Isolation Of Lectin-like Protein From Lactobacillus And The Study Of Characteristics

One strain lactobacillus PCL was isolated from sauerkraut juice by usingMRS media. The lactic acid paper chromatogram showed that the strain can producelactic acid. This bacteria was Gram positive, short bacilliform. 16S rDNAhomology between the strain and Lactobacillus plantarum strain p215 was 99ï¼….Rabbit erythrocyte agglutination test of its fermentation supernatant showedthat the strain can excrete Jectin-like substance into MRS media. After cen-trifuged Lactobacillus plantarum PCL fermentation broth, concentrated sup-ernatant and removed lactic acid, separated and purified protein by gelchromatography and ion exchange chromatography, the lectin-like protein wasobtained, a single band appeared in the Urea-SDS-Polyacrylamide gel electroph-oresis with the molecular weight about 85kDa.Some characters of L.plantarum PCL lectin-like protein have been studiedresults were as follows:1. L. plantarum PCL lectin-like protein appeared a rabbit erythrocyteagglutination activity of 2⁸ when its concentration was 0.068mg/mL and thesmallest agglutination concentration of 0.265μg/mL.2. The effect of temperature on agglutinating activity of L. plantarum PCLlectin-like protein was little, after 30 minutes water bath from 30℃toboiling water, the agglutinating activity was invariable.3. Several momosaccharide did not inhibit agglutinating activity ofL. plantarum PCL lectin-like protein, such as D(+)-Glucose, D(+)-Galactose,L-(+)-Ara-binose, D-Xylose, D-Sorbitol, L-(-)-Sorbose, D(+)-Mannose,α-L-Rhamnose Monohydrate, D-(-)-Fructose, but Dextrose(Anhydrous) andD-(-)-Arabinose could promote agglutinating activity in a certain extent.4. The effect of pH on agglutinating activity of L. plantarum PCL lectin-likeprotein varied along with different buffer solutions, acidic condition ofdibasic sodium phosphate-citrate buffer solution showed a little inhibit-tion, alkaline condition of Na₂HPO₄-KH₂PO₄(1/15mol/L) buffer solution presented a wake promotion, Na₂CO₃-NaHCO₃(0.1mol/L) buffer solution inalkaline range did not affect L.plantarum lectin-like protein.5. The four metal ions of Mn²⁺,Ca²⁺,Mg²⁺,K⁺ did not inhibit the agglutinatingactivity of L. plantarw PCL lectin-like protein.6. 10ï¼…(V/V) solution of methanol, ethanol, acetone, n-hexane, benzene methanol,ethyl acetate, isopropyl alcohol, glycol and 1ï¼…(V/V) solution ofβ-mercap-toethanol did not affect agglutinating activity of L.plantarum PCL lectin-like protein, butanol and isoamyl alcohol show some promotion to someextent.7. The fluoresceence spectra characteristic fluoresceence peak of L.plantarumPCL lectin-like protein under the excitation wavelength of 308mm was nearthe peak at 380hm, and fluoresceence intensity was larger. Therefore L.pl-antarum PCL lectin-like protein was rich in tyrptophan residues and mayshare Collagen-like special structure.8. Contrasting to several tectins, L.plantarum PCL lectin-like protein showedhigh temperature resistance, high agglutinating activity, a wide range ofpH adaptability, Ca²⁺, Mg²⁺ and several monosaccharide did not inhibit itsagglutinating activity.9. Inhibition of L. plantarum PCL lectin-like protein to HIV in vitro wasdetected by using MTT method, the result showed that EC₅₀ on HIV-1_â…¢B is0.079μg/mL and TI 28.34.10. By comparing with other lectins, the anti-HIV activity of L.plantarum PCLlectin-like protein was close to CA and EHA, and they were significantlyhigher than those in several other lectins. So L. plantarum PCL lectin-likeprotein presented a better inhibition on HIV, it is worthy of being furtherstudied.