2-Keto-D-gluconic acid and prodigiosin producing by a Serratia marcescens.

Microbial fermentation has become the main method to produce target compound. In this study, a 2-Keto-D-gluconic acid (2-KGA) producing mutant strain was obtained by mutation with rational screening methods. Meanwhile, prodigiosin was produced when the nitrogen source in the medium was changed to peptone and its fermentation conditions were evaluated to achieve high-efficient accumulation. The mutant strain SDSPY-136 was firstly identified as Serratia marcescens by morphological observation and 16S rDNA sequencing. The 2-KGA synthetic capacity of S. marcescens SDSPY-136 was evaluated by shake fermentation with 110 g/L glucose as substrates. For fermentation, 2-KGA yield, conversation rate and purity of SDSPY-136 reached 104.60 g/L, 95.10%, 99.11% in 72 h. The red pigment was extracted from the fermentation broth using acidic methanol and identified as prodigiosin by FT-IR. The optimal conditions were as follows: glycerol 20 g/L, peptone 20 g/L, MgSO415 g/L, pH 6.0, a 2% (v/v) inoculum, 30 °C and 200 rpm of shaking culture. Eventually, prodigiosin reached a yield of 9.89 g/Lafter shake culturing for 50 h under this condition. The mutant S. marcescens SDSPY-136 was shown to be promising for 2-KGA and prodigiosin production and a suitable object for prodigiosin metabolism research of S. marcescens.

Increased Phenolic Content and Enhanced Antioxidant Activity in Fermented Glutinous Rice Supplemented with Fu Brick Tea.

Glutinous rice-based foods have a long history are consumed worldwide. They are also in great demand for the pursuit of novel sensory and natural health benefits. In this study, we developed a novel fermented glutinous rice product with the supplementation of Fu brick tea. Using in vitro antioxidant evaluation and phenolic compounds analysis, fermentation with Fu brick tea increased the total phenolic content and enhanced the antioxidant activity of glutinous rice, including scavenging of 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) radical, 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) radical, and hydroxyl radical, ferric-reducing antioxidant power, and ferric ion reducing power and iron chelating capability. Besides, compared with traditional fermented glutinous rice, this novel functional food exhibited a stronger activity for protecting DNA against hydroxyl radical-induced oxidation damage. Quantitative analysis by HPLC identified 14 compounds covering catechins and phenolic acids, which were considered to be positively related to the enhanced antioxidant capability. Furthermore, we found that 80% ethanol was a suitable extract solvent compared with water, because of its higher extraction efficiency and stronger functional activities. Our results suggested that this novel fermented glutinous rice could serve as a nutraceutical food/ingredient with special sensory and functional activities.

Chemical characterization and evaluation of the antioxidants in Chaenomeles fruits by an improved HPLC-TOF/MS coupled to an on-line DPPH-HPLC method.

An improved method based on HPLC-TOF/MS was developed to catalog the antioxidants in five species of Chaenomeles (Mugua). Forty-four fractions from the Mugua extracts show appreciable levels of antioxidative activity in scavenging the stable free-radical 2,2-diphenyl-1-picrylhydrazyl and the hydroxyl radicals. Twelve major antioxidant's chemical structures are identified. Antioxidant activities differ between species, but intra-species level of antioxidants, regardless of their ripeness, are similar. C. sinensis has the highest antioxidant level. A rigorous quality control procedure was implemented to ensure accuracy of antioxidant quantification. This improved procedure can be used for rapid discovery of antioxidants in other plant extracts.

Exploring the Interactions of Soybean 7S Globulin with Gallic Acid, Chlorogenic Acid and (-)-Epigallocatechin Gallate.

In this study, the noncovalent interaction mechanisms between soybean 7S globulin and three polyphenols (gallic acid (GA), chlorogenic acid (CA) and (-)-epigallocatechin gallate (EGCG)) were explored and compared using various techniques. Fluorescence experiments showed that GA and EGCG had strong static quenching effects on 7S fluorescence, and that of CA was the result of multiple mechanisms. The interactions caused changes to the secondary and tertiary structure of 7S, and the surface hydrophobicity was decreased. Thermodynamic experiments showed that the combinations of polyphenols with 7S were exothermic processes. Hydrogen bonds and van der Waals forces were the primary driving forces promoting the binding of EGCG and CA to 7S. The combination of GA was mainly affected by electrostatic interaction. The results showed that the structure and molecular weight of polyphenols play an important role in their interactions. This work is helpful for developing products containing polyphenols and soybean protein.

A Multi-Scale Approach to Investigate Adhesion Properties of Pseudomonas aeruginosa PAO1 to Geotrichum candidum LG-8, a Potential Probiotic Yeast.

This study investigated properties of Pseudomonas aeruginosa PAO1 adhesion to Geotrichum candidum LG-8 cells in variable pH and salt conditions. The primary mechanism was revealed by multi-scale microscopy technics. The adhesion of PAO1 to the living fungus occurred within 1 h and was limited at concentrations of bile salts higher than 0.5%. The adhesion efficiency gradually increased to 58.1% with the pH increasing from 2.0 to 7.0 and then decreased to 48.2% at pH 9.0. However, the dead LG-8 has an advantage over the living ones to adhere PAO1 in same pH and bile salt conditions. Optical microscopy showed that both unsterilized and sterilized G. candidum LG-8 cells removed approximately one hundred fold bacteria in 4 h. Laser scanning confocal microscopy (LSCM) analysis indicated that polysaccharides of the fungus contributed to adhesion. Scanning electron microscopy (SEM) analysis proved that syrup-like EPS (extracellular polymeric substances) of LG-8 coating PAO1 was in part a mechanism. Atomic force microscopy (AFM) showed roughness of the LG-8 surface changed in the adhesion process. Furthermore, a pedestal-like structure of bacteria was observed by transmission electron microscopy (TEM) analysis, indicating that the bacteria were also actively involved in the adhesion process. G. candidum LG-8 is a potential candidate for the control of P. aeruginosa PAO1 in the food industry and immunodeficiency patients.

Effect of non-covalent and covalent complexation of (-)-epigallocatechin gallate with soybean protein isolate on protein structure and in vitro digestion characteristics.

The present study was aimed to investigate the effects of non-covalent and covalent interactions between soy protein isolate (SPI) and different concentrations (1, 2 and 5 mM) of (-)-epigallocatechin gallate (EGCG) regarding the structural and functional properties of the complex. The combination with EGCG caused changes in the secondary structure of SPI. The covalent complexes formed at low concentrations of EGCG tended to form a network structure. Compared with the SPI-EGCG non-covalent complexes, the covalent complexes exhibited higher thermal stability and oxidation resistance and a polyphenol-protective effect. In addition, the corresponding anti-digestive ability of the covalent complexes was strong and would therefore be more stable in the intestinal tract. The findings of this study provide a theoretical reference and research basis for the use of different SPI-polyphenol complexes as functional food ingredients or as bioactive materials.

Soy protein isolate -(-)-epigallocatechin gallate conjugate: Covalent binding sites identification and IgE binding ability evaluation.

The conjugate prepared from (-)-epigallocatechin gallate (EGCG) and soy protein isolate (SPI) under alkaline and aerobic conditions was analyzed using a Nano-LC-Q-Orbitrap-MS/MS technique. The sulfhydryl and free amino groups of SPI were involved in covalent binding. Fifty-one peptides were conjugated with EGCG. Fifty-nine modified sites were identified, located on Cys, His, Arg, and Lys, respectively. It is the first time to confirm that each of the two phenolic rings of EGCG contained a reactive site that bound to an amino acid residue. The amino acid residue reactivity, amino acid sequence and composition affected the EGCG binding site in SPI. Lys and Arg residues are the most likely sites for modification, and modification appears to reduce IgE binding. This study is helpful to elucidate the pattern of covalent binding of polyphenols to proteins in food systems and provides a theoretical basis for the directional modification of soy proteins with polyphenols.

Multistarter fermentation of glutinous rice with Fu brick tea: Effects on microbial, chemical, and volatile compositions.

A higher fermentation efficiency was achieved, using multistarter fermentation of glutinous rice supplemented with Fu brick tea (FGR-FBT), than when using traditional fermentation. The effects of multistarter fermentation on the microbial, chemical, and volatile compositions were determined. When FBT was incorporated during glutinous rice fermentation, increased population of yeasts and fungi, as well as enhanced α-amylase, proteinase and ß-glucosidase activities, were observed. Specific fungi were isolated and identified as Aspergillus spp., which are known to secrete extracellular enzymes that modify the chemical properties, including ethanol levels, pH, total acids, and total soluble solids. The aroma profile of fermented glutinous rice was studied in the absence and presence of FBT, using HS-SPME-GC-MS and the electronic-nose. This analysis indicated that 35 characteristic volatile compounds were only found in FGR-FBT. The results show that FBT can be added during the fermentation of food products to enhance microbial biotransformation and modify flavour metabolism.

An improved 2D-HPLC-UF-ESI-TOF/MS approach for enrichment and comprehensive characterization of minor neuraminidase inhibitors from Flos Lonicerae Japonicae.

Flos Lonicerae Japonicae(Jinyinhua) possesses clearing heat and detoxification activity, and has been used as a traditional Chinese medicine to treat influenza for many years. Due to the complex chemical composition and diverse content of Jinyinhua, especially the many trace ingredients, the effective components are unknown. In this study, an improved two-dimensional high performance liquid chromatography-ultrafiltration combined with electrospray ionization-time-of-flight/mass spectroscopy (ESI-TOF/MS) approach was designed and used for the enrichment, screening and characterization of minor neuraminidase inhibitors in Jinyinhua. In the first dimension, semi-prep-HPLC was employed for the preliminary separation of different polarity components and enrichment of low content components from Jinyinhua extract. In the second dimension, hydrophilic interaction liquid chromatography and reverse phase-HPLC were used to separate the different polar fractions, respectively. The fractions then underwent ultrafiltration and ESI-TOF/MS for the comprehensive screening and characterization of potential neuraminidase inhibitors. As a result, a total of 44 compounds were found to have neuraminidase inhibitory activity, and 22 of these compounds were preliminarily identified by accurate molecular weight and UV absorption data compared with standards and references. The activity of 16 of these compounds was verified by the neuraminidase inhibition assay. This study provides support for the rapid screening of minor neuraminidase inhibitors from complex natural medicines.

On-line screening and identification of free radical scavenging compounds in Angelica dahurica fermented with Eurotium cristatum using an HPLC-PDA-Triple-TOF-MS/MS-ABTS system.

Eurotium cristatum, a beneficial fungus isolated from Fuzhuan tea, was used to ferment Angelica dahurica for the first time. The antioxidant capacities of the extracts before and after fermentation were compared using ABTS, DPPH and FRAP assays. The results showed that the antioxidant capacities of the extracts acquired using organic solvents were greater after fermentation. Moreover, based on a comparison of the HPLC chromatograms, the chemical composition of Angelica dahurica changed substantially during fermentation. To further understand the changes in its antioxidant constituents, an on-line HPLC-PDA-Triple-TOF-MS/MS-ABTS system was employed. Twelve antioxidants belonging to three different classes were detected and identified, and their antioxidant capacities were preliminarily evaluated. The results indicated that the substances produced during the fermentation of Eurotium cristatum played important roles in enhancing the antioxidant capacity.

An in vitro AChE inhibition assay combined with UF-HPLC-ESI-Q-TOF/MS approach for screening and characterizing of AChE inhibitors from roots of Coptis chinensis Franch.

In this study, an in vitro acetylcholinesterase (AChE) inhibition assay based on microplate reader combined with ultrafiltration high performance liquid chromatography-electrospray quadrupole time of flight mass (UF-HPLC-ESI-Q-TOF/MS) was developed for the rapid screening and identification of acetylcholinesterase inhibitors (AChEI) from roots of Coptis chinensis Franch. Incubation conditions such as enzyme concentration, incubation time, incubation temperature and co-solvent was optimized so as to get better screening results. Five alkaloids including columbamine, jatrorrhizine, coptisine, palmatine and berberine were found with AChE inhibition activity in the 80% ethanol extract of C. chinensis Franch. The screened compounds were identified by HPLC-DAD-ESI-Q-TOF/MS compared with the reference stands and literatures. The screened results were verified by in vitro AChE inhibition assays, palmatine showed the best AChE inhibitory activities with IC50 values of 36.6µM among the five compounds. Results of the present study indicated that the combinative method using in vitro AChE inhibition assay and UF-HPLC-ESI-Q-TOF/MS could be widely applied for rapid screening and identification of AChEI from complex TCM extract.