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Partner 1c
CoNISMa, Consorzio Nazionale Interuniversitario
Scienze del Mare
DISTAM Università degli Studi di Milano
WP7 screening and characterization of the new
products with potential biotechnological
application
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2001 isolates collection
Strict anaerobes
aerobes
URANIA 25 BANNOCK
23 DISCOVERY 21 LATALANTE 23
URANIA BANNOCK
DISCOVERY 4 LATALANTE 2
6
95

• Transferred to Proteus
• Characterized for diversity
• Species identification
• Screened for activity

• Under isolation procedure

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Metabolic typing of aerobic isolates
Screening for

• Oil utilisation as unique carbon source
• Catechol utilization as unique carbon source
• Esterase activity on long chain satured fatty
  acids (Tween60)
• Esterase activity on long chain insatured fatty
  acids (tryoleine)
• Production of polysaccarides (acetone
  precipitation)
• Antibacterial activity against Sarcina lutea

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Oil utilization

• Inoculum preparation in M9 mineral broth with
  yeast extract as carbon source
• Cells wash, and resuspension in mineral medium
  (NaCl concentration optimum for each strain)
  with 3 crude oil as unique carbon source
• Incubation at 28C and growth revealing with INT
  (iodoniltetrazolium violet)

All the positive strains were able to grow on
catechol as unique carbon source, except 1
Bannock isolate
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Lipase-esterase activity on long chain satured
fatty acids
Most of the very positive strains were active
also when incubated at 15C
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Lipase-esterase activity on long chain unsatured
fatty acids
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Polysaccarides production

• Growth on rich liquid medium (PCB) with 5 NaCl
• Precipitation with 1 volume of acetone

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Antibacterial activity

• Growth on rich solid medium (PCA) with 3 NaCl
• Killing of cells with chloroform
• Overlay with Sarcina lutea culture, incubation at
  30C
• Screening of inhibition rings

Tested all 94 collection strains no
antibacterial activity found
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