As my experience most of the bio active metabolites come to the broth part. most of the fats and sugar contain in myceliya part. Broth can extract with EtOAc first and then crude can dissolve in MeOH. soluble part can further separate by Column Chromatography.
Hey,
Before your think to extract bioactivity compounds you need to think how cultivate the actinomycetes. I will give some examples here:
1 - Single cultivation (put you bacteria in a medium and grown, after that, fractionation process, bioassays and identification of chemical compounds). This method could be used in liquid medium or solid one! The chemical profile change and you could get access to different chemical compounds. The same will apply when you change the medium and the physical properties of the growing step.
2 - Co-culture: basically the idea is put two microbes to interacting each other during the growing step. This is amazing method because almost of microorganisms has activity genes silenced! Thus, we can turn on this genes and get new chemical profile.
3 - Metagenome: less than 1% of microorganisms could be cultivated in laboratorial conditions and/or until we develop a new way to growing this bugs. So, you can use the metagenome to get access to chemo-genetical information of the specific actinomyces group.
Have another methods deriving of these one, you can check better.
After that, choose your method to work of this guys, you can think about the isolation, bioassays and identification of the natural products from actinomyces!!
Good luck!
Hi,
There are two types of metabolite (i) intracellular and extracellular.
- For intracellular the biomass was harvested by centrifugation (5000 rpm) at 20C for 20 minutes, and then the mycelia were washed three times with sterile distilled water under aseptic conditions. The cells were then resuspended in a little amount of methanol and ground with the help of a pestle and mortar. Methanol was added to the ground cells in the ratio of 1:1 (w/v) and the mixture was shaken vigorously overnight; the extracts were then filtered through a blotting paper. The filtrates were then evaporated using a rotary evaporator at 50C.
- For extracellular the cell free fermented broth was extracted twice with ethyl acetate. After separation, the organic phase was dried over Na2SO4 (anhydrous). The extract was then concentrated in a rotary vacuum at 50C.
Ref: P. Saravana Kumar a, V. Duraipandiyan b, S. Ignacimuthu. Isolation, screening and partial purification of antimicrobial antibiotics from soil Streptomyces sp. SCA 7. Kaohsiung Journal of Medical Sciences (2014) 30, 435-446.
Thank you,
All the best!!!
- Badges
- Science topic
- Similar topics
- Mathematics
- Statistics
- Sampling Design
- Sample Size