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. 2009 Jul;40(3):623-30.
doi: 10.1590/S1517-838220090003000026. Epub 2009 Sep 1.

Production, purification and application of extracellular chitinase from Cellulosimicrobium cellulans 191

Luciana F Fleuri  1 , Haroldo Y Kawaguti, Hélia H Sato
Affiliations

Production, purification and application of extracellular chitinase from Cellulosimicrobium cellulans 191

Luciana F Fleuri et al. Braz J Microbiol. 2009 Jul.

Abstract

This study concerned the production, purification and application of extracellular chitinase from Cellulosimicrobium cellulans strain 191. In shaken flasks the maximum yield of chitinase was 6.9 U/mL after 72 h of cultivation at 25°C and 200 rpm. In a 5 L fermenter with 1.5 vvm aeration, the highest yield obtained was 4.19 U/mL after 168 h of fermentation at 25°C and 200 rpm, and using 3 vvm, it was 4.38 U/mL after 144 h of fermentation. The chitinase (61 KDa) was purified about 6.65 times by Sepharose CL 4B 200 gel filtration with a yield of 46.61%. The purified enzyme was able to lyse the cell walls of some fungi and to form protoplasts.

Keywords: Cellulosimicrobium cellulans; chitinase; fungal lysis; protoplasts.

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Figures

Figure 1
Figure 1
Chitinase production by C. cellulans strain 191 in a 5 L fermenter at 25°C, 200 rpm and 1.5 vvm: (♦) Chitinase activity; (▲) pH; (■) Cell mass.
Figure 2
Figure 2
Chitinase production by C. cellulans strain 191 in a 5 L fermenter at 25°C, 200 rpm and 3.0 vvm: (♦) Chitinase activity; (▲) pH; (■) Cell mass.
Figure 3
Figure 3
Purification of the chitinase obtained from C. cellulans strain 191 on a Sepharose CL4B200 column: (♦) Absorbance at 280 nm; (■) U chitinase/mL.
Figure 4
Figure 4
SDS-PAGE electrophoresis of the purified chitinase.
Figure 5
Figure 5
Fungi lysis and protoplast formation by purified chitinase.
See this image and copyright information in PMC

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