Traces Of Galactose Research Articles

A Novel Pectin-Like Glycopeptide Isolated from the Fruit of Fructus Mori Impedes Aggregation and Production of Aβ42.

The fruit of Fructus Mori is food and medicine, which has been demonstrated to have a significant neuroprotective effect. However, the effective constituent remains unknown. We speculate that the glycopeptide in the extract of the fruit has similar activity. To address this hypothesis, we isolated a novel pectin-like glycopeptide (FMP-6-S4) with a molecular weight of 11.23 kDa from the fruit. It contains about 20% of peptide comprising 17 amino acids and 80% glycan consisting of L-rhamnose (L-Rha), D-galactose (D-Gal), D-galacturonic acid (D-GalA), L-arabinose (L-Ara) and d-glucose (D-Glc) in molar ratios of 7.25:4.62:77.66:5.62:4.85. The backbone of the glycan part consisted of 1,4-linked α-D-GalpA and 1, 2-linked α-L-Rhap, while the branches were composed of hexenuronic acid (HexA) substituted at the C-3 position of partial galacturonic acid, and traces of galactose, glucose, and arabinose were substituted at the C-4 position of rhamnose. The in vitro experiments revealed that FMP-6-S4 might inhibit Aβ42 (β-amyloid peptides 42) aggregation and decrease Aβ42 production by modulating APP (amyloid precursor protein) processing.

Optimization of a newly established gas-chromatographic method for determining lactose and galactose traces: Application to Pecorino Romano cheese

In this work, we present and validate new gas-chromatographic with flame injection detector method to measure traces of lactose and galactose in cheese. Analytes are first extracted from the matrix by repeated treatments with a 40% (v/v) methanol/water solution, followed by their silanization with a 2:1 mixture of trimethylsilylimidazole and trimethylchlorosilane. Validation parameters show excellent limits of detection and quantification, and very good precision and trueness. The method was tested with a wide sampling of Protected Designation of Origin (PDO) Pecorino Romano sheep cheese (33 samples, different for geographic area of production, seasonality and ripening levels). For all of them, the concentration of lactose and galactose were always below the relevant limits of detection (i.e. 0.54 and 0.90 mg kg−1, respectively). On this basis, PDO Pecorino Romano sheep cheese is, according to both European Community and European Food Safety Authority guidelines, a “lactose free” foodstuff, useful in the dietary management of patients with galactosaemia and of people who are lactose intolerant.

Structural, functional, and biological properties of potato peel oligosaccharides

Enzymatic hydrolysis of water-soluble polysaccharides from potato peel waste (PPPW) generates low molecular weight oligosaccharides with a yield of 63%. The oligosaccharides generated from potato peel polysaccharides (OPPP) were purified by Superdex-30 column. The results showed the presence of 8 peaks (OPPP1–OPPP8). The identification of all the fractions by chromatography analysis (GC-FID) illustrated that the most prominent residues were glucose with traces of galactose, arabinose and rhamnose. Finally, mass spectrometry (MALDI-ToF-ToF) analysis showed that the generated oligosaccharides were heterogeneous and contained different degree of polymerization (DP). Indeed, the obtained oligosaccharides fractions OPPP3, OPPP4, OPPP5, OPPP6 and OPPP7 were compose of the following degree of polymerization DP5; DP4; DP2; DP1 and DP1, respectively. Potato peel oligosaccharides (OPPP) efficiency were tested using different concentrations in functional properties. The results showed good foaming and emulsion properties. This study also aimed to investigate the antioxidant activity of OPPP. The items explored included the DPPH radical-scavenging capacity (IC50 OPPP=2.5mg/mL), reducing power (OD: 0.622±0.032 at a concentration of 20mg/mL), β-carotene bleaching inhibition activity (45.335±3.653%), and also the ABTS radical scavenging activity (14.835±0.1%).These findings indicate that potato peel oligosaccharides have potent antioxidant activities. Hence, one can suggest that these oligosaccharides might contribute as additives in food, pharmaceutical and cosmetic preparations.

Lysinibacillus alkaliphilus sp. nov., an extremely alkaliphilic bacterium, and emended description of genus Lysinibacillus.

A novel aerobic, alkaliphilic, Gram-staining-positive, endospore-forming bacterium, strain OMN17T, was isolated from a typical sandy loam soil under long-term OMN fertilization (half organic manure N plus half mineral N fertilizer) in northern China and was subjected to a polyphasic taxonomic study. The best growth was achieved at 30 °C and pH 8-10 in medium containing 0.5% (w/v) NaCl. The cell-wall peptidoglycan of strain OMN17T was type A4α; (l-Lys-Gly-d-Asp) and the cell-wall sugars were ribose, traces of galactose and arabinose. The only respiratory quinone found in strain OMN17T was menaquinone 7 (MK-7). The major cellular fatty acids were iso-C15 : 0 and anteiso-C15 : 0. The major polar lipids were found to be phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. Phylogenetic analysis of strain OMN17T based on the 16S rRNA gene sequence showed that the strain was most closely related to Lysinibacillus halotolerans (97.8%), Lysinibacillus sinduriensis (97.5%), Lysinibacillus chungkukjangi (97.4%) and Lysinibacillus xylanilyticus (97.0%). The DNA-DNA hybridization results indicated that this strain was distinct from other species of the genus Lysinibacillus, the degree of relatedness being 21.8 ± 0.2% with L. halotolerans, 45.6 ± 1.8% with L. sinduriensis, 33.7 ± 1.2% with L. chungkukjangi and 23.7 ± 0.7% with L. xylanilyticus. The DNA G+C content of strain OMN17T was 38.1 mol%. The phenotypic, chemotaxonomic and genetic analyses identified strain OMN17T as a novel species of the genus Lysinibacillus, for which the name Lysinibacillus alkaliphilus sp. nov. is proposed. The type strain is OMN17T ( = DSM 28019T = CCTCC AB 2014073T). An emended description of the genus Lysinibacillus is also provided.

Structural characterization and antioxidant activity of a low-molecular polysaccharide from Dendrobium huoshanense

The present study aimed at investigating the structural features and antioxidant activities of a polysaccharide fraction (DHP1A) obtained from Dendrobium huoshanense, a precious herb medicine in China. DHP1A mainly consisted of mannose (Man), glucose (Glc) and a trace of galactose (Gal), with a molecular weight of 6700Da. Its backbone contained (1→4)-linked α-D-Glcp, (1→6)-linked α-D-Glcp and (1→4)-linked β-D-Manp, with a branch of terminal β-D-Galp. The in vitro antioxidant evaluation revealed that DHP1A had a remarkable inhibition effect on the FeCl2-induced lipid peroxidation. Furthermore, DHP1A pretreatment decreased the production of malondialdehyde (MDA), and restored the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx), as well as the level of glutathione (GSH) in the livers of CCl4-treated mice. These results suggested that DHP1A was a potential antioxidant component in D. huoshanense.

Antiinflammatory, antioxidant and cytotoxic actions of β-glucan-rich extract from Geastrum saccatum mushroom

The Geastrum saccatum a mushroom, native to Brazil, is produced under natural conditions in the unexplored reserve of Mata da Estrela-RN. This species has curative properties for eye infections and diseases such as asthma. The tissues of this mushroom contain carbohydrates, proteins, lipids, moisture and ashes in amounts of 42.3%, 37.05%, 9.01, 1.4% and 10.2%, respectively. An extract from this mushroom was characterized by chemical analyses and 13C and 1H NMR spectroscopy. It contains high amount of glucose and traces of galactose. The signal appearing at 103.5 ppm was assigned to C1 of β-glucose. The signals observed between 20 and 40 ppm suggest the presence of a glucan–protein compound. This glucan inhibited the lipid peroxidation at the dose of 0.27 mg/mL (59.1%) and it can protect cells against oxidative stress by scavenging of the hydroxyl (77%) and superoxide (88.4%) radicals at 0.27 mg/mL. The glucan (30 mg/kg) reduces the polymorphonuclear cell migration (57.6%). The ear edema induced by croton oil was inhibited by glucan (60.4% at 10 mg/kg) and by its association with diclofenac (5 mg/kg) (89.2%) or l-NAME (60 mg/kg) (86.23%). Histological analyses of the ear edema induced by croton oil in the presence of glucan (10, 30 or 50 mg/kg) showed a reduced degree of the polymorphonuclear cell migration. We concluded that the glucan has antioxidant, and antiinflammatory properties as well as its antiinflammatory effect are mediated by inhibition of both nitric oxide synthase (NOS) and cyclooxygenase (COX).

Studies of the Extracellular Glycocalyx of the Anaerobic Cellulolytic Bacterium Ruminococcus albus 7

Anaerobic cellulolytic bacteria are thought to adhere to cellulose via several mechanisms, including production of a glycocalyx containing extracellular polymeric substances (EPS). As the compositions and structures of these glycocalyces have not been elucidated, variable-pressure scanning electron microscopy (VP-SEM) and chemical analysis were used to characterize the glycocalyx of the ruminal bacterium Ruminococcus albus strain 7. VP-SEM revealed that growth of this strain was accompanied by the formation of thin cellular extensions that allowed the bacterium to adhere to cellulose, followed by formation of a ramifying network that interconnected individual cells to one another and to the unraveling cellulose microfibrils. Extraction of 48-h-old whole-culture pellets (bacterial cells plus glycocalyx [G] plus residual cellulose [C]) with 0.1 N NaOH released carbohydrate and protein in a ratio of 1:5. Boiling of the cellulose fermentation residue in a neutral detergent solution removed almost all of the adherent cells and protein while retaining a residual network of adhering noncellular material. Trifluoroacetic acid hydrolysis of this residue (G plus C) released primarily glucose, along with substantial amounts of xylose and mannose, but only traces of galactose, the most abundant sugar in most characterized bacterial exopolysaccharides. Linkage analysis and characterization by nuclear magnetic resonance suggested that most of the glucosyl units were not present as partially degraded cellulose. Calculations suggested that the energy demand for synthesis of the nonprotein fraction of EPS by this organism represents only a small fraction (<4%) of the anabolic ATP expenditure of the bacterium.

Degradation of polysaccharides and non-digestible oligosaccharides by Bacillus subtilis and Bacillus pumilus isolated from Soumbala, a fermented African locust bean (Parkia biglobosa) food Condiment

In order to select starter cultures for controlled fermentation of African locust beans, Bacillus subtilis and B. pumilus from Soumbala were investigated for their ability to degrade arabinogalactan, stachyose, raffinose and sucrose—the main polysaccharides and oligosaccharides in African locust bean. The methods used were liquefaction of gels of galactomannan for screening, pH measurement and High Performance Anion Exhange Chromatography – Pulsed Amperometric Detection (HPAEC–PAD) for detailed degradation study of the carbohydrates. For B. subtilis isolates, liquefaction of galactomannan gels was observed and HPAEC–PAD showed that arabinogalactan and stachyose were partially degraded while raffinose was fully degraded after 48 h of fermentation. Melibiose, fructose and traces of galactose were detected as residual sugars. For B. pumilus isolates much weaker degradation of the carbohydrates was observed. Two isolates of B. subtilis showed strong ability to degrade the oligosaccharides and they are seen as promising starter cultures for fermentation of African locust bean.

Isolation, purification and some structural features of the mucilaginous exudate from Musa paradisiaca

The water-soluble polysaccharides isolated from the vascular gel of Musa paradisiaca, were fractionated via anion exchange chromatography into four fractions. Fractionated polymers contained arabinose, xylose and galacturonic acid as major sugars, together with traces of galactose, rhamnose, mannose and glucose residues. Methylation analysis revealed the presence of a highly branched arabinoxylan with a significant amount of terminal arabinopyranosyl units and an arabinogalactan type I pectin. Periodate oxidation studies supported the results of methylation analysis.

Prominent 85-kDa oligomannosidic glycoproteins of rat brain are signal regulatory proteins and include the SHP substrate-1 for tyrosine phosphatases.

The glycoprotein component in rat brain reacting most strongly with Galanthus nivalis agglutinin (GNA) on western blots migrates as an 85-kDa band. GNA identifies mannose-rich oligosaccharides because it is highly specific for terminal alpha-mannose residues. After purification of this 85-kDa glycoprotein band by chromatography on GNA-agarose and preparative gel electrophoresis, binding of other lectins demonstrated the presence of fucose and a trace of galactose, but no sialic acid. Treatment with N-Glycanase or endoglycosidase H produced a 65-kDa band, indicating that it consisted of about one-fourth N-linked oligomannosidic carbohydrate moieties. High-performance anion-exchange chromatography and fluorescence-assisted carbohydrate electrophoresis indicated that the major carbohydrate moiety is a heptasaccharide with the structure Manalpha1-6(Manalpha1-3)Manalpha1-6(Manalpha1-3) Manbeta1-4Glc-NAcbeta1-4GlcNAc (Man5GlcNAc2). Determination of amino acid sequences of peptides produced by endoproteinase digestion demonstrated that this 85-kDa mannose-rich glycoprotein component contained the SHP substrate-1 for phosphotyrosine phosphatases and at least one other member of the signal-regulatory protein (SIRP) family. The unusually high content of oligomannosidic carbohydrate moieties on these receptor-like members of the immunoglobulin superfamily in neural tissue could be of functional significance for intercellular adhesion or signaling.

Isolation, purification, and properties of the exocellulase from Sporotrichum ( Chrysosporium) thermophile

The main extracellular exocellulase produced by Sporotrichum thermophile during growth on cellulose was isolated by ammonium sulfate precipitation, ion exchange chromatography, and adsorption-desorption on hydroxylapatite. The enzyme was identified on the basis of (a) its inability to liberate colored products from a dyed carboxymethyl cellulose (i.e., absence of endocellulase activity) and (b) its ability to hydrolyze the aglyconic bond of p-nitrophenyl-lactoside. The purified enzyme was homogeneous as judged by zone electrophoresis, SDS electrophoresis, electrofocalization, and titration curve. The molecular weight was 63,750 ± 4,500 and the pI was 4.52 ± 0.05. The enzyme was a glycoprotein containing 5.4% mannose and 1.7% glucose with traces of galactose and glucosamine. The enzyme was found to be very heat resistant and to have a low pH optimum (pH 3.5) on amorphous cellulose. Its activity was not affected by various cations, was only slightly inhibited by sulfhydryl reagents, but was sensitive to inhibition by ferric ions. The reactivity and mode of action on various substrates, as well as its cooperative action with other cellulolytic components from the same organism, support the evidence that the isolated exocellulase is a cellobiohydrolase (EC 3.2.1.91).

Characterization of vitellin and vitellogenin from Rhodnius prolixus: Identification of phosphorylated compounds in the molecule

The protein, phospholipid and carbohydrate components of vitellin and vitellogenin from Rhodnius prolixus were analyzed after being labelled by feeding the insects with blood enriched with 32P i. The carbohydrate moiety is comprised mainly of mannose residue (90%), but also contains glucose and traces of galactose and glucosamine. Both vitellin and vitellogenin are present in two different aggregational states that differ in molecular weight. Each of the individual aggregates releases, under denaturing conditions, the same four polypeptides with approximate molecular weights of 180, 158, 44 and 38 K daltons with no consistent stoichiometries. Analysis of the phosphorylated yolk protein after specific precipitation with serum against vitellin showed 62% of the total radioactivity in the phospholipid fraction and 38% in the glycoprotein fraction. Upon further analysis by thin layer chromatography, the following phospholipids were found: phosphatidylcholine (64.2%), phosphatidylethanolamine (30%), phosphatidic acid (4,4%), sphingomyelin (0.7%), phosphatidylserine (0.6%) and cardiolipin (0.2%). Analysis of the glycoprotein fraction showed that all four subunits were phosphorylated, part of the radioactivity being found as [ 32P]serine and [ 32P]mannose.

Carboxypeptidase Y from Saccharomyces cerevisiae conformational differences reflected in kinetic behaviour in water and deuterium oxide

The glycoenzyme carboxypeptidase Y (peptidyl--amnino-acid hydrolase, EC 3.4.16.1), from baker's yeast (British Fermentation Products Strain, Ng 72), of molecular weight 60000, had a protein portion closely similar to those in the literature for carboxypeptidase Y isolations from other yeast sources, but was 25.3 wt% carbohydrate (mannose 83.3% by wt., glucosamine 10.3% by wt. with traces of galactose and galactosamine). Circular dichroic spectra indicated that the enzyme lost its β-structure as the pH was lowered from 8.08 to 4.16. At p 2H 8.22 in 2H 2O media the conformation of this enzyme was different from that observed at pH 8.08. A tyrosine residue appeared to be perturbed by lowering the pH of the medium. Carboxypeptidase Y was rapidly, and essentially irreversibly, inactivated at low p 2H. The pH profile of k cat for the carboxypeptidase Y-catalysed hydrolysis of 4-nitrophenyltrimethylacetate showed two inflections at 45°C: one at p K app, ∼3.7 insensitive to temperature variation (ascribed to a carboxyl group), and one of p K app ∼5.7 markedly temperature-dependent and possibly caused by a histidine residue.

Biodegradation of wheat straw lignocarbohydrate complexes (LCC)

Hot water extractions of ground wheat straw were performed as part of a study to assess digestibility and utility as a substrate for ligninolytic fungi. The treatments gave solutions containing phenolics and carbohydrates. After 48 h reflux percolation or 3 h autoclaving, little additional carbohydrates or phenolics were extracted into the liquors. During percolation, the pH of the wet straw in the reflux thimble fell only slightly and it is difficult to attribute the extraction in this case to an autohydrolytic process catalyzed by organic acids liberated from the straw. Gel permeation chromatography enabled the separation of the extracts into fractions, several of which were free of reducing sugars but contained phenolics and bound carbohydrates. Rechromatography of these fractions failed to separate these two components, indicating a tight linkage between them. The bound sugars were identified as glucose, arabinose, xylose and traces of galactose, but uronic acids were absent. The high molecular size fraction was rich in glucose whereas lower molecular size fractions were richer in xylose and arabinose and poor in glucose.

Biochemical and serological characteristics of soluble yeast phase antigens of Histoplasma Capsulatum

Soluble antigens of whole yeast-phase cells were extracted with a 0.1 M phosphate buffer containing 0.1 M sodium chloride and 0.02% iodacetate. After being separated by differential filtration into fractions less than or greater than 50,000 daltons these antigens were purified by molecular sieve and chromatographic separations on ionic exchange resins. Two high molecular weight fractions obtained from diethylaminoethyl-cellulose (DEAE) at pH 8.0 and 7.0 with tris (hydroxymethyl) aminomethane (Tris) buffer were M antigens; those obtained at pH 4.0 and 4.0 with salt were H antigens. The four fractions had protein to carbohydrate ratios of 7.3, 14.0, 8.4, and 6.5 respectively, and all had essentially the same amino acid composition with no methionine and tyrosine and little histodine, arginine, phenylalanine and lysine. They had high concentrations of glucose, less mannose and traces of galactose. The low molecular weight fractions had the new complex "Y antigen", M antigen with protein to carbohydrate ratios of 1.4, 1.4 and 0.3 respectively. The amino acid and sugar composition of Y antigen strongly resembled the composition of the low molecular weight H and M antigens. Unlike the high molecular weight antigens, these low molecular weight antigens had methionine in relatively high concentrations; they had the same sugars as their respective high molecular weight counterparts. The yeast phase antigens differed from their respective mycelial counterparts in the following ways: glucose was the major sugar in the yeast phase with less amounts of mannose and traces of galactose, whereas in the mycelial antigens, mannose was the major sugar, with lesser amounts of galactose, and hexosamine. The H and M antigens of the yeast phase had high concentrations of glycine and alanine, whereas in the mycelial phase, these antigens had high concentrations of threonine and proline; the H and M antigens of the yeast phase had 5 to 16 times the protein to carbohydrate ratio observed for the same antigens of histoplasmin.

Cell-wall composition and structure of yeast cells and conjugation tubes of Tremella mesenterica.

Cell walls prepared from vegetative yeast cells and from hormone-induced conjugation tubes of the basidiomycete Tremella mesenterica had similar compositions. Evidence was found for 1,3-alpha-glucan (yeast 38%, tube 25%), 1,3-beta-1,6-beta-glucan (yeast 33%, tube 48%) and chitin (both less than 3%) in the walls. The walls also contained xylose (5 to 7%), mannose (6%), glucuronic acid (approx. 2%), and traces of galactose. Protein amounted to less than 2% of the wall weight. The cell capsule was very insoluble and could not be removed from the cell wall. The conjugation hormone did not appear to exert its effect on cell shape by causing gross changes in wall composition.

Lipids in plant tissue cultures IV. The characteristic patterns of lipid classes in callus cultures and suspension cultures

Lipids from callus cultures and suspension cultures of higher plants constitute 5 to 8% of the dry tissue's weight. The predominant lipid classes are the sterols, steryl esters, steryl glycosides and esterified steryl glycosides. Considerable amounts of a variety of sterylglycolipids, whose structures are not completely elucidated, are also present. Triglycerides and phospholipids occur in small proportions, whereas monogalactosyl diglycerides, digalactosyl diglycerides and sulfoquinovosyl diglycerides are present only in traces, if at all. β-Sitosterol is the predominant constituent sterol, stigmasterol and campesterol as well as a variety of as yet unidentified sterols occur in smaller proportions. The major constituent fatty acids are palmitic, oleic, linoleic and linolenic acids. Saturated very long-chain fatty acids are found in smaller proportions. Unusual fatty acids, such as epoxy acids, which occur in the seed lipids of certain plants, are not found in tissue cultures derived from these plants. Clucose and traces of galactose are the only sugars obtained by acid hydrolysis of the glycolipids occurring in plant tissue cultures.

Resistance of Escherichia coli to penicillins. IX. Genetics and physiology of class II ampicillin-resistant mutants that are galactose negative or sensitive to bacteriophage C21, or both.

Ampicillin-resistant mutants of class II are determined by a doubling of chromosomally and episomally mediated ampicillin resistance on agar plates. Several mutants were isolated from a female as well as from an Hfr strain. The mutants differed from each other in various properties such as response to colicin E2 and sodium cholate, response to the phages T4 and C21, and fermentation of galactose. By conjugation and transduction experiments, it was shown that mutations in at least four loci gave the class II phenotype. The mutations were found to be in the galU gene, the ctr gene, and two new genes close to mtl denoted lpsA and lpsB. The carbohydrate compositions of the lipopolysaccharides of the mutants were investigated and found to be changed compared to the parent strains. GalU mutants lacked rhamnose and galactose and had 11% glucose compared to the parent strain. The lpsA mutant also lacked rhamnose and had only traces of galactose and 58% glucose, whereas the lpsB mutant contained 14% rhamnose, traces of galactose, and 81% glucose compared to the parent strain.

A lipoglycoprotein from hemolymph of healthy and diseased larvae of the Japanese beetle, Popillia japonica

The major protein in the hemolymph of third-instar Popillia japonica larvae is a lipoglycoprotein (LGP). This protein was isolated from healthy larvae and larvae infected with Bacillus popilliae, a cause of milky disease. The LGP was separated by density gradient centrifugation and partially purified by exhaustive dialysis. During the course of milky disease, the concentration of LGP decreases markedly. Lipids associated with the LGP were extracted and characterized by thin-layer chromatography and gas-liquid chromatography. Fatty acids detected in significant amounts were palmitic (C 16), stearic (C 18), oleic (C 18:1), linoleic (C 18:2), and lignoceric (C 24). The relative concentration of palmitic acid was significantly reduced in LGP isolated from diseased larval hemolymph. Carbohydrate constituents released from LGP by hydrolysis were determined by gas-liquid chromatography of the trimethylsilylated derivatives. Mannose, glucose, and a trace of galactose were present in LGP from hemolymph of normal larvae; however, only trace quantities of any of these carbohydrates could be detected in the LGP from hemolymph of infected larvae. Amino acids present in the greatest amounts in the LGP were aspartic and glutamic acids. Appreciable quantities of tyrosine, leucine, and threonine were also present. The amino acid composition of LGP in hemolymph from healthy larvae did not differ significantly from that in diseased larvae.

Extracellular enzyme system utilized by the fungus Chrysosporium lignorum for the breakdown of cellulose : II. Separation and characterization of three cellulase peaks

The separation of three cellulase peaks (CMC activity) from culture solutions of the fungus Chrysosporium lignorum has been demonstrated on a DEAE-Sephadex A-50 column. The carbohydrate content in the different peaks decreased upon dialysis to 13, 10, and 7 percent. The carbohydrate moiety consisted of mannose and traces of galactose. The hydrolysis products of cellodextrins and dewaxed cotton fibers incubated with the enzyme peaks were in all cases cellobiose and glucose in the approximate proportions 3:1. It was found that the higher the carbohydrate content of the peak, the higher the transglucosidation caused by the peak. Strong indications for the existence of a C 1 enzyme in the culture solution of C. lignorum are reported.