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1.
Crit Rev Food Sci Nutr ; : 1-21, 2022 Nov 08.
Article in English | MEDLINE | ID: mdl-36345974

ABSTRACT

Zymomonas mobilis is a gram-negative facultative anaerobic spore, which is generally recognized as a safe. As a promising ethanologenic organism for large-scale bio-ethanol production, Z. mobilis has also shown a good application prospect in food processing and food additive synthesis for its unique physiological characteristics and excellent industrial characteristics. It not only has obvious advantages in food processing and becomes the biorefinery chassis cell for food additives, but also has a certain healthcare effect on human health. Until to now, most of the research is still in theory and laboratory scale, and further research is also needed to achieve industrial production. This review summarized the physiological characteristics and advantages of Z. mobilis in food industry for the first time and further expounds its research status in food industry from three aspects of food additive synthesis, fermentation applications, and prebiotic efficacy, it will provide a theoretical basis for its development and applications in food industry. This review also discussed the shortcomings of its practical applications in the current food industry, and explored other ways to broaden the applications of Z. mobilis in the food industry, to promote its applications in food processing.


Potential applications of Zymomonas mobilis in food industry summarized for the first time.Research status of Z. mobilis in food additive synthesis, fermentation applications, and probiotics are discussed in details.Future research perspectives of Z. mobilis in food industry further proposed.

2.
J Sci Food Agric ; 101(3): 927-936, 2021 Feb.
Article in English | MEDLINE | ID: mdl-32748962

ABSTRACT

BACKGROUND: A large number of digestates have not been fully utilized due to a lack of scientific, reasonable guidance, as well as imperfect technology. Hybrid giant Napier has great potential for use as a type of energy plant. As such, this study investigated the effects of digestate on the growth of a candidate energy crop and examined whether digestate was an ecologically viable means for soil restoration. RESULTS: The results showed that the total yields of all treatment groups receiving irrigation of digestate were higher (5.19-26.00%) than those of the control. The total phosphorus, total potassium, available nitrogen, available phosphorus, and available potassium content of the soil had also increased after digestate application, compared with the control. Urease activities for all treatments increased 15.28 to 69.44% more than that of the corresponding control. Soil dissolved organic matter (DOM) mainly contained humic-like and fulvic-like components through the application of digestate. More fluorescent components were also identified by two-dimensional correlation spectroscopy (2D-COS). These fluorescent components can improve the aromaticity and molecular weight of soil DOM so as to improve soil quality. CONCLUSIONS: Digestate improved not only the aboveground biomass accumulation, but also the chemical properties of the soil, which was an appropriate strategy for restoring soil quality and contributing to the sustainable development of marginal. The long-term impact of digestate application on soil quality will require additional long-term experiments. © 2020 Society of Chemical Industry.


Subject(s)
Pennisetum/chemistry , Soil/chemistry , Biomass , Fertilizers/analysis , Humic Substances/analysis , Nitrogen/analysis , Nitrogen/metabolism , Pennisetum/growth & development , Pennisetum/metabolism , Phosphorus/analysis , Phosphorus/metabolism , Potassium/analysis , Potassium/metabolism
3.
Biotechnol Bioeng ; 114(9): 1946-1958, 2017 09.
Article in English | MEDLINE | ID: mdl-28464284

ABSTRACT

Biosynthesis of advanced biofuels by engineered non-natural microorganisms has been proposed to be the most promising approach for the replacement of dwindling fossil fuel resources. Brevibacterium flavum (Bf) is a model brevibacterium aerobe which lacks basic and applied research that could enable this species to produce biofuels. There are no reports regarding engineering this microorganism to produce advanced alcohols before. Here, for the first time, we developed the bacterium as a novel biosynthetic platform for advanced alcohols production via the mutagenesis and engineering to produce 2-ketoacids derived alcohols. In order to enhance the strain's capability of producing advanced alcohols, we preferentially improved intrinsic metabolism ability of the strain to obtain improved expression host (IEH) via generating mutagenesis libraries by whole cell mutagenesis (WCM). The IEH was determined via screening out the mutant strain with the highest production of branched-chain organic acids (BCOA) using high throughput screening method.. Subsequently, a novel vector system for Bf was established, and the corresponding biosynthetic pathway of directing carbon flux into the target advanced alcohols was recruited to make the bacterium possess the capability of producing advanced alcohols and further enhance the production using the IEH. Specifically, we generated bioengineered strains that were able to synthesize up to the highest 5362 and 4976 mg/L isobutanol, 1945 and 1747 mg/L 2-methyl-1-butanol (2 MB), and 785.34 and 781 mg/L 3-methyl-1-butanol (3 MB) from pure glucose and duckweed substrates, respectively. Our findings confirmed the feasibility and potential of using Bf as a novel biosynthetic platform to generate advanced biofuels with glucose and inexpensive renewable feedstock-duckweed as a fermentation substrate. Biotechnol. Bioeng. 2017;114: 1946-1958. © 2017 Wiley Periodicals, Inc.


Subject(s)
Alcohols/metabolism , Biofuels/microbiology , Biosynthetic Pathways/physiology , Brevibacterium flavum/physiology , Keto Acids/metabolism , Metabolic Engineering/methods , Renewable Energy , Alcohols/isolation & purification , Genetic Enhancement/methods
4.
J Environ Manage ; 198(Pt 1): 70-74, 2017 Aug 01.
Article in English | MEDLINE | ID: mdl-28453987

ABSTRACT

Calcium-rich biochars (CRB) prepared through pyrolysis of crab shell at various temperatures were characterized for physicochemical properties and P removal potential. Elemental analysis showed that CRB was rich in calcium (22.91%-36.14%), while poor in carbon (25.21%-9.08%). FTIR, XRD and TG analyses showed that calcite-based CRB was prepared at temperature ≤600 °C, while lime-based CRB was prepared at temperature ≥700 °C. Phosphorus removal experiment showed that P removal efficiencies in 80 mg P/L phosphate solution and biogas effluent ranged from 26% to 11%, respectively, to about 100% and 63%, respectively, depending on the pyrolysis temperature of the resulting biochar. Specifically, compared to common used CaCO3 and Ca(OH)2, P removal potential of calcite-based CRB was much higher than that of CaCO3; while that of lime-based CRB was close to that of Ca(OH)2. These results suggested that CRB was competent for P removal/recovery from wastewater.


Subject(s)
Calcium , Charcoal , Phosphorus , Animals , Brachyura
5.
BMC Microbiol ; 16(1): 160, 2016 07 22.
Article in English | MEDLINE | ID: mdl-27448996

ABSTRACT

BACKGROUND: Currently, efficient screening methods for selection of desired bacterial phenotypes from large populations are not easy feasible or readily available due to the complicated physiological and metabolic networks of solventogenic clostridia. In this study, to contribute to the improvement of methods for predicting the butanol-producing ability of Clostridium beijerinckii based on starch substrate, we further investigate a simple, visualization screening method for selecting target strains from mutant library of Clostridium beijerinckii NCIMB 8052 by using trypan blue dye as an indicator in solid starch via statistical survey and validation of fermentation experiment with controlling pH. RESULTS: To verify an effective, efficient phenotypic screening method for isolating high butanol-producing mutants, the revalidation process was conducted based on Trypan Blue was used for visualization, and starch was used as the bacterial metabolic substrate. The availability of the screening system was further evaluated based on the relationship between characteristics of mutant strains and their α-amylase activities. Mutant clones were analyzed in detail based on their distinctive growth patterns and rate of fermentation of soluble starch to form butanol and were compared by statistical method. Significant correlations were identified between colony morphology and changes in butanol concentrations. The screening method was validated via statistical analysis for characterizing phenotypic parameters. The fermentation experiment of mutant strains with controlling pH value also demonstrated a positive correlation between increased α-amylase activity and increased solvent production by Clostridium beijerinckii was observed, and therefore indicated that the trypan blue dyeing method can be used as a fast method to screen target mutant strain for better solvent producers from, for instance, a mutant library. CONCLUSIONS: The suitability of the novel screening procedure was validated, opening up a new indicator of approach to select mutant solventogenic clostridia with improved fermentation of starch to increase butanol concentrations. The applicability can easily be broadened to a wide range of interesting microbes such as cellulolytic or acetogenic microorganisms, which produce biofuels from feedstock rich in starch.


Subject(s)
Butanols/metabolism , Clostridium beijerinckii/genetics , Clostridium beijerinckii/isolation & purification , Clostridium beijerinckii/metabolism , High-Throughput Screening Assays/methods , Solvents/metabolism , Biofuels , Clostridium beijerinckii/enzymology , Enzyme Activation , Enzyme Assays , Fermentation , Hydrogen-Ion Concentration , Metabolic Networks and Pathways/genetics , Mutagenesis , Mutation , Phenotype , Starch/metabolism , alpha-Amylases/metabolism
6.
Microb Cell Fact ; 15: 4, 2016 Jan 13.
Article in English | MEDLINE | ID: mdl-26758018

ABSTRACT

BACKGROUND: With the increasing global crude oil crisis and resulting environmental concerns, the production of biofuels from renewable resources has become increasingly important. One of the major challenges faced during the process of biofuel production is the low tolerance of the microbial host towards increasing biofuel concentrations. RESULTS: Here, we demonstrate that the ethanol tolerance of Zymomonas mobilis can be greatly enhanced through the random mutagenesis of global transcription factor RpoD protein, (σ(70)). Using an enrichment screening, four mutants with elevated ethanol tolerance were isolated from error-prone PCR libraries. All mutants showed significant growth improvement in the presence of ethanol stress when compared to the control strain. After an ethanol (9 %) stress exposure lasting 22 h, the rate of glucose consumption was approximately 1.77, 1.78 and 1.39 g L(-1) h(-1) in the best ethanol-tolerant strain ZM4-mrpoD4, its rebuilt mutant strain ZM4-imrpoD and the control strain, respectively. Our results indicated that both ZM4-mrpoD4 and ZM4-imrpoD consumed glucose at a faster rate after the initial 9 % (v/v) ethanol stress, as nearly 0.64 % of the initial glucose remained after 54 h incubation versus approximately 5.43 % for the control strain. At 9 % ethanol stress, the net ethanol productions by ZM4-mrpoD4 and ZM4-imrpoD during the 30-54 h were 13.0-14.1 g/l versus only 6.6-7.7 g/l for the control strain. The pyruvate decarboxylase activity of ZM4-mrpoD4 was 62.23 and 68.42 U/g at 24 and 48 h, respectively, which were 2.6 and 1.6 times higher than the control strain. After 24 and 48 h of 9 % ethanol stress, the alcohol dehydrogenase activities of ZM4-mrpoD4 were also augmented, showing an approximate 1.4 and 1.3 times increase, respectively, when compared to the control strain. Subsequent quantitative real-time PCR analysis under these stress conditions revealed that the relative expression of pdc in cultured (6 and 24 h) ZM4-mrpoD4 increased by 9.0- and 12.7-fold when compared to control strain. CONCLUSIONS: Collectively, these results demonstrate that the RpoD mutation can enhance ethanol tolerance in Z. mobilis. Our results also suggested that RpoD may play an important role in resisting high ethanol concentration in Z. mobilis and manipulating RpoD via global transcription machinery engineering (gTME) can provide an alternative and useful approach for strain improvement for complex phenotypes.


Subject(s)
Ethanol/pharmacology , Zymomonas/drug effects , Zymomonas/metabolism , Bacterial Proteins/genetics , Fermentation/drug effects , Mutation/genetics , Zymomonas/genetics
7.
Microb Cell Fact ; 15(1): 101, 2016 Jun 10.
Article in English | MEDLINE | ID: mdl-27287016

ABSTRACT

BACKGROUND: The cell growth and ethanol yield of Zymomonas mobilis may be detrimentally affected by salt stress frequently present in some biomass-based fermentation systems, leading to a decrease in the rate of sugar conversion to ethanol or other bioproducts. To address this problem, improving the salt tolerance of Z. mobilis is a desirable way. However, limited progress has been made in development of Z. mobilis with higher salt tolerance for some technical challenges in the past decades. Recently, transposon insertion mutant system has been widely used as a novel genetic tool in many organisms to develop mutant strains. In this study, Tn5-based transposon insertion mutagenesis system firstly used for construction of higher salt tolerance strain in Z. mobilis. RESULTS: Approximately 200 Z. mobilis ZM4 mutants were generated by using Tn5-based transposon mutagenesis system. The mutant strain ZMT2 with improved salt tolerance phenotype was obtained by screening on RM agar plates with additional 1 % NaCl. Strain ZMT2 was confirmed to exhibit better fermentation performance under NaCl stress than wild type of strain ZM4. The transposon insertion was located in ZMO1122 (himA) by genome walking. Discruption of himA gene showed that himA may play an important role in response to salt tolerance in Z. mobils. CONCLUSIONS: The mutant strain ZMT2 with a transposon insertion in himA gene of the genome showed obviously higher sugar conversion rate to ethonal under up to 2 % NaCl stress than did the wild ZM4 strain. Besides, ZMT2 exhibited shared fermentative capabilities with wild ZM4 strain under no or low NaCl stress. This report firstly showed that himA played a role in responding to NaCl stress. Furthermore, the result indicated that Tn5-based transposon mutagenesis system was a feasible tool not only for genetic engineering in Z. mobilis strain improvement, but also in tapping resistent genes.


Subject(s)
Salt Tolerance/genetics , Transposases/genetics , Zymomonas/genetics , Zymomonas/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Ethanol/metabolism , Genetic Engineering , Glucose/metabolism , Mutagenesis, Insertional , NAD/metabolism , Phenotype , Real-Time Polymerase Chain Reaction , Transposases/metabolism , Zymomonas/growth & development
8.
Appl Microbiol Biotechnol ; 99(12): 5363-71, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25895089

ABSTRACT

Furfural from lignocellulosic hydrolysates is the key inhibitor for bio-ethanol fermentation. In this study, we report a strategy of improving the furfural tolerance in Zymomonas mobilis on the transcriptional level by engineering its global transcription sigma factor (σ(70), RpoD) protein. Three furfural tolerance RpoD mutants (ZM4-MF1, ZM4-MF2, and ZM4-MF3) were identified from error-prone PCR libraries. The best furfural-tolerance strain ZM4-MF2 reached to the maximal cell density (OD600) about 2.0 after approximately 30 h, while control strain ZM4-rpoD reached its highest cell density of about 1.3 under the same conditions. ZM4-MF2 also consumed glucose faster and yield higher ethanol; expression levels and key Entner-Doudoroff (ED) pathway enzymatic activities were also compared to control strain under furfural stress condition. Our results suggest that global transcription machinery engineering could potentially be used to improve stress tolerance and ethanol production in Z. mobilis.


Subject(s)
Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Furaldehyde/metabolism , Sigma Factor/genetics , Sigma Factor/metabolism , Zymomonas/metabolism , Fermentation , Genetic Engineering , Zymomonas/genetics
9.
Appl Microbiol Biotechnol ; 99(13): 5739-48, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25935346

ABSTRACT

Furfural and acetic acid from lignocellulosic hydrolysates are the prevalent inhibitors to Zymomonas mobilis during cellulosic ethanol production. Developing a strain tolerant to furfural or acetic acid inhibitors is difficul by using rational engineering strategies due to poor understanding of their underlying molecular mechanisms. In this study, strategy of adaptive laboratory evolution (ALE) was used for development of a furfural and acetic acid-tolerant strain. After three round evolution, four evolved mutants (ZMA7-2, ZMA7-3, ZMF3-2, and ZMF3-3) that showed higher growth capacity were successfully obtained via ALE method. Based on the results of profiling of cell growth, glucose utilization, ethanol yield, and activity of key enzymes, two desired strains, ZMA7-2 and ZMF3-3, were achieved, which showed higher tolerance under 7 g/l acetic acid and 3 g/l furfural stress condition. Especially, it is the first report of Z. mobilis strain that could tolerate higher furfural. The best strain, Z. mobilis ZMF3-3, has showed 94.84% theoretical ethanol yield under 3-g/l furfural stress condition, and the theoretical ethanol yield of ZM4 is only 9.89%. Our study also demonstrated that ALE method might also be used as a powerful metabolic engineering tool for metabolic engineering in Z. mobilis. Furthermore, the two best strains could be used as novel host for further metabolic engineering in cellulosic ethanol or future biorefinery. Importantly, the two strains may also be used as novel-tolerant model organisms for the genetic mechanism on the "omics" level, which will provide some useful information for inverse metabolic engineering.


Subject(s)
Acetic Acid/metabolism , Adaptation, Biological , Drug Tolerance , Ethanol/metabolism , Furaldehyde/metabolism , Zymomonas/genetics , Zymomonas/metabolism , Anti-Bacterial Agents/metabolism , Lignin/metabolism , Metabolic Engineering , Zymomonas/drug effects
10.
J Environ Manage ; 157: 49-53, 2015 Jul 01.
Article in English | MEDLINE | ID: mdl-25881151

ABSTRACT

The surplus of manure phosphorus (P) with increasing livestock production might pose a risk of P loss to the environment due to the high mobility of P in manure. Thus, there is an increasing need to mitigate P loss from manure. This study aimed to investigate the effect of hydrothermal carbonization (HTC) on the immobilization of P in cow manure. The results demonstrated that the P content in cow manure was increased substantially by ∼20% after HTC, while the water-extractable P (WEP) and Mehlich-3-extractable P (MEP) in manure was reduced significantly by >80% and 50%, respectively. The decrease in P solubility might result from the increased apatite P (increased by >85%) and decreased soluble Ca (decreased by ∼50%) after HTC. These results suggested that HTC could be an efficient strategy to immobilize P in cow manure, thereby potentially mitigating the P loss problem from cow manure.


Subject(s)
Carbon/chemistry , Manure/analysis , Phosphorus/chemistry , Water Movements , Water Pollutants/chemistry , Animals , Cattle , Female , Solubility
11.
Front Med (Lausanne) ; 11: 1386797, 2024.
Article in English | MEDLINE | ID: mdl-38606152

ABSTRACT

Objective: To investigate the effects of perioperative general anesthesia (GA) and spinal anesthesia (SA) on postoperative rehabilitation in elderly patients with lower limb surgery. Methods: This retrospective propensity score-matched cohort study included patients aged 65 years or older who underwent lower limb surgery between January 1, 2020, and May 31, 2023. The GA and SA were selected at the request of the orthopedic surgeon, patient, and their family members. The main outcomes included the incidence of the patient's inability to self-care at discharge, postoperative complications including pulmonary infection, thrombus of lower extremity veins, infection of incisional wound and delirium, length of hospital stay, and incidence of severe pain in the first 2 days postoperatively. Results: In total, 697 patients met the inclusion criteria, and 456 were included in the final analysis after propensity score matching. In the GA and SA groups, 27 (11.84%) and 26 (11.40%) patients, respectively, could not care for themselves at discharge. The incidence rates did not differ between the groups (p = 0.884). In contrast, the incidence of postoperative complications (GA: 10.53% and SA: 4.39%; p = 0.013) and the length of hospital stay (GA: 16.92 ± 10.65 days and SA: 12.75 ± 9.15 days; p < 0.001) significantly differed between the groups. Conclusion: The choice of anesthesia is independent of the loss of postoperative self-care ability in older patients (>65 years) and is not a key factor affecting postoperative rehabilitation after lower limb surgery. However, compared with GA, SA reduces the incidence of postoperative complications and a prolonged hospital stay. Thus, SA as the primary anesthetic method is a protective factor against a prolonged hospital stay.

12.
Plant Physiol Biochem ; 210: 108619, 2024 May.
Article in English | MEDLINE | ID: mdl-38604013

ABSTRACT

Rhizosphere interactions from plant-soil-microbiome occur dynamically all the time in the "black microzone" underground, where we can't see intuitively. Rhizosphere metabolites including root exudates and microbial metabolites act as various chemical signalings involving in rhizosphere interactions, and play vital roles on plant growth, development, disease suppression and resistance to stress conditions as well as proper soil health. Although rhizosphere metabolites are a mixture from plant roots and soil microbes, they often are discussed alone. As a rapid appearance of various omics platforms and analytical methods, it offers possibilities and opportunities for exploring rhizosphere interactions in unprecedented breadth and depth. However, our comprehensive understanding about the fine-tuning mechanisms of rhizosphere interactions mediated by these chemical compounds still remain clear. Thus, this review summarizes recent advances systemically including the features of rhizosphere metabolites and their effects on rhizosphere ecosystem, and looks forward to the future research perspectives, which contributes to facilitating better understanding of biochemical communications belowground and helping identify novel rhizosphere metabolites. We also address challenges for promoting the understanding about the roles of rhizosphere metabolites in different environmental stresses.


Subject(s)
Plant Roots , Rhizosphere , Soil Microbiology , Plant Roots/microbiology , Plant Roots/metabolism , Agriculture/methods , Microbiota/physiology , Plants/metabolism , Plants/microbiology
13.
Thorac Cancer ; 14(20): 1946-1957, 2023 Jul.
Article in English | MEDLINE | ID: mdl-37277864

ABSTRACT

BACKGROUND: Cisplatin (DDP) treatment is one of the most predominant chemotherapeutic strategies for lung cancer patients. Circular RNAs (circRNAs) have been revealed to participate in the chemoresistance in lung cancer. Hence, the role and mechanism of circ_0010235 in cisplatin resistance in lung cancer was investigated. METHODS: Expression levels of circ_0010235, microRNA (miR)-379-5p and E2F transcription factor 7 (E2F7) were analyzed using quantitative reverse transcription PCR (qRT-PCR) and western blot. Cell DDP sensitivity, proliferation, apoptosis, invasion, and migration were detected by cell counting kit-8 assay, 5-ethynyl-2'-deoxyuridine (EDU) assay, flow cytometry and western blot, respectively. The binding interaction was verified using dual-luciferase reporter assay. A murine xenograft model was established to investigate effects in vivo. RESULTS: Circ_0010235 was highly expressed in DDP-resistant lung cancer tissues and cells. Knockdown of circ_0010235 elevated DDP sensitivity, constrained proliferation, invasion and migration as well as fostered apoptosis in DDP-resistant lung cancer cells. Moreover, circ_0010235 silencing boosted DDP sensitivity and impeded tumor growth in lung cancer in vivo. Mechanistically, circ_0010235 acted as a sponge for miR-379-5p to elevate the expression of its target E2F7. Rescue experiments showed that miR-379-5p inhibition attenuated circ_0010235 knockdown-evoked reduction on DDP resistance of DDP-resistant cancer cells. In addition, miR-379-5p re-expression elevated DDP sensitivity and suppressed the malignant phenotype of DDP-resistant lung cancer cells through miR-379-5p. CONCLUSION: Circ_0010235 knockdown reduced DDP resistance and tumor growth via miR-379-5p/ E2F7 axis in lung cancer, suggesting an effective therapeutic target for lung cancer patients.


Subject(s)
Lung Neoplasms , MicroRNAs , RNA, Circular , Animals , Humans , Mice , Apoptosis , Cell Count , Cell Proliferation , Cisplatin/pharmacology , Cisplatin/therapeutic use , Drug Resistance, Neoplasm/genetics , E2F7 Transcription Factor , Lung Neoplasms/drug therapy , Lung Neoplasms/genetics , MicroRNAs/genetics , RNA, Circular/genetics , RNA, Circular/metabolism
14.
Appl Microbiol Biotechnol ; 95(1): 189-99, 2012 Jul.
Article in English | MEDLINE | ID: mdl-22592554

ABSTRACT

Furfural from lignocellulosic hydrolysates is the prevalent inhibitor to microorganisms during cellulosic ethanol production, but the molecular mechanisms of tolerance to this inhibitor in Zymomonas mobilis are still unclear. In this study, genome-wide transcriptional responses to furfural were investigated in Z. mobilis using microarray analysis. We found that 433 genes were differentially expressed in response to furfural. Furfural up- or down-regulated genes related to cell wall/membrane biogenesis, metabolism, and transcription. However, furfural has a subtle negative effect on Entner-Doudoroff pathway mRNAs. Our results revealed that furfural had effects on multiple aspects of cellular metabolism at the transcriptional level and that membrane might play important roles in response to furfural. This research has provided insights into the molecular response to furfural in Z. mobilis, and it will be helpful to construct more furfural-resistant strains for cellulosic ethanol production.


Subject(s)
Furaldehyde/pharmacology , Gene Expression Profiling , Gene Expression Regulation, Bacterial/drug effects , Stress, Physiological , Zymomonas/physiology , Biotechnology , Ethanol/metabolism , Furaldehyde/metabolism , Genome, Bacterial , Hydrolysis , Lignin/metabolism , Oligonucleotide Array Sequence Analysis , Zymomonas/genetics , Zymomonas/growth & development
15.
Virol J ; 8: 126, 2011 Mar 18.
Article in English | MEDLINE | ID: mdl-21414233

ABSTRACT

BACKGROUND: Porcine circovirus type 2 (PCV2) is the causative agent of postweaning multisystemic wasting syndrome (PMWS), and porcine dermatitis and nephropathy syndrome (PDNS). It has caused heavy losses in global agriculture in recent decades. Rapid detection of PCV2 is very important for the effective prophylaxis and treatment of PMWS. RESULTS: A loop-mediated isothermal amplification (LAMP) assay was used to detect PCV2 in this study. Three pairs of primers were specially designed for recognizing eight distinct sequences of the ORF2 gene. This gene lies in the PCV2 virus genome sequence, and encodes the Rep protein that is involved in virus replication. Time and temperature conditions for amplification of PCV2 genes were optimized to be 55 min at 59°C. The analysis of clinical samples indicated that the LAMP method was highly sensitive. The detection limit for PCV2 by the LAMP assay was 10 copies, whereas the limit by conventional PCR was 1000 copies. The assay did not cross-react with PCV1, porcine reproductive and respiratory syndrome virus, porcine epidemic diarrhea virus, transmissible gastroenteritis of pigs virus or rotavirus. When 110 samples were tested using the established LAMP system, 95 were detected as positive. CONCLUSION: The newly developed LAMP detection method for PCV2 was more specific, sensitive, rapid and simple than before. It complements and extends previous methods for PCV2 detection and provides an alternative approach for detection of PCV2.


Subject(s)
Circovirus/isolation & purification , Nucleic Acid Amplification Techniques/methods , Porcine Postweaning Multisystemic Wasting Syndrome/virology , Animals , Circovirus/genetics , Porcine Postweaning Multisystemic Wasting Syndrome/diagnosis , Sensitivity and Specificity , Swine , Viral Proteins/genetics
16.
J AOAC Int ; 94(4): 1227-32, 2011.
Article in English | MEDLINE | ID: mdl-21919356

ABSTRACT

The validation of the anthocyanin synthase (ANS) gene as a carnation endogenous reference gene applicable both in classical and real-time PCR methods is a prerequisite for the development of PCR assays for genetically modified (GM) carnation detection. This is important due to the fact that GM carnation lines, developed by Florigene Pty Ltd, have been approved for commercialization. In this study, both methods were tested on 14 different carnation cultivars, and identical amplification products were obtained with all of them. No amplification products were observed with samples from 14 other plant species, which demonstrated that the system was specific to carnation. The results of Southern blot analysis confirmed that the ANS gene had a low copy number in the 10 tested carnation varieties. In qualitative and real-time PCR assays, the LOD values of 0.05 and 0.005 ng carnation DNA, respectively, were validated. Moreover, the real-time PCR system was validated with high PCR efficiency and linearity. Thus, the ANS gene had species specificity, low heterogeneity, and low copy number among the tested cultivars. These results provide evidence that the gene can be used as an endogenous reference gene of carnation, as well as in qualitative and quantitative PCR systems.


Subject(s)
Dianthus/genetics , Genes, Plant/genetics , Polymerase Chain Reaction/methods , Anthocyanins/genetics , Anthocyanins/metabolism , DNA, Plant/analysis , Gene Expression Regulation, Plant , Reproducibility of Results , Sensitivity and Specificity , Species Specificity
17.
Bioprocess Biosyst Eng ; 34(3): 305-14, 2011 Mar.
Article in English | MEDLINE | ID: mdl-20963445

ABSTRACT

The full-length cDNA encoding an acetylcholinesterase (AChE) was cloned and sequenced from the housefly, Musca domestica, by reverse transcriptase-polymerase chain reaction (RT-PCR). Sequence analysis revealed that this 2,076 bp sequence encodes a mature protein of 612 amino acids (67 kDa) and a 79 residue signal peptide. The amino acid sequence shared 52.8-81.4% identity with the AChE proteins of other insects. The cDNA sequence, which lacked the signal peptide was inserted into the vector pPIC9K and then introduced into strain GS115 of the yeast Pichia pastoris. The recombinant AChE protein was then expressed in P. pastoris strain GS115 by methanol induction. Site-directed mutagenesis of the A262G, Y327F, Y327D and I374D residues, either singly or in combination, was performed by reverse PCR. These mutants improved the catalytic activity and sensitivity to the organophosphate and carbamate insecticides. Although the sensitivity of other mutants was slightly increased, the results still showed that the sensitivity of triple mutant, GDD (A262G/Y327D/I374D), enhanced remarkably as much as 16 times for methomyl, 14 times for both carbofuran and chlorpyrifos, and ten times for parathion-methyl, compared to that of the wild-type. The results strongly suggested that these residues are the key structural elements controlling AChE enzyme catalytic activity and sensitivity to inhibition by insecticides. The AChE enzyme obtained by this method could be used to detect the organophosphate and carbamate insecticide residues in fruits and vegetables, a characteristic of great potential research and industrial application.


Subject(s)
Acetylcholinesterase/chemistry , Acetylcholinesterase/genetics , Food Contamination/analysis , Houseflies/enzymology , Pesticide Residues/analysis , Acetylcholinesterase/biosynthesis , Acetylcholinesterase/metabolism , Amino Acid Sequence , Animals , Carbamates/analysis , Cholinesterase Inhibitors/analysis , Houseflies/metabolism , Insecticide Resistance/genetics , Molecular Sequence Data , Mutagenesis, Site-Directed/methods , Organophosphates/analysis , Pichia/genetics , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism
18.
Chemosphere ; 279: 130592, 2021 Sep.
Article in English | MEDLINE | ID: mdl-34134411

ABSTRACT

The bioavailability of heavy metal and growth of hyperaccumulator are key factors controlling the phytoextraction of heavy metal from soil. In this study, the efficacy and potential microbial mechanisms of digestate application in enhancing Cd extraction from soil by Pennisetum hybridum were investigated. The results showed that digestate application significantly promoted the height, tiller number, and biomass yield of Pennisetum hybridum. The application also increased the activities of urease, sucrase, dehydrogenase, available Cd contents of rhizosphere soils (from 2.21 to 2.46 mg kg-1), and the transfer factors of Cd from root to shoot and leaf. Assuming three annual harvests, digestate application would substantially reduce time needed for Pennisetum hybridum to completely absorb Cd from soil-from 15-16 yr-10 yr. Furthermore, the results of microbial community diversity analysis showed that digestate irrigation was more facilitated for the growth of the predominant bacteria, which were Actinobacteria and Chloroflexi at phylum level, and Sphingomonas and Nitrospiraat genus level, which mainly have the functions of promoted plant growth and metal resistance. The results suggested that the enhanced phytoextraction of Cd by Pennisetum hybridum with digestate application might mainly attributed to the increased Cd bio-availability and the enhanced plant growth, indicating that an approach combining digestate and Pennisetum hybridum could be a promising strategy for remediating Cd-contaminated soils.


Subject(s)
Pennisetum , Soil Pollutants , Biodegradation, Environmental , Cadmium/analysis , Soil , Soil Pollutants/analysis
19.
Bioresour Technol ; 321: 124497, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33307481

ABSTRACT

Low ability of waste sewage sludge to degrade cellulose is observed due to its less cellulolytic bacteria content. The enrichment of sewage sludge in the absence or presence of carboxymethylcellulose (CMC) was conducted to improve anaerobic digestion (AD) of cellulose in this study. Compared to initial sewage sludge (IS), enriched sludge without CMC addition (ES) displayed 69.81% higher CH4 yield and about 1.7-fold greater anaerobic biodegradation of cellulose. In particular, bacterial and archaeal diversities in samples inoculated with ES were significantly altered, with Ruminiclostridium and Methanobacterium as the predominant genera. Enriched sludge with CMC addition (ESC) displayed enhanced methane production at initial cellulose fermentation but showed no distinct difference compared with the control after incubation 24 days. These findings suggest that enrichment of waste sewage sludge without CMC addition is more beneficial for promoting AD of cellulose, providing a novel insight for efficient energy utilization of lignocellulosic wastes.


Subject(s)
Cellulose , Sewage , Anaerobiosis , Archaea/metabolism , Bioreactors , Cellulose/metabolism , Methane
20.
ACS Omega ; 6(13): 8816-8828, 2021 Apr 06.
Article in English | MEDLINE | ID: mdl-33842753

ABSTRACT

This paper reports the mineralogical and geochemical compositions of C6 coal in the Late Permian Longtan Formation of the Wenjiaba Mine, Northern Guizhou in southwest (SW) China. The geochemical and mineralogical studies are the basis for the potential recovery of critical metals. The Longtan Formation, which is one of the major coal-bearing strata in SW China, contains dozens of coal seams. C6 coal is the main mineable coal seam in the Wenjiaba Mine and the whole coalfield. Proximate and ultimate analyses, inductively coupled plasma mass spectrometry (ICP-MS) and X-ray fluorescence (XRF) spectrometry on trace and major element concentrations, and X-ray diffraction and SEM-EDS analyses were carried out. Results suggest that this anthracite coal is characterized by low ash yield and medium sulfur content. The minerals are mainly composed of clay minerals (kaolinite, chlorite, illite, and mixed-layer illite/smectite), pyrite, and carbonates. Lithium is significantly enriched in C6 coal, with an average of 124 µg/g, and it has a higher concentration in the lower portion of the coal seam than that in the upper one. Strontium is significantly enriched in samples WJB-05 and WJB-06, with concentrations of 3030 and 4580 µg/g, respectively, but it is normal or just slightly enriched in other benches of C6 coal. Additionally, Cu, Nb, and Ta are slightly enriched in the coal. Lithium, dominantly hosted by kaolinite in C6 coals, has a recovery potential. Celestine is one of the major Sr-bearing minerals in C6 coal.

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