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Carbon Balance Studies on Selected Species of Marine Filamentous Fungi
Title
Carbon Balance Studies on Selected Species of Marine Filamentous Fungi.
Creator
Dias, Anthony P., Sguros, Peter L., Florida Atlantic University
Abstract/Description
Selected species of marine filamentous fungi, Halosphaeria mediosetigera, Humicola alopallonella, Culcitalna achraspora and Orbimyces spectabilis, grown in a semi-synthetic medium containing glucose 0.5, ammonium nitrate 0.24, tris(hydroxyrnethyl)aminomethane 0.12 and yeast extract 0.1% (Medium A) and modifications thereof have been studied with regard to cellular composition, carbon fate and growth kinetics. Cultures were aerated with sterile, carbon dioxide-free air and washed mycelia were...
Show moreSelected species of marine filamentous fungi, Halosphaeria mediosetigera, Humicola alopallonella, Culcitalna achraspora and Orbimyces spectabilis, grown in a semi-synthetic medium containing glucose 0.5, ammonium nitrate 0.24, tris(hydroxyrnethyl)aminomethane 0.12 and yeast extract 0.1% (Medium A) and modifications thereof have been studied with regard to cellular composition, carbon fate and growth kinetics. Cultures were aerated with sterile, carbon dioxide-free air and washed mycelia were analyzed for macroconstituent elementary composition, total carbon, protein, chitin and lipids. Culture filtrates were tested for total carbon, .extracellular carbohydrates and organic acids. Physiological profiles were obtained to ascertain and correlate the ·extent and rate of growth; and changes in pH, concentration of glucose . and anthronepositive materials. · Elementary analyses of washed fungal mycelia showed 41.0 to 49.0% carbon, 4.7 to 6.0% nitrogen and 5.4 to 6.5% · hydrogen. Optimal growth was accompanied by a concomitant fall in pH followed by a shift back to the alkaline side. Following autolysis at 8 days, almost one-third of the glucose substrate reappeared in the medium accompanied by smaller amounts of mannose with Halosphaeria. With Culcitalna, following exhaustion of glucose in the medium, autolysis was accompanied by the reappearance of glucose. Pentoses and a hexose other than glucose also were indicated by relative mobilities on thin layer chromatograms. The most profound pH changes were obtained with Humicola reaching a low of 2.4 in 15 days, but the pH at 45 days had stabilized around 5.3. Increasing the glucose concentration to 5.0% (Medium B) did not increase the mycelial yields to any substantial degree except for Halosphaeria. In all cases pH fluctuated around 4 between 10 and 25 days, hut the final pH was about 6. Production of mycelia was always accompanied by substantial utilization of the glucose in the medium. When the glucose concentration was raised to 5%, to force the buildup of metabolic products, and yeast extract omitted (Medium C), substantial amounts of glucose were utilized, but this was not manifested by parallel increases in growth. Carbon balance studies confirmed that pH changes were not due to the production of organic acids, but probably to preferential utilization of ailllllonium cations. The initial substrate carbon was found almost exclusively in the fungal mycelia and evolved carbon dioxide. Carbon recoveries obtained under standard conditions were 97.9, 97.4, 98.2 and 94.0% respectively for Halosphaeria mediosetigera, Humicola alopallonella, Culcitalna achraspora and Orbimyces spectabilis in the standard medium containing 0.5% glucose. In 5% glucose (Medium B) the recoveries were 93.0, 90.6 and 84.7% for the first three organisms mentioned. Time did not allow a complete investigation of the causes. of decreased carbon recoveries, but greater production of incompletely oxidized metabolites appeared probable. Economic coefficients were determined for three species in three media while determinations on the fourth species are in progress. Culcitalna gave values of 71 in Medium A, 40 in Medium B and 3 in Medium C; Halosphaeria gave 78 in A, 45 in B and 15 in C; Orbimyces gave 75 in A and 7 in C . The effect of omission of yeast extract and of elevated glucose concentration on the economic coeffic ients was pronounced . The only products detected under exaggerated carbon conditions were acetic acid in the culture broth of Culcitalna and a greenish pigment in the culture filtrate of Halospaeria. The analogously-suggested accumulation of 2-ketogluconic acid in the latter case could be indicative of the functioning of the hexose monophosphate shunt or another primary aerobic pathway.
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Date Issued
1970
PURL
http://purl.flvc.org/fau/fd/FA00000743
Subject Headings
Marine fungi
Format
Document (PDF)
EVIDENCE FOR THE EXISTENCE OF THE TRICARBOXYLIC ACID CYCLE AND GLYOXYLATE PATHWAY IN CERTAIN MARINE FILAMENTOUS FUNGI
Title
EVIDENCE FOR THE EXISTENCE OF THE TRICARBOXYLIC ACID CYCLE AND GLYOXYLATE PATHWAY IN CERTAIN MARINE FILAMENTOUS FUNGI.
Creator
VEMBU, DEVI T., Florida Atlantic University
Abstract/Description
Cellulolytic marine isolates, Culcitalna achraspora and Halosphaeria mediosetigera, were shaker-grown at 25 C in artificial seawater containing glucose, NH4NO3 , tris(hydroxymethyl) aminomethane and yeast extract, pH 7.5. Cultures were harvested in the late linear phase where whole cell endogenous respiration was maximal. Spectrophotometric determinations on acetone powder extracts revealed the presence of aconitase, NAD-linked isocitrate dehydrogenase, succinate dehydrogenase, fumarase,...
Show moreCellulolytic marine isolates, Culcitalna achraspora and Halosphaeria mediosetigera, were shaker-grown at 25 C in artificial seawater containing glucose, NH4NO3 , tris(hydroxymethyl) aminomethane and yeast extract, pH 7.5. Cultures were harvested in the late linear phase where whole cell endogenous respiration was maximal. Spectrophotometric determinations on acetone powder extracts revealed the presence of aconitase, NAD-linked isocitrate dehydrogenase, succinate dehydrogenase, fumarase, malate dehydrogenase and isocitratase. Culcitalna also showed NADP-linked isocitrate dehydrogenase activity. Manometric examinations of acetone powder and dialyzed sonic extracts were largely confirmatory. Oxidation of citric acid cycle intermediates by extracts were observed with Culcitalna for all but succinate with sonic extracts and for all but succinate and 2-oxoglutarate in acetone powder extracts. With Halosphaeria, neither extract could oxidize citrate or succinate, while acetone powder extracts were unable to degrade cis-aconitate and 2-oxoglutarate. Native mycelia were active only against malate. Attempts to permeate cell walls with ethyl esters of intermediates failed.
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Date Issued
1972
PURL
http://purl.flvc.org/fcla/dt/13478
Subject Headings
Marine fungi
Format
Document (PDF)
Soluble NAD- and NADP-Linked Oxidoreductases in Filamentous Marine Fungi: Adaptation of Fluorescent Screening Technique and Development of Supporting Methodology
Title
Soluble NAD- and NADP-Linked Oxidoreductases in Filamentous Marine Fungi: Adaptation of Fluorescent Screening Technique and Development of Supporting Methodology.
Creator
Rodrigues, Joanna, Sguros, Peter L., Florida Atlantic University
Abstract/Description
The metabolism of filamentous marine fungi has received little attention either from the specific or comparative standpoints. As part of an overall investigation of the carbon metabolism of this heterogeneous group of Ascomycetes and Deuteromycetes, a rapid, semi-quantitative method has been adapted to the screening of their NAD- and NADP-linked dehydrogenases. The Ascomycete, Halosphaeria mediosetigera (700), and Deteromycetes, Culcitalna achraspora (230) and Humicola alopallonella (710),...
Show moreThe metabolism of filamentous marine fungi has received little attention either from the specific or comparative standpoints. As part of an overall investigation of the carbon metabolism of this heterogeneous group of Ascomycetes and Deuteromycetes, a rapid, semi-quantitative method has been adapted to the screening of their NAD- and NADP-linked dehydrogenases. The Ascomycete, Halosphaeria mediosetigera (700), and Deteromycetes, Culcitalna achraspora (230) and Humicola alopallonella (710), were grown in a semi-synthetic, artificial sea water broth in submerged culture, from quantitatively standardized inocula, and harvested in the linear growth phase when respirometric data indicated maximum endogenous oxygen uptakes. Based upon dry weight, culture 700 gave QO2 maxima of 9.3-14.6 at 72 hr growth, while culture 710 values were 12.4-16.2 for the same time; culture 230 gave values of 13.8-18.4 at 96 hr growth. Harvested mycelia were washed, quantitated and subjected to sonic disintegration at 21 kHz for 20 minutes at 5-10 C in pH 7.0 phosphate buffer containing cysteine. Debris was removed by centrifuging at 12,000 X g at 4 C for 30 minutes and the resulting turbid, yellow supernatant fraction clarified by two successive 2 hr centrifugings at 4 C and 41,000 X g. Whatman #1 filter paper sheets, divided into 1 cm. squares, were pretreated with pH 8.0 Tris buffer containing MgCl2, and air-dried. The papers were spotted at the grid intersections with appropriate substrates. Cell extracts were mixed with NAD or NADP in Tris and the mixture superimposed upon the substrates. Within 2 hr the papers were viewed under ultraviolet light at 3600 A for spot fluorescence from the reduced coenzymes. In all extracts (5-10 mg protein per ml), dehydrogenases were indicated for glucose, glucose-6-phosphate, gluconate, 6-phosphogluconate, isocitrate and malate. Less definite were indications of oxidoreductases for galactose, lactate and glycerol. It is therefore indicated that these filamentous marine fungi utilize glucose carbon via the hexose monophosphate pathway and the tricarboxylic acid cycle.
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Date Issued
1970
PURL
http://purl.flvc.org/fau/fd/FA00000821
Subject Headings
Marine fungi, Dehydrogenases
Format
Document (PDF)
An undescribed fungal pathogen of reef-forming crustose coralline algae discovered in American Samoa
Title
An undescribed fungal pathogen of reef-forming crustose coralline algae discovered in American Samoa.
Creator
Littler, Mark M., Littler, Diane S., Harbor Branch Oceanographic Institute
Date Issued
1998
PURL
http://purl.flvc.org/FCLA/DT/3318897
Subject Headings
Pathogenic fungi, Coralline algae, Marine fungi
Format
Document (PDF)
EFFECT OF TEMPERATURE, HYDROGEN ION AND CERTAIN GROWTH SUBSTRATES ON THE ACTIVITY OF THE CELLULOLYTIC ENZYME COMPLEX IN SELECTED MARINE FUNGI
Title
THE EFFECT OF TEMPERATURE, HYDROGEN ION AND CERTAIN GROWTH SUBSTRATES ON THE ACTIVITY OF THE CELLULOLYTIC ENZYME COMPLEX IN SELECTED MARINE FUNGI.
Creator
JENSEN, JAMES ROBERT, Florida Atlantic University, Sguros, Peter L., Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description
Halosphaeria mediosetigera and Culcitalna achraspora have been studied to elucidate the mechanism by which they degrade cellulosic materials in the sea. H. mediosetigera (700 LC 1), a C1-less variant, was also employed in this work. Standardized shake-cultures were grown on cellulose and cellulose derivatives supplemented with NH4N03, tris(hydroxymethyl)aminomethane and yeast extract in artificial sea water (Lyman and Fleming), pH 7.5, at 25 C. Induced C1 and Cx enzyme and cellobiase...
Show moreHalosphaeria mediosetigera and Culcitalna achraspora have been studied to elucidate the mechanism by which they degrade cellulosic materials in the sea. H. mediosetigera (700 LC 1), a C1-less variant, was also employed in this work. Standardized shake-cultures were grown on cellulose and cellulose derivatives supplemented with NH4N03, tris(hydroxymethyl)aminomethane and yeast extract in artificial sea water (Lyman and Fleming), pH 7.5, at 25 C. Induced C1 and Cx enzyme and cellobiase activities were determined calorimetrically by correlation with cotton fiber weight loss, by the formation of reducing sugars from carboxymethylcellulose and by Glucostat, respectively. Optimum pH values for both Cx enzyme and cellobiase of C. achraspora were 6.0 at 37 C while those for H. mediosetigera (700 LC 1) were 5.0 and 6.0, respectively, at 37 C. In both cultures, Cx enzymes had a variable optimum temperature ( 37-50 C) at pH 6.0, depending on the length of the incubation period, while cellobiases had the same optimum temperature (50 C) at pH 6.0. Thermostability observations on Cx enzyme and cellobiase of C. achraspora showed almost complete deactivation at 100 C in 15 min for the former and 50 C in 15 min for the latter. Thermostability observations on Cx enzyme and cellobiase of H. mediosetigera (700 LC 1) showed almost complete deactivation at 45 C in 15 min for the former and at 50 C in 15 min for the latter. Data indicate that at least three enzymatic functions are involved in cellulose breakdown by these cultures.
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Date Issued
1971
PURL
http://purl.flvc.org/fcla/dt/13434
Subject Headings
Marine fungi, Cellulose--Biodegradation
Format
Document (PDF)
PRODUCTION, INDUCTION, AND RESOLUTION OF THE CELLULOLYTIC ENZYME COMPLEX IN SELECTED FILAMENTOUS MARINE FUNGI
Title
PRODUCTION, INDUCTION, AND RESOLUTION OF THE CELLULOLYTIC ENZYME COMPLEX IN SELECTED FILAMENTOUS MARINE FUNGI.
Creator
WHITE, JAMES L., JR., Florida Atlantic University
Abstract/Description
Cellulolytic enzymes were induced in the marine fungal isolates, Culcitalna achraspora, Halosphaeria mediosetigera, and Zalerion xylestrix by native cotton and related substrates. Marine environmental conditions were maintained in an artificial seawater medium. Induced c1 and ex enzymes and cellobiase were determined colorimetrically. Induction also occurred on cellobiose, but not on glucose and lactose. Permenter studies with glucose and with toilet tissue carbon sources gave close...
Show moreCellulolytic enzymes were induced in the marine fungal isolates, Culcitalna achraspora, Halosphaeria mediosetigera, and Zalerion xylestrix by native cotton and related substrates. Marine environmental conditions were maintained in an artificial seawater medium. Induced c1 and ex enzymes and cellobiase were determined colorimetrically. Induction also occurred on cellobiose, but not on glucose and lactose. Permenter studies with glucose and with toilet tissue carbon sources gave close correlations between mycelial dry weight, enzyme component activity, and cell nitrogen. In cotton substrate media, Tween 80 increased C1 activity by 148%, and Cx activity by 160%, on the average. Millipore ultrafiltration was the best technique for concentrating cellulase for Sephadex G-75 column chromatography, while conventional techniques were unsuccessful. Fractionation diagrams for all cultures were similar with 2 distinct peaks. The low molecular weight fraction contained mainly Cx enzyme activity, while the high molecular weight fraction contained C1 and Cx enzymes and cellobiase. These fungi possess a cellulase similar, but not identical, to that found in terrestrial species.
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Date Issued
1972
PURL
http://purl.flvc.org/fcla/dt/13537
Subject Headings
Marine fungi, Cellulose--Biodegradation
Format
Document (PDF)
Metabolites from the Marine-Derived Fungus Chromocleista sp. Isolated from a Deep-Water Sediment Sample Collected in the Gulf of Mexico
Title
Metabolites from the Marine-Derived Fungus Chromocleista sp. Isolated from a Deep-Water Sediment Sample Collected in the Gulf of Mexico.
Creator
Park, Young Chul, Gunasekera, Sarath P., Lopez, Jose V., McCarthy, Peter J., Wright, Amy E.
Date Issued
2006
PURL
http://purl.flvc.org/FCLA/DT/3164115
Subject Headings
Metabolites, Fungal metabolites, Marine fungi, X-ray crystallography, Marine sediments
Format
Document (PDF)