Purification , Characterization . mveus , and Crystallization of Two Types of Lipase from Rhizopus
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چکیده
DEAE-Toyopearl (1 pass) and CM-Toyopearl (2 passes). Lipase I consists of two polypeptide chains [a small peptide with sugar moiety (A-chain) and a large peptide of moleculaT weight 34,OOO (B-chain)]. Lipase II has a molecular weight of 30,OOO consisting of a single polypeptide chain. Lipase I appeared to be conyerted to Lipase II by limited proteQlysis by a specific protease a small amount of which is in the culture supernatant from Rh. niveus, because one of the peptides formed has the same N-terminal sequence and C-terminal amino acid as Lipase II, as well as the molecu]ar mass estimated by SDS-PAGE. Lipase I had a pH optimum of 6.0-6.5 and a temperature optimum of 350C, while, for Lipase II these values were pH 6.0 and 400C. Both enzymes were obtained in the crystalline state using the hanging drop method of yaper ditfusion and PEG as the precipitating agents.
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