The nematophagous fungus Verticillium chlamydosporium produces a chymoelastase-like protease which hydrolyses host nematode proteins in situ.
The nematophagous fungus Verticillium chlamydosporium secreted several proteases in submerged culture in which soya peptone was the sole carbon and nitrogen source. One protease, VCP1 (M(r) 33,000, pI 10.2), was purified 14-fold from culture filtrates to apparent homogeneity using preparative isoelectric focusing in free solution, and shown to rapidly hydrolyse the chymotrypsin substrate Suc-(Ala)2-Pro-Phe-pNA and elastin. VCP1 had a Km for Suc-(Ala)2-Pro-Phe-pNA of 4.3 x 10(-5) M and a kcat of 5.8 s-1. It was highly sensitive to PMSF and TPCK, but only moderately sensitive to chicken egg-white and soya bean trypsin inhibitors. VCP1 degraded a wide range of polymeric substrates, including Azocoll, hide protein, elastin, casein and albumin, and accounted for most of the non-specific protease activity detected in culture filtrates. The purified enzyme hydrolysed proteins in situ from the outer layer of the egg shell of the host nematode Meloidogyne incognita and exposed its chitin layer. VCP1 was secreted by several isolates of V. chlamydosporium and V. lecanii, pathogens of nematodes and insects respectively, but not plant-pathogenic species of Verticillium. These observations suggest that VCP1 or similar enzyme(s) may play a role in the infection of invertebrates.
Segers
R
R
Rothamsted Experimental Station, Entomology & Nematology Department, Harpenden, Hertfordshire, UK.
Butt
T M
TM
Kerry
B R
BR
Peberdy
J F
JF
D000418:Albumins / Q000378:metabolism; D001391:Azo Compounds / Q000378:metabolism; D002364:Caseins / Q000378:metabolism; D002918:Chymotrypsin / Q000378:metabolism; D003094:Collagen / Q000378:metabolism; D004549:Elastin / Q000378:metabolism; D010450:Endopeptidases / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D006868:Hydrolysis; D003904:Mitosporic Fungi / Q000201:enzymology*; D012697:Serine Endopeptidases / Q000737:chemistry*; D013379:Substrate Specificity
D000418:Albumins; D001391:Azo Compounds; D002364:Caseins; C028470:Azocoll; D003094:Collagen; D004549:Elastin; D010450:Endopeptidases; D012697:Serine Endopeptidases; C053722:chymoelastase; D002918:Chymotrypsin