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101.
102.
Summary -Amylase production with Bacillus amyloliquefaciens was investigated in two different aqueous two-phase systems and in polyethylene glycol (PEG) 600 solutions of different concentrations. The cells did not partition totally to the bottom phases of the aqueous two-phase systems, and the enzyme production was repressed in both systems as well as in PEG 600 solutions. Concomitantly, the cultivation time was prolonged, indicating an increased maintenance metabolism. The surface properties of cells grown in 200 g/kg PEG 600 were investigated by phase partitioning and compared to the surface properties of Bacillus subtilis, which under these conditions showed increased -amylase production. The cells of B. amyloliquefaciens partitioned to the top phase in a PEG-dextran system, whereas the cells of B. subtilis partitioned to the bottom phase. The results are discussed in relation to water activity, oxygen transfer rate and PEG-induced changes of the surface properties of the cells. The possible role of PEG as an uncoupler of the proton motive force at high concentrations is also discussed. 相似文献
103.
Max Lechner Herbert Märkl Friedrich Götz 《Applied microbiology and biotechnology》1988,28(4-5):345-349
Summary In a newly constructed one-vessel dialysis fermentor, a strain of Staphylococcus carnosus TM300 carrying the lipase secretion plasmid pLipPS1 was used to investigate exoenzyme and biomass production. The bacterial culture grows in an inner compartment of 21 volume, separated from a 101 nutrient broth compartment by a conventional dialysis membrane. In order to avoid substrate depletion and to prolong the growth phase, a highly concentrated nutrient broth was used. The biomass production reached 60 g cell dry weight/l. The increase in extracellular lipase concentration was directly coupled with the increase of cell mass and reached a value of 230 mg/l culture supernatant. Harvesting the cells in the late growth phase, the lipase content was about 30% of the total exoproteins in the supernatant. 相似文献
104.
Julie Anderson Kirihara John P. Hunsperger Walter C. Mahoney Joachim W. Messing 《Molecular & general genetics : MGG》1988,211(3):477-484
Summary A methionine-rich 10 kDa zein storage protein from maize was isolated and the sequence of the N-terminal 30 amino acids was determined. Based on the amino acid sequence, two mixed oligonucleotides were synthesized and used to probe a maize endosperm cDNA library. A fulllength cDNA clone encoding the 10 kDa zein was isolated by this procedure. The nucleotide sequence of the cDNA clone predicts a polypeptide of 129 amino acids, preceded by a signal peptide of 21 amino acids. The predicted polypeptide is unique in its extremely high content of methionine (22.5%). The maize inbred line BSSS-53, which has increased seed methionine due to overproduction of this protein, was compared to W23, a standard inbred line. Northern blot analysis showed that the relative RNA levels for the 10 kDa zein were enhanced in developing seeds of BSSS-53, providing a molecular basis for the overproduction of the protein. Southern blot analysis indicated that there are one or two 10 kDa zein genes in the maize genome. 相似文献
105.
106.
Interaction of gravi- and phototropic stimulation in the response of maize (Zea mays L.) coleoptiles 总被引:3,自引:0,他引:3
The influence of gravitropic stimulation upon blue-light-induced first positive phototropism for stimulations in the same (light source and center of gravity opposite to each other) and in opposing directions was investigated in maize cole-optiles by measuring fluence-response patterns. As a result of gravitropic counterstimulation, phototropic bending was transient with maximum curvature occurring 100 min after stimulation. On a horizontal clinostat, however, the seedlings curved for 20 h. Gravistimulation in the opposite direction acted additively upon blue-light curvature. Gravistimulation in the same direction as phototropic stimulation produced a complex behaviour deviating from simple additivity. This pattern can be explained by a gravitropically mediated sensitization of the phototropic reaction, an optimal dependence of differential growth on the sum of photo-and gravistimulation, and blue-light-induced inhibition of gravitropic curvature at high fluences. These findings indicate that several steps of photo-and gravitransduction are separate. Preirradiation with red light desensitized the system independently of applied gravity-treatment, indicating that the site of red-light interaction is common to both transduction chains.Abbreviations BL
blue light
- G+
stimulation by light and gravity in the same direction (i.e. light source and center of gravity opposite to each other)
- G-
stimulation by light and gravity in opposing directions 相似文献
107.
A serum free medium was developed, that could be used for the large scale propagation of various cell lines in bioreactors. The medium is based on a 1:1 mixture of Iscove's Modified Dulbecco's Medium and Ham's Medium F12, supplemented with transferrin, insulin and a BSA/oleic acid complex. Several myelomas, hybridomas derived from different myelomas and spleen cells, and other lymphoid and non-lymphoid cell lines were cultivated at growth rates comparable to those observed using serum-supplemented media. There was furthermore no reduction in the formation of products such as monoclonal antibodies or recombinant human interleukin-2.Abbreviations Ag8
Mouse myeloma cell line P3-X63-Ag8.653
- BME
Basal Medium Eagle
- BSA
Bovine Serum Albumin
- DMEM
Dulbecco's Modified Eagle's Medium
- EDTA
Ethylenediaminete-traacetic Acid
- e-PC
Phosphatidyl choline from egg yolk
- FCS
Fetal Calf Serum
- FGF
Fibroblast Growth Factor
- GHL
Glycyl-histidyl-lysine
- HDL
High Density Lipoprotein
- HPL
Human Plasma Lipid
- IF
1:1 mixture of IMDM and Ham's F12
- IMDM
Iscove's Modified Dulbecco's medium
- LDL
Low Density Lipoprotein
- NS1
Mouse myeloma cell line NSI-1-Ag4-1
- PBS
Phosphate Buffered Saline
- s-PC
Phosphatidylcholine from soy beans
- s-PE
Phosphatidylethanolamine from soy beans
- s-lecithin
lecithin from soy beans 相似文献
108.
Päivi Heikkilä Arvi I. Kahri Christian Ehnholm Petri T. Kovanen 《In vitro cellular & developmental biology. Plant》1988,24(9):936-942
Summary To define the role of endogenously synthesized cholesterol in the differentiation of adrenocortical cells in primary culture,
fetal rat adrenal cells were cultured in the presence of exogenous cholesterol (serum-supplemented medium) or in the absence
of it (serum-free medium or lipoprotein-free medium). Ultrastructurally the cells had features of glomerulosa cells: mitochondria
were oval or rod shaped with lamellar inner membranes. The amount of smooth endoplasmic reticulum was small, and lipid droplets
were few. When the cells were cultured in serum-free medium some intracytoplasmic vacuoles were seen. The undifferentiated
zona glomerulosa-like cells secreted low amounts of corticosterone and 18-OH-deoxycorticosterone (18-OH-DOC) in all three
media (serum-supplemented medium, serum-free medium, and lipoprotein-free medium). Stimulation of the adrenocortical cells
with ACTH induced the ultrastructural features of differentiated zona fasciculata-like cells. Mitochondrial inner membranes
were well developed in lipoprotein-free medium, but not in serum-free medium. The amount of intracellular lipids was increased
in both media devoid of cholesterol. In the ACTH stimulated cultures the presence of exogenous cholesterol resulted in increased
secretions of corticosterone and 18-OH-DOC. In the absence of an exogenous source of cholesterol, the amounts of steroids
secreted were only half of that secreted in the presence of serum-supplemented medium. Endogenously synthesized cholesterol
is sufficient for the morphologic differentiation of fetal rat adrenocortical cells under ACTH stimulation. However, without
exogenously provided cholesterol, the steroid production accounts only for half of the maximal output achieved using serum-supplemented
medium.
This work was supported by Finnish Culture Foundation. 相似文献
109.
Summary Leu-enkephalin containing secretory granules were demonstrated in axon terminals of immunogoldlabeled electron-microscopic sections of the sinus gland of three brachyuran crustaceans. These granules have a diameter of 120+-15 nm and differ in electron density from those located in adjacent terminals containing hyperglycemic or molt-inhibiting hormone. These neurohormones do not show co-localization with leu-enkephalin. The cross-reactivity of leu-enkephalin antiserum with met-enkephalin is less than 1%. The sinus glands of the three species examined show no immunoreactivity for FMRF-amide. A modulatory activity of endogenous enkephalin by paracrine mechanisms is suggested. 相似文献
110.
Chronic treatment of hamsters with estradiol for several months has previously been shown to decrease the specific content
of cytochrome P450 in the kidney, a target of hormonal carcinogenesis, but not in liver. The reason for this decrease in metabolic
enzyme activity is unknown and has been examined in this investigation. We now report that the decrease in specific content
of renal cytochrome P450 by 73% in response to estradiol was not affected by co-treatment with tamoxifen for 1 month. The
subcutaneous infusion of 250 μg/day estradiol for 7 days lowered renal cytochrome P450 by 71% from control values and was
therefore used for further mechanistic studies. This treatment decreased renal activities of estradiol 2- or 4-hydroxylase
by 77 to 80%, of 7-ethoxycoumarin-O-deethylase by 66% of control values, respectively, and completely eliminated aryl hydrocarbon
hydroxylase activities, whereas liver enzymes remained unaffected. After 7 days of infusion of estradiol, fluorescent products
of lipid peroxidation were more than doubled in hamster kidney but remained unchanged in liver. The possibility of enzyme
destruction by binding of estradiol 2,3-quinone to metabolizing enzymes was investigatedin vitro. In the presence of 2-hydroxyestradiol, cumene hydroperoxide, and microsomes, conditions known to favor the oxidation of
the steroid to quinone, the binding of catechol estrogen metabolite to microsomal protein increased 60 fold over control values
in the absence of cofactor. Purified rat liver cytochrome P450c also oxidized 2-hydroxyestradiol to 2,3-estradiol quinone.
The rate of oxidation was linear for the first 2–3 min, but thereafter decreased with time. Under these incubation conditions,
irreversible binding of catechol estrogen metabolite to cytochrome P450c increased for the first 2–3 min and then remained
at this plateau level. It was concluded that enzyme destruction by a reactive estrogen metabolite or by lipid peroxides may
be a major reason for the organ-specific decrease in cytochrome P450 enzymes in kidneys of estrogen-treated hamsters. 相似文献