Isolation and Characterization of a Serine Proteinase, Inactivating m-Subunit of Lactate Dehydrogenase, from<i>Penicillium citrinum</i>KE-1

Watazu, Yoshifumi; Nagamatsu, Katashi; Shirahase, Yasushi; Kaneda, Nobuaki; Murao, Sawao; Okabe, Hiroaki

doi:10.1271/bbb.58.745

“…The N-terniinal amino acid sequence of the 33 kD allergen of P. citrinum was determined to be ANVVQSNVP which showed a high degree of sequence homology to that oi' a serine proteinase from P. citrinum KE-1 (KE) [15]. Besides, this sequence was also found to be identical to the first nine amino acids of an alkaline serine proteinase of P. citrinum (PC) reported by Yamamoto et al | IO|.…”

Section: Discussionmentioning

confidence: 60%

Characterization of the 33‐kilodalton major allergen of Penicillium citrinum by using MoAbs and N‐terminal amino acid sequencing

Shen¹,

Lin²,

Liaw³

et al. 1997

Clin Experimental Allergy

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Purification and characterization of a 315 kDa keratinolytic subtilisin-like serine protease from Microsporum canis and evidence of its secretion in naturally infected cats

Mignon¹,

Swinnen²,

Bouchara³

et al. 1998

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A keratinolytic protease, secreted as the major component by a feline clinical isolate of Microsporum canis cultivated in a minimal medium containing cat keratin, was purified by affinity chromatography on bacitracin agarose and gel filtration. The apparent molecular mass of the enzyme was 31.5 kDa and the pI was 11.8. The enzyme was not glycosylated and its first 15 N-terminal amino acids showed numerous similarities with other fungal subtilisins. The optimum pH was around 9 while inactivation of the enzyme was reversible at pH 4, but not at pH 11. The enzyme was stable at 37 degrees C with an apparent optimum temperature around 55 degrees C. PMSF, soybean trypsin inhibitor (SBTI) and chymostatin strongly inhibited the proteinase. The highest affinity (Km of 0.37 mM) and physiological efficiency (k(cat)/Km) were obtained for the synthetic substrate N-Suc-Ala-Ala-Pro-Phe-p-nitroanilide. These results indicate that the keratinase belongs to the subtilisin-like serine protease family. Purified rabbit immunoglobulins G prepared against the keratinase and used in an immunohistochemical test allowed the detection of the keratinase produced by the fungus invading hair structures in naturally infected cats. The in vitro keratinolytic activity of the enzyme and its production in vivo suggest that it may contribute to pathogenicity.

show abstract

Purification and characterization of a 315 kDa keratinolytic subtilisin-like serine protease fromMicrosporum canisand evidence of its secretion in naturally infected cats

Mignon¹,

Swinnen²,

Bouchara³

et al. 1998

View full text Add to dashboard Cite

A keratinolytic protease, secreted as the major component by a feline clinical isolate of Microsporum canis cultivated in a minimal medium containing cat keratin, was purified by affinity chromatography on bacitracin agarose and gel filtration. The apparent molecular mass of the enzyme was 31.5 kDa and the pI was 11.8. The enzyme was not glycosylated and its first 15 N-terminal amino acids showed numerous similarities with other fungal subtilisins. The optimum pH was around 9 while inactivation of the enzyme was reversible at pH 4, but not at pH 11. The enzyme was stable at 37 degrees C with an apparent optimum temperature around 55 degrees C. PMSF, soybean trypsin inhibitor (SBTI) and chymostatin strongly inhibited the proteinase. The highest affinity (Km of 0.37 mM) and physiological efficiency (k(cat)/Km) were obtained for the synthetic substrate N-Suc-Ala-Ala-Pro-Phe-p-nitroanilide. These results indicate that the keratinase belongs to the subtilisin-like serine protease family. Purified rabbit immunoglobulins G prepared against the keratinase and used in an immunohistochemical test allowed the detection of the keratinase produced by the fungus invading hair structures in naturally infected cats. The in vitro keratinolytic activity of the enzyme and its production in vivo suggest that it may contribute to pathogenicity.

show abstract

Isolation and Characterization of a Serine Proteinase, Inactivating m-Subunit of Lactate Dehydrogenase, fromPenicillium citrinumKE-1

Cited by 3 publications

References 7 publications

Characterization of the 33‐kilodalton major allergen of Penicillium citrinum by using MoAbs and N‐terminal amino acid sequencing

Characterization of the 33‐kilodalton major allergen of Penicillium citrinum by using MoAbs and N‐terminal amino acid sequencing

Purification and characterization of a 315 kDa keratinolytic subtilisin-like serine protease from Microsporum canis and evidence of its secretion in naturally infected cats

Purification and characterization of a 315 kDa keratinolytic subtilisin-like serine protease fromMicrosporum canisand evidence of its secretion in naturally infected cats

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