Microbial Steroids

This volume provides practical experimental laboratory protocols for a wide range of steroid bioconversions. The chapters in this book cover topics such as bioconversions and chemical synthesis pathways; strain characterization; bioconversion from sterols to androstenedione and androstadienedione; steroid hydroxylations; biocatalysis; and downstream processes to purify steroid intermediates. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Comprehensive and thorough, Microbial Steroids: Methods and Protocols is a valuable resource for laboratory and industrial professionals. It is also useful for graduate students studying biotechnology, microbiology, genetics, and molecular biology.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt

Steroid Bioconversions.- Chemical Pathways of Corticosteroids, Industrial Synthesis from Sapogenins.- Genome Sequencing of Steroid Producing Bacteria with Illumina Technology.- Genome Sequencing of Steroid Producing Bacteria using Ion Torrent Technology and a Reference Genome.- RNA-Seq-Based Comparative Transcriptomics: RNA Preparation and Bioinformatics.- Intra- and Extra-Cellular Proteome Analyses of Steroid-Producer Mycobacteria.- Genetic Techniques for Manipulation of the Phytosterol Biotransformation Strain Mycobacteriumneoaurum NRRL B-3805.- Identification and Characterization of the genes and Enzymes Belonging to the Bile Acid Catabolic Pathway in Pseudomonas.- Selection of Biodegrading Phytosterol Strains.- Stigmasterol Removal by an Aerobic Treatment System.- Production and Biotransformation of Phytosterol Microdispersions to Produce 4-Androstene-3, 17-Dione.- -Sitosterol Bioconversion to Androstenedionein Microtiter Plates.- Bioconversion of Phytosterols into Androstenedione by Mycobacterium.- Scale-Up of Phytosterols Bioconversion into Androstenedione.- Bioconversion of Phytosterols into Androstadienedione by Mycobacterium smegmatis CECT 8331.- Process for Biotransformation of Androsta-4-ene-3, 17-Dione (4-AD) to Androsta-1, 4-Diene-3, 17-Dione (ADD).- Laboratory-Scale Hydroxylation of Steroids by P450BM3 Variants.- Obtaining of 11-Hydroxy and Rost-4-ene-3, 17-Dione from Natural Sterols.- Steroid 11-Alfa-Hydroxylation by the Fungi Aspergillus nidulans and Aspergillus ochraceus.- Biotransformation of DHEA into 7, 15-diOH-DHEA.- Stabilization of Enzymes by Using Thermophiles.- Biocatalysis of Steroids with Mycobacterium Sp. in Aqueous and Organic Media.- Downstream Process Synthesis for Microbial Steroids.- Analysis of Intermediates of Steroid Transformations in Resting Cells by Thin Layer Chromatography (TLC).- A Ligand-Binding Assay to Measure the Affinity and Specificity of Sterol-Binding Proteins In Vitro.