A novel fungal expression sys1em

Secretion of active calf chymosin from the filamentous fungus trichoderma reesei

A. Harkki, J. Uusitalo, M. Bailey, M. Penttila, J. K.C. Knowles

Research output: Contribution to journalArticleScientificpeer-review

106 Citations (Scopus)

Abstract

We have studied the expression of a bovine chymosin cDNA in Trichoderma ree- sei to test the ability of this novel fungal host vector system to express and secrete heterologous gene products. Four different expression plasmids were constructed to determine the optimum manner to fuse the chymosin cDNA to the promoter and the terminator of the major T. reesei cellu- lase gene, cellobiohydrolase I (cbhl). All four constructions, when transformed into Trichoderma, determined the secretion of a polypeptide corresponding in size to prochymosin and reacting with antichy- mosin antiserum. This polypeptide had a molecular weight indistinguishable from chymosin and showed milk clotting activity. In preliminary fermentation studies, one transformant secreted as much as 40 mg/1 of active chymosin.

Original languageEnglish
Pages (from-to)596-603
Number of pages8
JournalBio/Technology
Volume7
Issue number6
DOIs
Publication statusPublished - 1 Jan 1989
MoE publication typeA1 Journal article-refereed

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Chymosin
Trichoderma
Polypeptides
Fungi
Genes
Electric fuses
Fermentation
Complementary DNA
Molecular weight
Cellulose 1,4-beta-Cellobiosidase
Peptides
Cellulase
Immune Sera
Milk
Plasmids
Molecular Weight

Cite this

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title = "A novel fungal expression sys1em: Secretion of active calf chymosin from the filamentous fungus trichoderma reesei",
abstract = "We have studied the expression of a bovine chymosin cDNA in Trichoderma ree- sei to test the ability of this novel fungal host vector system to express and secrete heterologous gene products. Four different expression plasmids were constructed to determine the optimum manner to fuse the chymosin cDNA to the promoter and the terminator of the major T. reesei cellu- lase gene, cellobiohydrolase I (cbhl). All four constructions, when transformed into Trichoderma, determined the secretion of a polypeptide corresponding in size to prochymosin and reacting with antichy- mosin antiserum. This polypeptide had a molecular weight indistinguishable from chymosin and showed milk clotting activity. In preliminary fermentation studies, one transformant secreted as much as 40 mg/1 of active chymosin.",
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A novel fungal expression sys1em : Secretion of active calf chymosin from the filamentous fungus trichoderma reesei. / Harkki, A.; Uusitalo, J.; Bailey, M.; Penttila, M.; Knowles, J. K.C.

In: Bio/Technology, Vol. 7, No. 6, 01.01.1989, p. 596-603.

Research output: Contribution to journalArticleScientificpeer-review

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