Xin Lü

Improvement of simultaneous determination of neutral monosaccharides and uronic acids by gas chromatography

Although pre-column derivatization with n-propylamine and acetic anhydride combined with the gas chromatograph was a useful method for the analysis of monosaccharide composition, failure often occurs because of lack of detail information on the mechanism as well as the operating key point in derivatization process. In this study, the key points in the derivatization (lactonization time, the amount of n-propylamine and acetic anhydride) were investigated and optimized to improve the method. Under the optimal conditions, the derivatives of seven neutral monosaccharides and two uronic acids were simultaneouly obtained, after which they were well separated and detected by GC. It was also found that all derivatives of monosaccharides were stable even stored for 20days. The linearity, sensitivity, precision, reproducibility and recovery rate of the improved method were evaluated. Thereafter, five polysaccharide samples from different sources were analyzed to validate the improved method.

Purification and characterization of a novel bacteriocin produced by Lactobacillus crustorum MN047 isolated from koumiss from Xinjiang, China

The growing emergence of antibiotic-resistant bacteria in the food industry needs to be controlled with effective antimicrobials. In this study, bacteriocin MN047 A (BMA) was found to have antibacterial activity against multidrug-resistant bacteria. It was produced by Lactobacillus crustorum MN047, which was first isolated from koumiss, a traditional fermented dairy product from Xinjiang Autonomous Region, China. It was purified by ammonium sulfate precipitation, ion-exchange chromatography, and reversed-phase chromatography. It had a low molecular mass of 1,770.89 Da according to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and the sequence was identified as QLPWQILGIVAGMFQA by liquid chromatography-tandem mass spectrometry analysis and MASCOT searching. It was proteinaceous in nature: the bacteriocin was digested by protease but not by α-amylase or lipase. It showed broad pH toleration (pH 2-11), good thermostability, and good storage stability. It had a broad inhibitory spectrum, including both gram-positive and gram-negative bacteria. Growth curve and time-kill kinetics indicated that it was bactericidal to the indicator strains, and this finding was verified by scanning electron microscope and transmission electron microscope after treatment with BMA. As well, BMA halted the growth of Staphylococcus aureus and Escherichia coli in the G1 and G2/M phases according to cell-cycle analysis by flow cytometry, indicating that BMA had comprehensive inhibitory effects against foodborne pathogens.

Isolation of nitrite-degrading strains from Douchi and their application to degrade high nitrite in Jiangshui : Isolation of nitrite-degrading strains from Douchi

BACKGROUND Excessive nitrite in food is potentially harmful to human health because of carcinogenic effects caused by its nitroso-derivatives. Douchi, which widely distributed throughout the country, is a traditional solid fermented soybean food with low nitrite content. RESULTS In this study, bacteria which can degrade nitrite were isolated from Douchi and identified from their 16S rDNA sequences. Acinetobacter guillouiae, Acinetobacter bereziniae, Bacillus subtilis, Bacillus tequilensis, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus aryabhattai and Bacillus methylotrophicus were selected. It was shown that all strains were able to degrade nitrite to some extent, including Bacillus subtilis NDS1, which was able to degrade 99.41% of nitrite. The enzyme activities of these strains were determined at 24 and 48 h and were shown to correspond with their nitrite degradation rates. The strains were used to inoculate Jiangshui, a kind of traditional fermented vegetable from northwest China that often has a high nitrite content. Of the strains tested, Bacillus subtilis NDS1, Bacillus tequilensis NDS3, Acinetobacter bereziniae NDS4, Bacillus subtilis NDS6, and Bacillus subtilis NDS12 were able to degrade nitrite in Jiangshui more rapidly, with Acinetobacter bereziniae NDS4 degrading almost all nitrite in 48 h compared with 180 h of control. CONCLUSION These results indicate that the selected strains have potential to be used as nitrite-degrading agents in food.

Biodegradation of lignin by Pseudomonas sp. Q18 and the characterization of a novel bacterial DyP-type peroxidase

Lignin valorization can be obtained through cleavage of selected bonds by microbial enzymes, in which lignin is segregated from cellulose and hemicellulose and abundant phenolic compounds can be provided. In this study, Pseudomonas sp. Q18, previously isolated from rotten wood in China, was used to degrade alkali lignin and raw lignocellulosic material. Gel-permeation chromatography, field-emission scanning electron microscope, and GC–MS were combined to investigate the degradation process. The GC–MS results revealed that the quantities of aromatic compounds with phenol ring from lignin increased significantly after incubation with Pseudomonas sp. Q18, which indicated the degradation of lignin. According to the lignin-derived metabolite analysis, it was proposed that a DyP-type peroxidase (PmDyP) might exist in strain Q18. Thereafter, the gene of PmDyP was cloned and expressed, after which the recombinant PmDyP was purified and the enzymatic kinetics of PmDyP were assayed. According to results, PmDyP showed promising characteristics for lignocellulosic biodegradation in biorefinery.