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Intergeneric fusant development using chitinase preparation of Rhizopus stolonifer NCIM 880

Fungal chitinase have tremendous applications in biotech industries, with this approach we focused on extracellular chitinase from Rhizopus stolonifer NCIM 880 for the formation of fungal protoplasts. The maximum chitinase production reached after 24 h at 2.5% colloidal chitin concentration in prese...

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Autores principales: Sonawane, Kailas D., Dandagal, Narayan R., Naikwadi, Akibjaved G., Gurav, Piyush T., Anapat, Samar V., Nadaf, Naiem H., Jadhav, Deepak B., Waghmare, Shailesh R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5108735/
https://www.ncbi.nlm.nih.gov/pubmed/27844458
http://dx.doi.org/10.1186/s13568-016-0287-8
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author Sonawane, Kailas D.
Dandagal, Narayan R.
Naikwadi, Akibjaved G.
Gurav, Piyush T.
Anapat, Samar V.
Nadaf, Naiem H.
Jadhav, Deepak B.
Waghmare, Shailesh R.
author_facet Sonawane, Kailas D.
Dandagal, Narayan R.
Naikwadi, Akibjaved G.
Gurav, Piyush T.
Anapat, Samar V.
Nadaf, Naiem H.
Jadhav, Deepak B.
Waghmare, Shailesh R.
author_sort Sonawane, Kailas D.
collection PubMed
description Fungal chitinase have tremendous applications in biotech industries, with this approach we focused on extracellular chitinase from Rhizopus stolonifer NCIM 880 for the formation of fungal protoplasts. The maximum chitinase production reached after 24 h at 2.5% colloidal chitin concentration in presence of starch as an inducer. Chitinase was extracted efficiently at 65% cold acetone concentration and then purified by using DEAE-Cellulose column chromatography. Purified chitinase having molecular weight 22 kDa with single polypeptide chain was optimally active at pH 5.0 and temperature 30 °C. The purified chitinase revealed kinetic properties like Km 1.66 mg/ml and Vmax 769 mM/min. Crude chitinase extract efficiently formed protoplasts from A. niger, A. oryzae, T. viride and F. moniliforme. The formed protoplasts of A. niger and T. viride showed 70 and 66% regeneration frequency respectively. Further, intergeneric fusants were developed successfully and identified at molecular level using RNA profiling. Thus, this study could be useful for strain improvement of various fungi for biotechnological applications.
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spelling pubmed-51087352016-12-07 Intergeneric fusant development using chitinase preparation of Rhizopus stolonifer NCIM 880 Sonawane, Kailas D. Dandagal, Narayan R. Naikwadi, Akibjaved G. Gurav, Piyush T. Anapat, Samar V. Nadaf, Naiem H. Jadhav, Deepak B. Waghmare, Shailesh R. AMB Express Original Article Fungal chitinase have tremendous applications in biotech industries, with this approach we focused on extracellular chitinase from Rhizopus stolonifer NCIM 880 for the formation of fungal protoplasts. The maximum chitinase production reached after 24 h at 2.5% colloidal chitin concentration in presence of starch as an inducer. Chitinase was extracted efficiently at 65% cold acetone concentration and then purified by using DEAE-Cellulose column chromatography. Purified chitinase having molecular weight 22 kDa with single polypeptide chain was optimally active at pH 5.0 and temperature 30 °C. The purified chitinase revealed kinetic properties like Km 1.66 mg/ml and Vmax 769 mM/min. Crude chitinase extract efficiently formed protoplasts from A. niger, A. oryzae, T. viride and F. moniliforme. The formed protoplasts of A. niger and T. viride showed 70 and 66% regeneration frequency respectively. Further, intergeneric fusants were developed successfully and identified at molecular level using RNA profiling. Thus, this study could be useful for strain improvement of various fungi for biotechnological applications. Springer Berlin Heidelberg 2016-11-14 /pmc/articles/PMC5108735/ /pubmed/27844458 http://dx.doi.org/10.1186/s13568-016-0287-8 Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Original Article
Sonawane, Kailas D.
Dandagal, Narayan R.
Naikwadi, Akibjaved G.
Gurav, Piyush T.
Anapat, Samar V.
Nadaf, Naiem H.
Jadhav, Deepak B.
Waghmare, Shailesh R.
Intergeneric fusant development using chitinase preparation of Rhizopus stolonifer NCIM 880
title Intergeneric fusant development using chitinase preparation of Rhizopus stolonifer NCIM 880
title_full Intergeneric fusant development using chitinase preparation of Rhizopus stolonifer NCIM 880
title_fullStr Intergeneric fusant development using chitinase preparation of Rhizopus stolonifer NCIM 880
title_full_unstemmed Intergeneric fusant development using chitinase preparation of Rhizopus stolonifer NCIM 880
title_short Intergeneric fusant development using chitinase preparation of Rhizopus stolonifer NCIM 880
title_sort intergeneric fusant development using chitinase preparation of rhizopus stolonifer ncim 880
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5108735/
https://www.ncbi.nlm.nih.gov/pubmed/27844458
http://dx.doi.org/10.1186/s13568-016-0287-8
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