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1.
P. Jangbua K. Laoteng P. Kitsubun M. Nopharatana A. Tongta 《Letters in applied microbiology》2009,49(1):91-97
Aims: This study aims to maximize the yield of gamma-linolenic acid by a filamentous fungus, Mucor rouxii , using low cost production by solid-state fermentation.
Methods and Results: We optimized substrate types and culture conditions including inoculum size and temperature. The optimal growth of M. rouxii was found in the cultures inoculated with 5 × 105 spores g−1 substrate. The fungal cells grew well on rice bran and soy bean meal, whereas a lower biomass was found in the cultures grown on polished rice, broken rice and spent malt grain. The GLA content was highly accumulated in rice bran ferment and its maximal content of about 6 g kg−1 fermented mass was observed in the 5th-day culture grown at 30°C. However, the GLA content in the rice bran ferment was not enhanced by low temperature.
Conclusions: The GLA production of M. rouxii could be enhanced by optimizing the agricultural by-product substrates and culture condition.
Significance and Impact of the Study: Low cost GLA production process was achieved, and fermented product containing GLA can be incorporated into feed additives without further oil extraction to alternate the expensive plant oils. 相似文献
Methods and Results: We optimized substrate types and culture conditions including inoculum size and temperature. The optimal growth of M. rouxii was found in the cultures inoculated with 5 × 10
Conclusions: The GLA production of M. rouxii could be enhanced by optimizing the agricultural by-product substrates and culture condition.
Significance and Impact of the Study: Low cost GLA production process was achieved, and fermented product containing GLA can be incorporated into feed additives without further oil extraction to alternate the expensive plant oils. 相似文献
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To understand the relationship between fatty acid metabolism and the growth morphology of Mucor rouxii, fatty acid profiling was studied comparatively in cells grown under conditions which included different atmospheric conditions or the addition of phenethyl alcohol (PEA). The significant difference in fatty acid profiles from M. rouxii grown under aerobic or anaerobic conditions was not found to be directly related to morphological growth. Oxygen limitation, which induced the formation of pure multipolar budding yeasts, led to a decrease in long-chain fatty acids-- particularly unsaturated fatty acids-- and an increase in medium-chain saturated fatty acids, a finding which contrasted with the aerobic cultures, including mycelia and PEA-induced bipolar budding cells. High levels of C18 : 1Delta(9) were found in aerobic yeast cultures with additional PEA when compared to that in the aerobically grown mycelia. The identification of unusual fatty acids in Mucor in response to alcoholic and hypoxic stresses - including odd-numbered fatty acids and 7-hydroxy dodecanoic acid (7-OH C12 : 0) in addition to the more common fatty acids - implied that an important role existed for these unusual fatty acids. 相似文献
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Purification and properties of two endochitosanases from Mucor rouxii implicated in its cell wall degradation 总被引:1,自引:0,他引:1
Abstract From autolysed cultures of Mucor rouxii , two chitosanases, A and B, were purified to electrophoretic homogeneity. Apparent M r values of 76 000 and 58 000 and p I values of 4.9 and 4.7 were determined for A and B, respectively. Both chitosanases showed a high specificity for chitosan and chitosan derivatives. They had optimum activities at pH 5.0 and at temperatures of 55°C and 50°C for A and B, respectively. Enzyme A was inhibited by acetate ions and enzyme B by high substrate concentration. Both enzymes showed an endo-splitting type of activity, and the end product of chitosan degradation contained a mixture of dimer, trimer and higher molecular mass oligomers of glucosamine. Glucosamine oligosaccharides were poorly hydrolysed by these enzymes. Both enzymes extensively degraded the chitosan extracted from M. rouxii cell walls. 相似文献
6.
Fabiana M de Almeida Agda K.B Lúcio Maria de Lourdes T.M Polizeli João A Jorge Héctor F Terenzi 《FEMS microbiology letters》1997,155(1):73-77
Two different trehalose-hydrolysing activities, known as acid or non-regulatory trehalases, and neutral or regulatory trehalases, have been recognised in a number of fungal species. The true role of these apparently redundant hydrolases remained obscure for many years. However, recent evidence suggests that neutral trehalases would be specialised in the mobilisation of cytosolic trehalose, while acid trehalases would only hydrolyse extracellular trehalose. Results obtained with Mucor rouxii, a Zygomycete initially thought to posses only neutral trehalase activity, reinforced this hypothesis. M. rouxii grows efficiently in trehalose as the sole carbon source. Trehalose-grown or carbon-starved cells exhibit a high trehalase activity of optimum pH 4.5, bound to the external surface of the cell wall, in contrast with the neutral (pH 6.5) trehalase, which occurs in the cytosol. Other differences between the neutral and the acid trehalases are the temperature optimum (35°C and 45°C, respectively) and thermal stability (half-life of 2.5 min and 12 min at 45°C, respectively). The neutral trehalase, but not the acid trehalase, is activated in vitro by cAMP-dependent phosphorylation, stimulated by Ca2+, and inhibited by EDTA. It shows maximal activity at germination and decreases as growth proceeds. In contrast the activity of the acid trehalase is totally repressed in glucose-grown cultures and increases upon exhaustion of the carbon source, and is strongly induced by extracellular trehalose. 相似文献
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Molecular weight distribution of chitosan isolated from Mucor rouxii under different culture and processing conditions 总被引:1,自引:0,他引:1
The isolation of chitosan from a fungal source offers the potential of a product with controlled physicochemical properties not obtainable by the commercial chemical conversion of crustacean chitin. A variety of culture and processing protocols using Mucor rouxii were studied for their effects on biomass yield and chitosan molecular weight. Weight-averaged molecular weight determined by gel permeation chromotography ranged from 2.0 x 10(5) to approximately 1.4 x 10(6) daltons. The chitosan yield ranged from 5% to 10% of total biomass dry weight and from 30% to 40% of the cell wall. Of the culture parameters studied, length of incubation and medium composition effected biomass production and molecular weight. Modification of the processing protocol, including the type and strength of acid, and cell wall disruption in acid prior to refluxing were used to optimize the efficiency of chitosan extraction.The degree of deacetylation of fungal and commercial chitosans was compared using infrared spectrometry, titration, and first derivative of UV absorbance spectrometry. The chitosan obtained directly from the fungal cell wall had a higher degree of deacetylation than commercial chitosan from the chemical conversion process. 相似文献
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Everardo Lopez-Romero Eduardo Monzon Jose Ruiz-Herrera 《FEMS microbiology letters》1985,30(3):369-372
Abstract The free sterol composition of chitosomes and whole yeast cells of Mucor rouxii was determined by gas-liquid chromatography (GLC). The results obtained showed that, in contrast to whole cells, chitosomes are particularly rich in ergosterol: 87% vs. 29% of total free sterols. Three other sterols were present in varying amounts in both chitosomes and whole cells. These were identified by GLC as cholesterol, stigmasterol and sitosterol. These results indicate that the yeast cells preferentially accumulate ergosterol in chitosomes providing these microvesicles with a unique composition that may be important for the structure of the organelle. 相似文献
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We investigated whether cellular responses to various stress conditions are regulated in synchronization with the ultradian rhythm of respiratory-fermentative metabolism which is coupled to the cell cycle rhythm in continuous cultures of the yeast Saccharomyces cerevisiae. The cellular resistance to heat oscillated with a peak at the late respiro-fermentative phase, which approximately corresponds to the unbudding period of the cell cycle. Cellular resistance to H(2)O(2) and that to the superoxide-generating agent menadione oscillated in the same phase as that of heat resistance. The resistance to cadmium and that to 1-chloro-2,4-dinitrobenzene, an uncoupler of energy metabolism in mitochondria, both oscillated with a peak advanced by about 80 degrees relative to that of heat resistance, approximately covering the respiro-fermentative phase. Thus, cellular resistance to various stresses in S. cerevisiae oscillated in synchronization with the metabolic oscillation in the continuous culture. 相似文献
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A soluble NAD-dependent alcohol dehydrogenase (ADH) activity was detected in mycelium and yeast cells of wild-type Mucor rouxii. In the mycelium of cells grown in the absence of oxygen, the enzyme activity was high, whereas in yeast cells, ADH activity
was high regardless of the presence or absence of oxygen. The enzyme from aerobically or anaerobically grown mycelium or yeast
cells exhibited a similar optimum pH for the oxidation of ethanol to acetaldehyde (∼pH 8.5) and for the reduction of acetaldehyde
to ethanol (∼pH 7.5). Zymogram analysis conducted with cell-free extracts of the wild-type and an alcohol-dehydrogenase-deficient
mutant strain indicated the existence of a single ADH enzyme that was independent of the developmental stage of dimorphism,
the growth atmosphere, or the carbon source in the growth medium. Purified ADH from aerobically grown mycelium was found to
be a tetramer consisting of subunits of 43 kDa. The enzyme oxidized primary and secondary alcohols, although much higher activity
was displayed with primary alcohols. K
m values obtained for acetaldehyde, ethanol, NADH2, and NAD+ indicated that physiologically the enzyme works mainly in the reduction of acetaldehyde to ethanol.
Received: 11 March 1999 / Accepted: 14 July 1999 相似文献
12.
Mirko Trutnau Nicole Suckale Gerlinde Groeger Thomas Bley Jelka Ondruschka 《Engineering in Life Science》2009,9(6):437-443
Chitosan is a major structural component of fungal cell walls and has diverse medical and other applications. However, cost‐effective culture and extraction methods for fungi need to be developed. Therefore, Mucor rouxii was grown on YPG‐media in both submerged batch and semi‐continuous cultures. Chitosan was extracted from the mycelia to explore strategies to enhance yields and production rates. As observed in earlier studies, M. rouxii is able to adapt to shear stress when cultured semi‐continuously. Modeling the hyphal growth of batch experiments shows that the mycelia were ruptured by shear forces within a short cultivation time shown by a decreased hyphal length. However, an increasing chitosan content was observed with an increasing cultivation period in semi‐continuous cultures, which is an indication for the adaption to shear stress. Semi‐continuous culture resulted in the highest contents of extractable chitosan. The results and models of hyphal growth, including tip extension and branching, suggest that repeated batch cultures may be optimal for chitosan production. 相似文献
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J. C. du Preez M. Immelman J. L. F. Kock S. G. Kilian 《World journal of microbiology & biotechnology》1997,13(1):81-87
The effect of different initial acetic acid concentrations on the growth of and lipid and gamma-linolenic acid (GLA) production
byMucor circinelloides CBS 203.28 was determined in a 14 litre stirred tank reactor operated in a fedbatch, pH-stat mode with acetic acid as carbon
source and pH titrant. Increased acetic acid concentrations in the culture resulted in a significant increase in the crude
oil content of the biomass. By contrast, all the other parameters such as the biomass concentration, GLA and oil yield on
acetic acid, the GLA content of the biomass and oil, the growth rate and volumetric rate of GLA production decreased with
an increase in acetic acid concentration. The best results were obtained with acetic acid at 2 g/1, which gave 39.8 mg GLA/g
biomass and 15.6% GLA in the neutral lipid fraction, amounting to 340 mg GLA/1 culture. A decrease in the glyco- and phospho-lipid
fractions during the cultivation coincided with an increase in the neutral lipid fraction. The GLA content of the biomass
remained within rather narrow limits of 3.5% to 4% of the biomass, irrespective of the oil content of the biomass. The fatty
acid profile was not greatly affected by the acetic acid concentration. The hyphae of the fungus were characterized by the
accumulation of large intracellular oil droplets and some septa delimited the hyphae. 相似文献
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J. C. du Preez M. Immelman S. G. Kilian 《World journal of microbiology & biotechnology》1996,12(1):68-72
The Fischer-Tropsch reaction water, which contains C2 to C5 monocarboxylic acids, generated as a co-product of the Sasol industrial oil-from-coal process, constitutes a potential cheap carbon substrate for the production of gamma-linolenic acid (GLA) by selced Mucor species. Three strains of Mucor were each grown in an air-lift reactor operated in a fed-batch, pH-stat mode under N-limitation with a mixture of C2 to C5 monocarboxylic acids as both pH titrant and carbon source. The production of GLA from this substrate was evaluated. Growth typically resulted in the rapid assimilation of acetic, n-butyric and n-valeric acids. Although propionic, iso-butyric and iso-valeric acids were assimilated to varying degrees, these acids accumulated in the culture. Mucor circinelloides CBS 203.28 gave the best results in that it assimilated 36% to 100% of each acid, had a biomass yield coefficient of 0.3 (calculated on acids utilized), and contained 28% crude oil, 84% of which comprised neutral lipids with a GLA content of 14.4%, giving 33 mg GLA/g biomass. GLA accumulation coincided with a decrease in the stearic-acid content of the neutral-lipid fraction. The results were comparable with previous results obtained with acetic acid and glucose as sole carbon sources, demonstrating the feasibility of producing GLA from the above mixture of organic acids. 相似文献
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The effect of various culture conditions on growth, lipid production and fatty acid composition in Mucor rouxii and Mucor sp.1b were studied. Total lipid production was higher in media containing potassium nitrate for both the cultures (30%) and cultures grown on plant seed oil produced more than 44% lipid. Among the carbon sources tested, γ-linolenic acid (GLA) production was maximal in cultures grown on glucose. The major fatty acids produced by these two cultures were palmitic, stearic and oleic acids. Levels of GLA in M. rouxii and M. sp.1b was in the range of 3–17% under different culture conditions. Lactose was a poor promoter for biomass and lipid production in both cultures. No GLA was found in fungal cultures grown on sesame oil. The optimal conditions for the production of GLA was standardised in these cultures. 相似文献
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An acid protease was purified from the mycelial form of Mucor rouxii by a method which involved salt and acid precipitation, gel filtration and anion-exchange chromatography. The enzyme had a molecular mass of 16,000 Da. Its optimum pH was 4.0, maximal activity was obtained at 50°C, and it was inactivated at 70°C. It was not affected by leupeptin or N -p-tosyl-L-lysine chloromethyl ketone (TLCK) but diazoacetyl-DL-norleucine methyl ester (DNME) in the presence of Cu2+ and more noticeably pepstatin A, strongly inhibited the activity. This acid protease did not activate zymogenic chitin synthetase from the fungus, but brought about its inactivation even at low concentrations and after short periods of incubation time.Abbreviations TLCK
N -p-tosyl-L-lysine chloromethyl ketone
- DNME
diazoacetyl-DL-norleucine methyl ester
- TCA
trichloroacetic acid
- SDS
sodium dodecyl sulfate 相似文献
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The distribution of F-actin microfilaments and microtubules was analyzed in germinating sporangiospores of Mucor rouxii by labeling with rhodamine-tagged phalloidin and by immunofluorescence microscopy. The transition from isodiametrical to apical growth was accompanied by a switch from uniform distribution of F-actin patches to a polarized accumulation of F-actin material at the germ tube tips. Immunoblotting of cell-free extracts of M. rouxii with a monoclonal anti-porcine -tubulin antibody (TU-01) disclosed two discrete bands of -tubulin suggesting the existence of two -tubulin genes in this fungus. Immunofluorescence microscopy of germinating cells stained with the same antibody revealed an elaborate network of cytoplasmic microtubules that persisted during the entire germination process and extended into the apex of the germ tube. Although their precise roles remain undetermined, the observed arrangement of cytoskeletal elements during germination is consistent with their presumed involvement in cell wall morphogenesis: the long axial microtubules serving as long-distance conveyors of wall-building vesicles to the apical region while the concentrated F-actin patches mark the participation of microfilaments in the zone of intense vesicle exocytosis at the hyphal apex.Abbreviations DAPI
4,6-diamidino-2-phenylindole
- DTT
dithiothreitol
- EGTA
Ethylene glycol-bis (beta-aminoethyl ether)
- N,N,N,N
tetraacetic acid
- F-actin
Filamentous actin
- MES
2-(N0morpholino)-ethanesulfonic acid
- PIPES
Piperazine-N,N-bis-2-ethanesulfonic acid
- PMSF
Phenyl-methylsulphonyl fluoride
- TBS
Tris-buffered saline 相似文献
20.
1,3‐propanediol is an important chemical widely used in polymer production. In this study, two strains, Zygosacharomyces rouxii JL2011 and Klebsiella pneumoniae S6, were used as a mixed culture for 1,3‐propanediol production directly from glucose. Two important parameters including inoculation time of K. pneumoniae S6 at stage of mixed culture and initial cell ratio of Z. rouxii JL2011 to K. pneumoniae S6 in mixed fermentation were optimized in culture flasks. In those experiments, the best results were obtained with a yield of 6.8 g/L 1,3‐propanediol from glucose when K. pneumoniae S6 was inoculated after 48 h in the culture of Z. rouxii JL2011 by mixed culture of Z. rouxii JL2011 and K. pneumoniae S6 with initial cell ratio of 1:200. In a 7‐L bioreactor, the maximum 1,3‐propanediol production could reach up to 15.2 g/L. Thus, this study presents an effective process for 1,3‐propanediol microbial production from glucose by using mixed culture of Z. rouxii JL2011 and K. pneumoniae S6. This work does not only demonstrate a new way to produce 1,3‐propanediol from a low‐cost feedstock, but may also make a valuable contribution to the development of a cost‐effective fermentation based on renewable resources. 相似文献