Screening of chitin deacetylase from Mucoralean strains (Zygomycetes) and its relationship to cell growth rate.
Chitin deacetylase (CDA) is an enzyme that catalyzes the hydrolysis of acetamine groups of N-acetyl-D: -glucosamine in chitin, converting it to chitosan in fungal cell walls. In the present study, the activity in batch culture of CDA from six Mucoralean strains, two of them wild type, isolated from dung of herbivores of Northeast Brazil, was screened. Among the strains tested, Cunninghamella bertholletiae IFM 46114 showed a high intracellular enzyme activity of 0.075 U/mg protein after 5 days of culture, and a wild-type strain of Mucor circinelloides showed a high intracellular enzyme activity of 0.060 U/mg protein, with only 2 days of culture, using N-acetylchitopentaose as substrate. This enzyme showed optimal activity at pH 4.5 in 25 mM glutamate-sodium buffer at 50 degrees C, and was stable over 1 h preincubation at the same temperature. The kinetic parameters of CDA did not follow Michaelis-Menten kinetics, but rather Hill affinity distribution, showing probable allosteric behavior. The apparent K(HILL) and Vmax of CDA were 288+/-34 nmol/l and 0.08+/-0.01 U mg protein(-1) min(-1), respectively, using N-acetylchitopentaose as substrate at pH 4.5 at 50 degrees C.
Amorim
R V S
RV
Laboratório de Imunopatologia Keizo Asami (LIKA), Universidade Federal de Pernambuco, Av. Professor Moraes Rego s/n, Cidade Universitária, 50 670-901, Recife, PE, Brazil.
Ledingham
W M
WM
Fukushima
K
K
Campos-Takaki
G M
GM
D000581:Amidohydrolases / Q000378:metabolism*; D020093:Cunninghamella / Q000201:enzymology* / Q000254:growth & development*; D004789:Enzyme Activation; D007218:Industrial Microbiology; D007700:Kinetics; D008828:Microbiological Techniques; D009089:Mucor / Q000201:enzymology* / Q000254:growth & development*
D000581:Amidohydrolases; C019866:chitin deacetylase