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A moderately cold active, extracellular alkaline protease producing bacterium was isolated from a fresh water lake. The isolate was found to be a gram-positive, rod shaped organism later identified as Bacillus cereus MTCC 6840. The bacterium produced the maximum amount of enzyme when allowed to grow for 24 h at temperature 25º and pH 9.0. Among a variety of substrates used, fructose as a carbon source and a combination of yeast extract and peptone as nitrogen source, supported the maximum protease production by the organism (120 U/ml). Fe++ and Co++ stimulated the enzyme activity whereas Ca++, Cu++, K+, Mg++ and Mn++ inhibited it to different extents. The protease was found to be highly stable in the presence of NaCl, SDS and acetone. Treatment with EDTA and PMSF resulted in the considerable loss of enzyme activity. The enzyme was found to be optimally active at pH 9.0 and temperature 20ºC. Uma bactéria produtora de protease alcalina extracelular, moderadamente ativa no frio, foi isolada da água de um lago. Trata-se de um bacilo Gram positivo, identificado como Bacillus cereus MTCC6840. A maior produção da enzima foi em 24h a 25ºC e pH 9,0. A produção máxima de protease (120 U/ml) ocorreu quando foi utilizada frutose como fonte de carbono e uma combinação de extrato de levedura com peptona como fonte de nitrogênio. Fe++ e Co++ estimularam a atividade da enzima, enquanto Ca++, Cu++, K+,Mg++ e Mn++ tiveram efeito inibitório, com intensidades diferentes. A protease permaneceu estável na presença de NaCl, SDS e acetona. O tratamento com EDTA e PMSF causou uma significativa perda na atividade. A enzima apresentou atividade ótima em pH 9,0 e temperatura de 20ºC

brazilian journal of microbiology

Production of moderately halotolerant, SDS stable alkaline protease from Bacillus cereus MTCC 6840 isolated from lake Nainital, Uttaranchal state, India