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US 7,951,561 B2Granted
Method for the preparation of κ-carrageenase
AssigneeCouncil of Scientific and Industrial Research
InventorsYasmin Najmuddin Khambhaty, Kalpana Haresh Mody, Bhavanath Jha
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Abstract
Halophilic marine gram negative bacteria, isolated from decayed algae, hydrolyzed κ-carrageenan. To maximize κ-carrageenase production in the cost effective manner, a novel medium was defined having minimum components and their optimum concentration by statistical optimization method. Hence there evolved a novel medium composition for enhanced κ-carrageenase production by a new halotolerant marine bacterium, Pseudomonas spp.
Claims4
A method for the preparation of κ-carrageenase, said method comprising the steps of:
- (i) growing a halotolerant marine bacterium, Pseudomonas sp., accession number MTCC 5261, in a liquid medium comprising carrageenan, yeast extract, sodium chloride, K2HPO4 and KH2PO4 in a concentration ratio ranging from 0.1:1.0:50:1:0.5 to 3.0:1.0:15.0:0.5:0.05, respectively, for 16-72 hours at 15-500° C. to obtain a culture;
- (ii) centrifuging the culture obtained from step (i) at 5000-8000 rpm for 20-40 min. to obtain a cell free extract as a supernatant;
- (iii) treating the supernatant obtained from step (ii) with ammonium sulphate at a temperature in the range of 3-15° C. and aging the mixture for a period in the range of 12-36 hours followed by centrifugation at 5000-8000 rpm at 3-15° C. to obtain a pellet containing κ-carrageenase;
- (iv) suspending the pellet obtained from step (iii) in Tris-HCl buffer solution to obtain a suspension containing κ-carrageenase;
- and
- (v) dialyzing the suspension obtained in step (iv) for removing adhered ammonium sulphate followed by gel filtration to obtain purified κ-carrageenase.
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