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Simultaneous Detection of Common Food-borne Bacterial Pathogens in Innocuous Poultry and Seafood by Multiplex PCR Assay

Microbiology.1992;0(03):-.

To develop a multiplex PCR assay to detect Enterohemorrhagic Escherichia coli (EHEC), Vibrio parahaemolyticus (VP), Salmonella spp. and Listeria monocytogenes(LM) in innocuous poulthy and seafood simultaneously, four pairs of primers were designed from hemolysis gene (hlyAB) of EHEC, toxR gene of VP, invA gene of Salmonella spp. and iap gene of LM. They were detected by conventional PCR and multiplex PCR assay. DNA sequencing was used to examine PCR products. This multiplex PCR method was a highly specific, rapid and sensitive method which could simultaneously detect EHEC, VP, Salmonella spp. and LM. The detection level was 10~(3)cfu/mL.

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Optimization of Culture in Vacuum Freeze-drying Lactobacillus

Jianyou ZHANG

Microbiology.1992;0(01):-.

Starter is one of the most important factors in the yoghourt producing.The using of vacuum freeze-drying starter can greatly improve the quality of products and avoid some technical problems.The best combination of S.t and L.b were mushroom juice 14mL、carrot juice 10mL、maize liquid 0.6mL、Vc 0.2g and beer 10mL、tomato 10mL、carrot juice 15mL、mushroom juice 16mL respectively,the best cultivate time was also determined.

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Stable Coexistence of Five Bacterial Strains as a Community in an Immobilized Continuous Culture System

Shengping XUE

Microbiology.1992;0(01):-.

Five kinds of probiotic bacteria are immobilized and cultured continuously in simulated intestinal systems with corn fiber as the carrier.The biomass and the colonization in biofilm are investigated by selective culture methods and scanning electron microscope.Bifidobacteriurn longum TQ21-2-2,Lactobacillus acidophile CICC06005,Clostridum butyric TO-A,Bacillus mesentericus TO-A and Enterococcus faecalis T-110 can coexist stably.The biomass of five probiotics in the immobilized phase is greater than that of liquid in the continuous shake flask reactor.Five kinds of probiotic bacteria formed biofilm on corn fiber which be observed by SEM.Continuous culture of immobilized cells could be used better model than that of liquid, the model can be used to study for microecology and microecologocal agent.

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Purification and Properties of the a-amylase from a Bacillus sp.WS-3L

Yan LONG

Microbiology.1992;0(01):-.

An extracellular?-amylase(AmyL)from a Bacillus sp WS-3L was purified 345 fold and had a recovery of 15.5%.The amylase was capable of hydrolyzing starch to yield a series of maltooligosaccharides. It was optimally active at 45℃and pH values around 6.5 and showed stability at the temperature below 40℃and pH 7.0-8.0.The amylase was inhibited by Cu~(2+)、NH_4~+、Ag~+、Hg~+ and EDTA、SDS.Michaelist constants(K_m)of the AmyL for were 2.81 mg/mL、8.37 mg/mL、1.80 mg/mL, and maximum velocity(V_(max))of the enzyme for soluble starch,amylose,amylopectin were 11.67?tmol/(min.mL)、10.00?mol/(min?mL)、13.33?mol/(min.mL)respectively.It was suggested that amylopectin is the better hydrolysis substrate for the enzyme.It was observed that the adsorption and digestion of the enzyme on different raw starches was remarkably different.Raw corn starch exhibited high adsorption of the enzyme.It was suggested that the highly stable enzyme was able to be obtained and applied very fast by corn starch chromatography.

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A Rapid and Efficient Method for Directed Screening of Lipase-producing Burkholderia cepacia Complex with Organic Solvent Tolerance from Plant Rhizosphere

Zhengyu SHU ; Ruifeng LIN ; Huan JIANG ; Yanfeng ZHANG ; Jianzhong HUANG

Microbiology.1992;0(06):-.

Lipase from Burkholderia cepacia complex is one of the most versatile biocatalyst and is used widely in many biotechnological application fields including detergent additives, the resolution of racemic compounds, etc. Based on the known whole genomic information of B. cepacia, both ampicillin and kanamycin were added to the TB-T media, the traditional selective media, to screen B. cepacia complex strains from rhizosphere soil samples. The single colonies on the plates with the modified TB-T media were then qualitatively determined the ability to produced the extracellular lipase in the rhodamine B-olive oil agar plates. Thirty-five strains of lipolytic pseudo-B. cepacia complex were isolated and the positive rate of lipolytic bacteria was 65%. Among them, 15 pseudo-B. cepacia complex strains with the tolerance to benzene, n-hexane and n-heptane at the concentration of 10% (V/V) were selected and identified by the recA gene sequence. All of the 15 lipolytic bacteria belonged to the B. cepacia complex.

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On the Technique for L-cystine Conversion by Pseudomonas putida TS1138

Rongqiao HE

Microbiology.1992;0(01):-.

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Expression of Tissue Inhibitor of Metalloproteinase-2(TIMP-2) in Pichia pastoris

Xunyou YAN

Microbiology.1992;0(01):-.

To construct human tissue inhibitor of metalloproteinase-2 and express it in P.pastoris.The sequence was designed according to the amino acid sequence of the TIMP-2 and high-level expression sequences of P.pastoris.We gained the TIMP-2 gene by artificially synthesis method.The gene was cloned into pPIC9 to construct a recombinant vector pPIC9-T2.The vector was transformed into P.pastoris by electroporation,through phenotype selection and induced with methanol we get the expression cell.Separation,purification and bioactivity analysis of the expressed products was performed.

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CONSTANT REGIONS OF QUANTUM YIELDS OF GIBBERELLIN AND HYPOCRELLIN A

Jun XU

Microbiology.1992;0(06):-.

Determination of the quantum yields of Gibberellin, Hypocrellin A and phenol showed that there were respectively ranges of excitation wavelengths for the relative constant of quantum yields of them. The quantum yields of Gibberellin excited by its own multiple excitaion wavelengths (285 and 410 nm) equaled each other. The quantum yields were similar by excitation of 585 nm and 625 nm with determination of the emission at 640 nm for Hypocrellin A. The emission at 690nm with Hypocrellin A would disappear after it had stood at room temperature for about one month, it suggests that it should be convenient to determine the quantum yield, at a shorter wavelength as for the excitation maximum, of a fluorophore whose concentration was low, weak quantum yield and wavelengths of the excitation and emission locating nearly, with a strong Reilagh scattering; and that the 690 nm emission of Hypocrellin A should be emitted by the groups which were subjected to the effects of physical or chemical disturbences.

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Isolation and Identification of Marine Actinomycete JMC06001 Exhibiting Strong Antibacterial Activity

Yiguang CHEN

Microbiology.1992;0(01):-.

One slightly halophilic marine actinomycete strain JMC 06001 was isolated from a saline mud sample collected from the Island Naozhou in the South China Sea,near Zhanjiang,a city of southern China. The fermentation broth of strain JMC 06001 strongly inhibited the growth of Staphylococcus aureaus,Sarcina lutea,Bacillus subtilis,Escherichia coli,Micrococcus luteus and Proteus vulgaris.The combination of morphology,physiological and biochemical characteristics,chemotaxonomic data and 16S rRNA gene sequence analysis supported the view that strain JMC 06001 belong to a known species of the genus Steptomyces, S.peucetius.The strain studied grown well on most media tested,with white aeriel hyphae and pale yellow to pale brown substance hyphae.Yellow diffusible pigments were produced on oatmeal agar(ISP 3), potato extract agar and glucose/asparagines agar,and pale brown to deep brown diffusible pigments were produced on yeast extract/malt extract agar(ISP 2),glycerol/asparagine agar(ISP5),peptone-yeast ext-Fe agar(ISP 6)and nutrient agar.Growth occurred at 4℃～40℃and pH 6.0～9.0,with optimum growth at 28℃and pH 7.0.The tolerant range of NaCl was 0～1.5 mol/L,with best growth occurring in media containing 0.2～0.5 mol/L NaC1.

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Industrial Brewing Yeast with High-glutathione Production and Low-ADH II Enzyme Activity

Bingsheng QIU

Microbiology.2008;0(08):-.

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