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Biodeep May 17-19, 2004 Milano

Biodeep May 17-19, 2004 Milano. “Nature has more imagination than our dreams”. Contents. Strains and clones transferred Screening for industrial partners Esterases Acylases Proteases Biodeep Database Communications Conclusions. Strains transferred.

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Biodeep May 17-19, 2004 Milano

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  1. Biodeep May 17-19, 2004 Milano “Nature has more imagination than our dreams”

  2. Contents • Strains and clones transferred • Screening for industrial partners • Esterases • Acylases • Proteases • Biodeep Database • Communications • Conclusions

  3. Strains transferred • Milan University (Dr. T. Brusa) 1. Strains studied means that we tried at least once to culture them 2. Strains cultured means strains that have been cultured in our lab (and therefore stored in collection) and can be used for future work

  4. Strains transferred • Essex University (Dr. A. Sass) 1. Strains studied means that we tried at least once to culture them 2. Strains cultured means strains that have been cultured in our lab (and therefore stored in collection) and can be used for future work 3. The 19 strains of the 1st batch which could not be cultured in our lab have been sent a second time. The 40 strains of batch 3 are anaerobes (stricts or facultatives).

  5. Strains transferred • LLM (Dr. F. Morel) 1. Strains studied means that we tried at least once to culture them 2. Strains cultured means strains that have been cultured in our lab (and therefor stored in collection) and can be used for future work

  6. Strains transferred • Messina University (Dr. G. D’auria) No synthetic media 1. Strains studied means that we tried at least once to culture them 2. Strains cultured means strains that have been cultured in our lab (and therefor stored in collection) and can be used for future work 3. See next slide 4. All the strains excepted the ones growing in P10 medium (petrol based medium)

  7. Strains transferred • Cultures from Univ. Messina • Enrichment cultures and pure strains • No synthetic media defined (from the water point of view) in the case of batch 1 and 2 • We tried to grow in batch 1 and 2 the stains in a defined medium • The growth conditions defined for the 22 strains cultured in our lab are available • The last 66 strains from batch 3 grow in ONR medium

  8. Strains transferred • Review (3) 1. Strains studied means that we tried at least once to culture them 2. Strains cultured means strains that have been cultured in our lab and can be used for future work 3. Pure strains as well as enrichment cultures New reception of strains from last cruise ? Who, When ?

  9. Clones transferred • clones from TUB (Dr. P. Golyshin) • 11 genomic fragments encoding 11 different esterases • Corresponding (partial or FL) sequences have been provided with some genomic fragments • 6 ORF subcloned at Protéus. Expression successful • 2 genomic fragments encoding 2 amylases. No activity detected

  10. Esterase for Personal care • Screening for substrate specificity in synthesis reactions for personal care application • No microbial screening due to reaction constraints • Screening of the different recombinant clones • One enzyme validated for substrate specificity in hydrolysis (O.2). Trials in synthesis (after subcloning and expression scaling up) failed.

  11. Esterase for NME synthesis • Screening for regio-specificity in hydrolysis reaction for the synthesis of a novel chiral pharmaceutical molecule • Microbial screening and molecular screening in progress • 2 positive strains for the collection coming from the University of Messina • Cloning of the corresponding genes to be done

  12. Acylases for Personal care • Screening for an enzyme able to catalyze formation of amide bond for personal care application • Microbial screening in hydrolysis and synthesis • 9 positive strains in hydrolysis (Essex, Messina and LMM) and 3 synthesis (Essex, Messina) • End of screening in progress (Univ. Milano) • Cloning of the corresponding genes to be done

  13. Proteases for research • Protéus is in touch with a company developing research tools for molecular biology for the development of Proteases • Discussions in progress

  14. Clones collection Cloning of esterases and acylases in progress within the frame of industrial programs

  15. Biodeep database • Protéus • Design of the data sheet model • Initial frame of the database obtained from Dr.G. D’auria • Upgrade of this database and data insertion performed at Proteus • University of Milano • Insertion of the database on the Biodeep web site for all the partners DATABASE

  16. Communications • Our industrial partners • Congress : • IBC Biocatalysis Conference (Boston, USA) • CPHI (Francfort, Germany) • BioCat 2003, (Barcelona, Spain) • Informex (Las Vegas , USA) • ...etc...

  17. Conclusions and Outlook • Current status of the Biodeep microbial collection :454 strains (or mixed cultures) • Current status of the Biodeep clones collection : 15 cloned genomic fragments containing enzymatic activities (8 ORF subcloned) • Valorization in international congress : more than 15 presentations • 4 different industrial programs involving the Biodeep microbial or clones collection to date • Valorization will continue at Protéus

  18. …Thanks for your attention. www.proteus.fr

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