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Original Articles

Activity of protease extracted from rice-rhizosphere soils under double cropping of rice and wheat

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Pages 597-603 | Received 28 Oct 1994, Accepted 15 Dec 1994, Published online: 04 Jan 2012
 

Abstract

Protease-active extracts were prepared from 2 rice-rhizosphere and non-rhizosphere soil samples 3 proteolytic isolates from the soil samples which were taken from water logged paddy fields (Gray Lowland Soils) under double cropping of rice and wheat that had been treated with chemical fertilizer (CF) or organic manure (OM). The activities were assayed and characterized using an artificial substrate, benzyloxycarbonyl-L-phenylalanyl-L-tyrosyl-L-leucine (ZFTL). The protease-active extracts from the rhizosphere and non-rhizosphere of the CF and OM soils (abbreviated as CFR, CFN, OMR, and OMN) showed an optimum pH between 7 and 8. The extracts hydrolyzed ZFTL to Tyr-Leu and Leu; generally, Tyr-Leu was the main reaction product. Activities of all the samples were inhibited by EDTA and p-chloromercuribenzoic acid (PCMB) but not by pepstatin and phenylmethanesulfonyl fluoride (PMSF). These results indicate that the main components of the ZFTL-hydrolases in CFR, CFN, 0MR, and OMN were essentially similar.

Optimum pH for the activity of the protease extracted from a Gram negative rod (MNB003)., two actinomycetous isolates (MNA014., MRA017) was about 8, 8, and 8.5, respectively. Extract from the culture of MNB003 hydrolyzed ZFTL to Tyr-Leu., Leu,., the protease activity was inhibited by EDTA., PCMB. Protease activities of the extracts from cultures of the two actinomycetes were inhibited by EDTA but not by PCMB. The extract from culture of MNA014 produced mainly Tyr-Leu from ZFTL while MRA017 produced Leu. Characteristics of the protease activity from cultures of MNB003., MNA014 were similar to those of the enzyme extracted from a paddy field soil under single cropping (Takeuchi., Hayano 1994: Soil Sci. Plant Nutr., 40, 691-695) and from an Andosol under tomato cultivation (Hayano et al. 1987: Biol. Fertil. Soils, 4, 179-183), respectively.

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