The Occurrence of a Sulfuric Acid Ester of Choline in the Mycelium of a Strain of Penicillium chrysogenum

Abstract

added to an excess of methylmagnesium iodide over a period of five minutes with rapid stirring. The mixture was hydrolyzed by adding ice and dilute hydrochloric acid. The organic layer was dried and evaporated on the steam-bath. The colorless oil which remained after evaporation was crystallized from petroleum ligroin. The melting point was 75-77', yield 1.02 g. This compound was not further investigated. Two meso Forms of 2,5-Diphenyl-4-methyl-4-tetrahydro-pyrano1.-Eighty ml. of a benzene solution containing 1.1 g. of cis-2,6-diphenyl-4-tetrahydr~pyrone~ was added over a period of 20 min. to a solution of an excess of methylmagnesium iodide in ether which was being stirred rapidly. The reaction was allowed to proceed for 20 min. and hydrolyzed with ice and diluted HC1. The organic layer was dried and evaporated on a steam-bath and the residue chromatographed on the Magnesol-Celite column. A fraction of 20 mg. melting a t 67-69', which we believe to be the meso-1-compound, was followed by a larger fraction of meso-2-compound weighing 140 mg. which melted from 142-145'. This latter fraction proved to be identical with the material from the t-butyl alcohol-benzaldehyde reaction by mixed melting point.2,6-Diphenyl-J ,5-dimethyl-4-ethyl-4-tetrahydropyranol.--An excess of ethylmagnesium bromide in 180 ml. of ether was added to 11 g. of 2,6-diphenyl-3,5-dimethyl-4-tetra-hydropyrone8 dissolved in ether. After reaction, the material was hydrolyzed with NH,Cl solution and the product recovered and crystallized from petroleum ligroin; yield, 9 g., m.p. 177-178'. This substance is identical with that from the acid condensation; the mixed melting point showed no depression.

Sulfate ester formation and hydrolysis: a potentially important yet often ignored aspect of the sulfur cycle of aerobic soils.

“…Similar results (0.2 to 0.3% of the dry weight) were reported for A. sydowi (273) and Penicillium chrysogenum (39,238). In contrast with various Pseudomonas species that form this ester (see below), choline sulfate was not detected in culture filtrates of the choline sulfate-producing fungi (39,104,238,273). The formation of the ester in fungi required the initial formation of 3'-phosphoadenylyl sulfate (131, 232), with the subsequent transfer of S042to choline catalyzed by choline sulfokinase (232).…”

“…Even when grown under conditions in which the sulfate for choline sulfate must be derived endogenously from taurine, the ester can accumulate in the mycelia of Aspergillus nidulans to a concentration of 0.6% of the dry weight (233). Similar results (0.2 to 0.3% of the dry weight) were reported for A. sydowi (273) and Penicillium chrysogenum (39,238). In contrast with various Pseudomonas species that form this ester (see below), choline sulfate was not detected in culture filtrates of the choline sulfate-producing fungi (39,104,238,273).…”

Choline Sulphate in Fungi

“…N+( CH,),, in the sulphur metabolism of fungi is still a matter of controversy. This sulphate ester has been isolated from -4spergoillus sydowi (Woolley & Peterson, 1937) and Peiiicillium chrysogenum (De Flines, 1955; Stevens & Vohra, 1955) in yields greater than 0.2% of the dry weight of the mycelium; similar high concentrations have also been found in lichens of the Rocella spp. (Lindberg, 1955a) and the red alga Gelida cartilaginurn (Lindberg, 19556).…”

The formation of choline O-sulphate by Pseudomonas C12B and other Pseudomonas species

“…The higher fungi can accumulate large quantities (0.2-1.5 % dry wt.) of choline 0-sulphate (De Flines, 1955;Stevens & Vohra, 1955;Takebe, 1960) and very little of it occurs extracellularly . These data and other evidence (see also Takebe & Yanagita, 1959;Takebe, 1960) prompted to suggest that choline 0sulphate acts as a storage form of sulphate in these fungi.…”

“…Choline 0-sulphate has a wide distribution in nature and has been detected in high concentration in plants (Nissen & Benson, 1961), lichens (Lindberg, 1955a;Harper & Letcher, 1966), red algae (Lindberg, 1955b), and in spores and mycelia of the higher fungi (Woolley & Peterson, 1937;De Flines, 1955;Stevens & Vohra, 1955;Kaji & McElroy, 1958;Takebe, 1960;Ballio et al, 1960). The ester was not detected in fungi representative of the Mucorales and Endomycetales or in various bacteria ofdiverse genera (including Pseudomonas) when these microorganisms were grown in chemically undefined culture media .…”