2. Outline:
Why look at marine derived bacteria for future
medicine?
Streptomyces CP54-7 and isolating A80915A
Significance and future goals
3. Actinomyces as a source of
medicine
Olano, C., Méndez, C., & Salas, J. A. (2009). Antitumor Compounds from Marine
Actinomycetes. Marine Drugs, 7(2), 210–248. doi:10.3390/md7020210
Doxorubicin –
Chemotherapeutic
Miyaoka, R., Hosokawa, M., Ando, M., Mori, T., Hamaguchi, H., & Takeyama, H. (2014). In Situ
Detection of Antibiotic Amphotericin B Produced in Streptomyces nodosus Using Raman
Microspectroscopy. Marine Drugs, 12(5), 2827–2839. doi:10.3390/md12052827
Amphotericin B – Antifungal
4. Why look towards the ocean?
Multidrug resistant
organisms
Huge diversity
Unexplored potential
http://www.f.u-
tokyo.ac.jp/~fukuyama/project2000.htm
http://barcelona-
ocean.com/training/course/ecology-marine-
microorganisms
https://www.superstock.com/stock-
photos-images/824-94770
5. Methodology
Hypothesis:
Marine obligate actinomycetes have
the potential to produce novel
compounds that can be further
developed to treat cancer and other
human diseases.
1. Isolate bacteria
from sediment
samples
2. Culture isolated
strains in growth
media
3. Extract chemical
compounds
produced by
bacteria in growth
media
4. Separate the
crude extract into
pure compounds
5. Analyze the data
CP54-7
1. High yielding strain
2. Interesting
chemical profile
9. Purification of CP54-7A
100% Divided into 7 Fractions
F6 - 210.94mg
80% Divided into 11 fractions
F11 - 88.12mg
6
11
100% MeOH
80% MeOH
10. Isolation of A80915A
These appeared to be the same compound based on
previous H-NMR and RT-HPLC chemical profile
100% F6 - 10.18mg
80% F11 - 3.2mg
100% MeOH crude profile
50-100% 20 min ACN
80% MeOH purification
80-100% 20 min MeOH
12. 2D NMR Analysis - COSY
Correlations
Shows how nearby protons
are interacting with each
other
A.
A.
B.
B.
Click to add text
H-NMR 1
H-NMR2
13. 2D NMR Analysis - HSQC
Correlations
Shows how adjacent C-H
are interacting with each
other
H-NMR
C-NMR
C.
D.
C.
D.
14. 2D NMR Analysis - HMBC
Correlations
Shows you how neighboring
C-H bonds are interacting
from multiple bond lengths
C-NMR
H-NMR
E.
E2. E1.
E3-4
F.
F1
F2
F3
15. Figure 3B - A80915A induces apoptosis
of HCT-116 cells, analyzed using FACS
16. Future goals:
Continue separation of other organic
compounds in crude extract
If novel compounds are found, elucidate the
structure and subject the compound to
biochemical assays
Figure out the mechanism of action for these
novel compounds
17. Acknowledgements
Special thanks to:
Dr. Taro Amagata, Dr. Carmen Domingo, Dr. Raymond
Esquerra, Dr. Tester Baird and Dr. Frank Bayliss
My lab mates: Stephanie Gee, Eric Yip, Peishan Li, Juliana
Thai, Duyen Vo, Jorge Rivas and John Flores
The Department of Chemistry & Biochemistry and
Department of Biology
NIH-MARC Grant No: 5T34GM008574-19