I want to transform some Trichoderma strains, previously I used Glucanex, but it is no longer available in Mexico, has anyone tried the Aspergillus lysing enzyme mixture or any other? That I can easily get in Mexico. Thanks.
2/23/23
Dear Claudia,
I'll try to answer your question. Several years ago, I was attempting to isolate genomic DNA from yeast (S. cerevisiae). I needed a "gentle" way to lyse the cells so as to not denature the DNA. I used an enzyme preparation from Zymo Research (Tuftin, CA, https://www.zymoresearch.com). The enzyme was Zymolyase, from Arthrobacter luteus. It contains several glucanases that act on the glucans in yeast cell walls. It worked very well with the yeast. In the presence of an osmotic stabilizer (sorbitol or sucrose), the enzyme converted the yeast cells into spheroplasts. Without the stabilizer, the the enzyme caused complete lysis of the cells & release of DNA.
Glucan is a component of many types of fungal cell walls, so it may work on your Trichoderma. You can go to Zymo's web site and check this out. Below is some information on the enzyme. You can contact Zymo and ask if the enzyme will work on Trichoderma. If not, they may have a suitable substitute for you.
I hope this information helps you. Good luck w/ your research!
Bill Colonna Iowa State University, Ames, Iowa, USA [email protected]
HIGHLIGHTS
- 100 T Equivalent: Prepared from Arthrobacter lutes. Essential enzyme activities are β-1,3-glucanase and β-1,3-glucan laminaripentao-hydrolase.
- Convenient: Provided lyophilized along with a storage buffer for reconstitution.
- Efficient Cell Wall Digestion: Supplied storage buffer has been optimized to confer maximum levels of enzymatic activity.
DESCRIPTION
Digestion of yeast and fungal cell walls is necessary for many experimental procedures including spheroplasting, immunofluorescence, transformation, protein purification, and others. The use of lytic enzymes like Zymolyase is routinely used for digestion. The Zymolyase from Zymo Research is prepared from Arthrobacter luteus, lyophilized, and packaged with a resuspension buffer. The buffer has been optimized to confer maximal levels of enzymatic activity. The main activities of the enzyme are β-1,3 glucanase and β-1,3-glucan laminaripentao-hydrolase, which hydrolyze glucose polymers at the β-1,3-glucan linkages releasing laminaripentaose as the principal product. Optimal Zymolyase activity is at 30° - 37°C; lytic activity ceases at higher temperatures. R-Zymolyase includes 0.5 U/µl RNase A when reconstituted. Susceptible fungal genera: Asbya, Candida, Debaryomyces, Eremothecium, Endomyces, Hansenula, Hanseniaspora, Kloekera, Kluyveromyces, Lipomyces, Metschikowia, Pichia, Pullularia, Saccharomyces, Saccharomyces, Saccharomycopsis, Schizosaccahromyces, and Torulopsis.
TECHNICAL SPECIFICATIONS
Activityβ-1,3-glucan laminaripentao-hydrolase and β-1,3-glucanase (trace amount of protease, ca. 1.5 units per 10 µl). DNase and RNase: none detectable.Concentration5 U/µlDeactivationLytic activity is lost in 5 minutes at 60°CEnzyme SourceArthrobactor luteusFormatLyophilized enzyme provided w/ storage bufferOptimum pH & TemppH 7.5, 35°C (lysis of viable yeast), pH 6.5, 45°C (hydrolysis of yeast glucan).Product StorageZymolyase is stable for over 1 year at -20°C or many years below -70°C.Protein Concentration~10-15 mg/mlSusceptible Fungal Genera:Asbya, Candida, Debaryomyces, Eremothecium, Endomyces, Hansenula, Hanseniaspora, Kloekera, Kluyveromyces, Lipomyces, Metschikowia, Pichia, Pullularia, Saccharomyces, Saccharomyces, Saccharomycopsis, Schizosaccahromyces, TorulopsisUnit DefinitionOne lytic unit is defined as catalyzing a 10% decrease in optical density at 800 nm (OD800) at 30°C in 30 minutes.UsageSpheroplast/Protoplast formation; Yeast cell fusion; Yeast transformation; Lysis of fungi
PRODUCT FAQ
CITATIONS
W.J. Colonna thank you very much, I'm going to check the enzyme preparation you said.
Greetings.
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