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. 1990 Apr;56(4):837–843. doi: 10.1128/aem.56.4.837-843.1990

Purification and Characterization of an Extracellular Acid Proteinase from the Ectomycorrhizal Fungus Hebeloma crustuliniforme

Hong Zhu 1,†,*, Da-Cheng Guo 1, Bruce P Dancik 1
PMCID: PMC184309  PMID: 16348169

Abstract

Hebeloma crustuliniforme produced an extracellular acid proteinase in a liquid medium containing bovine serum albumin as the sole nitrogen source. The proteinase was purified 26-fold with 20% activity recovery and was shown to have a molecular weight of 37,800 (as indicated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and an isoelectric point of 4.8 ± 0.2. The enzyme was most active at 50°C and pH 2.5 against bovine serum albumin and was stable in the absence of substrates at temperatures up to 45°C and pHs between 2.0 and 5.0. Pepstatin A, diazoacetyl-dl-norleucine methylester, metallic ions Fe2+ and Fe3+, and phenolic acids severely inhibited the enzyme activity, while antipain, leupeptin, N-α-p-tosyl-l-lysine chloromethyl ketone, and trypsin inhibitor inhibited the activity moderately. The proteinase hydrolyzed bovine serum albumin and cytochrome c rapidly compared with casein and azocasein but failed to hydrolyze any of the low-molecular-weight peptide derivatives tested.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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