Analysis of Pickled Cucumber Products, Based on Microbial Diversity and Flavor Substance Detection.

Changes to the microbial community during pickled cucumber fermentation were studied using the 16S rDNA technique. The changes of volatile organic compounds (VOCs) during pickled cucumber fermentation were studied by gas chromatograph-ion mobility spectrometry. At the phylum level, Cyanophyta and Proteobacteria were the dominant flora in the natural fermentation group, and Firmicutes were the dominant flora in the added-bacteria fermentation group. At the generic level, the addition of Lactobacillus led to changes in the community of the bacteria in the added-bacterial fermentation group and decreased the species abundance of other bacteria. In total, 75 volatile organic compounds were identified from naturally fermented pickled cucumber, and 60 volatile organic compounds were identified from fermented pickled cucumber with bacterial addition. The main metabolites were esters, aldehydes, acids, alcohols, ketones, alkanes, nitriles, and alkenes. These metabolites will bring their unique aroma components to the pickled cucumber. Metabolomic analysis of the O2PLS model showed that Weissella and Lactobacillus were closely and positively correlated with nine alcohols, six esters, five aldehydes, four acids, three ketones, and one pyrazine. Pseudomonas and norank_f_Mitochondria show a close positive correlation with four kinds of alcohols, two kinds of esters, one kind of aldehyde, and one kind of nitrile.

Response characteristics of flax retting liquid addition during chicken manure composting: Focusing on core bacteria in organic carbon mineralization and humification.

To explore the applicability of flax retting liquid (FRL) addition, the physicochemical properties, microbial community structure and function, carbon conversion and humus (HS) formation were assessed during chicken manure (CM) aerobic composting. Compared with the control group, the addition of FRL increased the temperature at thermophilic phase, while the microbial mass carbon content (MBC) in SCF and FRH groups raised to 96.1 ± 0.25 g/Kg and 93.33 ± 0.27 g/Kg, respectively. Similarly, FRL also improved the concent of humic acid (HA) to 38.44 ± 0.85 g/Kg, 33.06 ± 0.8 g/Kg, respcetively. However, fulvic acid (FA) decreased to 30.02 ± 0.55 g/Kg, 31.4 ± 0.43 g/Kg, respectively and further reduced CO2 emissions. FRL influenced the relative abundance of Firmicutes at thermophilic phase and Ornithinimicrobium at maturity phase. Additionally, FRL strengthen the association among flora and reduce the number of bacteria, which was negative correlated with HA and positive with CO2 during composting. These findings offer powerful technological support for improving agricultural waste recycling.

Analysis of Microbial Diversity and Metabolites in Sauerkraut Products with and without Microorganism Addition.

The microbial compositions and metabolites of fermented sauerkraut with and without the addition of microorganisms have been compared. The OTU clustering, nonvolatile compounds, volatile compounds and associations between bacterial taxa and metabolites were analyzed by 16S rRNA high-throughput sequencing technology, ultra performance liquid chromatography (UPLC), gas chromatography ion mobility mass spectrometry (GC-IMS) and the O2PLS model studies. The results showed that at the phylum level, the microbial species in the four sauerkraut types consisted mainly of the phyla Firmicutes and Proteobacteria, but different modes of microbial addition formed their own unique microbial communities. There were significant differences in the microbial communities among different northeast China sauerkraut samples, and different microbial communities exerted similar effects to inhibit Firmicutes production. At the genus level, sauerkraut without added microorganisms had the lowest microbial diversity. A total of 26 amino acids and 11 organic acids were identified and were more abundant in nonmicrobially fermented sauerkraut; 88 volatile organic compounds were identified in the 4 types of sauerkraut, with the microbially fermented sauerkraut being richer in alcohols, esters and acids. Different brands of sauerkraut contain their own unique flavor compounds. Cystine and tyrosine, ascorbic acid and acetic acid, and alcohols and esters are closely related to a wide range of microorganisms in sauerkraut. Elucidating the correlations among microbiota and metabolites will help guide future improvements in sauerkraut fermentation processes.

Aerobic composting with sauerkraut fermentation waste water: Increasing the stability and complexity of bacterial community and changing bacterial community assembly processes.

Aerobic composting renders the sauerkraut fermentation waste water harmless while adding soluble nutrients. Unravelling the bacterial community assembly processes, changes in community robustness and community cohesion and the relationship between them under composting treatment of sauerkraut fermentation waste water is an interesting topic. Sauerkraut fermentation waste water was used for composting, which increased bacterial linkages, community robustness, competitive behaviour during warming periods and cooperative behaviour during cooling periods, and the control of community assembly processes shifts from deterministic processes (variable selection) to stochastic processes (decentralised limitation). At the same time, the influence of community robustness and community cohesion on community assembly processes was increased. Community cohesion and robustness were significantly correlated with community function. These results indicate that community robustness and community cohesion are critical for the biological handling of hazardous substances.

Characterization of Dextran Biosynthesized by Glucansucrase from Leuconostoc pseudomesenteroides and Their Potential Biotechnological Applications.

Glucansucrase was purified from Leuconostoc pseudomesenteroides. The glucansucrase exhibited maximum activity at pH 5.5 and 30 °C. Ca2+ significantly promoted enzyme activity. An exopolysaccharide (EPS) was synthesized by this glucansucrase in vitro and purified. The molecular weight of the EPS was 3.083 × 106 Da. Fourier transform infrared (FT-IR) and nuclear magnetic resonance (NMR) spectroscopy showed that the main structure of glucan was 97.3% α-(1→6)-linked D-glucopyranose units, and α-(1→3) branched chain accounted for 2.7%. Scanning electron microscopy (SEM) observation of dextran showed that its surface was smooth and flaky. Atomic force microscopy (AFM) of dextran revealed a chain-like microstructure with many irregular protuberances in aqueous solution. The results showed that dextran had good thermal stability, water holding capacity, water solubility and emulsifying ability (EA), as well as good antioxidant activity; thus it has broad prospects for development in the fields of food, biomedicine, and medicine.

Aerobic composting of chicken manure with penicillin G: Community classification and quorum sensing mediating its contribution to humification.

Chicken manure containing antibiotics is a hazardous biological waste. The purpose of our study was to investigate how different concentrations of penicillin G alter the bacterial community to affect humification during aerobic composting of chicken manure. The effect of quorum sensing on the bacterial community was also evaluated. Penicillin G mainly affects low fold changes (within 4) for low-abundance (within 200) microbial genera. We found that the bacterial community cooperated to regulate humus and humic acid synthesis. The microbial genera that make up the bacterial community are different, but each bacterial community may have the same ecological function. Quorum sensing affects humic acid synthesis by regulating carbohydrate metabolism and amino acid metabolism in bacterial communities through mechanisms such as the pentose phosphate pathway and the shikimate pathway. This work presents an understanding of the impact of quorum sensing on the collaboration between bacterial communities during composting.

Analysis of the structure and properties of dextran produced by Weissella confusa.

An EPS produced by Weissella confusa H2 was purified through Sephadex G-100, and the preliminary structure characteristics and biological activities of H2 EPS were analyzed. Molecular mass of purified H2 EPS was 2.705 × 106 Da as measured with gel permeation chromatography (GPC). Composition of monosaccharides, nuclear magnetic resonance (NMR) spectroscopy spectroscopy and fourier transform infrared (FT-IR) showed that the EPS was a linear homopolysaccharide, mainly constituted of glucose and it is suggested that the EPS was dextran with α-(1 â†’ 6) glycosidic bonds and a few α-(1 â†’ 3) branches. Atomic force micrograph (AFM) and scanning electron microscopy (SEM) analysis of dextran further revealed sheets branched microstructure anchored with many irregular protuberances in aqueous solution. The XRD pattern reflected non-crystalline amorphous nature. In addition, the solubility, water-holding capacity, thermal property, rheological property and heavy metal chelating activity of the purified H2 dextran were determined. The dissolution percentage and water holding capacity of the dextran were 98.78 ± 1.37% and 426.03 ± 7.26%, respectively. The dextran exhibited good hydrophilicity, thermal stability and heavy metal chelating activity. Rheological studies exhibited rotational speed, pH, temperature, metal ions solutions dependent semiviscous nature. These results support its use as an additive in the food and environmental protection fields.

Optimization of cultivation strategy and medium for bacteriocin activity of Enterococcus faecium HDX-2.

A lactic acid bacteria (LAB) isolated from pickled Chinese cucumber was screened for bacteriocin production. The strain was identified to be Enterococcus faecium HDX-2. Based on the Plackett-Burman (PB) experiment, glucose, Ca2+, and initial pH were found to be the most significant parameters of bacteriocin production. Afterward, effects of the three main parameters on bacteriocin activity were further investigated by central composite design (CCD) and the optimum composition was glucose 22 g/L, Ca2+ 0.6 mM, and initial pH 7.2. Optimum results showed that bacteriocin activity was increased to 1337.60 ± 36.71 AU/mL, 2.23-fold higher than in MRS medium without parameters optimization. The bacteriocin also showed significant antimicrobial activity against Listeria monocytogenes in meat and cheese model system.

Detection of the microbial diversity and flavour components of northeastern Chinese soybean paste during storage.

A combination of 16S rDNA and GC-IMS was used to study the changes in the composition of microorganisms and volatile organic compounds (VOCs) during the storage of northeastern Chinese soybean paste. Firmicutes and Actinobacteriota dominated the microbial communities of the soybean paste at the phylum level, bacterial profiles of different samples were different at genus level. Fifty-one VOCs were identified from soybean paste, most of which existed in the early storage stage. Most esters and alcohols decreased with the extension of the storage time, while acids and pyrazines accumulated in the later period of storage. Esters, alcohols, acids and aldehyde compounds are the key substances in the volatile components of soybean paste, which give the soybean paste the sour, sweet, rose, mushroom and smoky flavor characteristics. The biomarker Bacillus-velezensis in soybean paste is directly related to ester features; Kroppenstedtia, Sporolactobacillus-nakayamae, and Corynebacterium-stationis are positively associated with the biosynthesis of aldehydes.

Purification and characterization of an exopolysaccharide from Leuconostoc lactis L2.

An exopolysaccharide (EPS)-producing bacterium was isolated from the homemade fermentation product of hami melon (Cucumis melo var. saccharinus) and identified as Leuconostoc lactis L2 mainly based on 16S rDNA sequence analysis and the API 50 CHL test. The monosaccharide composition of Leu. lactis L2 EPS was determined to be glucose by gas chromatography (GC), and its molecular weight was 3.7 × 103 kDa by high-performance size exclusion chromatography (HPSEC). The EPS microscopic characteristics were compact, branched and tubular under scanning electron microscopy (SEM). Atomic force microscopy (AFM) showed a smooth and consistent polymeric matrix. The EPS was a linear polymer composed of α-(1 → 6)-linked D-pyranose residues, based on the Fourier transform infrared (FT-IR), 1H, 13C nuclear magnetic resonance (NMR) and heteronuclear single quantum coherence (HSQC) spectra. This EPS exhibited a high water solubility index (WSI) and water holding capacity (WHC) (91.90 ±â€¯2.45% and 509.45 ±â€¯28.59%, respectively), as well as good emulsifying properties with many organic solvents. These results collectively suggest the applicable potential of this EPS in the food, feed and pharmaceutical fields.

Monoraphidium sp. HDMA-20 is a new potential source of α-linolenic acid and eicosatetraenoic acid.

BACKGROUND: ω-3 polyunsaturated fatty acids (PUFAs) are synthesized from α-Linolenic acid (ALA, C18:3ω3) and play important roles in anti-inflammatory and antioxidant responses in mammal cells. ALA is an essential fatty acid which cannot be produced within the human body and must be acquired through diet. The purpose of this study was to evaluate the potential of a novel microalgal strain (HDMA-20) as a source of ω-3 PUFAs including ALA and eicosatetraenoic acid (ETA, C20:4ω3). METHOD: Phylogenetic Neighbor-Joining analysis based on 18S ribosomal DNA sequence was used to identify the microalga strain HDMA-20. Autotrophic condition was chosen to cultivate HDMA-20 to reduce the cultivation cost. GC-MS was used to determine the fatty acid composition of HDMA-20 lipid. RESULTS: A microalgal strain (HDMA-20) from Lake Chengfeng (Daqing, Heilongjiang province, China) was found to accumulate high content of ω-3 PUFAs (63.4% of total lipid), with ALA and eicosatetraenoic acid (ETA, C20:4ω3) accounting for 35.4 and 9.6% of total lipid, respectively. Phylogenetic analysis based on 18S ribosomal DNA sequences suggested that the HDMA-20 belonged to genus Monoraphidium (Selenastraceae, Sphaeropleales) and its 18S rDNA sequence information turned out to be new molecular record of Monoraphidium species. The biomass productivity and lipid content of HDMA-20 were also investigated under autotrophic condition. The biomass productivity of HDMA-20 reached 36.3 mg L- 1 day- 1, and the lipid contents was 22.6% of dry weight. CONCLUSION: HDMA-20 not only represent an additional source of ALA, but also a totally new source of ETA. The high content of ω-3 PUFAs, especially ALA, of HDMA-20, makes it suitable as a source of nutrition supplements for human health. In addition, HDMA-20 exhibited good properties in growth and lipid accumulation, implying its potential for cost-effective ω-3 PUFAs production in future.

Isolation of a novel strain of Monoraphidium sp. and characterization of its potential for α-linolenic acid and biodiesel production.

α-Linolenic acid (ALA) is an essential fatty acid which cannot be synthesized de novo in mammals and must be ingested regularly in the diet. In this study, a microalgal strain named HDMA-11 was isolated from Lake Ming, China, and was found to accumulate a high ALA content (39.2% of total lipids). Phylogenetic neighbor-joining analysis indicated that HDMA-11 belongs to the genus Monoraphidium (Selenastraceae, Sphaeropleales) and its 18S ribosomal DNA sequence seemed to be a new molecular record of a Monoraphidium species. The fatty acid profiles, biomass productivity and lipid content of HDMA-11 were also investigated in autotrophic conditions. The high levels of polyunsaturated fatty acids in HDMA-11, especially ALA, make it suitable as a source of nutritional supplementation for human health. Furthermore, HDMA-11 exhibited good properties for biodiesel production, characterized by high lipid content (28.5% of dry weight), moderate biomass productivity (31.5 mg L-1 day-1) and a promising lipid profile.

[Optimization of xylose fermentation for ethanol production by Candida shehatae HDYXHT-01].

Plackett-Burman (PB) design and central composite design (CCD) were applied to optimize of xylose fermentation for ethanol production by Candida shehatae HDYXHT-01. The PB results showed that (NH4)2SO4, KH2PO4, yeast extract and inoculum volume were the main affecting factors. With ethanol productivity as the target response, the optimal fermentation was determined by CCD and response surface analysis (RSM). The optimal fermentation conditions were (NH4)2SO4 1.73 g/L, KH2PO4 3.56 g/L, yeast extract 2.62 g/L and inoculum volume 5.66%. Other fermentation conditions were xylose 80 g/L, MgSO47H20 0.1 g/L, pH 5.0 and 250 mL flask containing 100 mL medium and cultivated at 30 degrees C for 48 h and the agitation speed was 140 r/min. Under this fermentation conditions, ethanol productivity was 26.18 g/L, which was 1.15 times of the initial.

[Fermentation optimization by response surface methodology for enhanced production of beta-glucosidase of Aspergillus niger HDF05].

In order to obtain high beta-glucosidase productivity, we optimized the fermentation parameters for beta-glucosidase production by Aspergillus niger HDF05. First, we screened the important parameters by Plackeet-Burman design. Second, we used the path of steepest ascent to approach to the biggest response region of parameters affecting beta-glucosidase production. Then, we obtained the optimal parameters by central composite design and response surface analysis. We developed a quadratic polynomial equation for predicting beta-glucosidase production level. The results showed that the important parameters were temperature, packing volume, concentrations of wheat bran and (NH4)2SO4. The optimal fermentation parameters were as follows: temperature 28 degrees C, packing volume 71.4 mL/250 mL, wheat bran 36 g/L and (NH4)2SO4 5.5 g/L. Under the optimal conditions, we obtained the maximum enzyme activity of 60.06 U/mL, with an increase of 23.9% compared to the original fermentation parameters. During enzymatic hydrolysis of acid-pretreated corncob, addition of beta-glucosidase from Aspergillus niger HDF05 greatly reduced the inhibition caused by cellobiose, and the hydrolysis yield was improved from 66.7% to 80.4%.

Statistical optimization of xylitol production from corncob hemicellulose hydrolysate by Candida tropicalis HDY-02.

The statistical experimental designs were adopted to optimize the culture medium in xylitol production by Candida tropicalis HDY-02 with corncob hemicellulose hydrolysate as substrate. In the first step, Plackett-Burman design was used for screening the important variables. KH(2)PO(4), yeast extract, (NH(4))(2)SO(4) and MgSO(4)·7H(2)O were found to significantly affect xylitol yield. In the second step, central composite design (CCD) was used to determine the optimum level of each of the significant variables. A second-order polynomial was determined by the multiple regression analysis of the experimental data. The interactive effects of yeast extract and MgSO(4)·7H(2)O on xylitol yield of C. tropicalis HDY-02 were determined to be significant. The validation experimental was consistent with the prediction model. The optimum combinations for xylitol yield were 5 gl(-1) (NH(4))(2)SO(4), 1.3 gl(-1) KH(2)PO(4), 4.6 gl(-1) yeast extract and 0.6 gl(-1) MgSO(4)·7H(2)O. Under these optimal conditions, the continuous fed-batch experiments could produce xylitol of 58 gl(-1) with a yield of 0.73 g g(-1) xylose.

[Construction of integrative vector for xylulokinase gene and its overexpression in Saccharomyces cerevisiae].

OBJECTIVE: An integrative vector of Saccharomyces cerevisiae for xylulokinase gene expression was constructed to overexpress xylulokinase activity. METHODS: On the basis of plasmid p406ADH1, 4 components were integrated, which were KanR gene as G418 resistant marker, ADH1 terminator fragment, xylulokinase gene from Saccharomyces cerevisiae W5 and 18S rDNA sequence for homologous recombination. After enzyme digestion and ligation, high copy recombinant expression vector pCXS-RKTr was constructed. pCXS-RKTr was linearized and transferred into Saccharomyces cerevisiae W5 , then xylulokinase activity was detected to determine the expression of pCXS-RKTr. RESULTS: Xylulokinase gene located on pCXS-RKTr was highly expressed in W5. The xylulokinase activity was 2. 87 times of the original strain. CONCLUSION: An integrative vector of industry strain Saccharomyces cerevisiae is successfully constructed and xylulokinase gene of Saccharomyces cerevisiae itself was over expressed by this vector. This intergrative vector can efficiently raise the xylulokinase activity of Saccharomyces cerevisiae. This system laid a foundation for the construction of gene engineering Saccharomyces cerevisiae strain which can ferment xylose to ethanol.

Paracin 1.7, a bacteriocin produced by Lactobacillus paracasei HD1.7 isolated from Chinese cabbage sauerkraut, a traditional Chinese fermented vegetable food.

OBJECTS: To purify and characterize bacteriocin produced by L. paracasei HD1.7. METHODS: Paracin 1.7 was purified by chromatography and its molecular weight was measured by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Antibacterial activity was measured by using the agar-well diffusion method. RESULTS: The bacterial strain for the fermentation was identified as Lactobacillus subsp. paracasei. Paracin 1.7 had the activity of inhibiting the growth of other bacteria. Maximum production of Paracin 1.7 was in the stationary phase. Paracin 1.7 can be well purified with Cation exchange chromatography, Sephedex (G50, G25, G10) gel chromatography and high performance liquid chromatography (HPLC) on C18 column and its determined molecular weight was about 11 kDa by Tricine-SDS-PAGE. Paracin 1.7 shows a broad spectrum of activities against various strains in the genera of Proteus, Bacillus, Enterobacter, Staphylococcus, Escherichia, Lactobacillus, Microccus, Pseudomonas, Salmonella and Saccharomyces, some of which belong to food borne pathogenic bacteria. Although Paracin 1.7 displayed stability toward heat and acidic pH, it was sensitive to several proteolytic enzymes. The inhibitory activities remain well after stored at 4 degrees C for 4 months; the inhibitory activity declined only 4.19%. CONCLUSION: Paracin 1.7 can be a potential food preservative on the basis of its antibacterial characters.

Sugarcane bagasse mild alkaline/oxidative pretreatment for ethanol production by alkaline recycle process.

In order to decrease the alkali and water consumptions in the sugarcane bagasse alkaline/oxidative pretreatment for ethanol production, an alkaline recycle process was carried out. Two recycles of NaOH/H2O2 pretreatment did not decrease the pretreatment and enzymatic hydrolysis efficiencies and the consumptions of NaOH and water would be saved by 26% and 40%, respectively. A simultaneous saccharification and fermentation (SSF) culture with pretreated bagasse as substrate was developed giving 25 g ethanol l(-1) with a yield of 0.2 g g(-1) bagasse and productivity of 0.52 g l(-1) h(-1).

Fermentation of pretreated sugarcane bagasse hemicellulose hydrolysate to ethanol by Pachysolen tannophilus.

Sugarcane bagasse hemicellulose hydrolysates, pretreated by either over-liming or electrodialysis and, supplemented with nutrient materials, were fermented to ethanol using Pachysolen tannophilus DW06. Compared with detoxification by over-liming, detoxification by electrodialysis decreased the loss of sugar and increased the acetic acid removal, leading to better fermentability. A batch culture with electrodialytically pretreated hydrolysate as substrate was developed giving 21 g ethanol l(-1) with a yield of 0.35 g g(-1) sugar and productivity of 0.59 g l(-1) h(-1).