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Figure 1 | BMC Biotechnology

Figure 1

From: A new chitinase-like xylanase inhibitor protein (XIP) from coffee (Coffea arabica) affects Soybean Asian rust (Phakopsora pachyrhizi) spore germination

Figure 1

Evaluation of chitinolytic activity of a new protein from Coffea arabica (CaclXIP) after heterologous expression. (A) SDS-PAGE (12.5% acrylamide gel) of the different fractions obtained from CaclXIP expression. 1. Proteins from the culture supernatant inoculated with P. pastoris bearing an empty vector; 2. Proteins from the culture supernatant inoculated with P. pastoris bearing pGAPZα-B/caclxip; 3. CaclXIP eluted from ion exchange chromatography with 350 mM NaCl. Proteins were precipitated from 200 μL of each fraction with 4 volumes of cold acetone, solubilized in 50 μL of sample buffer and 10 μL were load on gel. A protein with molecular weight around 32 kDa was purified by ion exchange chromatography. The gel was stained with Coomassie Blue. (B) Chitinolytic assay. Each sample from (A) was dialyzed against reaction buffer (80 mM sodium phosphate buffer pH 6.8) and five micrograms of each sample were used per assay. Numbers 1, 2 and 3 are as described in (A). The same assay was performed at pH 4.8, 5.8, 7.8, and 8.8, without any significant variation.

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