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Fragmentation rate constants, which can be used to estimate the tensile strength of fungal hyphae, were used to elucidate
relationships between morphological changes and addition of fatty acids during cephalosporin C production in Acremonium chrysogenum M35. The number of arthrospores increased gradually during fermentation, and, in particular, was higher in the presence of
rice oil, oleic acid or linoleic acid than in their absence. Because supplementation of rice oil or fatty acids increased
cephalosporin C, we concluded that differentiation to arthrospores is related to cephalosporin C production. To estimate the
relative tensile strengths of fungal hyphae, fragmentation rate constants (k
frag) were measured. When rice oil, oleic acid, or linoleic acid were added into medium, fragmentation rate constants were higher
than for the control, and hyphal tensile strengths reduced. The relative tensile strength of fungal hyphae, however was not
constant presumably due to differences in physiological state. 相似文献
3.
Antibiotic Synthesis and Morphological Differentiation of Cephalosporium acremonium 总被引:8,自引:2,他引:6
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In submerged cultures, Cephalosporium acremonium exists in four morphological forms: hyphae, arthrospores, conidia, and germlings. The phase of hyphal differentiation into arthrospores coincides with the maximum rate of β-lactam antibiotic synthesis. Furthermore, arthrospores, separated by density-gradient centrifugation, possess 40% greater antibiotic-producing activity than any other morphological cell type. In a series of mutants, each with an increased potential to produce β-lactam antibiotics, differentiation into arthrospores was proportional to the increased titer of these antibiotics. Thus, arthrospores exhibit enhanced synthesis of β-lactam antibiotics and appear to be a determining factor in high-yielding mutants. Since a non-antibiotic-producing mutant readily differentiated into arthrospores, antibiotic synthesis and cellular differentiation are not obligately related. 相似文献
4.
Biosynthesis of penicillin N and cephalosporin C: Antibiotic production and other features of the metabolism of a Cephalosporium sp 总被引:2,自引:2,他引:0
Brenda Smith S. C. Warren G. G. F. Newton E. P. Abraham 《The Biochemical journal》1967,103(3):877-890
1. The production of penicillin N and cephalosporin C by two mutants of a Cephalosporium sp. has been studied with cultures grown in a chemically defined medium and with suspensions of washed mycelium in water or a buffered salt solution. 2. Antibiotic synthesis began at an early stage of growth and its rate per unit weight of mycelium appeared to pass its maximum as morphological changes were occurring in young hyphae. This rate subsequently declined, but rapid production could continue after net growth had ceased. 3. In a series of shake-flask fermentations in the growth medium, increases in the yield of penicillin N above the mean were correlated with much smaller increases in the yield of cephalosporin C and vice versa. 4. In suspensions of washed mycelium, moderate decreases in the efficiency of aeration increased the yield of penicillin N and decreased that of cephalosporin C. A similar result normally followed the addition of methionine to the suspension fluid, and in both cases there was usually an increase in the yield of the two antibiotics combined. 5. The apparent intracellular concentrations of the antibiotics were much lower than those attained extracellularly and also much lower than those of most of the amino acids in the intracellular pool. No detectable amount of [(14)C]penicillin N added to the extracellular fluid was found to enter the mycelium. 6. Very small amounts of peptide material whose behaviour was similar to that of the sulphonic acid of delta-(alpha-amino-adipoyl)cysteinylvaline on paper electrophoresis at pH1.8 were found in extracts of the mycelium that had been oxidized with performic acid. 6-Aminopenicillanic acid and 7-aminocephalosporanic acid were not detected. 7. Ultrasonic treatment of the mycelium resulted in rapid fragmentation of mycelial chains, rupture of many individual cells, and the liberation of amino acids and other substances into the medium. 8. Ultrasonically treated preparations synthesized penicillin N and cephalosporin C rapidly after a lag of 12hr. Antibiotic synthesis was accompanied by the growth of hyphae from swollen mycelial fragments and by the re-establishment of permeability barriers resulting in the uptake of amino acids from the medium. 相似文献
5.
The effect of the method of methionine addition, growth-limiting carbon source (glucose vs sucrose), and culture growth rate on cephalosporin C production was investigated in a Cephalosporium acremonium defined medium fed batch fermentation. Batch addition of methionine, at a concentration of 3 g/L, prior to the start of a fed sucrose fermentation was found to interfere with the ability of the culture to utilize this sugar, thus limiting growth and decreasing cephalosporin C production. Batch methionine addition had no effect on glucose-limited cultures. Concurrent exponential feeding of methionine with sucrose improved both culture growth and productivity. Under the control of identical carbon source limiting feed profiles, sucrose was observed to support greater cephalosporin C production than glucose. Optimal cephalosporin C production in a C. acremonium defined medium fed batch fermentation was obtained through controlling culture growth during the rapid growth phase at a relatively low level with respect to mumax (mu approximately 0.036 h-1) until achieving a desired cell mass with a concurrent sucrose and methionine feed, followed by maintaining relatively vigorous growth (mu approximately 0.01 h-1) with sucrose for the duration of the fermentation. 相似文献
6.
Summary Lysine added to a defined medium at a concentration of 1 mg/ml enhances the capacity of Cephalosporium acremonium to produce cephalosporin C. The lysine effect was accompanied by a delayed differentiation to arthrospores followed by more extensive mycelial fragmentation. Higher lysine concentrations reduced the synthesis of cephalosporin C. The effect of lysine on cephalosporin C synthesis was not strain related and was not elicited by the other amino acids tested. Both lysine stimulation and inhibition were evidenct using resting cells. The lysine inhibition of cephalosporin C synthesis with resting cells was relieved by supplementing the medium with 1 to 2 mg/ml -aminoadipic acid. This reversal supports the hypothesis that lysine restricts the availability of -aminoadipic acid for -lactam biosynthesis. 相似文献
7.
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9.
Changes in Cellular Fatty Acid Composition of Cephalosporium acremonium during Cephalosporin C Production
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Young-Sun Sohn Keun-Cheol Lee Young-Hwan Koh Gwang-Hoon Gil 《Applied microbiology》1994,60(3):947-952
Cephalosporium acremonium was cultivated in fermentation medium containing sucrose or methyl oleate as a carbon source for cephalosporin C production. The level of antibiotic production was 48 g of cephalosporin C per liter under optimum conditions when methyl oleate was used. The C18:1 (oleic acid) methyl ester appeared to be utilized faster than the C18:2 (linoleic acid) methyl ester in fermentation broth. Physiological characteristics of C. acremonium were investigated by determining the fatty acid composition of the total cellular free lipid. Significant changes in cellular fatty acid composition occurred during inoculum cultivation and fermentation. The percentage of C18:1 increased from 19.1 to 38.5%, but the percentage of C18:2 decreased from 56.7 to 36.1%, and there was an increase in pH during inoculum cultivation. The cellular fatty acid composition of C. acremonium grown in fermentation medium containing methyl oleate (methyl oleate medium) was significantly different from that in fermentation medium containing sucrose (sucrose medium). The major fatty acids detected were C16:0 (palmitic acid), C18:1, and C18:2. In methyl oleate medium, the ratio of C18:1 to C18:2 increased from 0.34 to 1.37, while the cell morphology changed from hyphae to arthrospores and conidia. In contrast, in sucrose medium, the ratio of C18:1 to C18:2 decreased from 0.70 to 0.43, and most of the cells remained hyphal at the end of fermentation. We observed that hyphae contained a higher proportion of C18:2 but arthrospores and conidia contained a higher proportion of C18:1. 相似文献
10.
Association of Yersinia enterocolitica with the manufacture of cheese and occurrence in pasteurized milk. 总被引:9,自引:4,他引:5
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D A Schiemann 《Applied microbiology》1978,36(2):274-277
The effects of several physical and chemical agents on the survival of Trichophyton mentagrophytes arthrospores were investigated. Although arthrospores of this dermatophyte were highly resistant to chilling and freezing, they were extremely susceptible to moderate heat (above 50 degrees C) and desiccation. This high susceptibility could be significantly reduced when they were dried in the presence of exogenous proteins. These arthrospores were markedly susceptible to glutaraldehyde. They appeared to be significantly more resistant than their hyphal counterparts to common antimycotics such as clotrimazole, griseofulvin, miconazole nitrate, and nystatin. Clinical and epidemiological implications of these observations are discussed. 相似文献
11.
Teijeira F Ullán RV Guerra SM García-Estrada C Vaca I Martín JF 《The Biochemical journal》2009,418(1):113-124
The cluster of early cephalosporin biosynthesis genes (pcbAB, pcbC, cefD1, cefD2 and cefT of Acremonium chrysogenum) contains all of the genes required for the biosynthesis of the cephalosporin biosynthetic pathway intermediate penicillin N. Downstream of the cefD1 gene, there is an unassigned open reading frame named cefM encoding a protein of the MFS (major facilitator superfamily) with 12 transmembrane domains, different from the previously reported cefT. Targeted inactivation of cefM by gene replacement showed that it is essential for cephalosporin biosynthesis. The disrupted mutant accumulates a significant amount of penicillin N, is unable to synthesize deacetoxy-, deacetyl-cephalosporin C and cephalosporin C and shows impaired differentiation into arthrospores. Complementation of the disrupted mutant with the cefM gene restored the intracellular penicillin N concentration to normal levels and allowed synthesis and secretion of the cephalosporin intermediates and cephalosporin C. A fused cefM-gfp gene complemented the cefM-disrupted mutant, and the CefM-GFP (green fluorescent protein) fusion was targeted to intracellular microbodies that were abundant after 72 h of culture in the differentiating hyphae and in the arthrospore chains, coinciding with the phase of intense cephalosporin biosynthesis. Since the dual-component enzyme system CefD1-CefD2 that converts isopenicillin N into penicillin N contains peroxisomal targeting sequences, it is probable that the epimerization step takes place in the peroxisome matrix. The CefM protein seems to be involved in the translocation of penicillin N from the peroxisome (or peroxisome-like microbodies) lumen to the cytosol, where it is converted into cephalosporin C. 相似文献
12.
M.James C. Crabbe 《FEMS microbiology letters》1988,56(1):71-77
Abstract Yeast-like blastosporic growth, lowered levels of isopenicillin N synthetase and deacetoxycephalosporin C synthetase/hydroxylase, and lowered antibiotic production were induced in Cephalosporium acremonium strain C.O. 728 by inclusion of cysteine or glutathione in the growth medium. These effects were similar to those observed under normal conditions with strain M 8650/4, which had higher levels of glutathione reductase and protein disulphide reductase than strain C.O. 728. The addition of cytochalasins A and B slightly increased yeast-like growth in C.O. 728, although levels of biosynthetic enzyme activity and antibiotic secretion were not significantly affected. The Ca2+ ionophore A23187 inhibited C.O. 728 growth, slowed penicillin secretion into the medium, and totally inhibited cephalosporin secretion. These results show that maintenance of Ca2+ levels and the absence of thiols are important for antibiotic production in Cephalosporium acremonium . 相似文献
13.
Masazumi Matsumura Toshiomi Yoshida Hisaharu Taguchi 《Applied microbiology and biotechnology》1982,16(2-3):114-118
Summary DL-seleno-methionine resistant mutants of Cephalosporium acremonium were isolated which have an enhanced capacity to utilized sulfate for the synthesis of cephalosporin C. Of these mutants, one designated as SMR-I3 produced three-fold more cephalosporin C from sulfate than its parent CW19. Mutant SMR-I3 required less dl-methionine for maximal synthesis of cephalosporin C, but an excess of dl-methionine inhibited the synthesis of the antibiotic. Furthermore, the mutant accumulated excessive methionine in the amino acid pool and possessed superior activity for sulfate uptake. These observations indicate that in the mutant SMR-I3, the biosynthesis of methionine from sulfate is very active and excess methionine becomes available for the synthesis of cephalosporin C. 相似文献
14.
不同补料控制方式发酵生产头孢菌素C的性能比较 总被引:1,自引:0,他引:1
在7 L发酵罐下,对利用顶头孢霉菌(Cephalosporins acremonium)发酵生产头孢菌素C(CPC)过程的最优底物流加工艺进行了研究。提出了一种新式硫铵豆油耦联型的硫铵流加策略。该控制策略可将发酵液中的氨态氮浓度控制在3 6 g/L之间,同时满足了发酵前期细胞生长与CPC合成对氮源和硫源的需求,促进了顶头孢霉菌菌丝分化,为发酵后期的CPC高效生产奠定了前期基础。比较了CPC合成期内间歇、匀速和DO-Stat自动流加3种不同豆油流加方式的发酵性能。研究发现,耦联使用硫铵/后程通富氧空气DO-Stat法进行硫铵和豆油的同时补料和CPC发酵,可将碳源浓度与溶解氧浓度DO同时控制于适中水平,使CPC合成以高浓度和低副产物积累的方式进行,最终CPC浓度和得率分别达到35.77 g/L和13.3%。主代谢副产物脱乙酰氧头孢菌素C(DAOC)的积累量和DAOC/CPC分别仅有0.178 g/L和0.5%。 相似文献
15.
The origin of desacetylcephalosporin C in cephalosporin C fermentation broths was investigated. Esterase activity was detected in cell-free extracts of Cephalosporium acremonium, but these extracts failed to deesterify cephalosporin C. When cephalosporin C was added to sterile and inoculated fermentation media, the antibiotic decayed at nearly identical rates. The formation of desacetylcephalosporin C during the fermentation was measured by quantitative chromatography and by the incorporation of valine-1-(14)C into the molecule. The rate constants obtained from the results of these experiments were equivalent to those for the decay of cephalosporin C in sterile and inoculated media. The data demonstrate that desacetylcephalosporin C is produced by nonenzymatic hydrolysis of cephalosporin C. 相似文献
16.
C R Barrera 《Journal of bacteriology》1983,155(2):886-895
The formation of arthrospores in Mucor rouxii was studied by transmission and scanning electron microscopy and light microscopy. The arthrospores formed in a random manner in terminal and internal regions of the hyphae. The earliest appearance of the arthrospores was seen by scanning electron microscopy as compartments delineated by double ridges. These ridges probably corresponded to the site of septal wall formation. The elongated compartments varied considerably in size. As the arthrospores matured, they tended to separate as a result of a gradual change in the shape of the arthrospores to a nearly spherical form and also as the result of simultaneous degradation of the outermost cell wall layer. The mature arthrospores were surrounded by a complex cell wall consisting of at least three distinct layers in addition to the original hyphal cell wall. Crystal-like structures were seen in the cytoplasm of some of the arthrospores in addition to the usual organelles such as mitochondria, nuclei, and ribosomes. Septum formation by centripetal cell wall growth from the lateral hyphal wall was documented by transmission electron microscopy. However, evidence was also found which suggested that not all internal cell wall development in the fungal hyphae during arthrosporogenesis necessarily led to the formation of mature arthrospores. 相似文献
17.
Protoplast yields from the fungus Acremonium diospyri, prepared using Novozym 234, were affected by culture age, osmotic stabilizer and its concentration, and incubation time. Protoplasts were released by extension through pores in small hyphal fragments produced by enzyme action on the mycelium, and regeneration was similar to that reported for other fungi. Stabilized protoplasts were shown to synthesize carotenoids on exposure to light, but their carotenoid composition differed from that of mycelium incubated under the same conditions. 相似文献
18.
Three strains of Cephalosporium acremonium with different potential of cephalosporin C production show differences in the levels of sulphur amino acid metabolic enzymes. The regulation of these enzymes is also different in the strains. In the superior producing strain two enzymes directly involved in cysteine synthesis are the least susceptible to repression by methionine. In this strain cystathionine metabolizing enzymes seem to favour cysteine synthesis. Antibiotic producing strains, in contrast to the non-producing strain, are highly sensitive to chromate. 相似文献
19.
Rustom S. Mody Valiollah Heidarynejad Alka M. Patel Dr. Prafulla J. Dave 《Current microbiology》1990,20(2):95-103
Mutants deficient in the biosynthesis of prodigiosin have been obtained by treatingSerratia marcescens with high doses of ultraviolet radiation. Mutants were selected on the basis of the color characteristics of their colonies when grown on peptone glycerol medium. New types of mutants, with unusual blocks in the biosynthetic pathway of prodigiosin, were obtained. All the mutants were classified under a new scheme on the basis of the syntrophic pigmentation characteristic and infrared spectroscopic analysis of their pigment. By these criteria mutants could be distinguished into eight distinct classes. Classes I to III include mutants of the three classes (M1, B3, and B1) reported previously [Morrison, DA (1966) J Bacteriol 91:1599–1604] and several new ones. Mutants blocked in the methylamylpyrrole (MAP) arm of the bifurcated pathway were assigned to class I. A class II mutant was distinguished by its inability to synthesize methoxybipyrrolecarboxyaldehyde (MBC), but was able to produce norprodigiosin. Class III mutants were deficient in the synthesis of hydroxybipyrrolecarboxaldehyde (HBC). Double mutants were obtained with defects in the expression of both MBC and MAP and were assigned to class IV. Mutants of class V were unable to synthesize HBC and MAP, but could form MBC when furnished with exogenous HBC. Class VI and VII mutants were defective in the synthesis of all three precursors, but differed in their ability to perform the coupling step. Finally, a mutant of class VIII was found to produce the three intermediates, but was deficient in prodigiosin or norprodigiosin biosynthesis, indicative of a defect in the enzymatic condensation of MAP with the bipyrroles MBC and HBC. The anomalous pattern of syntrophism among certain interclass mutants suggests that the physiology of pigment formation inS. marcescens is quite complex. 相似文献