Study On Cellulase Production By Oil Camellia Testa Fermention

The camellia testa would adversely affect the quality of squeezed oil-camellia oil,thus the teste was separated in the processing. Lignocellulose is the main component of the oil-camellia testa, a substrate for microorganism to product cellulose enzyme. The sources of cellulase is very extensive and the use of microbial fermentation method is the most convenient way to product cellulase.This study was designed to ferment oil-camellia testa for producing cellulase by using of Trichoderma koningii. In the study the component of oil-camellia testa was determined by chemical analysis and the structure of oil-camellia testa was observed by scanning electron microscope(SEM). Through single factor experiments the effects of different growth conditions for Trichoderma koningii were compared. The process parameters of fermentative production of cellulase were optimized by using response surface method. The factors that influence the activity of cellulase were determined by measuring the relative activity of cellulase at different temperatures, pH and metal ions,1) The chemical analysis results indicated that moisture, ash, crude fat, crude protein, reducing sugar, camellia saponin, hemicellulose and cellulose content were11.07%,1.02%,2.53%,1.59%,10.68%,22.00%,17.32%and31.35%in oil-camellia testa, respectively.2) SEM result showed that the cell-wall of oil-camellia testa was mainly composed of hemicellulose,cellulose and lignin. The structure of cell-wall showed layers’structure, with hemicellulose in the inner layer, cellulose in the outer and lignin in the inter-layer structure between hemicellulose and cellulose.3) Single-factor test was first adopted to determine the effects of facters on CMCase activity. The optimum fermenting condition of the factors were determined by response surface method. The optimal fermentation condition was as follows:raw material pretreatment was alkaline treatment; nitrogen source was0.2%(NH4)2SO4; initial pH was5.8; inoculation dosage was5%; volume of liquid was22mL in100mL conical flask; fermentation day was5d. Under this optimal condition, the CMCase enzyme activity was179.15U increased by24.52%.4) In this paper,the β-glucosidase activity, CMCase and the filter paper activity curves under different conditions of temperature and pH were measured as well as the three type of enzyme relative activity to study the stability after the pretreatment in different temperatures, pH and concentrations of metal ions for2h. Finally, the integrated curve of the three type of enzyme activity,the optimum enzyme activity temperature is50-60℃, optimum pH of4-5, the stored enzyme solution temperature was20-30℃, the enzyme solution stored pH was3-6. In crude enzyme solution,β-glucosidase metal ion activator were:Co2+, Mn2+, Fe3+, Ca2+, Ba2+, and inhibitor were Cu2+, Al3+, Mg2+; CMCase metal ion activator were:Co2+, Mn2+, Ca2+, Ba2+, Al3+, Fe3+, Mg2+, and inhibitor were Cu2+, K+; filter paper activity of the metal ion activator were:Co2+, Mn2+, and Fe3+, Ba2+, Al3+, Mg2+, K+, Ca2+was activated at a suitable concentration, inhibitor was Cu2+.