Purification and partial characterisation of tentatively classified acid phosphatase from the earthworm Eisenia veneta

In order to use leakage of lysosomal acid phosphatase (AP) as a biomarker o f stress to earthworms. more information about AP's in earthworms are neede d. This paper describes the details about tentatively classified APs in the earthworm Eisenia veneta. Two isoenzymes (enzyme I and II) of acid phospha tase (AP) and one alkaline phosphatase (enzyme III) from the earthworm E. v eneta were separated by gel filtration. All three enzymes were further puri fied and concentrated on a Con A Sepharose 4B column. Enzyme I was inhibite d by tartrate, showed an optimal pH range between 4.0 and 5.0 and was assum ed to be of lysosomal origin. Enzyme II was the major enzyme showing the hi ghest activity of the three enzymes. It was expected to be a lysosomal AP u nder physiological conditions. Enzyme II had a molecular mass 113 kDa and w as composed of apparently identical polypeptide chains of 36 kDa each. This enzyme was inhibited by tartrate, showed an optimal pH in the range 6.0-7. 5 and was slowly degraded at temperatures above 40 degrees C. Enzyme III is not inhibited by tartrate and has a pH-optimum > 9. The subcellular locati on under physiological conditions was assumed to be the cytosol. (C) 2000 E lsevier Science Inc. All rights reserved.