Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 25
Filter
Add more filters










Publication year range
1.
J Fungi (Basel) ; 10(3)2024 Mar 20.
Article in English | MEDLINE | ID: mdl-38535236

ABSTRACT

Nucleotide substitutions have played an important role in molecular evolution, and understanding their dynamics would contribute to genetic studies. Related research with defined DNA sequences lasted for decades until whole-genome sequencing arose. UV radiation (UVR) can generate base changes and other genetic variations in a short period of time, so it would be more meaningful to explore mutations caused by UVR from a genomic perspective. The monokaryon enoki strain WT583 was selected as the experimental material in this study because it can spontaneously produce large amounts of oidia on PDA plates, and the monokaryons originating from oidia have the same genotype as their mother monokaryon. After exposure to UV radiation, 100 randomly selected mutants, with WT583 as the reference genome, were sent for genome sequencing. BWA, samtools, and GATK software were employed for SNP calling, and the R package CMplot was used to visualize the distribution of the SNPs on the contigs of the reference genome. Furthermore, a k-mer-based method was used to detect DNA fragment deletion. Moreover, the non-synonymous genes were functionally annotated. A total of 3707 single-base substitutions and 228 tandem mutations were analyzed. The immediate adjacent bases showed different effects on the mutation frequencies of adenine and cytosine. For adenine, the overall effects of the immediate 5'-side and 3'-side bases were T > A > C > G and A > T > G > C, respectively; for cytosine, the overall effects of the immediate 5'-side and 3'-side bases were T > C > A > G and C > T > A > G, respectively. Regarding tandem mutations, the mutation frequencies of double-transition, double-transversion, 3'-side transition, and 5'-side transition were 131, 8, 72, and 17, respectively. Transitions at the 3'-side with a high mutation frequency shared a common feature, where they held transversions at the 5'-side of A→T or T→A without covalent bond changes, suggesting that the sequence context of tandem motifs might be related to their mutation frequency. In total, 3707 mutation sites were non-randomly distributed on the contigs of the reference genome. In addition, pyrimidines at the 3'-side of adenine promoted its transversion frequency, and UVR generated DNA fragment deletions over 200 bp with a low frequency in the enoki genome. The functional annotation of the genes with non-synonymous mutation indicated that UVR could produce abundant mutations in a short period of time.

2.
J Glob Antimicrob Resist ; 35: 216-222, 2023 12.
Article in English | MEDLINE | ID: mdl-37797810

ABSTRACT

OBJECTIVES: The study aimed to characterize the quinolone resistance of Salmonella enterica serovar Typhimurium and its monophasic variant (Salmonella enterica serovar 1,4,[5],12:i:-) isolated from food and patients in China. METHODS: All of the isolates were assessed for quinolone susceptibility via the broth microdilution method. Then, the isolates were checked for mutations within quinolone resistance-determining regions of gyrA, gyrB, parC, and parE and were examined for plasmid-mediated quinolone resistance genes. RESULTS: High rates of resistance to nalidixic acid in the S. Typhimurium (70.7%) and S. 1,4,[5],12:i:- (41.9%) isolates were observed, and a considerable proportion of isolates with reduced susceptibility to ciprofloxacin and levofloxacin were also detected. The high frequency of mutations in GyrA (60.8%) and a variety of genes (aac[6']-Ib-cr [23.2%], oqxAB [19.2%], qnrS [13.6%], and qnrA [3.2%]) conferring quinolone resistance in these Salmonella isolates were noteworthy. Lastly, the isolates carrying qnrS for transferability and transmission of the quinolone resistance were analysed by conjugation. Multiple locus variable-number tandem repeat analysis profiles indicated that some qnrS-positive isolates were clonally related, whilst the other isolates were genetically divergent. This suggested that both clonal spread of resistant strains and horizontal transmission of the plasmid-mediated resistance genes contributed to the dissemination of qnrS-positive Salmonella isolates. CONCLUSION: This study highlights the prevalence of quinolone-resistant S. Typhimurium and S. 1,4,[5],12:i:- in China, posing a threat to public health.


Subject(s)
Quinolones , Salmonella enterica , Humans , Quinolones/pharmacology , Salmonella typhimurium/genetics , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Serogroup , Salmonella enterica/genetics
3.
J Environ Manage ; 331: 117341, 2023 Apr 01.
Article in English | MEDLINE | ID: mdl-36689861

ABSTRACT

Identifying the leakage channel and the influencing range is essential for controlling the environmental risks of leachate from the tailings pond. The investigation of leachate pollution in tailings pond has the defect of focusing only on the scope of tailings pond in recent studies. This study innovatively built a comprehensive investigation and accurate verification system for leachate leakage of tailings pond integrated with the aeromagnetic survey, ground penetrating radar, hydrochemistry and isotope coupling methods. Geophysical exploration found that among the four fault zones, and the F1 was the channel for leachate to recharge the groundwater 2.53 km away from the tailings pond. The fissures inside the tailings pond were connected with the natural fissures outside, forming a leachate migration channel. The hydrochemistry and isotope characteristics showed that the groundwater far away from the tailings pond were polluted by arsenic containing leachate, which verified the geophysical exploration results. The significant correlation between arsenic and SO2-4 concentration indicated that arsenic in leachate originated from the oxidation release of sulfide minerals (i.e., arsenopyrite). This study sheds light on the comprehensive investigation of leachate leakage in the tailings pond. This development method also provides guidance for environmental risk identification of other contaminated sites.


Subject(s)
Arsenic , Ponds , Environmental Pollution , Oxidation-Reduction , Environmental Monitoring/methods
4.
Food Sci Nutr ; 10(10): 3444-3452, 2022 Oct.
Article in English | MEDLINE | ID: mdl-36249961

ABSTRACT

Oudemansiella raphanipes (OR) is a commercial mushroom which possesses high nutritional value and excellent and unique flavors. In this study, various agricultural wastes were utilized as substitute materials in the low-cost and high-yield production of mycelia biomass and polysaccharides by liquid fermentation. The sawdust, wheat bran, apple pomace, sugarcane, and corn particles were employed to cultivate OR, using the potato dextrose broth as control. Additionally, a preliminary characterization and in vitro antioxidant activities of partial purified OR polysaccharides were investigated. The substrate of sugarcane was suitable for mycelia growth of OR, with high yield of mycelia biomass and polysaccharides content. In vitro antioxidant activity assays demonstrated that OR polysaccharides could effectively scavenge 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 1,1-diphenyl-2-picrylhydrazyl radicals. OR polysaccharides had configuration as revealed by Fourier transform infrared, and was mainly composed of fucose (Fuc), rhamnose (Rha), arabinose (Ara), galactose (Gal), glucose (Glc), xylose (Xyl), mannose (Man), ribose (Rib), and galacturonic acid (Gal-UA), with mass percentages of 3.29%, 0.64%, 1.09%, 16.03%, 72.69%, 0.56%, 3.18%, 0.93%, and 1.59%, respectively. This study may offer support for decreasing the cost of OR polysaccharides production and dealing with these agricultural wastes.

5.
PLoS One ; 16(7): e0254070, 2021.
Article in English | MEDLINE | ID: mdl-34252147

ABSTRACT

Soft rot disease is a major pathogenic bacteria of Fuding areca taro and has caused serious losses. This study aims to screen biocontrol bacterial against soft rot disease. A total of 53 bacterial strains were isolated from the rhizosphere soil, nine of which exhibited good biocontrol effect against the pathogenic bacteria of soft rot disease as seen in antagonistic screening of biocontrol bacteria from corm in vitro. Strains were selected by physical and chemical experiments, biocontrol effect tests in vivo, molecular sequencing, morphological observation and field tests. Four strains including CAB-L005, CAB-L012, CAB-L014, and CAB-L022 exhibited strong antagonistic effects. On the basis of the sequence homology of 16S rRNA genes, the similarity between strain CAB-L005 and Bacillus tropicus was 100%, that between strain CAB-L012 and Bacillus subtilis was 99%, and that between strain CAB-L014 and Bacillus tequilensis was 100%, and similarity between strain CAB-L022 and Bacillus cereus was 100%. The isolated bacteria demonstrated good biocontrol effects in field experiments. In this study, four strains with good biocontrol application value were isolated and identified, providing a foundation for biocontrol against soft rot disease in areca taro.


Subject(s)
Bacteria/isolation & purification , Biological Control Agents , Colocasia/microbiology , Plant Diseases/microbiology , Bacteria/growth & development , Colocasia/genetics , Fermentation , Hydrogen-Ion Concentration , Phylogeny , Temperature
6.
Bioprocess Biosyst Eng ; 44(10): 2109-2120, 2021 Oct.
Article in English | MEDLINE | ID: mdl-34047828

ABSTRACT

ε-Poly-L-lysine (ε-PL) is a naturally-occurring L-lysine homopolymer having a broad-spectrum antimicrobial activity and used widely as a food preservative. In the present study, the combined use of immobilization and in situ product removal (ISPR) was evaluated for the production of ε-PL by Streptomyces ahygroscopicus GIM8. Results showed that ε-PL production in the flask cultures decreased from 0.84 to 0.38-0.56 g/L upon immobilization on loofah sponge with different amounts (0.5-3 g in 50 mL medium in a flask). By applying continuous ISPR to the immobilized flask cultures, ε-PL production as high as 3.51 g/L was obtained compared to 0.51 g/L of the control. A satisfactory titer of 1.84 g/L ε-PL could also be achieved with intermittent ISRP (three cycles of ISPR operation during cultivation). Further investigation showed that low levels of ε-PL retained in the broth appeared to favor its biosynthesis. In the repeated-batch fermentation in a 5 L immobilized bioreactor, with continuous ISPR, the final average ε-PL concentration and productivity were 3.35 g/L and 0.797 g/L/day, respectively, and 3.18 g/L and 0.756 g/L/day for the alternative (intermittent ISPR), in comparison to 1.16 g/L and 0.277 g/L/day with no ISPR usage. In the fed-batch fermentation with immobilized cells, the combined use of intermittent ISPR and extra nutrient feeding increased ε-PL concentration and productivity up to 24.57 g/L and 9.34 g/L/day. The fermentation processes developed could serve as an effective approach for ε-PL production and, moreover, the combination could greatly simplify downstream processing for ε-PL separation and purification.


Subject(s)
Fermentation , Polylysine/biosynthesis , Streptomyces/metabolism , Bioreactors , Culture Media , Glucose/metabolism
7.
PLoS One ; 15(12): e0244749, 2020.
Article in English | MEDLINE | ID: mdl-33382761

ABSTRACT

Ganoderma lucidum (Fr.) Krast, commonly known as "Lingzhi" in Chinese, is a medicinal mushroom that is rich in biologically active substances. Polysaccharides and triterpenoids are the two major components responsible for the bioactivity of this fungus. In the present study, the ultrasonic-assisted co-extraction (UACE) of polysaccharides and triterpenoids from G. lucidum was optimized using response surface methodology with a desirability function, with the equal importance for the two components. Following single factor experiments, the optimal conditions were determine as ultrasonic power of 210 W, extraction temperature of 80C, ratio of liquid to solid of 50 mL/g, and 100 min extraction time, using aqueous ethanol (50%, v/v) as the extracting solvent. Under the optimal conditions, the extraction yields of polysaccharides and triterpenoids reached 0.63% and 0.38%, respectively. On the basis of the scavenging capacity of 2,2-diphenyl-1-picrylhydrazyl and evaluation of reducing power, the antioxidant capacities of the polysaccharides obtained by optimal UACE process were higher than those of polysaccharides extracted using traditional hot water extraction, whereas the triterpenoid-rich extracts showed antioxidant activities similar to those obtained using the ethanol maceration method. The present study is the first report on the simultaneous extraction of polysaccharides and triterpenoids from G. lucidum. The developed UACE process could be useful in preparation of a polysaccharide- and triterpenoid-rich ingredient that holds great promise for application in the Ganoderma industry.


Subject(s)
Antioxidants/analysis , Drugs, Chinese Herbal , Polysaccharides/analysis , Reishi/chemistry , Triterpenes/analysis , Ultrasonics
8.
Sci Total Environ ; 713: 136385, 2020 Apr 15.
Article in English | MEDLINE | ID: mdl-31955074

ABSTRACT

This study investigated the prevalence and levels of Salmonella contamination of retail raw poultry meat in China, and examined serovar distribution and antimicrobial susceptibility profiles of the recovered isolates. In total, 664 poultry meat samples were collected from retail markets in 39 cities across China. Salmonella was isolated from 249 (37.5%) samples, including 190 (36.7%) chicken, 48 (40.7%) duck and 11 (39.2%) pigeon samples. The most probable number (MPN) values of 36.1% of the positive samples ranged from 0.3 to 10 MPN/g, with three samples exceeding 110 MPN/g. Among the 667 Salmonella isolates, 35 serovars and 42 multilocus sequence typing patterns were identified. Predominant serovars included Salmonella enterica serovar Enteritidis (32.7%), Salmonella enterica serovar Indiana (14.2%) and Salmonella enterica serovar Typhimurium (11.9%), while two novel STs were identified (ST7352 and ST7612). Except for one unnamed strain (4,12:d:-), all of the identified serovars have previously been linked to human infections. Antimicrobial susceptibility testing of the 318 non-duplicate isolates revealed that only 5 (1.6%) were susceptible to all 22 tested antimicrobials, while 191 (60.1%) exhibited resistance to at least three classes of antimicrobials. The highest levels of resistance were observed for nalidixic acid (72.3%), followed by ampicillin (55.3%) and streptomycin (48.7%). Of particular concern was the detection of highly multidrug-resistant Salmonella enterica serovar Indiana isolates, most (84.1%) of which showed co-resistance to ciprofloxacin and ceftriaxone. Overall, our findings showed a high prevalence of Salmonella contamination of retail raw poultry meat, which could expose consumers to multidrug-resistant isolates. This study provides comprehensive data for evaluation of new control measures for Salmonella contamination of poultry.


Subject(s)
Poultry , Animals , Anti-Bacterial Agents , China , Drug Resistance, Multiple, Bacterial , Humans , Meat , Microbial Sensitivity Tests , Prevalence , Serogroup
9.
Food Sci Biotechnol ; 28(4): 1135-1145, 2019 Aug.
Article in English | MEDLINE | ID: mdl-31275713

ABSTRACT

To facilitate Ganoderma lucidum submerged cultivation and achieve high productivity, four fine powder solid substrates incorporated with different nitrogen-rich supplements were utilized to grow the fungus and as solid seed for its submerged culture. Of the four solid seeds, the soybean meal solid seed gave the highest biomass (10.73 g/L) and exopolysaccharide (EPS) (1.22 g/L), higher than those (8.36 g/L biomass and 0.44 g/L EPS) obtained with mycelial liquid seed. The optimal level of soybean meal supplementation was 20% (w/w) for production of the solid seed. Following single factor experiments, levels of three selected process variables were optimized as: the moisture content of solid seed, 70%; inoculum size, 0.8 g/flask; and rotary speed, 160 rpm. These conditions were validated experimentally with improved EPS yield of 1.33 g/L. The developed solid seed can be conveniently used for G. lucidum submerged culture with improved EPS productivity.

10.
Mycobiology ; 47(1): 97-104, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30988993

ABSTRACT

Mushroom cultivation has gained increased attention in recent years. Currently, only four types of spawn, including sawdust spawn, grain spawn, liquid spawn, and stick spawn, are commonly available for mushroom cultivation. This limited spawn diversity has led to difficulty in selecting suitable inoculum materials in some cultivation. In this study, three small blocks of lignocellulosic agro-wastes and one block of a synthetic matrix were prepared as support for growing Pleurotus ostreatus in liquid medium. Mycelium-adsorbed blocks were then evaluated for their potential as block spawn for fructification. Our results indicated that the edible fungus was adsorbed and abundantly grew internally and externally on loofah sponge and synthetic polyurethane foam (PUF) supports and also has the ability to attach and grow on the surface of sugarcane bagasse and corncob supports. The mycelia of P. ostreatus adhered on corncob exhibited the highest metabolic activity, while those on the PUF showed the least activity. Mycelial extension rates of block spawns made of agro-waste materials were comparable to that of sawdust spawn, but the block spawn of PUF showed a significantly lower rate. No significant differences in cropping time and yield were observed among cultivations between experimental block spawns and sawdust spawns. Moreover, the corncob block spawn maintained its fruiting potential during an examined period of 6-month storage. The developed block spawn could be practically applied in mushroom cultivation.

11.
Food Sci Biotechnol ; 27(4): 1129-1136, 2018 Aug.
Article in English | MEDLINE | ID: mdl-30263843

ABSTRACT

To facilitate Ganoderma lucidum submerged culture and obtain high productivity, a fine powder of wheat bran was used to grow the fungus for solid-state fermentation and as solid seed for its submerged cultures. The results indicated that the optimal inoculum size was low, being 0.75 g in 250 mL-sized flasks containing 80 mL medium. The maximal exopolysaccharide concentration and biomass produced was 0.74 and 14.71 g/L, respectively, which is considerably higher than that obtained with the commonly used mycelial pellet liquid seed (0.47 and 8.56 g/L, respectively). The EPS and biomass productivity of the solid seed cultures decreased only slightly, even after a 6-month storage period. EPS produced showed higher antioxidant activity compared with that produced in the liquid seed cultures. The developed solid seed can serve as a ready-to-use inoculum for long-term use in G. lucidum submerged culture for the hyperproduction of highly bioactive EPS and biomass.

12.
PLoS One ; 13(4): e0196287, 2018.
Article in English | MEDLINE | ID: mdl-29694432

ABSTRACT

Ganoderma lucidum is a medicinal mushroom that has been widely used in East Asia for the treatment of various diseases. The pharmacological activity of this fungus is primarily attributable to the polysaccharides and triterpenoids. In this study, to obtain the fruit bodies with improved content of active constituents, we examined the effect of salicylic acid (SA) and calcium ion on the biosynthesis of polysaccharides and triterpenoids by spraying the chemicals during the fruiting. To explore the underlying mechanisms for the variation, the transcripts of related genes involved in the polysaccharide and triterpenoid biosynthesis were measured. Results showed that Ca2+ had no effect on production of polysaccharides and triterpenoids, whereas SA increased triterpenoid content by 23.32%, compared to the control, but it had little influence on polysaccharide production. Interestingly, the combined induction increased polysaccharide and triterpenoid content by 9.02% and 13.61%, respectively, compared to the control. Under Ca2+ induction, the transcript of ugp gene in the polysaccharide biosynthetic pathway up-regulated in all three stages (mycelium, primordium, and fruit body), while pgm and gls gave no response in the mycelium and primordium stages, and up-regulated in the fruit body stage. Differently, six key triterpenoid biosynthetic genes including hmgr, hmgs, mvd, fps, sqs, and ls did not respond to the induction. In the case of SA and combined induction, pgm and ugp were up-regulated in all three stages, while gls showed an increased expression in the primordium stage and no response in other stages. The six triterpenoid biosynthetic genes were up-regulated in all three stages. The present study provides a useful approach to producing G. lucidum fruit bodies with high polysaccharide and triterpenoid content. This is important to the G. lucidum industry.


Subject(s)
Polysaccharides/metabolism , Reishi/metabolism , Signal Transduction , Triterpenes/metabolism , Calcium/pharmacology , Chromatography, High Pressure Liquid , Fungal Proteins/genetics , Fungal Proteins/metabolism , Polysaccharides/analysis , RNA, Fungal/isolation & purification , RNA, Fungal/metabolism , Reishi/chemistry , Reishi/growth & development , Salicylic Acid/pharmacology , Signal Transduction/drug effects , Triterpenes/analysis , Up-Regulation/drug effects
13.
Indian J Microbiol ; 56(4): 516-521, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27784952

ABSTRACT

Tissue isolation from mushrooms is frequently practiced by both researchers and growers to isolate new and improved strains. In the present study, we designed a simple and convenient device for precise tissue isolation and therefore investigated the effect of tissue size on mycelial growth of seven mushroom species. The developed device consists of a cutting needle and a transfer needle. The cutting needle was used to obtain circular tissue plugs having a height up to 3 mm and variable diameters (2-5 mm) from mushroom fruit bodies. The transfer needle was a stainless steel round rod (1.5 mm in diameter) with a blade-like end. It can be used for collecting mushroom tissue when the cutting needle fails to extract it. With the aid of these devices, precise tissue isolation was achieved. Plate cultures demonstrated that tissue size had little effect on mycelium extension for Lentinula edodes (the winter shiitake), Hypsizygus marmoreus, and Agrocybe aegerita, but influenced the aerobic mycelium density. For Pleurotus ostreatus, Pleurotus eryngii, and Volvariella volvacea, large tissue plugs produced faster mycelial growth and higher aerobic mycelium density compared with small ones. On the contrary, small plugs from the tissue of the flower shiitake and Agaricus bisporus favored mycelial growth. The present study revealed that the preferable tissue size for mycelial growth varies among mushroom species, and the developed device is expected to greatly facilitate the isolation of new and improved mushroom strains.

14.
PLoS One ; 11(1): e0147531, 2016.
Article in English | MEDLINE | ID: mdl-26814481

ABSTRACT

A photo-medical capsule that emits blue light for Helicobacter pylori treatment was described in this paper. The system consists of modules for pH sensing and measuring, light-emitting diode driver circuit, radio communication and microcontroller, and power management. The system can differentiate locations by monitoring the pH values of the gastrointestinal tract, and turn on and off the blue light according to the preset range of pH values. Our experimental tests show that the capsule can operate in the effective light therapy mode for more than 32 minutes and the wireless communication module can reliably transmit the measured pH value to a receiver located outside the body.


Subject(s)
Helicobacter Infections/therapy , Helicobacter pylori/radiation effects , Light , Phototherapy/methods , Electrodes , Equipment Design , Gastrointestinal Tract/physiology , Humans , Hydrogen-Ion Concentration , Phototherapy/instrumentation , Wireless Technology
15.
Food Sci Biotechnol ; 25(4): 1109-1113, 2016.
Article in English | MEDLINE | ID: mdl-30263382

ABSTRACT

This study aimed to classify a collection of Enterobacter sakazakii (E. sakazakii) strains previously identified from powdered infant formula (PIF) to species level by recN gene sequencing and biochemical testing to determine the distribution of Cronobacter species in China and investigate the strain diversity by cellular fatty acid (CFA) analysis. Of 24 E. sakazakii isolates, 23 were identified as C. sakazakii and one was C. malonaticus. The 23 C. sakazakii isolates showed the same CFA profiles. The C. malonaticus isolate was discriminated from the C. sakazakii isolates by the significant difference in the amounts of C12:0, C14:0, and C17:0 cyclo acids. These results showed that C. sakazakii and C. malonaticus were the common Cronobacter species distributed in PIF in China and that the isolates of the two species exhibited different CFA profiles. These findings are of value for epidemiological investigations and provide an alternative method for confirming various Cronobacter spp.

16.
Wei Sheng Wu Xue Bao ; 55(6): 725-31, 2015 Jun 04.
Article in Chinese | MEDLINE | ID: mdl-26562997

ABSTRACT

OBJECTIVE: To assess the effect of cellular activity on ε-poly-1-lysine (ε-PL) biosynthesis and thereby to rationally improve the production, we studied the cellular activity, ε-PL formation and other parameters cross flask fermentation by Streptomyces ahygroscopicus. METHODS: Laser scanning confocal microscopy and a colorimetric method were used to determine cellular activity using BacLight Live/Dead and 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) as viable stains. To enhance the activity of the cells in the ε-PL production period, yeast extract was added. RESULTS: During ε-PL submerged fermentation in flasks, most cells were active in the growth period (0 - 16 h); cells had metabolic activity in the growth and earlier ε-PL production periods between 0 and 30 h fermentation. Almost no activity was detected after 48 h fermentation when no ε-PL was produced. The improved fermentation achieved 2. 24 g/L ε-PL from 1.04 g/L. CONCLUSION: Biosynthesis of ε-PL can be boosted by up-regulating cell activity in its production phase.


Subject(s)
Polylysine/biosynthesis , Streptomyces/metabolism , Bioreactors/microbiology , Culture Media/metabolism , Fermentation , Streptomyces/growth & development
17.
J Basic Microbiol ; 55(2): 172-9, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25291434

ABSTRACT

ϵ-Poly-L-lysine (ϵ-PL) is an L-lysine homopolymer with strong antimicrobial activity, which is generally produced by Streptomyces strains. ϵ-PL is only produced under acidic conditions in liquid culture, and to improve the current understanding of ϵ-PL biosynthesis, the present study was undertaken to investigate the effects of ϵ-PL on its producer Streptomyces ahygroscopicus GIM8, under acidic and neutral conditions. The results indicated that a neutral pH favored ϵ-PL adsorption onto the cells, whereas minimal adsorption occurred at pH 4.0, the maximum pH for ϵ-PL production. At pH 7.0, small amounts of ϵ-PL caused considerable ATP leakage from the cells, which showed increased membrane permeability. Conversely, ATP leakage was inhibited by ϵ-PL at pH 4.0. Transmission electron microscopy investigation indicated that the cytoplasmic membrane was the primary site of ϵ-PL activity at pH 7.0, and that cell shape was maintained. Metabolic activity profiles revealed that ϵ-PL decreased cellular metabolic activity at a relatively low rate at pH 7.0. However, the toxic effect was significantly enhanced at pH 4.0. Based on these data, a mechanism for the effect of ϵ-PL on ϵ-PL-producing cells under neutral and acidic conditions is proposed. Additionally, acidic conditions may potentially be required for ϵ-PL biosynthesis in liquid culture because low pH can increase membrane permeability and prevent binding of ϵ-PL onto cells, both of which favor the secretion of the ϵ-PL produced by the cells into the broth. This research contributes to the current understanding of ϵ-PL biosynthesis.


Subject(s)
Polylysine/biosynthesis , Streptomyces/metabolism , Adenosine Triphosphate/metabolism , Adsorption , Cell Membrane/metabolism , Hydrogen-Ion Concentration , Microscopy, Electron, Transmission , Polylysine/toxicity , Streptomyces/ultrastructure
18.
J Microbiol Biotechnol ; 25(3): 358-65, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25269813

ABSTRACT

ε-Poly-L-lysine (ε-PL) is a homopolymer of L-lysine molecules connected between the ε amino and alpha carboxyl groups. This polymer is currently used as a natural preservative in food. Insufficient biomass is a major problem in ε-PL fermentation. Here, to improve cell growth and ε-PL productivity, various nitrogen-rich nutrients were supplemented into flask cultures after 16 h cultivation, marking the onset of ε-PL biosynthesis. Yeast extract, soybean powder, corn powder, and beef extract significantly improved cell growth. In terms of ε-PL productivity, yeast extract at 0.5% (w/v) gave the maximum yield (2.24 g/l), 115.4% higher than the control (1.04 g/l), followed by soybean powder (1.86 g/l) at 1% (w/v) and corn powder (1.72 g/l) at 1% (w/v). However, supplementation with beef extract inhibited ε-PL production. The optimal time for supplementation for all nutrients examined was at 16 h cultivation. The kinetics of yeast-extract-supplemented cultures showed enhanced cell growth and production duration. Thus, the most commonly used two-stage pH control fed-batch fermentation method was modified by omitting the pH 5.0-controlled period, and coupling the procedure with nutrient feeding in the pH 3.9-controlled phase. Using this process, by continuously feeding 0.5 g/h of yeast extract, soybean powder, or corn powder into cultures in a 30 L fermenter, the final ε-PL titer reached 28.2 g/l, 23.7 g/l, and 21.4 g/l, respectively, 91.8%, 61.2%, and 45.6% higher than that of the control (14.7 g/l). This describes a promising option for the mass production of ε-PL.


Subject(s)
Fermentation , Polylysine/biosynthesis , Streptomyces/growth & development , Streptomyces/metabolism , Batch Cell Culture Techniques , Biomass , Hydrogen-Ion Concentration , Nitrogen/metabolism
19.
Wei Sheng Yan Jiu ; 42(4): 615-8, 2013 Jul.
Article in Chinese | MEDLINE | ID: mdl-24024375

ABSTRACT

OBJECTIVE: Salmonella isolates from meat in Guangdong Province were characterized to determine their phenotype and serotype, to investigate contamination status of meats by Salmonella and provide scientific basis for detection and further control. METHODS: 209 meat samples were detected according to methods GB 4789.4--2010 National food safety standard Food microbiological examination: Salmonella. The API 20E system was used to identify isolates. Serotyping of Salmonella isolates was performed by slide agglutination method using antisera to Salmonella. RESULTS: According to the detection results of 209 meat samples, forty-six Salmonella strains were isolated from 42 meat samples, the positive rate was 20. 10%. The majority of positive samples were fresh meat, 69.23% in duck, 37.14% in chicken, 20.00% in beef and 16.67% in pock. The strains identified as Salmonella by the API 20E were distributed in 7 profile numbers with two of them being predominant (6704752 and 6704552). The most common serotypes were Derby (21.74%), Typhimurium (10.87%), Enteritidis (8.70%), Tshiongwe (8.70%), Indiana (8.70%), Weltevreden (8.70%). CONCLUSION: Salmonella present in retail meats were common and phenotypically diverse in Guangdong Province.


Subject(s)
Food Contamination/analysis , Meat/microbiology , Salmonella/isolation & purification , Animals , Cattle , Chickens , China , Ducks , Salmonella/classification , Salmonella enteritidis/isolation & purification , Salmonella typhimurium/isolation & purification , Serotyping , Swine
20.
J Microbiol Biotechnol ; 22(9): 1218-23, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22814495

ABSTRACT

Epsilon-poly-L-lysine (epsilon-PL), produced by Streptomyces or Kitasatospora strains, is a homo-poly-amino acid of Llysine, which is used as a safe food preservative. The present study investigates the combined use of cell immobilization and in situ adsorption (ISA) to produce epsilon-PL in shaken flasks. Loofah sponge-immobilized Streptomyces ahygroscopicus GIM8 produced slightly more epsilon-PL than those immobilized on synthetic sponge, and sugarcane bagasse. Moreover, loofah sponge supported the maximum biomass. Hence, loofah sponge was chosen for cell immobilization. Meanwhile, the ion-exchange resin D152 was employed for ISA. The loofah sponge-immobilized cells produced 0.54 +/- 0.1 g/l epsilon-PL, which significantly increased to 3.64 +/- 0.32 g/l after combining with ISA through the addition of resin bags. The free cells with ISA using the dispersed resin yielded 2.73 +/- 0.26 g/l of epsilon-PL, an increase from 0.82 +/- 0.08 g/l. These data illustrate that the proposed combination method improved production most significantly compared with either immobilization or ISA only. Moreover, the immobilized cells could be repeatedly used and an epsilon-PL total amount of 8.05 +/- 0.84 g/l was obtained. The proposed combination method offers promising perspectives for epsilon-PL production.


Subject(s)
Cells, Immobilized/metabolism , Polylysine/biosynthesis , Streptomyces/metabolism , Adsorption , Biomass , Cells, Immobilized/cytology , Equipment Reuse , Glucose/metabolism , Hydrogen-Ion Concentration , Luffa , Polylysine/analysis , Streptomyces/cytology
SELECTION OF CITATIONS
SEARCH DETAIL
...