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Proteinase activity in potato plants

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Abstract

Several vegetative tissues of potato plants were screened for proteinase activity. Both endopeptidase and exopeptidase activities were investigated using gelatin and L-amino acid-4-nitroanilides (benzoyl-L-arginine-4-nitroanilide/BAPA, glutaryl-L-phenyl-alanine-4-nitroanilide/GLUPHEPA, alanine-4-nitro-anilide/APA, leucine-4-nitroanilide/LPA, and benzoyl-L-tyrosine-4-nitroanilide/BTPA) as substrates. Leaves and rootes were found to contain the highest levels of endopeptidase activity; lesser activities were detected in flower petals, sprouts, and tubers. Three different types of proteinases, L-BAPAase (serine proteinase), APAase (thiol proteinase), and BTPAase (sensitive to reducing agents), were characterized in various physical and chemical properties. Their temperature optima were determined to be 25° (L-BAPAase) and 40° (BTPAase, APAase) respectively; their pH optimum was between 8.6 and 9.0, their isoelectric points were between pH 4.25 and 6.0, and their molecular weight was estimated 70,000 (L-BAPAase, APAase) and between 150,000–250,000 (BTPAase). The trypsin-like activity against L-BAPA was inhibited by diisopropylfluorophosphate and by tosyllysine-chloromethyl ketone, but not by trypsin inhibitors from potato and legume.

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Abbreviations

APA:

alanine-4-nitroanilide

BAPA:

benzoyl-L-arginine-4-nitroanilide

BTPA:

benzoyl-L-tyrosine-4-nitroanilide

DFP:

diisopropylfluorophosphate

DMF:

dimethyl formamide

EDTA:

ethylenedinitrilotetraacetic acid

GLUPHEPA:

glutaryl-L-phenylalanine-4-nitroanilide

LPA:

leucine-4-nitroanilide

PHMB:

p-hydroxy-mercuribenzoate

PI-I:

potato chymotrypsin inhibitor I

PPI:

potato proteinase leaf

PPr:

potato proteinase root

PPt:

potato proteinase tuber

PVP:

polyvinylpyrrolidone

TLCK:

tosyl-L-lysinechloromethyl ketone

TPCK:

tosyl-L-phenylalanyl chloromethane

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Authors and Affiliations

  1. Institut für Lebensmittelchemie, Technische Universität München, D-8046, Garching, Federal Republic of Germany

    K. Santarius & H. -D. Belitz

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  1. K. Santarius
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  2. H. -D. Belitz
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Santarius, K., Belitz, H.D. Proteinase activity in potato plants. Planta 141, 145–153 (1978). https://doi.org/10.1007/BF00387881

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