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Pseudomonas fluorescens strain PfG32R, a potential biocontrol agent against soilborne pathogens, frequently loses its antifungal activity and ability
to produce enzymes. To characterize genetically the instability of these bacterial functions, we analyzed gacS and gacA genes of PfG32R and three spontaneous mutants of PfG32R (NR1, NR9, and ASW6), which had lost their ability to produce proteases
and antifungal activities. The gacS and gacA sequences of PfG32R had 77%–89% and 78%–87% homology, respectively, with several known gacS and gacA homologues. All three spontaneous mutants were subjected to complementation analysis. Introduction of clones containing an
intact gacS of PfG32R and another P. fluorescens strain Pf-5 as well as strain tea632 of P. syringae pv. theae complemented all three mutants restoring protease and antifungal activities, indicating a mutation in gacS. In sequencing analysis, the mutants had a deletion or change in amino acids in the conserved sensor kinase domains of GacS.
The three mutants maintained both their antibacterial activity against Ralstonia solanacearum and Clavibacter michiganensis ssp. michiganensis and siderophore production, indicating that they are not controlled by the GacS/GacA system.
The sequences reported in this article have been deposited in the DDBJ database under accession numbers AB219364 and AB219365. 相似文献
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饲料中添加蛋白酶AG对鲤鱼鱼种生长和蛋白质消化酶活性的影响 总被引:2,自引:0,他引:2
通过2个试验研究了饲料中添加蛋白酶AG对鲤鱼生长和前肠蛋白质消化酶活性的影响.试验Ⅰ选用540尾均重11.7 g的鲤鱼,随机均分为6组,随机选取3组分别饲喂鱼粉含量为10%、15%、20%的3种等蛋白基础饲料,另外3组分别饲喂添加有175 mg/kg蛋白酶AG的以上3种饲料,试验期60 d.试验Ⅱ选取120尾均重48.7 g的鲤鱼,随机分为2组,一组饲喂鱼粉含量6%的基础饲料为对照组,另外一组饲喂添加175 mg/kg蛋白酶AG的基础饲料为试验组,试验期为30 d.试验Ⅰ结果表明:摄食10%鱼粉饲料和20%鱼粉 175 mg/kg蛋白酶AG饲料的鲤鱼分别具有最低和最高的增重率;在10%鱼粉饲料中添加蛋白酶AG显著提高了鱼体增重率(P<0.05),但在15%、20%鱼粉饲料中添加蛋白酶AG,对鱼体增重率没有显著影响.试验Ⅱ结果表明:相比于对照组,试验组鲤鱼增重率提高了6.4%(P<0.05),饲料系数降低了5.4%(P<0.05).试验Ⅰ和试验Ⅱ中,添加蛋白酶AG对鱼体肌肉水分、粗脂肪、粗蛋白质含量均无显著影响(P>0.05);对消化酶活性的测定表明,在鱼粉含量为10%、6%的基础饲料中添加蛋白酶AG,可显著提高前肠组织蛋白酶活性和食糜蛋白酶活性(P<0.05),但在鱼粉含量为15%、20%的基础饲料中添加蛋白酶AG,对前肠组织蛋白酶活性和食糜蛋白酶活性没有影响.综上所述,在鱼粉含量较低的饲料中添加蛋白酶AG,可提高鲤鱼消化道蛋白酶活性,改善生长性能. 相似文献
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5.
不同生长阶段斑节对虾消化酶活性变化 总被引:3,自引:0,他引:3
通过对斑节对虾不同生长阶段蛋白酶、脂肪酶及淀粉酶活性变化的分析,探讨了斑节对虾在生长过程中对蛋白质、脂肪和碳水化合物的消化利用。结果表明,根据消化酶活性,可将斑节对虾生长分为两个阶段。第1阶段,在体长10~12.5 cm阶段之前,随着个体的增大蛋白酶和脂肪酶活性不断升高;第2阶段,在12.5 cm之后,蛋白酶和脂肪酶活性呈现出一定程度的下降。肝胰腺、胃、肠3种不同消化器官的消化酶活性在不同生长期也不一致,大致上蛋白酶活性从高到低依次为:肝胰腺>胃>肠;脂肪酶活性:肠>肝胰腺>胃;淀粉酶活性:肠>胃>肝胰腺。 相似文献
6.
A method for fractionating sorghum proteins using extraction solvents and techniques designed to obtain polymeric protein structures (especially disulfide linked) was developed. Extraction and separation conditions were optimized in terms of completeness of protein extraction, sample stability, and analytical resolution. After pre-extraction of albumins and globulins, a 3-step sequential procedure involving no reducing agents was applied to ground whole sorghum flour. The three fractions obtained represented proportionally different protein polymer contents and molecular weight distribution as evidenced by comparative size exclusion chromatography. Protein composition also varied among the extracts with differences in kafirin composition and non-kafirin proteins detected in the fractions by RP-HPLC and SDS-PAGE analysis. The ability to quantify and further characterize sorghum polymeric protein complexes will be useful for additional studies linking protein structures with functionality and digestibility and variations for these properties within diverse sorghum germplasm. 相似文献
7.
Pyrophosphate (140 mM, pH 7.1) extracts of two arable soils and one pasture soil were ultrafiltrated separating the extracted material into three fractions: AI with nominal molecular weight (nmw) > 100 kD, AII with nmw between 10 kD and 100 kD and R with nmw < 10 kD. Protease activity was determined in the fractions by using three different substrates: N-benzoyl-l-argininamide (BAA), specific for trypsin; N-benzyloxy-carbonyl-l-phenylalanyl l-leucine (ZPL), specific for carboxypeptidases; and casein, essentially a non-specific substrate. The derivative fractions were also analysed for their amino acid N and humic (HA) and fulvic (FA) acid contents. The organic matter of extracts and derivative fractions obtained from the pasture soil was analysed by isoelectric focusing (IEF) and that of fractions analysed by pyrolysis gas chromatography (Py-GC). Activities of the extract were monitored for their thermal stability and those of the extract and derivative fractions for their optimal pH.Due to the mechanical disintegrating action of sodium pyrophosphate over the humic substances during the fractionation process the amount of total organic C and FA in the fractions was ranked as R > AII > AI. The lowest amino acid N/organic C was found in the R fraction, whereas AII fraction was rich in humic acids, carbohydrates and amino acid N and AI fraction showed the lowest carbohydrate content. At least 70% of the total BAA- and ZPL-hydrolysing activity was associated to particles with nmw higher than 10 kD and at least 30% of these activities were present in particles with nmw higher 100 kD. Casein-hydrolysing activity was quite evenly distributed among the three fractions (AI, AII and R). The extracted protease-organic complexes were resistant to thermal denaturation and some of them showed optimal activity at pH values higher than 10 as a result of the polyanionic characteristics of the humic material surrounding enzyme molecules and of the presence of alkaline protease. Comparison of data obtained in Py-GC analyses and in protease activity suggests that BAA-hydrolysing activity was associated to a highly condensed humic matter and ZPL-hydrolysing activity to less resistant humic substances, while at least some of the extracted casein-hydrolysing activity was present as glyco-proteins not associated to humus. BAA-hydrolysing activity was probably inhibited by fresh organic matter of carbohydrate origin whereas lignin derived organic matter probably inhibited ZPL- and casein-hydrolysing activity. 相似文献
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A review of cyanobacterial odorous and bioactive metabolites: Impacts and management alternatives in aquaculture 总被引:2,自引:0,他引:2
Juliette L. Smith Greg L. Boyer Paul V. Zimba 《Aquaculture (Amsterdam, Netherlands)》2008,280(1-4):5-20
Increased demand has pushed extensive aquaculture towards intensively operated production systems, commonly resulting in eutrophic conditions and cyanobacterial blooms. This review summarizes those cyanobacterial secondary metabolites that can cause undesirable tastes and odors (odorous metabolites) or are biochemically active (bioactive metabolites) in marine and freshwater, extensive and intensive aquaculture systems. For the scope of this paper, biochemically active metabolites include (1) toxins that can cause mortality in aquaculture organisms or have the potential to harm consumers via accumulation in the product (hepatotoxins, cytotoxins, neurotoxins, dermatoxins, and brine shrimp/molluskal toxins), (2) metabolites that may degrade the nutritional status of aquaculture species (inhibitors of proteases and grazer deterrents) or (3) metabolites that have the potential to negatively affect the general health of aquaculture species or aquaculture laborers (dermatoxins, irritant toxins, hepatotoxins, cytotoxins). Suggestions are made as to future management practices in intensive and extensive aquaculture and the potential exposure pathways to aquaculture species and human consumers are identified. 相似文献
10.
Soil enzymes are linked to microbial functions and nutrient cycling in forest ecosystems and are considered sensitive to soil
disturbances. We investigated the effects of severe soil compaction and whole-tree harvesting plus forest floor removal (referred
to as FFR below, compared with stem-only harvesting) on available N, microbial biomass C (MBC), microbial biomass N (MBN),
and microbial biomass P (MBP), and dehydrogenase, protease, and phosphatase activities in the forest floor and 0–10 cm mineral
soil in a boreal aspen (Populus tremuloides Michx.) forest soil near Dawson Creek, British Columbia, Canada. In the forest floor, no soil compaction effects were observed
for any of the soil microbial or enzyme activity parameters measured. In the mineral soil, compaction reduced available N,
MBP, and acid phosphatase by 53, 47, and 48%, respectively, when forest floor was intact, and protease and alkaline phosphatase
activities by 28 and 27%, respectively, regardless of FFR. Forest floor removal reduced available P, MBC, MBN, and protease
and alkaline phosphatase activities by 38, 46, 49, 25, and 45%, respectively, regardless of soil compaction, and available
N, MBP, and acid phosphatase activity by 52, 50, and 39%, respectively, in the noncompacted soil. Neither soil compaction
nor FFR affected dehydrogenase activities. Reductions in microbial biomass and protease and phosphatase activities after compaction
and FFR likely led to the reduced N and P availabilities in the soil. Our results indicate that microbial biomass and enzyme
activities were sensitive to soil compaction and FFR and that such disturbances had negative consequences for forest soil
N and P cycling and fertility. 相似文献