Isolation & identification of Novel microorganisms for the Treatment

Carla Harkin
Dr Wolfram Brück
Dr Catherine Lynch
Dr Denis McCrudden
Development of efficient, economical and
environmentally-friendly pre-treatment of crustacean shell
wastes to extract chitinous material (Raw material
sources: Europe/Asia/Africa).
Stabilisation of chitinous material for transfer to project
partners (UMB, INS, TUM, FhG).
Development of assays for constant quality control.
Raw material Prep
Optimisation of
chemical chitin
Identification of
novel microorganisms
Optimisation of
microbial chitin
7. QC Analysis
Chemical extraction methods- costly and a hazard to the
Biological extraction methods- more cost effective while posing
less cause for environmental concern.
Microbial chitin extraction methods:
Direct use of enzymes (Brück et al, 2011; Jo et al, 2011).
Fermentation (Jung et al, 2005; Jung et al, 2007; Rao et al, 2000).
Initial optimisation, adapting methods from literature.
Identification of novel microorganisms for fermentation.
Raw material
Shaking Incubator
& drying
(Jung et al., 2005; Jung et al., 2007a,b; Oh et al., 2007)
Change of Inoculum
Isolation from soil samples
Screening for useful
Small scale fermentation
Large scale fermentation
 Demineralisation–acid production.
Replicated fermentation method on micro scale.
Fig 1: pH indication after addition of Methyl Red
Indicator. L-r: negative yellow, negative orange,
positive intense red.
 Deproteinisation–protease production.
Casein agar plates.
Colorimetric protease assay to quantify activity
(Vishwanatha et al, 2009).
Fig 2: Produciton of zone of proteolysis
in casein agar.
No. of Organisms
Protease producing
Acid producing
Protease & Acid producing
• Identification of isolates; PCR, sequencing- BLAST
• Characterization of isolates: biochemical tests, staining
• Continue to meet chitinous material production targets.
• Continuation of fermentation experiments using raw
crustacean waste from Ireland, Indonesia and Tunisia.
With select cultures, the aim is to scale-up fermentations
to pilot scale to investigate the reality of industrial large
scale fermentations in the breakdown of crustacean waste.
Development of an economic, environmentally friendly,
waste reducing facility.
Profit-generating business opportunities for Irish Seafood
Brück, W., M., Slater, J. W., Carney, B. F., 2011. Chitin and chitosan from marine
organisms. Chitin, Chitosan, Oligosaccharides and their derivatives. Boca Raton: CRC
Vishwanatha, T., Spoorthi, N., Reena, V., Divyashree B.C., Siddalingeshwara K.G., Karthic,
J., Sudipta K.M., 2010. Screening of substrates for protease production from Bacillus
lichenformis. International Journal of Engineering Science and Technology (2)11, pp.
Jung, W.J., Jo, G. H., Kuk, J. H., Kim, K. Y., Park, R. D., 2005. Extraction of chitin from
red crab shell waste by cofermentation with Lactobacillus paracasei subsp. tolerans KCTC3074 and Serratia marcescens FS-3. Applied Microbiology and Biotechnology (71) pp. 234237.
Jung, W.J., Jo, G. H., Kuk, J. H., Kim, Y. J., Oh, K. T., Park, R. D., 2007. Production of
chitin from red crab shell waste by successive fermentation with Lactobacillus paracasei
KCTC-3074 and Serratia marcescens FS-3. Carbohydrate Polymers (68) pp. 746-750.
Oh, K. T., Kim, Y. J., Nguyen, V. N., Jung, W.J., Park, R.D. 2007. Demineralization of crab
shell waste by Pseudomonas aeruginosa F722. Process Biochemistry. (42) pp. 1069–1074.
Jo, G.H., Park, R.D., Jung, W.J., 2011. Enzymatic production of chitin from crustacean
shell waste. Chitin, Chitosan, Oligosaccharides and their derivatives. Boca Raton: CRC
Rao, M. S., Muñoz, J., Stevens, W. F., 2000. Critical factors in chitin production by
fermentation of shrimp biowaste. Applied Microbiology and Biotechnology (54) pp. 808813.
Thank you very much for your attention.

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