NDLI: Carboxymethylcellulase and β-Glucosidase Secretion by Protoplasts of Trichoderma reesei

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Carboxymethylcellulase and β-Glucosidase Secretion by Protoplasts of Trichoderma reesei

Content Provider	Semantic Scholar
Author	Kolar, Harry R. Mischak, Harald Kammel, W. P. Kubicek, Christian Peter
Copyright Year	1985
Abstract	Novozym 234, a commercially available enzyme from Trichoderma harzianum, has been used to prepare protoplasts from Trichoderma reesei QM 9414. Optimal conditions were: 20 h old mycelium, 0.9 M-KCl as the osmotic stabilizer. 0.1% (w/v) Novozym 234 and 18 h incubation at 28°C. To prevent damage to the protoplasts during isolation the incubation mixtures were agitated by vibration, so avoiding shaking or stirring. More than 95% of the protoplasts were viable and were also metabolically intact as shown by their ability to take up [14C]leucine and incorporate it into cellular protein. Freshly prepared protoplasts could be induced by sophorose to produce and secrete carboxymethylcellulase and β-glucosidase. Optimal conditions were: 0.9 M-KCl as osmotic stabilizer, phosphate buffer pH 6.0, 7 mM-sophorose and 107 protoplasts ml-1. Separation of the secreted proteins by fast protein liquid chromatography revealed at least seven peaks of carboxymethylcellulase activity and β-glucosidase activity, indicating that some multiplicity of the forms of these enzymes is already present at the secretion stage.
Starting Page	1339
Ending Page	1347
Page Count	9
File Format	PDF HTM / HTML
DOI	10.1099/00221287-131-6-1339
Alternate Webpage(s)	https://www.microbiologyresearch.org/docserver/fulltext/micro/131/6/mic-131-6-1339.pdf?accname=guest&checksum=94B622F3D09B9756C713A43B6AB0BAE7&expires=1546266128&id=id
Alternate Webpage(s)	http://www.microbiologyresearch.org/docserver/fulltext/micro/131/6/mic-131-6-1339.pdf?accname=guest&checksum=2A501AC912BA2203384ABD8DF363265C&expires=1545285987&id=id
Alternate Webpage(s)	https://doi.org/10.1099/00221287-131-6-1339
Volume Number	131
Language	English
Access Restriction	Open
Content Type	Text
Resource Type	Article

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