Gel Filtration Chromatography Combined with Bradford Method for Determination of Total Residual Protein in Ferment Antibiotics

Aim A novel method has been developed for evaluation of the levels of total residual protein in antibiotics produced by fermentation using gel filtration chromatography (GFC) combined with Bradford assay based on determination of residual protein in lincomycin hydrochloride. Methods The chromatographic conditions were SuperdexTM peptide column, 0.01 mol*L-1 phosphate buffer solution as mobile phase, and flow rate of 1 mL·min-1. Five hundred microliters of lincomycin hydrochloride solution (3 g of lincomycin hydrochloride dissolved in 10 mL of mobile phase) was injected into the chromatograph and the eluted solution was collected between 6 min and 14.5 min (protein eluted from column within this period), and the residual content of total protein in the eluted solution was assayed using Bradford assay method. Results The average recovery was more than 90% for bovine serum albumin, the calibration equation for the range of 0-12 μg·mL-1 of protein was y=-0.002 4x2+0.064 2x+0.002 9, r2=0.999 9, RSD=0.1%-0.9%, and the LOD and LOQ were 3 and 10 ng·mL-1 of protein, respectively. Conclusion The novel method for determining the residual protein in ferment antibio-tics is simple, rapid, and precise.

Application of Bis(2-Ethylhexyl) Sulfosuccinate(AOT) /Isooctane Reverse Micellar System in Extraction of Sesame Residue Proteins

[ Objective] To investigate the factors affecting sesame residue protein extraction and find the optimum extraction conditions, [Method ] Sesame residue proteins were extracted with bis（2-ethylhexyl）sulfosuocinate （AOT）/isooctane reverse micellar system. Orthogonal test was used to screen the optimum combination of four factors including sesame residue concentration, pre-extraction pH value, potassium chloride concentration and reverse extraction time. The yield of extracted sesame residue proteins was the indicator to evaluate the efficiency of process. E Result1 The optimum extraction conditions are as follows： sesame residue concentration： 0.06 g/ml; pre-extraction pH value, 3.0; potassium chloride concentration, 1.5 mol/L; and reverse extraction time, 20 min. In the optimum extraction conditions, the extraction rate was up to 86%. [ Coaclusion] Extraction of sesame residue proteins can be well performed with AOT/isooctane reverse micellar system.

PHYLETIC RELATIONSHIP OF PROTEIN STRUCTURES BASED ON SPATIAL PREFERENCE OF RESIDUES

A distance measure that infers to indicate the evolutionary relationship of protein structures has been developed based on spatial preference factors of residues. The spatial preference factor is a reflection of the environment of residues in tertiary structure. Compared with the phyletic relationships derived from sequence homologies and three-dimensional structures, we find that the two lines of evolution are similar in general. This approach is applied to a group of glins here.

Evaluation of nutritive value and in vitro rumen fermentation gas accumulation of de-oiled algal residues

Background：Algae are widely recognized for their high oil content and for exponentially accumulating biomass with particular potential to provide single cell protein for human consumption or animal feed.It is believed that along with biodiesel from algae,the high protein de-oiled algal residue may become an alternative feed supplement option in the future.This study was conducted to investigate de-oiled algal residue obtained from the common Chlorella species,Thalassiosira weissflogii,Selenarstrum capricornutum,Scenedesmus sp.,and Scenedesmus dimorphus for assessment as potential feed supplements for ruminants by comparing with soybean（Glycine max） meal and alfalfa（Medicago sativa） hay.Results：With the exception of T.weissflogii,algal residue had higher concentrations of Cu,Zn,and Mn and lower concentration of Ca,Mg,and K than soybean meal and alfalfa hay.The algal residue CP（crude protein）concentrations ranged from 140 to 445 g/kg DM and varied among the de-oiled residues.In vitro rumen fermentation gas accumulation curves indicated that algal biomass degradation potential was less than that of soybean meal or alfalfa hay by up to 41.7%.The gas production curve,interpreted with a dual pool logistic model,confirmed that the fraction sizes for fast fermenting and slow fermenting of de-oiled algal residues were smaller than those in soybean meal and alfalfa hay,and the fermenting rate of the fractions was also low.Conclusions：Inferior in vitro rumen gas accumulation from the five de-oiled algal residues suggests that these algal byproducts are less degradable in the rumen.