemistry courses, and in particular young scientists working at academic and industrial laboratories, for independently solving synthetic problems and creating proposals for the synthesis of complex structures.

This book covers the state-of-the-art research on molecular biology assays and molecular techniques enabled or enhanced by microfluidic platforms. Topics covered include microfluidic methods for cellular separations and single cell studies, droplet-based approaches to study protein expression and forensics, and microfluidic in situ hybridization for RNA analysis. Key molecular biology studies using model organisms are reviewed in detail. This is an ideal book for students and researchers in the microfluidics and molecular biology fields as well as engineers working in the biotechnology industry. This book also: Reviews exhaustively the latest techniques for single-cell genetic, epigenetic, metabolomic, and proteomic analysis Illustrates microfluidic approaches for inverse metabolic engineering, as well as analysis of circulating exosomes Broadens readers' understanding of microfluidics convection-based PCR technology, microfluidic RNA-seq, and microfluidics for robust mobile diagnostics

Artificial organs may be considered as small-scale process plants, in which heat, mass and momentum transfer operations and, possibly, chemical transformations are carried out. This book proposes a novel analysis of artificial organs based on the typical bottom-up approach used in process engineering. Starting from a description of the fundamental physico-chemical phenomena involved in the process, the whole system is rebuilt as an interconnected ensemble of elemental unit operations. Each artificial organ is presented with a short introduction provided by expert clinicians. Devices commonly used in clinical practice are reviewed and their performance is assessed and compared by using a mathematical model based approach. Whilst mathematical modelling is a fundamental tool for quantitative descriptions of clinical devices, models are kept simple to remain focused on the essential features of each process. Postgraduate students and researchers in the field of chemical and biomedical engineering will find that this book provides a novel and useful tool for the analysis of existing devices and, possibly, the design of new ones. This approach will also be useful for medical researchers who want to get a deeper insight into the basic working principles of artificial organs.

This book provides a critical, carefully researched, up-to-date summary of membranes for membrane bioreactors. It presents a comprehensive and self-contained outline of the fundamentals of membrane bioreactors, especially their relevance as an advanced water treatment technology. This outline helps to bring the technology to the readers' attention, and positions the critical topic of membrane fouling as one of the key impediments to its more widescale adoption. The target readership includes researchers and industrial practitioners with an interest in membrane bioreactors.

This textbook teaches advanced undergraduate and first-year graduate students in Engineering and Applied Sciences to gather and analyze empirical observations (data) in order to aid in making design decisions. While science is about discovery, the primary paradigm of engineering and "applied science" is design. Scientists are in the discovery business and want, in general, to understand the natural world rather than to alter it. In contrast, engineers and applied scientists design products, processes, and solutions to problems. That said, statistics, as a discipline, is mostly oriented toward the discovery paradigm. Young engineers come out of their degree programs having taken courses such as "Statistics for Engineers and Scientists" without any clear idea as to how they can use statistical methods to help them design products or processes. Many seem to think that statistics is only useful for demonstrating that a device or process actually does what it was designed to do. Statistics courses emphasize creating predictive or classification models - predicting nature or classifying individuals, and statistics is often used to prove or disprove phenomena as opposed to aiding in the design of a product or process. In industry however, Chemical Engineers use designed experiments to optimize petroleum extraction; Manufacturing Engineers use experimental data to optimize machine operation; Industrial Engineers might use data to determine the optimal number of operators required in a manual assembly process. This text teaches engineering and applied science students to incorporate empirical investigation into such design processes. Much of the discussion in this book is about models, not whether the models truly represent reality but whether they adequately represent reality with respect to the problems at hand; many ideas focus on how to gather data in the most efficient way possible to construct adequate models. Includes chapters on subjects not often seen together in a single text (e.g., measurement systems, mixture experiments, logistic regression, Taguchi methods, simulation) Techniques and concepts introduced present a wide variety of design situations familiar to engineers and applied scientists and inspire incorporation of experimentation and empirical investigation into the design process. Software is integrally linked to statistical analyses with fully worked examples in each chapter; fully worked using several packages: SAS, R, JMP, Minitab, and MS Excel - also including discussion questions at the end of each chapter. The fundamental learning objective of this textbook is for the reader to understand how experimental data can be used to make design decisions and to be familiar with the most common types of experimental designs and analysis methods.

This book addresses the important clinical problem of accurately diagnosing osteoporosis, and analyzes how Bone Turnover Markers (BTMs) can improve osteoporosis detection. In her research, the author integrated microfluidic technology with electrochemical sensing to embody a reaction/detection chamber to measure serum levels of different biomarkers, creating a microfluidic proteomic platform that can easily be translated into a biomarker diagnostic. The Osteokit System, a result of the integration of electrochemical system and microfluidic chips, is a unique design that offers the potential for greater sensitivity. The implementation, feasibility, and specificity of the Osteokit platform is demonstrated in this book, which is appropriate for researchers working on bone biology and mechanics, as well as clinicians.

The biorefinery, integration of processes and technologies for biomass conversion, demands efficient utilization of all components. Hydrothermal processing is a potential clean technology to convert raw materials such as lignocellulosic and aquatic biomass into bioenergy and high added-value compounds. This book aims to show fundamental concepts and key technological developments that enabled industrial application of hydrothermal processing. The scope of this book is primarily for scientists working in the biorefinery field as well as engineers from industry and potential investors in biofuels. Therefore, the information in this book will provide an overview of this technology applied to lignocellulosic materials and aquatic biomass, and especially new knowledge. Critically, this book brings together experts in the application of hydrothermal processes on lignocellulosic and aquatic biomass.

This book presents the latest advances in and current research perspectives on the field of urban/industrial solid waste recycling for bio-energy and bio-fuel recovery. It chiefly focuses on five main thematic areas, namely bioreactor landfills coupled with energy and nutrient recovery; microbial insights into anaerobic digestion; greenhouse emission assessment; pyrolysis techniques for special waste treatment; and industrial waste stabilization options. In addition, it compiles the results of case studies and solid waste management perspectives from different countries.

This book provides an overview of the fundamentals and reference values for Ca stable isotope research, as well as current analytical methodologies including detailed instructions for sample preparation and isotope analysis. As such, it introduces readers to the different fields of application, including low-temperature mineral precipitation and biomineralisation, Earth surface processes and global cycling, high-temperature processes and cosmochemistry, and lastly human studies and biomedical applications. The current state of the art in these major areas is discussed, and open questions and possible future directions are identified. In terms of its depth and coverage, the current work extends and complements the previous reviews of Ca stable isotope geochemistry, addressing the needs of graduate students and advanced researchers who want to familiarize themselves with Ca stable isotope research.

This Volume presents key microscopy and imaging methods for revealing the structure and ultrastructure of environmental and experimental samples, of microbial communities and cultures, and of individual cells. Method adaptations that specifically address problems concerning the hydrophobic components of samples are highlighted and discussed. The methods described range from electron microscopy and light and fluorescence microscopy, to confocal laser-scanning microscopy, and include experimental set-ups for the analysis of interfacial processes like microbial growth and activities at hydrocarbon:water interfaces, biofilms and microbe:mineral interfaces. Three forms of fluorescence in situ hybridization - CARD-FISH, MAR-FISH and Two-pass TSA-FISH - are described for the ecophysiological analysis of functionally active microbes in samples. The methods presented will enable readers to obtain an ultrastructural picture of, and identify the key functional microbes in, samples under investigation. This in turn will constitute a key framework for the interpretation of information from other experimental approaches, such as physicochemical analyses and genomic investigations. Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.

The combustion properties of organic materials are used to assess their safety specifications. This knowledge is necessary to avoid potentially disastrous fires. The experimental determination of the combustion properties of a new organic compound is laborious and sometimes even impossible. This book describes methods for the determination and prediction of the combustion properties of organic compounds, along with some examples and exercises.

This Volume presents relevant single-cell and single-molecule approaches in the study of microbes producing and utilizing hydrocarbons and lipids. While generically applicable for all microorganisms, the approaches described are, wherever possible, adapted to the field of study of hydrocarbon and lipid microbiology. The methods include basic procedures for isolating single cells by means of microfluidics and flow cytometry, and their cultivation in arrays as pure clones; for isolating, amplifying and sequencing single-cell genomes and transcriptomes; and for analysing single-cell metabolomes by means of Raman spectroscopy. Single-molecule approaches include the use of protein:fluorescent dye fusions for protein localization and methods for the production of cell division protostructures and lipid monolayers. Methods for the functional analysis of single cells include detection of metabolically active (protein-synthesizing) cells in environmental samples by bioorthogonal non-canonical amino acid tagging, Raman spectroscopy combined with stable isotope labelling and fluorescent in situ hybridisation, and visualization of single cells participating in gene transfer activity. Lastly, protocols are presented for single-cell biotechnological applications, including biofuel production. Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.

This thesis reports studies on the substrate specificity of crucial ketosynthase (KS) domains from trans-AT Polyketide Synthases (PKSs). Using a combination of electrospray ionisation-mass spectrometry (ESI-MS) and simple N-acetyl cysteamine (SNAC) substrate mimics, the specificity of a range of KS domains from the bacillaene and psymberin PKSs have been succsessfully studied with regard to the initial acylation step of KS-catalysis. In addition, the ability to alter the substrate tolerance of KS domains by simple point mutations in the active site has been demonstrated. A series of acyl-ACPs have been synthesised using a novel methodology and employed to probe the substrate specificity of both KS domains and the previously uncharcterised acyl hydrolase domain, PedC. KS-catalysed chain elongation reactions have also been conducted and monitored by ESI-MS/MS. All KS domains studied exhibited higher substrate specificity at the elongation step than in the preceeding acylation step. Furthermore, a mechanism of reversible acylation is proposed using the PsyA ACP1-KS1 di-domain. The findings in this thesis provide important insights into mechanisms of KS specificity and show that mutagenesis can be used to expand the repertoire of acceptable substrates for future PKS engineering.

This Volume describes methods for cultivating hydrocarbon-producing and -consuming microbes, covering compounds in a range of states - gaseous (e.g. methane), liquid (e.g. alkanes of intermediate molecular weight) and solid (e.g. many PAHs and asphaltene). It also examines the cultivation of aerobic and anaerobic hydrocarbon degraders using a range of electron acceptors (e.g. oxygen, nitrate, sulphate, metals, (per)chlorate), and a separate chapter is devoted to explaining the cultivation of methanogens. Special attention is given to: high-throughput cultivation, growing microbes as biofilms, and cultivating fastidious microbes, as well as the preservation of microbial pure cultures and consortia. Accordingly, this Volume will be of value to anyone embarking on the selective enrichment and cultivation of novel microorganisms. Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.

This book introduces the most recent innovations in natural polymer applications in the food, construction, electronics, biomedical, pharmaceutical, and engineering industries. The authors provide perspectives from their respective range of industries covering classification, extraction, modification, and application of natural polymers from various sources in nature. They discuss the techniques used in analysis of natural polymers in various systems incorporating natural polymers as well as their intrinsic properties.

This Volume presents protocols for investigating the genetic, metabolic and ecological potential and functional analysis of microbial communities. Methods are described for the creation and bioinformatic assessment of metagenomic and metatranscriptomic libraries, and for metaproteomic analyses, which provide important insights into the metabolic potential and interactions of community members. These in turn lead to specific hypotheses concerning the functional contributions of individual populations in the community, which may be investigated by the stable isotope probing approaches described in this Volume, making it possible to identify those community members primarily responsible for particular functions. Methods for the direct extraction of proteins from environmental samples for sequencing and activity tests are presented, providing a broad overview of prevailing metabolic activities and of the types of microbe involved in them. Protocols for the analysis of nutrient flow through microbial communities and for the modelling of dynamic physiological interactions in communities are also provided. Lastly, the book presents a protocol for the quantitative assessment of permissiveness for the transfer of conjugative plasmids, important agents of physiological change and evolution in microbial communities. Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.