Cold−adapted marine bacteria producing extracellular hydrolytic enzymes are important for their industrial application and play a key role in degradation of particulate or ganic matter in their natural environment. In this work, members of a previously−obtained protease−producing bacterial collection isolated from different marine sources from Potter Cove (King George Island, South Shetlands) were taxonomically identified and screened for their ability to produce other economically relevant enzymes. Eighty−eight proteolytic bacterial isolates were grouped into 25 phylotypes based on their Amplified Ribosomal DNA Restriction Analysis profiles. The sequencing of the 16S rRNA genes from representative isolates of the phylotypes showed that the predominant culturable protease−producing bacteria belonged to the class Gammaproteobacteria and were affiliated to the genera Pseudomonas , Shewanella , Colwellia , and Pseudoalteromonas , the latter being the predominant group (64% of isolates). In addition, members of the classes Actinobacteria, Bacilli and Flavobacteria were found. Among the 88 isolates screened we detected producers of amylases (21), pectinases (67), cellulases (53), CM−cellulases (68), xylanases (55) and agarases (57). More than 85% of the isolates showed at least one of the extracellular enzymatic activities tested, with some of them producing up to six extracellular enzymes. Our results confirmed that using selective conditions to isolate producers of one extracellular enzyme activity increases the probability of recovering bacteria that will also produce additional extracellular enzymes. This finding establishes a starting point for future programs oriented to the prospecting for biomolecules in Antarctica.

Accidental oil spills at open sea is a common environmental problem. They lead to degradation of sea and shoreline life. In the last ten years there has been an increased interest in bioremediation using the enzymatic activity of the naturally occurring microorganisms. In this work the potential of mixed microbial cultures for biodegradation of crude oil in seawater and sand has been examined. Artificial seawater supplemented with nitrogen and phosphorus was inoculated with cultures isolated from refinery sludge. The same cultures were used for experiments in sand polluted by 5% (v/w) of crude oil. These experiments were performed in sterile and semi-natural (not sterile) conditions to see the degradation potential of isolated cultures, their growth characteristics and possible antagonisms between supplemented microorganisms and natural microflora. During the experiments the oxygen demand, number of bacteria (cfu) and optical density (OD660 ) were monitored. After 14 days of cultivation, the concentration of total petroleum hydrocarbons (TPH) in all samples was measured. All tested cultures had a potential for degradation of hydrocarbons in seawater and sand. After two weeks of experiment, loss of hydrocarbons in seawater polluted with crude oil was between 56.8% (A2 culture) and 64.4% (Al culture). The most effective culture for bioaugmentation of seawater does not have to be the best solution for bioaugmcntation of sand. In sand the best degraders in sterile and semi-natural conditions were found in the mixed cultures isolated from Corinth refinery sludge. For this culture concentration of hydrocarbons in sterile sand was 73.2% lower than in control sample and in non-sterile sand 70.5% lower than in control (sterile sand) without bioaugmentation. Finally, the addition of seawater and fertilizers to sand had also a positive influence on contaminants degradation by naturally occurring microorganisms (48%). Experiments performed with different environments (seawater and sand) and under different conditions (sterilized material and semi-natural conditions) confirmed that cultures should be tested in semi-natural conditions especially when indigenous microflora cans posse's high degradation potential. Allochtonie cultures, very active in sterile conditions, after inoculation to natural environment can even slow down the degradation.