Read more
This detailed volume explores experimental laboratory procedures for a wide range of steroid bioconversions. After an overview on the current trends and perspectives, the book continues with sections covering microbial screening and synthetic biology applied to microorganisms able to catabolize sterols, methods on strain characterization, including omics and biochemical analyses, methods of fermentation and biocatalysis for steroids production, as well as a chapter on the medical use of glucocorticoids in cancer patients. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Microbial Steroids: Methods and Protocols, Second Edition serves as an ideal reference source for laboratory and industrial professionals, as well as for students in a wide array of biological disciplines.
List of contents
Current Trends and Perspectives in Microbial Bioconversions of Steroids.- Isolation of Environmental Bacteria Able to Degrade Sterols and/or Bile Acids: Determination of Cholesterol Oxidase and Several Hydroxysteroid Dehydrogenase Activities in Rhodococcus, Gordonia, and Pseudomonas putida.- Selection of Biodegrading Phytosterol Strains.- Identification and Characterization of Some Genes, Enzymes, and Metabolic Intermediates Belonging to the Bile Acid Aerobic Catabolic Pathway from Pseudomonas.- Targeted Mutagenesis of Mycobacterium Strains by Homologous Recombination.- RNA Preparation and RNA-Seq Bioinformatics for Comparative Transcriptomics.- Bidimensional Analyses of the Intra- and Extra-Cellular Proteomes of Steroid-Producer Mycobacteria.- Fungal Sterol Analyses by Gas Chromatography - Mass Spectrometry Using Different Derivatives.- Cholesterol Assay Based on Recombinant Cholesterol Oxidase, ABTS, and Horseradish Peroxidase.- Measuring the Interaction of Sterols and Steroids with Proteins by Microscale Thermophoresis.- Cultivation of Mycolicibacterium spp. Mutants in Miniaturized and High-Throughput Format to Characterize Their Growth, Phytosterol Conversion Ability, and Resistance to the Steroid Products.- beta-Sitosterol Bioconversion in Small-Scale Devices: From Microtiter Plates to Microfluidic Reactors.- Biocatalysis of Steroids by Mycobacterium sp. in Aqueous and Organic Media.- Scale-Up of Phytosterols Bioconversion into Androstenedione.- Bioconversion of Phytosterols into Androstenedione by Mycolicibacterium.- Selective Microbial Conversion of DHEA into 7 - -DHEA.- Production of 11 -Hydroxysteroids Derivatives by Corynebacterium glutamicum Expressing the Rhizopus oryzae Hydroxylating System.- Obtaining of 24-Norchol-4-Ene-3,22-Dione from Phytosterol with Mutants of Mycolicibacterium neoaurum.- Stabilization of Enzymes by Using Thermophiles.- Stigmasterol Removal by an Aerobic Treatment System.- Glucocorticoid Effect in Cancer Patients.
