2.1.8.2. Absorbency Under Load (AUL) - BADA

In this solution, protein, lipids and other organic soluble matters were hydrolysized and
solubilized. Insoluble alkaline materials (IAM) in this solution were then separated by
centrifugation at 10000 rpm for 10 min.
The unfolded protein solution was neutralized at pH 7 by HCl 1 M, and then the filtered Protein
solution was precipitated by adding ammonium sulfate with ratio 1:4 (w/v). The precipitated fungi
protein was then separated by filter vacuum and washed 3 times with 70% aqueous ethanol
solution. Drying process was occurred in freeze dryer so prepared protein was ready to further
processing. Figure 15 shows a schematic diagram of basic steps of protein extraction from
zygomycetes fungi.
2.1.2. Separation and Purification Process of Protein
2.1.2.1.Clarification and concentration of Zygomycetes Protein
All biological systems (e.g., Cell culture or animal and plant sources) produce a complex mixture
of proteins, waste materials, growth media, extra cellular and intra cellular products in which the
target molecule is normally a small quantity. The first step of purification of target proteins is often
a removal of the particulate matter in the process stream. The process by which this removal is
attained called clarification.
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Much laboratory scale clarification is achieved by centrifugation of small volumes of process
stream. Definitely centrifugation is based on the difference in density between particulate materials
and the liquid where the particles are contained. Centrifuges can be used to remove particulates
material down to 0.5 μm in size before to filtration for preparation prior to determination by
analytical instruments such as chromatography and UV. Spectrophotometer.
After clarification many process use a concentration step to increase the quality of the protein as a
feed stream for subsequent process. This concentration step can be performed by chemical or
physical methods. A common technique in the laboratory is solvent extraction, which is usually
known as liquid-liquid extraction and can be done in the aqueous or solvent phase. The process is
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