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1.
China Journal of Chinese Materia Medica ; (24): 1073-1078, 2021.
Article in Chinese | WPRIM | ID: wpr-879006

ABSTRACT

The study aiming at exploring the potassium-dissolving capacity of rhizosphere potassium-dissolving bacteria from diffe-rent sources and screen the strains with high potassium-dissolving ability, so as to lay a theoretical foundation for cultivation and quality improvement of Paris polyphylla var. yunnanensis sources. The rhizosphere soil of 10 wild and transplanted species from Yunnan, Sichuan and Guizhou provinces was used as the research object. Potassium-dissolving bacteria were isolated and purified, and their potassium-dissolving capacity was determined by flame spectrophotometry, and identified by physiological, biochemical and molecular biological methods. Twenty-six potassium-dissolving bacteria were purified and 13 were obtained from wild and transplanted strains respectively. It was found through the determination of potassium-dissolving capacity that the potassium-dissolving capacity of 26 strains was significantly different, and the mass concentration of K~+ in the fermentation broth were 1.04-2.75 mg·L~(-1), the mcentration of potassium were 0.01-1.82 mg·L~(-1). The strains were identified as Bacillus, Agrobacterium rhizome and Staphylococcus by physiological, biochemical and 16 S rDNA molecular methods, among them Bacillus amylolyticus(4 strains) was the dominant bacterium of Bacillus. The physiology and biochemistry of rhizosphere potassium-dissolving bacteria in P. polyphylla var. yunnanensis rhizosphere were diffe-rent, and the living environment were different, so the potassium-dissolving capacity also changed. Strain Y4-1 with the highest potassium decomposability was Bacillus amylolytic with a potassium increase of 1.82 mg·L~(-1). The potassium-dissolving ability and the distribution of potassium-dissolving bacteria were different in various habitats. The screening of potassium-dissolving bacteria provided a new strain for the preparation of microbial fertilizer. It is expected that B. amyloidococcus Y4-1 can be used as an ideal strain to cultivate mycorrhizal seedlings of P. polyphylla var. yunnanensis.


Subject(s)
China , Liliaceae , Paenibacillus , Potassium , Rhizosphere , Soil
2.
Electron. j. biotechnol ; 47: 43-50, sept. 2020. tab, graf
Article in English | LILACS | ID: biblio-1253024

ABSTRACT

BACKGROUND: Rice sheath blight (caused by Rhizoctonia solani) and tobacco mosaic virus are very important plant diseases, causing a huge loss in global crop production. Paenibacillus kribbensis PS04 is a broad-spectrum biocontrol agent, used for controlling these diseases. Previously, extracellular polysaccharides (EPS) from P. kribbensis PS04 had been purified and their structure was inferred to be fructosan. This study aimed to evaluate the effects of exogenous EPS treatment on plant­pathogen interactions. RESULTS: Plant defense genes such as phenylalanine ammonia-lyase, catalase, chitinase, allene oxide synthase, and PR1a proteins were significantly induced by exogenous EPS treatment. Moreover, subsequent challenge of EPSpretreated plants with the pathogens (R. solani or tobacco mosaic virus) resulted in higher expression of defenseassociated genes. Increased activities of defense-associated enzymes, total phenols, and flavonoids were also observed in EPS pretreated plants. The contents of malondialdehyde in plants, which act as indicator of lipid peroxidation, were reduced by EPS treatment. CONCLUSIONS: This study comprehensively showed that EPS produced from P. kribbensis PS04 enhances disease resistance in plants by the activation of defense-associated genes as well as through the enhancement of activities of defense-related enzymes.


Subject(s)
Plant Diseases/immunology , Rhizoctonia/pathogenicity , Tobacco Mosaic Virus/pathogenicity , Paenibacillus/immunology , Plant Diseases/microbiology , Polysaccharides, Bacterial , Pest Control, Biological , Host-Pathogen Interactions , Paenibacillus/genetics , Disease Resistance/genetics , Real-Time Polymerase Chain Reaction , Fructose/analogs & derivatives
3.
Rev. argent. microbiol ; 51(1): 77-80, mar. 2019. graf
Article in English | LILACS | ID: biblio-1041818

ABSTRACT

Cohnella is a highly cellulolytic bacterial genus, which can be found in a variety of habitats. The aim of this study was to assess its presence in the digestive tract of termite species collected in North-eastern Argentina: Nasutitermes aquilinus, N. corniger and Cortaritermes fulviceps. Gut homogenates were incubated with cellulosic substrate for bacterial growth. Bacterial 16S rDNA was partially amplified using new primers for Cohnella spp. and cloned. Sequences obtained showed highest similarity (97.2-99.9%) with those of Cohnella spp. previously reported from diverse environments. Phylogenetic analysis tended to group the clones according to their host species and sampling sites. These results indicate the association of Cohnella-related intestinal symbionts with three common Neotropical termites. Their potential industrial application encourages further research.


Cohnella es un género de bacterias celulolíticas que puede ser encontrado en una variedad de hábitats. El propósito de este estudio fue registrar su presencia en el tracto digestivo de termitas (Nasutitermes aquilinus, N. corniger y Cortaritermes fulviceps) colectadas en el noreste argentino (NEA). Se incubaron homogenados de intestinos en sustrato celulósico para multiplicar las bacterias. Utilizando nuevos cebadores para Cohnella spp., se amplificó una porción del ADN ribosomal 16S bacteriano, el cual fue posteriormente clonado. Las secuencias obtenidas mostraron su mayor porcentaje de similitud (97,2-99,9%) con Cohnella spp., previamente reportadas en diversos ambientes. El análisis filogenético tendió a agrupar a los clones de acuerdo a la especie hospedante y al sitio de muestreo. Estos resultados indican que especies de termitas frecuentes en el NEA albergan simbiontes intestinales relacionados con el género Cohnella. Las potenciales aplicaciones industriales de estos microorganismos animan a profundizar los estudios.


Subject(s)
Isoptera/microbiology , Paenibacillus/isolation & purification , Paenibacillus/growth & development , Bacterial Growth/analysis , Sequence Analysis, DNA/methods , Gastrointestinal Tract/microbiology
4.
Pesqui. vet. bras ; 38(10): 1913-1917, out. 2018. tab, ilus
Article in English | LILACS, VETINDEX | ID: biblio-976384

ABSTRACT

Amorimia septentrionalis is an important sodium monofluoroacetate (MFA) containing plant that causes sudden death in ruminants in northeastern Brazil. MFA degrading bacteria are being used in the prevention against poisoning by this plant. The aim of this study was to evaluate if goats which had per os received MFA degrading bacteria remained resistant when exposed to natural poisoning by A. septentrionalis. Eighteen goats were randomly distributed into three groups: the goats of Group 1 previously received, during 40 days, a solution containing the bacteria Ralstonia sp. and Burkholderia sp., those goats in the Group 2 received the bacteria Paenibacillus sp. and Cupriavidus sp. and goats from Group 3 did not receive any bacteria. After the administration period, during 60 days, the animals of all groups were released to graze on a one hectare paddock, with significant amount of A. septentrionalis. They were observed daily for the spontaneous consumption of A. septentrionalis leaves and the occurrence of clinical signs of poisoning or sudden death. Goats from all groups consumed significant amounts of A. septentrionalis during the experimental period. Goats that did not receive MFA-degrading bacteria (Group 3) became sick and died from the 25th to the 27th day of the experiment, whereas the goats of the groups that received MFA-degrading bacteria showed only clinical sings when A. septentrionalis regrowth after the 55th day of the experiment. The days elapsed from field observation to death of Group 3 goats (25.5±0.9 days) were significantly lower (p<0.05) than Group 1 (58.6±1.3 days) and Group 2 (57.8±1.5 days). Thus, it can be concluded that administration of MFA degrading bacteria increases the resistance to natural poisoning by A. septentrionalis.(AU)


Amorimia septentrionalis que contém monofluoroacetato de sódio (MFA) é responsável pela ocorrência de mortes súbitas em ruminantes no nordeste do Brasil. Bactérias degradadoras desse composto estão sendo utilizadas na prevenção contra a intoxicação por essa planta. O objetivo deste estudo foi avaliar se caprinos que receberam, via oral, bactérias degradadoras de MFA permaneciam resistentes quando expostos a intoxicação natural por A. septentrionalis. Dezoito caprinos foram divididos em três grupos, os caprinos do Grupo 1 receberam anteriormente, durante 40 dias, uma solução contendo as bactérias Ralstonia sp. e Burkholderia sp., os do Grupo 2 receberam, também por 40 dias as bactérias Paenibacillus sp. e Cupriavidus sp. e os do Grupo 3 não receberam nenhuma bactéria. Após o período de administração, durante 60 dias, os animais de todos os grupos foram soltos para pastar em um piquete de um hectare, que apresentava uma quantidade significativa da planta. Diariamente eles foram observados quanto ao consumo espontâneo das folhas de A. septentrionalis e quanto à presença de sinais clínicos de intoxicação ou morte. Os caprinos de todos os grupos consumiram quantidades significantes da planta durante o período experimental. Os caprinos que não receberam as bactérias degradantes de MFA (Grupo 3) adoeceram e morreram entre o 25º e o 27º dia de experimento, enquanto que os que receberam as bactérias degradantes de MFA (Grupo 1 e 2) só apresentaram sinais clínicos no 55º dia de experimento, o que coincidiu com a rebrota da planta. Os dias transcorridos desde a observação a campo até a morte dos caprinos do Grupo 3 (25,5±0,9 dias) foram significativamente menores (p<0,05) que os do Grupo 1 (58,6±1,3 dias) e do Grupo 2 (57,8±1,5 dias). Com isso pode-se concluir que a administração de bactérias degradadoras de MFA aumenta à resistência a intoxicação natural por A. septentrionalis.(AU)


Subject(s)
Animals , Plant Poisoning/therapy , Plant Poisoning/veterinary , Bacteria/enzymology , Ruminants , Malpighiaceae/poisoning , Fluoroacetates/antagonists & inhibitors , Burkholderia , Ralstonia , Cupriavidus , Paenibacillus
5.
Braz. j. microbiol ; 48(4): 656-670, Oct.-Dec. 2017. tab, graf
Article in English | LILACS | ID: biblio-889178

ABSTRACT

ABSTRACT This study aimed to explore the effects of two siderophore-producing bacterial strains on iron absorption and plant growth of peanut in calcareous soil. Two siderophore-producing bacterial strains, namely, YZ29 and DZ13, isolated from the rhizosphere soil of peanut, were identified as Paenibacillus illinoisensis and Bacillus sp., respectively. In potted experiments, YZ29 and DZ13 enhanced root activity, chlorophyll and active iron content in leaves, total nitrogen, phosphorus and potassium accumulation of plants and increased the quality of peanut kernels and plant biomass over control. In the field trial, the inoculated treatments performed better than the controls, and the pod yields of the three treatments inoculated with YZ29, DZ13, and YZ29 + DZ13 (1:1) increased by 37.05%, 13.80% and 13.57%, respectively, compared with the control. Based on terminal restriction fragment length polymorphism analysis, YZ29 and DZ13 improved the bacterial community richness and species diversity of soil surrounding the peanut roots. Therefore, YZ29 and DZ13 can be used as candidate bacterial strains to relieve chlorosis of peanut and promote peanut growth. The present study is the first to explore the effect of siderophores produced by P. illinoisensis on iron absorption.


Subject(s)
Arachis/growth & development , Arachis/microbiology , Bacillus/metabolism , Paenibacillus/metabolism , Iron/metabolism , Arachis/metabolism , Arachis/chemistry , Seeds/growth & development , Seeds/metabolism , Seeds/microbiology , Seeds/chemistry , Soil/chemistry , Soil Microbiology , Bacillus/isolation & purification , Bacillus/classification , Bacillus/genetics , Biological Transport , Siderophores/metabolism , Plant Roots/microbiology , Paenibacillus/isolation & purification , Paenibacillus/classification , Paenibacillus/genetics , Rhizosphere , Agricultural Inoculants/metabolism
6.
Laboratory Medicine Online ; : 201-205, 2017.
Article in Korean | WPRIM | ID: wpr-51168

ABSTRACT

Paenibacillus urinalis was first isolated from the urine of a woman in 2008, and was reported to be a contaminant. Here, we report 5 cases of P. urinalis isolated over 5 months at a tertiary hospital. Using an API kit, 4 cases were classified as Cellulomonas species. Owing to the low reliability of API kit results and Gram stain results indicating gram variable bacilli for few specimens, MALDI-TOF MS and 16S rRNA gene sequencing were performed for identification. The last case showed Gram variable bacilli, and therefore, based on previous experience, 16S rRNA gene base sequence analysis was carried out without an additional API kit. All isolated strains were confirmed to be P. urinalis, and were judged to be contaminants. As for Gram variable bacteria, the use of current biochemical identification systems may lead to misidentification as other bacteria, which may cause unnecessary or improper use of antibiotics. Moreover, whereas most of the Paenibacillus species are reported to be contaminants, some of them are being reported as sources of infection. Therefore, more accurate identification will be necessary in the future. Accordingly, it is expected that accurate identification of this genus will help clinical physicians make decisions regarding appropriate treatment and use of antibiotics.


Subject(s)
Female , Humans , Anti-Bacterial Agents , Bacteria , Base Sequence , Cellulomonas , Genes, rRNA , Paenibacillus , Tertiary Care Centers
7.
Electron. j. biotechnol ; 19(1): 29-32, Jan. 2016. ilus
Article in English | LILACS | ID: lil-781167

ABSTRACT

Background: Microbial-induced remediation of Zn2+ pollution based on the capture and utilization of carbon dioxide was investigated. In this study, carbon dioxide was absorbed and transformed into carbonate ions under the enzymatic action of Paenibacillus mucilaginosus, which was being utilized to mineralize Zn2+. Results: The compositional and morphological properties of the precipitations were studied using Fourier transform infrared spectroscopy (FTIR), energy dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), and scanning electron microscopy (SEM). The thermal properties of the precipitates were investigated by thermogravimetric-differential scanning calorimetry (TG-DSC). The FTIR results confirmed that the functional groups of the precipitates were CO3² − and OH−. The XRD and EDS patterns showed that basic zinc carbonate could be obtained successfully by Microbial-induced remediation. The SEM micrographs demonstrated that the precipitates were in the nanometer range with sizes of 100-200 nm and were sphere-like in shape. Conclusions: The TG-DSC results showed that weight loss of the precipitates occurred around 253°C. The FTIR and TG-DSC results were in accord with the XRD and EDS results and proved again that the precipitates were basic zinc carbonate. This work thus demonstrates a new method for processing Zn2+ pollution based on the utilization of carbon dioxide.


Subject(s)
Zinc/metabolism , Carbon Dioxide/metabolism , Environmental Restoration and Remediation/methods , Paenibacillus , Spectrometry, X-Ray Emission , Thermogravimetry , X-Ray Diffraction , Spectroscopy, Fourier Transform Infrared , Biomineralization
8.
Braz. j. microbiol ; 46(4): 977-989, Oct.-Dec. 2015. tab, graf
Article in English | LILACS | ID: lil-769669

ABSTRACT

Abstract A total of 48 endophytic bacteria were isolated from surface-sterilized tissues of the medicinal plant Lonicera japonica, which is grown in eastern China; six strains were selected for further study based on their potential ability to promote plant growth in vitro (siderophore and indoleacetic acid production). The bacteria were characterized by phylogenetically analyzing their 16S rRNA gene similarity, by examining their effect on the mycelial development of pathogenic fungi, by testing their potential plant growth-promoting characteristics, and by measuring wheat growth parameters after inoculation. Results showed that the number of endophytic bacteria in L. japonica varied among different tissues, but it remained relatively stable in the same tissues from four different plantation locations. Among the three endophytic strains, strains 122 and 124 both had high siderophore production, with the latter showing the highest phosphate solubilization activity (45.6 mg/L) and aminocyclopropane-1-carboxylic acid deaminase activity (47.3 nmol/mg/h). Strain 170 had the highest indoleacetic acid (IAA) production (49.2 mg/L) and cellulase and pectinase activities. After inoculation, most of the six selected isolates showed a strong capacity to promote wheat growth. Compared with the controls, the increase in the shoot length, root length, fresh weight, dry weight, and chlorophyll content was most remarkable in wheat seedlings inoculated with strain 130. The positive correlation between enzyme (cellulose and pectinase) activity and inhibition rate on Fusarium oxysporum, the IAA production, and the root length of wheat seedlings inoculated with each tested endophytic strain was significant in regression analysis. Deformity of pathogenic fungal mycelia was observed under a microscope after the interaction with the endophytic isolates. Such deformity may be directly related to the production of hydrolytic bacterial enzymes (cellulose and pectinase). The six endophytic bacterial strains were identified to be Paenibacillus and Bacillus strains based on the results of 16S rRNA gene sequencing analysis and their physiological and biochemical characteristics. Results indicate the promising application of endophytic bacteria to the biological control of pathogenic fungi and the improvement of wheat crop growth.


Subject(s)
Bacillus/classification , Bacillus/genetics , Bacillus/growth & development , Bacillus/isolation & purification , Bacillus/metabolism , Bacillus/microbiology , China/classification , China/genetics , China/growth & development , China/isolation & purification , China/metabolism , China/microbiology , Endophytes/classification , Endophytes/genetics , Endophytes/growth & development , Endophytes/isolation & purification , Endophytes/metabolism , Endophytes/microbiology , Indoleacetic Acids/classification , Indoleacetic Acids/genetics , Indoleacetic Acids/growth & development , Indoleacetic Acids/isolation & purification , Indoleacetic Acids/metabolism , Indoleacetic Acids/microbiology , Lonicera/classification , Lonicera/genetics , Lonicera/growth & development , Lonicera/isolation & purification , Lonicera/metabolism , Lonicera/microbiology , Molecular Sequence Data/classification , Molecular Sequence Data/genetics , Molecular Sequence Data/growth & development , Molecular Sequence Data/isolation & purification , Molecular Sequence Data/metabolism , Molecular Sequence Data/microbiology , Paenibacillus/classification , Paenibacillus/genetics , Paenibacillus/growth & development , Paenibacillus/isolation & purification , Paenibacillus/metabolism , Paenibacillus/microbiology , Phylogeny/classification , Phylogeny/genetics , Phylogeny/growth & development , Phylogeny/isolation & purification , Phylogeny/metabolism , Phylogeny/microbiology , Plant Roots/classification , Plant Roots/genetics , Plant Roots/growth & development , Plant Roots/isolation & purification , Plant Roots/metabolism , Plant Roots/microbiology , Siderophores/classification , Siderophores/genetics , Siderophores/growth & development , Siderophores/isolation & purification , Siderophores/metabolism , Siderophores/microbiology , Triticum/classification , Triticum/genetics , Triticum/growth & development , Triticum/isolation & purification , Triticum/metabolism , Triticum/microbiology
9.
Braz. j. microbiol ; 46(2): 367-375, Apr-Jun/2015. tab, graf
Article in English | LILACS | ID: lil-749711

ABSTRACT

The pH of the culture medium directly influences the growth of microorganisms and the chemical processes that they perform. The aim of this study was to assess the influence of the initial pH of the culture medium on the production of 11 low-molecular-weight organic acids and on the solubilization of calcium phosphate by bacteria in growth medium (NBRIP). The following strains isolated from cowpea nodules were studied: UFLA03-08 (Rhizobium tropici), UFLA03-09 (Acinetobacter sp.), UFLA03-10 (Paenibacillus kribbensis), UFLA03-106 (Paenibacillus kribbensis) and UFLA03-116 (Paenibacillus sp.). The strains UFLA03-08, UFLA03-09, UFLA03-10 and UFLA03-106 solubilized Ca3(PO4)2 in liquid medium regardless of the initial pH, although without a significant difference between the treatments. The production of organic acids by these strains was assessed for all of the initial pH values investigated, and differences between the treatments were observed. Strains UFLA03-09 and UFLA03-10 produced the same acids at different initial pH values in the culture medium. There was no correlation between phosphorus solubilized from Ca3(PO4)2 in NBRIP liquid medium and the concentration of total organic acids at the different initial pH values. Therefore, the initial pH of the culture medium influences the production of organic acids by the strains UFLA03-08, UFLA03-09, UFLA03-10 and UFLA03-106 but it does not affect calcium phosphate solubilization.


Subject(s)
Acinetobacter/metabolism , Carboxylic Acids/metabolism , Culture Media/chemistry , Paenibacillus/metabolism , Phosphates/metabolism , Rhizobium tropici/metabolism , Acinetobacter/growth & development , Acinetobacter/isolation & purification , Fabaceae/microbiology , Hydrogen-Ion Concentration , Paenibacillus/growth & development , Paenibacillus/isolation & purification , Rhizobium tropici/growth & development , Rhizobium tropici/isolation & purification , Root Nodules, Plant/microbiology
10.
Infection and Chemotherapy ; : 64-67, 2015.
Article in English | WPRIM | ID: wpr-221777

ABSTRACT

We report the first case of bacteremia by a novel Paenibacillus species, Paenibacillus pasadenensis, from a 55-year-old male patient with acute respiratory distress syndrome, following a microsurgical clipping procedure of a ruptured intracranial aneurysm. The bacterium was identified using 16S rRNA gene sequencing analysis, which was applied because current conventional methods employed in the clinical microbiology laboratory proved unsuccessful. Since this bacterium was first identified in 2006 and has never been reported elsewhere, we believe this report can provide practitioners with useful insight on the pathogenicity of this species.


Subject(s)
Humans , Male , Middle Aged , Bacteremia , Genes, rRNA , Intracranial Aneurysm , Paenibacillus , Respiratory Distress Syndrome , RNA, Ribosomal , Virulence
11.
Chinese Journal of Biotechnology ; (12): 1401-1407, 2015.
Article in Chinese | WPRIM | ID: wpr-337480

ABSTRACT

Auxotrophic strains of N1-37 (Phe-) and N2-27 (His-), screened from mutations of Paenibacillus polymyxa JSa-9 previously, were used as the parent strains to screen high-producing LI-F antibacterial lipopeptide fusion strain through protoplast fusion with polyethylene glycol as a promote agent. Fusion strain F5-15 was obtained. Then the product of LI-F antibacterial lipopeptide was quantified by HPLC, and the difference of expression of the key genes of lipopeptide synthase between wild strain JSa-9 and the fusion strain was analyzed by real-time PCR. LI-F antibacterial lipopeptide yield of the fusion strain F5-15 was 3.1-fold of the original strain JSa9's, and the expression levels of the target genes were 10.48, 2.48, 2.1 and 11.8 fold of the initial strain JSa-9, respectively.


Subject(s)
Anti-Bacterial Agents , Chromatography, High Pressure Liquid , Lipopeptides , Paenibacillus , Metabolism , Protoplasts , Metabolism , Real-Time Polymerase Chain Reaction
12.
Mycobiology ; : 158-166, 2014.
Article in English | WPRIM | ID: wpr-729230

ABSTRACT

In this study, bacterial strains were isolated from soils from 30 locations of Samcheok, Gangwon province. Of the isolated strains, seven showed potential plant growth promoting and antagonistic activities. Based on cultural and morphological characterization, and 16S rRNA gene sequencing, these strains were identified as Paenibacillus species. All seven strains produced ammonia, cellulase, hydrocyanic acid, indole-3-acetic acid, protease, phosphatase, and siderophores. They also inhibited the mycelial growth of Fusarium oxysporum f. sp. radicis-lycopersici in vitro. The seven Paenibacillus strains enhanced a range of growth parameters in tomato plants under greenhouse conditions, in comparison with non-inoculated control plants. Notably, treatment of tomato plants with one identified strain, P. polymyxa SC09-21, resulted in 80.0% suppression of fusarium crown and root rot under greenhouse conditions. The plant growth promoting and antifungal activity of P. polymyxa SC09-21 identified in this study highlight its potential suitability as a bioinoculant.


Subject(s)
Ammonia , Cellulase , Crowns , Fusarium , Genes, rRNA , Hydrogen Cyanide , Solanum lycopersicum , Paenibacillus , Plants , Plasmodiophorida , Siderophores , Soil
13.
Braz. j. microbiol ; 45(1): 193-197, 2014. ilus, tab
Article in English | LILACS, VETINDEX | ID: biblio-1469605

ABSTRACT

In screening the culturable endoglucanase-producing bacteria in the rhizosphere of Rhizophora mangle, we found a prevalence of genera Bacillus and Paenibacillus. These bacteria revealed different activities in endoglucolysis and biofilm formation when exposed to specific NaCl concentrations, indicating modulated growth under natural variations in mangrove salinity.


Subject(s)
Bacillus , Sodium Chloride , Glucans , Paenibacillus , Rhizophoraceae , Rhizosphere
14.
Chinese Journal of Biotechnology ; (12): 98-108, 2014.
Article in Chinese | WPRIM | ID: wpr-242408

ABSTRACT

By engineering the subsite +1 of cyclodextrin glycosyltransferase (CGTase) from Paenibacillus macerans, we improved its maltodextrin specificity for 2-O-D-glucopyranosyl-L-ascorbic acid (AA-2G) synthesis. Specifically, we conducted site-saturation mutagenesis on Leu194, Ala230, and His233 in subsite +1 separately and gained 3 mutants L194N (leucine --> asparagine), A230D (alanine --> aspartic acid), and H233E (histidine --> glutamic acid) produced higher AA-2G yield than the wild-type and the other mutant CGTases. Therefore, the 3 mutants L194N, A230D, and H233E were further used to construct the double and triple mutations. Among the 7 obtained combinational mutants, the triple mutant L194N/A230D/H233E produced the highest AA-2G titer of 1.95 g/L, which was increased by 62.5% compared with that produced by the wild-type CGTase. Then, we modeled the reaction kinetics of all the mutants and found a substrate inhibition by high titer of L-AA for the mutants. The optimal temperature, pH, and reaction time of all the mutants were also determined. The structure modeling indicated that the enhanced maltodextrin specificity may be related with the changes of hydrogen bonding interactions between the side chain of residue at the three positions (194, 230 and 233) and the substrate sugars.


Subject(s)
Ascorbic Acid , Chemistry , Glucosyltransferases , Genetics , Metabolism , Hydrogen Bonding , Kinetics , Mutagenesis, Site-Directed , Paenibacillus , Polysaccharides , Chemistry , Protein Engineering , Substrate Specificity , Temperature
15.
Neonatal Medicine ; : 69-73, 2014.
Article in Korean | WPRIM | ID: wpr-43775

ABSTRACT

Paenibacillus spp. are gram-positive, rod-shaped, facultative anaerobic bacteria found in nature and rarely cause diseases in humans. We report our experience with Paenibacillus-induced sepsis complicated with pneumatocele in a very low birth weight male infant with a gestational age of 29 weeks and 5 days and a birth weight of 1,380 g, who was born by cesarean section with because of preterm labor and premature rupture of membrane. On day 12 after admission, the patient presented oxygen desaturation without apnea and fever. We identified pleural effusion on chest radiography and diagnosed pneumatocele on low-dose chest computed tomography. An empirical antibiotic was administered to treat the infection. The patient's blood culture revealed gram-positive rods, and Paenibacillus spp. was identified using16s rRNA sequencing.


Subject(s)
Female , Humans , Infant , Male , Pregnancy , Apnea , Bacteria, Anaerobic , Birth Weight , Cesarean Section , Fever , Gestational Age , Gram-Positive Rods , Infant, Very Low Birth Weight , Membranes , Obstetric Labor, Premature , Oxygen , Paenibacillus , Pleural Effusion , Radiography , Rupture , Sepsis , Thorax
16.
Rev. argent. microbiol ; 45(4): 257-61, dic. 2013.
Article in Spanish | LILACS, BINACIS | ID: biblio-1171796

ABSTRACT

American foulbrood (AFB) is a bacterial disease caused by the spore-forming, grampositive bacterium Paenibacillus larvae, which affects honeybee broods worldwide. The aim of this work was to compare the Epsilometer test (Etest) to the agar dilution method for testing a collection of 22 P. larvae strains to tetracycline by using MYPGP and Iso- Sensitest agars. Results showed that a categorical agreement of 100


was found when using Iso-Sensitest, while a categorical agreement of 86.36


was found (with 3 minor errors) when MYPGP was tested. In conclusion, the Etest could be a rapid and reliable method for testing MIC values of tetracycline in P. larvae only when used in combination with Iso-Sensitest agar. Nevertheless, these results should be confirmed with future studies involving a larger number of isolates.


Subject(s)
Anti-Bacterial Agents/pharmacology , Paenibacillus/drug effects , Tetracycline/pharmacology , Bees/microbiology , Animals , Gram-Positive Bacterial Infections , Microbial Sensitivity Tests/methods
17.
Chinese Journal of Biotechnology ; (12): 1234-1244, 2013.
Article in Chinese | WPRIM | ID: wpr-242486

ABSTRACT

We studied the mutation effect of subsites -3(Lys47), -7(146-152), and cyclization center (Tyr195) in active domain on product specificity of alpha-cyclodextrin glucanotransferase (alpha-CGTase) from Paenibacillus macerans sp. 602-1. The Lys47 was replaced by Thr47 and Tyr195 by Ile195, and the amino acids from 146 to 152 were replaced by Ile (named as delta6). All these mutant alpha-CGTases were actively expressed in E. coli BL21. Compared with the wild-type alpha-CGTase, the starch-degrading activities of all the mutant enzymes were declined. For mutant Y195I, the percentage of alpha-CD was decreased from 68% to 30%, and beta-CD was raised from 22.2% to 33.3%. Interestingly, gamma-CD was increased from 8.9% to 36.7% and became the main product, while the actual yield was increased from 0.4 g/L to 1.1 g/L. Mutant K47T and delta6 still produced alpha-CD as main product though the percentage of beta- and gamma-CD increased. Purified Y195I CGTase showed similar optimum temperature with the wild-type alpha-CGTase, but its optimum pH shifted from 5.0 to 6.0 with better pH stability. In summary, mutant Y195I CGTase has the potential to produce gamma-CD as the main product.


Subject(s)
Escherichia coli , Genetics , Metabolism , Glucosyltransferases , Genetics , Metabolism , Mutant Proteins , Genetics , Metabolism , Mutation , Paenibacillus , Recombinant Proteins , Genetics , gamma-Cyclodextrins , Metabolism
18.
Mycobiology ; : 244-251, 2012.
Article in English | WPRIM | ID: wpr-729698

ABSTRACT

In vitro and greenhouse screening of seven rhizobacterial isolates, AB05, AB10, AB11, AB12, AB14, AB15 and AB17, was conducted to investigate the plant growth promoting activities and inhibition against anthracnose caused by Colletotrichum acutatum in pepper. According to identification based on 16S rDNA sequencing, the majority of the isolates are members of Bacillus and a single isolate belongs to the genus Paenibacillus. All seven bacterial isolates were capable of inhibiting C. acutatum to various degrees. The results primarily showed that antibiotic substances produced by the selected bacteria were effective and resulted in strong antifungal activity against the fungi. However, isolate AB15 was the most effective bacterial strain, with the potential to suppress more than 50% mycelial growth of C. acutatum in vitro. Moreover, antibiotics from Paenibacillus polymyxa (AB15) and volatile compounds from Bacillus subtilis (AB14) exerted efficient antagonistic activity against the pathogens in a dual culture assay. In vivo suppression activity of selected bacteria was also analyzed in a greenhouse with the reference to their prominent in vitro antagonism efficacy. Induced systemic resistance in pepper against C. acutatum was also observed under greenhouse conditions. Where, isolate AB15 was found to be the most effective bacterial strain at suppressing pepper anthracnose under greenhouse conditions. Moreover, four isolates, AB10, AB12, AB15, and AB17, were identified as the most effective growth promoting bacteria under greenhouse conditions, with AB17 inducing the greatest enhancement of pepper growth.


Subject(s)
Anti-Bacterial Agents , Bacillus , Bacillus subtilis , Bacteria , Colletotrichum , DNA, Ribosomal , Fungi , Mass Screening , Paenibacillus , Plants , Plasmodiophorida , Sprains and Strains
19.
The Korean Journal of Physiology and Pharmacology ; : 225-230, 2012.
Article in English | WPRIM | ID: wpr-728096

ABSTRACT

We investigated the effects of beta-glucan purified from Paenibacillus polymyxa JB115 on the viability and proliferation of splenocytes. Splenocytes play a critical role in host immunity. MTT assays and trypan blue exclusion tests revealed that beta-glucan significantly promoted the viability and proliferation of splenocytes over a range of concentrations. However, there was no specific subset change. beta-glucan protected splenocytes from cytokine withdrawal-induced spontaneous cell death. For further mechanistic studies, ELISA assay revealed that beta-glucan enhanced the expression of anti-apoptotic molecules and interleukin 7 (IL-7), a cytokine critical for lymphocyte survival. We also investigated the IL-2 dependency of beta-glucan-treated splenocytes to determine if treated cells could still undergo clonal expansion. In flow cytometric analysis, beta-glucan induced increased levels of the activation marker CD25 on the surface of splenocytes and beta-glucan-treated splenocytes showed higher proliferation rates in response to IL-2 treatment. This study demonstrates that beta-glucan can enhance the survival of splenocytes and provides valuable information to broaden the use of beta-glucan in research fields.


Subject(s)
Animals , Mice , Cell Death , Dependency, Psychological , Diminazene , Enzyme-Linked Immunosorbent Assay , Interleukin-2 , Interleukin-7 , Lymphocytes , Paenibacillus , Plasmodiophorida , Trypan Blue
20.
Chinese Journal of Biotechnology ; (12): 185-195, 2011.
Article in Chinese | WPRIM | ID: wpr-324564

ABSTRACT

To enhance the thermostability and storage stability of alpha-cyclodextrin glycosyltransferase (a-CGTase), we added specific chemical additives into the preparation of alpha-CGTase, and studied the effect of additives on the storage stability of alpha-CGTase at different temperatures. Then we measured the protein structure of CGTase in the far UV (200-250 nm) and near UV (250-320 nm) ranges respectively by Circular dichroism (CD) spectra under high temperature and analyzed the relationship between thermostability and protein structure. The results indicated that the addition of selected additives (gelatin, glycerin, CaCl2 and PEG400) enhanced the thermostability of alpha-CGTase dramatically. After 45 days, the preparation of alpha-CGTase still had 100% of the enzyme activity with different additives superimposed at the optimum concentration at 40 degrees C. The CD spectra of alpha-CGTase showed that glycerin could protect the secondary and the tertiary structure of the CGTase under high temperature and therefore the enzyme maintained its high activity. Chemical additives can improve the stability of alpha-CGTase significantly and they preserve the enzyme activity by protecting its secondary structure.


Subject(s)
Enzyme Stability , Escherichia coli , Genetics , Metabolism , Glucosyltransferases , Chemistry , Genetics , Glycerol , Chemistry , Paenibacillus , Recombinant Proteins , Chemistry , Genetics
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