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1.
Cell-free extracts of Penicillium urticae (NRRL 2159A), and its Pat- mutants, J2, J1, and S11, were found to contain significant NADP-dependent isoepoxydon dehydrogenase activity. This reversible interconversion of the epoxides (-)-phyllostine and (+)-isoepoxydon occurred optimally at pH 5.8 and was completely inhibited by 1 mM p-chloromercuribenzoate (PCMB). The cytosol enzyme possessed specificity for both substrate and cofactor since neither (+)-epoxydon, an epimer of (+)-isoepoxydon, nor NADH was utilized. Cell extracts of the parent and of mutant J2, which is blocked before the epoxides in the patulin pathway, were found to convert phyllostine and isoepoxydon to a number of unknown metabolites which appeared as yellow spots on thin-layer chromatograms after spraying with a chromogenic reagent. Extracts of mutant J1 were unable to carry out this conversion, while whole cells of mutant S11 accumulated what appeared to be these same 'yellow' compounds. Since PCMB-treated extracts of J2 converted phyllostine but not isoepoxydon to these new metabolites, phyllostine appeared to be their more immediate precursor. The relative positions of isoepoxydon and phyllostine in the patulin pathway are discussed. 相似文献
2.
Penicillium urticae (NRRL 2159A) was grown in culture broth containing 1 muCi of [1-14C-A1acetate to produce [14C]patulin. [14C]patulin was purified from the broth and added to apple cider. After the patulin concentration of the cider was adjusted to 30 mug/ml with unlabeled patulin, the cider was subjected to various charcoal treatments. [14C]patulin was completely removed by shaking the cider with 20 mg of activated charcoal per ml and by eluting the cider through a 40- to 60-mesh charcoal column. Activated charcola at 5 mg/ml reduced patulin in naturally contaminated cider to nondetectable levels. 相似文献
3.
A patulin-negative mutant (J1) of Penicillium urticae (N.R.R.L. 2159A) was known to accumulate about 100mg per litre quantities of the 5,6-epoxygentisyl quinone, (-)-phyllostine and another metabolite (UIII). Both were derived from acetate and hence were polyketides. Purified UIII (m.p. 53 degrees C, [alpha](32) (D)+206 degrees , lambda(methanol) (max.) 240nm; epsilon 3806 litre.mol(-1).cm(-1)) was characterized as a partially reduced derivative of (-)-phyllostine and was found to be a diastereoisomer of the known phytotoxin, (+)-epoxydon. Hence its designation as (+)-iso- or epi-epoxydon. From (1)H n.m.r. and c.d. data the stereochemistry of the epoxide ring in (+)-isoepoxydon was determined to be identical with that in (+)-epoxydon (i.e. R,R) but the configuration of the secondary alcohol at C-4 was S rather than R as in (+)-epoxydon. Isoepoxydon (compound UIII) is therefore (4S,5R,6R)-5,6-epoxy-4-hydroxy-2-hydroxymethylcyclohex-2-en-1-one. The boat conformation in which the C-4 hydroxy group is axial is preferred. In the range of 1mm to 5mm, the antibiotic activity of (+)-isoepoxydon against Bacillus subtilis sp. was 56% of that obtained with patulin. Over a period of 1 to 3h, [(14)C]isoepoxydon was efficiently converted into patulin by a shake culture of the parent strain of P. urticae. The precursor relationship of isoepoxydon to patulin was confirmed by feeding unlabelled isoepoxydon (1mm) to a washed-cell suspension of a mutant (J2) in which, over a period of 3 to 5h, a better than 60% conversion into patulin was attained. The enzymic relationship between isoepoxydon and phyllostine and their positions in the late portion of the patulin biosynthetic pathway are discussed. 相似文献
4.
Submerged cultures of Penicillium urticae (NRRL 2159A) produced the antibiotics patulin and griseofulvin when grown in a glucose-nitrate medium. A high concentration of calcium (i.e., 68 mM) inhibited the production of both antibiotics while stimulating conidiogenesis. Conidial mutants that were defective in an early stage of conidiogenesis produced markedly less patulin, even under growth conditions that favored secondary metabolism. A mutant which lacked the ability to produce the patulin pathway metabolites m-cresol, toluquinol, m-hydroxybenzyl-alcohol, m-hydroxybenzaldehyde, gentisaldehyde, gentisyl alcohol, gentisic acid and patulin, as well as the pathway enzyme m-hydroxybenzyl-alcohol dehydrogenase, still produced yields of conidia that were equivalent to or greater than those of the parent strain. Other mutants which were blocked at later steps of the patulin pathway also produced conidia. These results indicate that patulin and the other related secondary metabolites noted above are not a prerequisite to conidiogenesis in P. urticae. Environmental and developmental factors such as calcium levels and conidiogenesis do, however, indirectly affect the production of patulin pathway metabolites. 相似文献
5.
Factors affecting the production of [14C]patulin from [1-14C]acetate by replacement cultures of Penicillium patulum have been investigated. Incorporation of [1-14C]acetate into patulin reached a maximum with 6- to 8-day-old cultures incubated at 28 degrees C for 8 h in a replacement medium containing 0.1 M glucose, inorganic salts, and undiluted [1-14C]acetate. The specific activity of [14C]patulin obtained from this method was 34 mCi/mmol when 0.5 mCi of [1-14C]acetate was supplied to the replacement medium. 相似文献
6.
Penicillic acid-negative mutants were obtained from a color mutant derived from Penicillium cyclopium NRRL 1888 through N-methyl-N'-nitro-N-nitrosoguanidine treatment. One mutant (SK2N6) accumulated 6-methyl-1,2,4-benzenetriol, which was not previously known to be a metabolite of P. cyclopium, in addition to orsellinic acid and orcinol. The radioactivity of [1-14C]acetic acid was rapidly incorporated into 6-methyl-1,2,4-benzenetriol in a culture of P. cyclopium SK2N6. Moreover, the radioactivity of [14C]6-methyl-1,2,4-benzenetriol was efficiently incorporated into penicillic acid in a culture of P. cyclopium NRRL 1888. These data indicate that 6-methyl-1,2,4-benzenetriol is a precursor for penicillic acid biosynthesis. The results on the addition of 1,4-dihydroxy-6-methoxy-2-methylbenzene, 6-methoxy-2-methylbenzoquinone(1,4), and 1-O-methylorcinol to a culture of P. cyclopium SK2N6 indicated that only the former two compounds are converted to penicillic acid. Thus, a new portion of the penicillic acid biosynthetic pathway is proposed. 相似文献
7.
6-Methyl-1,2,4-benzenetriol, a new intermediate in penicillic acid biosynthesis in Penicillium cyclopium.
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Penicillic acid-negative mutants were obtained from a color mutant derived from Penicillium cyclopium NRRL 1888 through N-methyl-N'-nitro-N-nitrosoguanidine treatment. One mutant (SK2N6) accumulated 6-methyl-1,2,4-benzenetriol, which was not previously known to be a metabolite of P. cyclopium, in addition to orsellinic acid and orcinol. The radioactivity of [1-14C]acetic acid was rapidly incorporated into 6-methyl-1,2,4-benzenetriol in a culture of P. cyclopium SK2N6. Moreover, the radioactivity of [14C]6-methyl-1,2,4-benzenetriol was efficiently incorporated into penicillic acid in a culture of P. cyclopium NRRL 1888. These data indicate that 6-methyl-1,2,4-benzenetriol is a precursor for penicillic acid biosynthesis. The results on the addition of 1,4-dihydroxy-6-methoxy-2-methylbenzene, 6-methoxy-2-methylbenzoquinone(1,4), and 1-O-methylorcinol to a culture of P. cyclopium SK2N6 indicated that only the former two compounds are converted to penicillic acid. Thus, a new portion of the penicillic acid biosynthetic pathway is proposed. 相似文献
8.
A hemA mutant of Escherichia coli containing a multicopy plasmid which complemented the mutation excreted 5-aminolevulinic acid (ALA) into the medium. [1-14C]glutamate was substantially incorporated into ALA by this strain, whereas [2-14C]glycine was not. Periodate degradation of labeled ALA showed that C-5 of ALA was derived from C-1 of glutamate. The synthesis of ALA by two sonicate fractions which had been processed by gel filtration and dialysis, respectively, was dependent on glutamate, ATP, NADPH, tRNA(Glu), and pyridoxal phosphate. tRNA(Glu) stimulated ALA synthesis in a concentration-dependent manner. Pretreatment with RNase reduced this stimulation. The amino acid sequence of the cloned insert, derived from the nucleotide sequence (J.-M. Li, C. S. Russell, and S. D. Cosloy, J. Cell Biol. 107:617a, 1988), showed no homology with any ALA synthase sequenced to date. These results suggest that E. coli synthesizes ALA by the C5 pathway from the intact five-carbon chain of glutamate. 相似文献
9.
During growth in the presence of tracer amounts of exogenously supplied alpha-keto[1-14C]glutarate (AKG) or alpha-keto [5-14C]glutarate, cytochrome-containing Bacteroides fragilis strain 2044 and Bacteroides vulgatus strain 8482 incorporated extremely small amounts of radioactivity into cell macromolecules and protoheme. Under identical conditions, Bacteroides "l" strain 7CM and Bacteroides buccae strain J1 incorporated substantial label from [5-14C]AKG, but not [1-14C]AKG, into cellular macromolecules and protoheme. Bacteroides succinogenes strain S85 incorporated radioactivity from both [1-14C]AKG and [5-14C]AKG into cell macromolecules, but only label from [5-14C]AKG appeared in protoheme. Selenomonas ruminantium strain HD1 and Butyrivibrio fibrisolvens strain D1, both of which are devoid of cytochromes, incorporated substantial label from both [1-14C]AKG and [5-14C]AKG into cell macromolecules, but failed to incorporate label from either position into protoheme. Bacteroides ruminicola sp. brevis strain GA33 incorporated label from both [1-14C]AKG and [5-14C]AKG into both cell macromolecules and protoheme. A substantial portion of the heme synthesized by this organism may be formed by the "plant" pathway involving the intact use of the AKG carbon skeleton. Major differences exist in the manner and extent of AKG utilization among cytochrome-containing anaerobes and between these organisms and bacteria devoid of cytochromes obtained from similar environments. 相似文献
10.
A crude extract that catalyzes the epoxidation of toluquinol and gentisyl alcohol was isolated from cultures of Penicillium patulum. About 60% of the activity sedimented from crude extract upon centrifugation at 105,000g for 2 h, and at 30,000g for 30 min after precipitation with 30% ammonium sulfate and resuspension in buffer. The quinone epoxide phyllostine, a product of gentisyl alcohol epoxidation, has previously been shown to be an intermediate in the biosynthesis of patulin [Sekiquchi, J., & Gaucher, G. M. (1978) Biochemistry 17, 1785-1791] and was shown to be further converted to neopatulin by the extract. The epoxide product of toluquinol, desoxyphyllostine (2-methyl-5,6-epoxy-1,4-benzoquinone), has not been reported previously from fungal cultures. Its structure was confirmed by GC-mass spectrometry and proton and 13C NMR. Its CD spectrum showed the same shape and signs as that of phyllostine, indicating that it too is an enzymatic product with a similar absolute configuration. Whereas chemical epoxidation of toluquinone and gentisyl quinone occurs with hydrogen peroxide, the enzymatic epoxidation utilized oxygen and the hydroquinone. The epoxidation was inhibited by 1,10-phenanthroline, EDTA, and p-(chloromercuri)benzenesulfonic acid and by degassing with nitrogen, but no inhibition was observed with KCN, catalase, or CO. The apparent Km's were similar for the two substrates (0.17 mM for toluquinol, 0.24 mM for gentisyl alcohol), with both substrates showing inhibition at 1.0 mM. The rate of desoxyphyllostine formation was more than 10 times that of phyllostine formation at equivalent substrate concentrations. Gentisaldehyde was not a substrate for the enzyme.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
11.
Biosynthesis of a mycobacterial lipopolysaccharide. Incorporation of [14C]acyl groups by whole cells in vivo 总被引:1,自引:0,他引:1
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1. Mycobacterium phlei (A.T.C.C. 356) cells were incubated with (14)C-labelled short-chain fatty acids and the 6-O-methylglucose-containing lipopolysaccharides that became esterified with radioactive acyl groups were isolated. The pattern of labelling of these lipopolysaccharides with the different acyl groups, the effects of different conditions on labelling patterns, and the kinetics of the turnover of (14)C-labelled acyl groups were studied. 2. The labelling patterns are summarized as follows. [1-(14)C]Acetate was incorporated into all of the acyl groups. [1-(14)C]Propionate led to labelling of propionate and succinate, while [1-(14)C]isobutyrate was incorporated mostly as such, along with a trace amount in iso-octanoate. 3. Under the conditions of the experiments, [1-(14)C]acetate was rapidly incorporated into succinyl (3-carboxypropionyl) and octanoyl groups, whereas the acetyl groups themselves were labelled more slowly. Radioactivity in propionyl and succinyl groups, originating from [1-(14)C]propionate, attained maximum values and then gradually decreased in both. Incorporation of [1-(14)C]isobutyrate proceeded slowly but reached a plateau and remained constant. While n-butyrate is not a normal constituent of methyl-glucose-containing lipopolysaccharides, it was incorporated as such when n-[1-(14)C]-butyrate was supplied in the medium. 4. [1-(14)C]Acetyl groups were readily displaced by unlabelled acetate. On the other hand, the specific radioactivity of the succinyl group continued to increase during a 3h incubation with unlabelled succinate. Propionyl and succinyl groups, labelled by [1-(14)C]propionate, were displaced slowly by unlabelled propionate or succcinate. The isobutyryl group of the lipopolysaccharides did not turn over, in contrast to the results obtained with the other acyl substituents. 相似文献
12.
1. The mechanism of regeneration of glycine during the growth of Pseudomonas AM1 on C(1) compounds has been investigated by brief incubation of bacterial suspensions with [2,3-(14)C(2)]succinate and observing the incorporation of radioactivity into various metabolites. 2. With the wild-type organism growing on methanol, radioactivity appeared rapidly in glycine and tricarboxylic acid-cycle intermediates, but there was a relatively slow labelling of serine and phosphorylated compounds. Serine became labelled predominantly in the C-2 position. 3. The proportion of radioactivity incorporated into glycine at earliest times was greatly diminished when succinate-grown cells were used. 4. Radioactivity was also incorporated from [2,3-(14)C(2)]succinate into glycine and serine by methanol-grown mutant 20S, which lacks phosphoserine phosphohydrolase. Both the glycine and serine were labelled mainly in C-2. 5. The formation of predominantly [2-(14)C]serine from [2,3-(14)C(2)]succinate in wild-type Pseudomonas AM1, and of [2-(14)C]serine and [2-(14)C]glycine in the mutant lacking the phosphorylated pathway from succinate to serine, is taken as strong evidence for a mechanism of glycine regeneration involving cleavage of a C(4) skeleton between C-2 and C-3, rather than by a direct combination of two C(1) units derived from the growth substrate. 6. The cleavage mechanism is quantitatively more significant during growth on methanol than on succinate. 相似文献
13.
1. Methods are described for the isolation of radioactively pure thiamine from yeast and its degradation on a small scale to its cyclic components. 2. A degradation of the pyrimidine ring and a thin-layer method for the separation of thiamine, its derivatives and pyrimidine and thiazole residues are described. 3. [(14)C]Formate is more effectively incorporated into the pyrimidine residue than into the thiazole residue, whereas the reverse is true with l-[Me-(14)C]methionine. 4. Experiments with [Me-(14)C,(35)S]methionine demonstrate that methionine provides an intact unit for the biosynthesis of the thiazole ring. 5. [6-(14)C]Orotic acid is insignificantly incorporated into the pyrimidine residue of thiamine. 6. Experiments with [1-(14)C]- and [2-(14)C]-acetate indicate that it is incorporated as a unit into the thiazole residue, but that only C-2 is incorporated into the pyrimidine residue. 7. l-[U-(14)C]Alanine is also effectively incorporated into the thiazole residue. 8. These results are discussed in relation to possible pathways of biosynthesis of the two ring components of the thiamine molecule. 相似文献
14.
The formation of lipid-linked sugars as intermediates in glycoprotein synthesis in rabbit mammary gland 总被引:5,自引:3,他引:2
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1. The incorporation of d-[1-(14)C]mannose, d-[2-(3)H]mannose and N-acetyl-d-[1-(14)C]-glucosamine into glycoproteins and lipid-linked intermediates of mammary explants obtained from lactating rabbits was studied. The amount of radioactivity incorporated into lipid-linked intermediates was very low compared with the incorporation into protein. Most of the radioactivity incorporated into the chloroform/methanol-soluble fraction was present as neutral lipid. Radioactivity from d-[2-(3)H]mannose was incorporated mainly into the fatty acid moiety, whereas radioactivity from d-[1-(14)C]mannose and N-acetyl-d-[1-(14)C]glucosamine was present in the glycerol moiety of triacylglycerol. 2. The labelled lipid-linked intermediate that was soluble in chloroform/methanol/water (10:10:3, by vol.) was partially characterized and was found to exhibit properties characteristic of an oligosaccharide linked to lipid via a pyrophosphate bridge. It migrated largely as a single zone of radioactivity on t.l.c. and was eluted from a column of DEAE-cellulose acetate as a single peak by 50mm-ammonium acetate. 3. The oligosaccharide moiety was released from the lipid by mild acid hydrolysis. The size of the oligosaccharide was estimated by paper chromatography to be 10 or 11 monosaccharide units. 4. d-[1-(14)C]Mannose was incorporated largely into glycopeptides with molecular weights in the range 40000-80000, as determined by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. Label from N-acetyl-d-[1-(14)C]glucosamine was incorporated into a glycopeptide with an electrophoretic mobility identical with that of rabbit casein (mol.wt. 32000) as well as into glycopeptides of higher molecular weight. 5. Approx. 50% of the total radioactivity in the protein labelled from N-acetyl-d-[1-(14)C]glucosamine was present as galactosamine, a component of the carbohydrate portion of rabbit casein. No labelled galactosamine was present in the lipid-linked oligosaccharide labelled from N-acetyl-d-[1-(14)C]glucosamine. It thus appears that the lipid-linked oligosaccharide is not involved in the glycosylation of casein. 相似文献
15.
A medium chain length fatty acid, [1-(14C)] lauric acid (12:0) was administered to the detached leaves of Artemisia and was incorporated into major lipids, including phospholipids and galactolipids. [1-(14C)]12:0 was elongated and desaturated into linolenic acid (18:3). In detached leaves of both Artemisia and Arabidopsis thaliana ecotype Columbia, radioactivity from [14C]18:3 was incorporated into jasmonic acid (JA) and methyl jasmonate (MJ). Higher amounts of [14C]JA were measured in Artemisia than Arabidopsis leaves. In Artemisia, [14C]JA was actively metabolized into [14C]MJ. Extracts prepared from the leaves of Artemisia, exhibited higher in vitro JA methyltransferase activity than those from Arabidopsis. 相似文献
16.
The types of unsaturated fatty acids found in platelet phospholipids must be regulated by a series of controls which include specificity for activation and acylation as well as modification of circulating fatty acids by platelets prior to incubation into phospholipids. In this study we show that washed human platelets not only incorporate [1-14C]6,9,12-18:3, [1-14C]6,9,12,15-18:4, [1-14C]5,8,11-20:3, [1-14C]5,8,11,14-20:4, and [1-14C]5,8,11,14,17-20:5 into their phospholipids but also chain elongate each of these acids with subsequent acylation of the chain elongated products into phospholipids. Platelets incubated alone with 1-14C-labeled 5,8,11-20:3, 5,8,11,14-20:4, 5,8,11,14,17-20:5, 7,10,13,16,19-22:5, or 4,7,10,13,16,19-22:6 incorporated each of these acids into individual phosphoglycerides with phosphatidylinositol having the highest specific activity followed by phosphatidylcholine with phosphatidylserine approximately equal to phosphatidylethanolamine. The incorporation specificity of 4,7,10,13,16,19-22:6 was atypical since it was a relatively poor substrate for acylation into all phospholipids except phosphatidylethanolamine. The 20-carbon acids were better substrates for incorporation into phospholipids than were the 22-carbon compounds. Simultaneous incubation of 10 microM [1-14C]5,8,11,14-20:4 with increasing levels (5 to 15 microM) of each of the above five other 1-14C-labeled acids showed a concentration-dependent increase in the amount of the second fatty acid incorporated into platelet phospholipids. Dietary fat modification thus has the potential of increasing the plasma pool of 22-carbon acids for incorporation into platelets. In addition the activation of 20-carbon eicosanoid precursors by the high affinity platelet activating enzyme (Wilson, D. B., Prescott, S. M. and Majerus, P. W. (1982) J. Biol. Chem. 257, 3510-3515) will yield an acyl-CoA for both acylation and chain elongation followed by subsequent incorporation of 22-carbon acids into phosphoglycerides. 相似文献
17.
Adenosine utilization in cordycepin-sensitive mutants of Saccharomyces cerevisiae. 总被引:1,自引:1,他引:0
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A purine-requiring, wild-type yeast strain was cordycepin resistant and failed to grow in medium containing adenosine; in contrast, a cordycepin-sensitive mutant (also purine requiring) grew well in medium containing adenosine. The cordycepin-sensitive mutant incorporated [8-14C]adenosine at nine times the wild-type rate, and adenosine completely fulfilled the purine requirement of the cells. Exogenous adenosine rapidly entered the mutant cells, apparently as free nucleoside, and was phosphorylated; uptake displayed concentration-dependent saturation kinetics (Km, 6 mM). Within 10 min 14C radioactivity was being incorporated into nucleic acids. 相似文献
18.
A study on the tricarboxylic acid cycle and the synthesis of acetylcholine in the lobster nerve 总被引:5,自引:3,他引:2
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1. The biosynthetic origin of the amide substituent of N-(alpha-hydroxyethyl)lysergamide has been studied. 2. [1-(14)C]Acetate, [(14)C]formate, [2-(14)C]mevalonic acid lactone, [2-(14)C]indole, dl-[3-(14)C]tryptophan, dl-[3-(14)C]serine, dl-[2-(14)C]alanine and [2-(14)C]pyruvate were efficiently incorporated into the alkaloid, but not dl-[1-(14)C]alanine or [1-(14)C]pyruvate. 3. Only the dl-[2-(14)C]alanine- and [2-(14)C]pyruvate-derived alkaloid contained appreciable radioactivity in the amide substituent. 4. l-[(15)N]Alanine-derived alkaloid was shown to be specifically labelled in the amide nitrogen. However, l-[(14)C,(15)N]alanine was found to be incorporated into the methylcarbinolamide substituent with an appreciable increase in the (15)N/(14)C ratio, suggesting that alanine is not the direct precursor of this moiety. 相似文献
19.
Our previous studies (Watson, J. A., Havel, C. M., Lobos, D. V., Baker, F. C., and Morrow, C. J. (1985) J. Biol. Chem. 260, 14083-14091) suggested that a matabolite, distal to isopentenyl 1-pyrophospate (IPP), served as a regulatory signal for sterol-independent modulation of Kc cell 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity. This report summarizes efforts to localize the potential source of the post-IPP regulatory signal molecule. We found no direct correlation between mevalonate-mediated suppression of Kc cell HMG-CoA reductase activity and the rates of [1-14C]-, [3-14C]-, [5-14C]-, or [5-3H]mevalonate incorporation into either carbon dioxide, neutral lipids, water, or water-soluble isopentenoid pyrophosphate esters. [1-14C]Mevalonate's rate of conversion to 14CO2 (a measure of total isopentenyl 1-pyrophosphate synthesis) was minimally 5-fold greater than that for neutral isopentenoid lipid synthesis (measured with either [5-3H]-, [3-14C]-, or [5-14C]mevalonate). However, [5-3H]mevalonate's rate of conversion into [3H]H2O (measure of shunted mevalonate carbon) was equivalent or greater than that measured for neutral isopentenoid lipid synthesis. [5-14C]Mevalonate radioactivity was incorporated into macromolecules and n-fatty acids. Kc cell extracts (100,000 X g supernatant fluid) readily oxidized alcohols with the following activity sequence: geraniol = nerol greater than farnesol = dimethylallyl alcohol greater than geranylgeraniol, isopentenyl alcohol, and allyl alcohol. Oxidation required NAD, and ethanol was not a substrate. We conclude that (a) Kc cells shunted a significant fraction (greater than or equal to 40%) of their post-IPP carbon to prenols for oxidative catabolism and (b) that shunted mevalonate carbon may play a significant role in the mevalonate-mediated regulation of Kc cell HMG-CoA reductase activity. 相似文献
20.
Manganese and antibiotic biosynthesis. I. A specific manganese requirement for patulin production in Penicillium urticae 总被引:1,自引:0,他引:1
The effect of trace metal nutrition on the functioning of the patulin biosynthetic pathway in submerged cultures of Penicillium urticae (NRRL 2159A) was examined by both chromatographic and enzymological means. Comprehensive metal ion analysis showed generally low levels of contaminating metal ions in media components. Of eight metal ions examined, only manganese strongly influenced secondary metabolite production. In control cultures or cultures deficient in calcium, iron, cobalt, copper, zinc, or molybdenum, pathway metabolites appeared in the medium at about 25 h after inoculation. The first pathway-specific metabolite, 6-methylsalicylic acid, accumulated only transiently before being converted to patulin whose concentration steadily increased. In manganese-deficient cultures, however, 6-methylsalicylic acid continued to accumulate, with only minor amounts of patulin being produced. Additionally, a marker enzyme for the pathway showed only 0-20% of control activity. Clear dose responses (patulin versus manganese) were found in different media, with no effect on growth yield. Addition of manganese to depleted cultures at 18, 26, or 36 h resulted in increasing marker enzyme activity and patulin concentrations. It is concluded that manganese exerts a specific, positive effect on patulin biosynthesis and may in some way control the section of the patulin pathway occurring after 6-methylsalicylic acid. 相似文献