Optimization of Cocktail Composition and Up-scale Fermentation Process Development of Four Skin Commensal Bacterial Strains

  • Ahmad Baikuni . Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmacy, Universitas Indonesia, Depok, West Java, 16424, Indonesia
  • Meidy Richky Wanyodiharjo . Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmacy, Universitas Indonesia
  • Harry Ardiansyah . Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmacy, Universitas Indonesia
  • Fathan Luthfi Hawari . Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmacy, Universitas Indonesia
  • Amarila Malik . Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmacy, Universitas Indonesia

Abstract

Analyzing the actual microbial composition in the skin microbiome is riveting to create a potential source
of active substances in skincare products. Our previous study isolated four bacterial strains from Javanese male and female skin samples, Staphylococcus hominis MBF12–19J, Staphylococcus warneri MBF02–19J, Bacillus subtilis MBF10–19J, and Micrococcus luteus MBF05–19J. This study aimed to determine the composition of those strains in a bacterial cocktail and optimize the condition for up-scale laboratory production of the cocktail. To assess the ability of bacteria to coexist and live in cocktail communities, simultaneous monitoring was performed using the Deferred Growth Inhibition Assay (DGIA) method and real-time PCR for DNA copy number measurement. Results showed the best composition based on even distribution and cell growth viability was at the ratio 1.5:1:0.5:0.5 of Micrococcus luteus MBF05-19J, Bacillus subtilis MBF10-19J, Staphylococcus warneri MBF02-19J, Staphylococcus hominis MBF12-19J, which is equivalent to DNA copy number/mL 1.209 x 1024 CFU : 2.484 x 1041 CFU : 2.645 x 1041 CFU : 9.041 x 1035 CFU, respectively. The optimum growth incubation time of individual bacterial cultures for an up-scale 2-L bio-fermentor mixture was as follows; Micrococcus luteus MBF05-19J = 21 hrs; Bacillus subtilis MBF10-19J = 7 hrs; Staphylococcus warneri MBF02-19J = 17 hrs; Staphylococcus hominis MBF12-19 = 15 hrs. For up-scale conditions, the fermentation incubation time was 3 hours at 37oC, agitation 50 RPM, and aeration 5% dissolved oxygen.

References

1. Eisenstein M. The skin microbiome. Nature. 2020;588(7838):S209.

2. Nakatsuji T, Chiang H, Jiang SB, Nagarajan H, Gallo RL, Diego S, et al. HHS Public Access. 2013;1–16.

3. Beri K. Skin microbiome & host immunity: Applications in regenerative cosmetics & transdermal
drug delivery. Futur Sci OA. 2018;4(6).

4. Khayyira AS, Rosdina AE, Irianti MI, Malik A. Simultaneous profi ling and cultivation of the skin
microbiome of healthy young adult skin for the development of therapeutic agents. Heliyon [Internet]. 2020;6(4):e03700. Available from: https://doi.org/10.1016/j.heliyon.2020.e03700

5. Pauer H, Glatthardt T, Ferreira N V, Ferreira RBR, Antunes LCM. Bioactive Molecules of the Human Microbiome. Microbiome Metabolome Diagnosis, Ther Other Strateg Appl Elsevier. 2019;115–25.

6. Azevedo AC, Bento CBP, Ruiz JC, Queiroz M V., Mantovani HC. Distribution and genetic diversity of bacteriocin gene clusters in rumen microbial genomes. Appl Environ Microbiol. 2015;81(20):7290–304.

7. Hofer A, Legat FJ , Gruber-Wacker nagel A, Quehenberger F, Wolf P. Topical liposomal DNArepair
enzymes in polymorphic light eruption. Photochem Photobiol Sci. 2011;10(7):1118–28.

8. Jimenez M, Langer R, Traverso G. Microbial therapeutics : New opportunities for drug delivery.
2019;216(5):1005–9.

9. Kato S, Haruta S, Cui ZJ, Ishii M, Igarashi Y. Network relationships of bacteria in a stable mixed culture. Microb Ecol. 2008;56(3):403–11.

10. Ngo DH, Vo TS. An updated review on pharmaceutical properties of gamma-aminobutyric acid. Molecules. 2019;24(15).

11. Uehara E, Hokazono H, Sasaki T, Yoshioka H, Matsuo N. Effects of GABA on the expression
of type I collagen gene in normal human dermal fibroblasts. Biosci Biotechnol Biochem [Internet].
2017;81(2):376–9. Available from: http://dx.doi.org/10.1080/09168451.2016.1238296

12. Ghoul M, Mitri S. The Ecology and Evolution of Microbial Competition. Trends Microbiol [Internet]. 2016;24(10):833–45. Available from: http://dx.doi.org/10.1016/j.tim.2016.06.011

13. Moran JC, Crank EL, Ghabban HA, Horsburgh MJ. Deferred Growth Inhibition Assay to Quantify
the Effect of Bacteria-derived Antimicrobials on Competition. 2016;(September):1–5.

14. Lee C, Kim J, Shin SG, Hwang S. Absolute and relative QPCR quantification of plasmid copy number in Escherichia coli. J Biotechnol. 2006;123(3):273–80.

15. Kilic A, Basustaoglu AC. Double triplex realtime PCR assay for simultaneous detection of
Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus hominis, and Staphylococcus haemolyticus and determination of their methicillin resistance directly from positive blood c. Res Microbiol. 2011;162(10):1060–6.

16. Iwase T, Seki K, Shinji H, Mizunoe Y, Masuda S. Development of a real-time PCR assay for the
detection and identifi cation of Staphylococcus capitis, Staphylococcus haemolyticus and Staphylococcus warneri. J Med Microbiol. 2007;56(10):1346–9.

17. Fajardo-cavazos P, Schuerger AC, Nicholson WL. Exposure of DNA and Bacillus subtilis Spores to Simulated Martian Environments : Use of Quantitative PCR ( qPCR ) to Measure Inactivation Rates of DNA to Function as a Template Molecule. 2010;10(4).

18. Red L. Penentuan Waktu Inkubasi Optimum Terhadap Aktivitas Bakteriosin. 2014;3(4):52–6.

19. Beller HR, Goh EB, Keasling JD. Genes involved in long-chain alkene biosynthesis in Micrococcus Luteus. Appl Environ Microbiol. 2010;76(4):1212–23
Published
2022-04-30
How to Cite
BAIKUNI, Ahmad et al. Optimization of Cocktail Composition and Up-scale Fermentation Process Development of Four Skin Commensal Bacterial Strains. JURNAL ILMU KEFARMASIAN INDONESIA, [S.l.], v. 20, n. 1, p. 113-119, apr. 2022. ISSN 2614-6495. Available at: <http://jifi.farmasi.univpancasila.ac.id/index.php/jifi/article/view/1224>. Date accessed: 30 nov. 2022. doi: https://doi.org/10.35814/jifi.v20i1.1224.
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