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Evaluation of sonication treatment and buffer composition on rumen bacteria protein extraction and carboxymethylcellulase activity
Author(s) -
Prauchner Carlos A,
Kozloski Gilberto V,
Farenzena Roberta
Publication year - 2013
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.5959
Subject(s) - sonication , incubation , rumen , chemistry , tonicity , chromatography , extraction (chemistry) , residue (chemistry) , composition (language) , bacteria , food science , biochemistry , biology , linguistics , philosophy , fermentation , genetics
BACKGROUND The methodological procedures for studying the fibrolytic activity of rumen bacteria are not clearly established. In this study the efficiency of sonication treatment and buffer composition (i.e. buffer varying in tonicity or pH ) on the level of protein extraction from the residue of forage samples incubated in the rumen of a grazing steer and the effect of buffer composition or CaCl 2 concentration on the carboxymethylcellulase ( CMCase ) activity of the released protein were evaluated.RESULTS The amount of protein released from the residue of incubation was higher ( P < 0.05) for the sonicated material and increased linearly with increasing buffer pH ( P < 0.05). The CMCase activity of the released protein was not improved by sonication treatment, whereas it was higher ( P < 0.05) for hypotonic than for hypertonic buffer. Both linear and quadratic effects ( P < 0.05) of buffer pH on CMCase activity were significant, with CMCase activity being maximal at pH 5.4–6.1. CMCase activity was higher ( P < 0.05) at a CaCl 2 concentration of 1 mmol L −1 compared with lower values.CONCLUSION Although sonication treatment increases the amount of protein extracted from rumen bacteria adhered to the residue of incubation, the CMCase activity of the released protein might be measured without sonication treatment and should be carried out with a hypotonic buffer solution that includes a calcium source. When pH is not a treatment factor, the buffer pH should be between 5.5 and 6. © 2012 Society of Chemical Industry