Main description:

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.

Contents:

Preface...
Table of Contents...
Contributing Authors...

1. Steroid Bioconversions
Marina V. Donova

2. Chemical Pathways of Corticosteroids, Industrial Synthesis from Sapogenins
Ignacio Herraiz

3. Genome Sequencing of Steroid Producing Bacteria with Illumina Technology
Victoria Y. Shtratnikova, Mikhail I. Schelkunov, and marina V. Donova

4. Genome Sequencing of Steroid Producing Bacteria using Ion Torrent Technology and a Reference Genome
Alberto Sola-Landa, Antonio Rodriguez-Garcia, Carlos Barreiro, and Rosario Perez-Redondo

5. RNA-Seq-Based Comparative Transcriptomics: RNA Preparation and Bioinformatics
Antonio Rodriguez-Garcia, Alberto Sola-Landa, and Carlos Barreiro

6. Intra- and Extra-Cellular Proteome Analyses of Steroid-Producer Mycobacteria
Carlos Barreiro, Alejandro Morales, Ines Vazquez-Iglesias, and Alberto Sola-Landa

7. Genetic Techniques for Manipulation of the Phytosterol Biotransformation Strain Mycobacteriumneoaurum NRRL B-3805
Jessica K. Loraine and Margaret C. M. Smith

8. Identification and Characterization of the genes and Enzymes Belonging to the Bile Acid Catabolic Pathway in Pseudomonas
Jose M. Luengo and Elias R. Olivera

9. Selection of Biodegrading Phytosterol Strains
Maria-Angelica Mondaca, Maricel Vidal, Soledad Chamorro, and Gladys Vidal

10. Stigmasterol Removal by an Aerobic Treatment System
Soledad Chamorro, Claudia Xavier, Victor Hernandez, Jose Becerra, and Gladys Vidal

11. Production and Biotransformation of Phytosterol Microdispersions to Produce 4-Androstene-3, 17-Dione
Rodrigo A. Mancilla, Roberto Pavez-Diaz, and Andrea Munoz-Orrego

12. -Sitosterol Bioconversion to Androstenedione in Microtiter Plates
Marco P.C. Marques and Pedro Fernandez

13. Bioconversion of Phytosterols into Androstenedione by Mycobacterium
Kjell D. Josefsen, Anna Nordborg, and Havard Sletta

14. Scale-Up of Phytosterols Bioconversion into Androstenedione
Sonia Martinez-Camara, Esther Bahillo, Jose-Luis Barredo, and Marta Rodriguez-Saiz

15. Bioconversion of Phytosterols into Androstadienedione by Mycobacterium smegmatis CECT 8331
Julia Garcia-Fernandez, Igor Martinez, Lorena Fernandez-Cabezon, Carmen Felpeto-Santero, Jose-Luis Garcia, and Beatriz Galan

16. Process for Biotransformation of Androsta-4-ene-3, 17-Dione (4-AD) to Androsta-1, 4-Diene-3, 17-Dione (ADD)
Surya Prakash and Abhay Bajaj

17. Laboratory-Scale Hydroxylation of Steroids by P450BM3 Variants
Sabrina Hoebenreich, Martin Spinck, and Nathalie Nett

18. Obtaining of 11 -Hydroxy and Rost-4-ene-3, 17-Dione from Natural Sterols
Dmitry Dovbnya, Sergey Khomutov, Vyacheslav Kollerov, Marina Donova

19. Steroid 11-Alfa-Hydroxylation by the Fungi Aspergillus nidulans and Aspergillus ochraceus
Lidia Ortega-de los Rios, Jose M. Luengo, and Jose M. Fernandez-Canon

20. Biotransformation of DHEA into 7 , 15 -diOH-DHEA
Hui Li, Jin Sun, and Zhenghong Xu

21. Stabilization of Enzymes by Using Thermophiles
Ana-Luisa Ribeiro, Mercedes Sanchez, Aurelio Hidalgo, and Jose Berenguer

22. Biocatalysis of Steroids with Mycobacterium Sp. in Aqueous and Organic Media
Carla C. C. R. de Carvalho and Pedro Fernandes

23. Downstream Process Synthesis for Microbial Steroids
Fabian B. Thygs and Juliane Merz

24. Analysis of Intermediates of Steroid Transformations in Resting Cells by Thin Layer Chromatography (TLC)
Govinda Guevara, Julian Perera, and Juana-Maria Navarro-Llorens

25. A Ligand-Binding Assay to Measure the Affinity and Specificity of Sterol-Binding Proteins In Vitro
Rabih Darwiche and Roger Schneiter