Marvin O. Bagby

Hydroxy fatty acids through hydroxylation of oleic acid with selenium dioxide/tert.-butylhydroperoxide

Oleic acid was hydroxylated in the allylic positions with the selenium dioxide/tert.-butylhydroperoxide system to give 8-hydroxy-9(E)-octadecenoic acid, 11-hydroxy-9(E)-octadecenoic acid and the novel 8,11-dihydroxy-9(E)-octadecenoic acid. This is a viable method for obtaining hydroxy fatty acids. The unsaturated hydroxy acids were hydrogenated with the hydrazine/air system to give the cor-responding saturated products. 8,11-Dihydroxyoctadecanoic acid thus obtained is also a novel compound. The saturated and unsaturated dihydroxy products were obtained aserythro/threo isomers as determined by nuclear magnetic resonance.

Vegetable-oil-based printing ink formulation and degradation

Abstract Printing ink vehicles that did not require petroleum components were prepared from modified vegetable oil. The apparent average molecular weight ( M W ) of vehicles made from representative vegetable oils, such as soybean, sunflower, cottonseed, safflower and canola oils were compared by gel permeation chromatography (GPC), and a correlation between viscosity and apparent M W of these vehicles was established. Physical properties of the formulated inks meet or exceed the industry standards for lithographic and letterpress newsprint applications. Elimination of petroleum-based resin and reduced pigment requirements, due to the light vehicle color, should provide a competitively priced alternative to petroleum-based inks. These ink vehicles along with the commercial ones were subjected to biodegradation. Results show that the vegetable-oil vehicles degrade faster and more completely than commercial soy- or mineral-oil-based vehicles. Fermentations were allowed to proceed for 5, 12 and 25 days. Both mono- and mixed cultures of microorganisms, which are commonly found in soil, were used. Greater increase in biodegradation with time was observed for our vehicles than the commercial vehicles. In 25 days, commercial mineral-oil-based vehicles degrade 17–27%, while commercial partial soy-oil-based vehicles degrade 58–68% and our 100% soy-oil-based vehicles degrade 82–92%.

10(R)-Hydroxystearic acid production by a novel microbe, NRRL B-14797, isolated from compost

SummaryA bacterium, NRRL B-14797, isolated from composted manure, converted oleic acid exclusively to 10(R)-hydroxystearic acid in 3-day batch cultures. 9(Z)-Unsaturated fatty acids in a lipid extract from soybean soapstock were also hydrated effectively. Aerobic bioconversions by isolate B-14797 were compared with those byPseudomonas B-2994 andNocardia 5767, which produce mixtures of 10-hydroxy- and 10-ketostearic acids. The results of studies with resting cells and cell-free extracts were consistent with action of a hydratase and absence of secondary alcohol dehydrogenase in strain B-14797.

Microbial oxidation of cumene by octane-grown cells

Previously, we reported that eight glucose-grown microbial cultures out of 1229 screened oxidize the alkyl side-chain of 2-phenylpropane (cumene) stereospecifically. Now, we have adapted these cultures to grow on n-octane and found that their cumene oxidation activities increased more than 30 times. We also found an additional 11 cultures (ten bacteria, one actinomycete) that oxidized cumene when grown on octane but not on glucose. In general, octane-grown cells were more active in cumene oxidation than glucose-grown cells. Rhodococcus rhodochrous NRRL B-2153 showed the best conversion yield (2-phenyl-1-propanol plus 2-phenyl-1-propionic acid was 5.5%) at 25°C, pH 8.0, 250 rpm, and 12 h of reaction. Structures of the reaction products were confirmed by gas chromatography (GC)/mass spectrometry and GC/infrared analyses. Products contained 84% ee (enantiomeric excess) of the R(−) isomer, as analyzed with a GC cyclodextrin chiral column. Strain B-2153 oxidized alkylbenzenes in the following order of reaction rate: ethylbenzene >amylbenzene > butylbenzene > cumene > propylbenzene > sec-butylbenzene. tert-Butylbenzene was not oxidized.

7,10-Dihydroxy-8(E)-Octadecenoic acid: Stereochemistry and a novel derivative, 7,10-Dihydroxyoctadecanoic acid

Abstract7,10-Dihydroxy-8(E)-octadecenoic acid (obtained by bioconversion of oleic acid with the bacterial strain PR3) was hydrogenated with hydrazine hydrate under air in ethanolic solution to give a novel compound, 7,10-dihydroxy-octadecanoic acid. The absolute configuration of both compounds was determined with the aid of circular dichroism to be R,R. A different purification method for 7,10-dihydroxy-8(E)-octadecenoic acid is described.

Allylic mono- and di-hydroxylation of isolated double bonds with selenium dioxide–tert-butyl hydroperoxide. NMR characterization of long-chain enols, allylic and saturated 1,4-diols, and enones

Selenium dioxide with tert-butyl hydroperoxide as re-oxidant was used in the allylic hydroxylation of isolated double bonds in straight-chain hydrocarbons. This was shown for mono-unsaturated fatty acids, esters and alcohols. Either allylic position was hydroxylated individually or both positions reacted to give dihydroxy isomers, affording numerous novel hydroxy compounds. Yields of monohydroxy compounds in which the OH group is between the double bond and C-1 were usually higher than those in which the OH group is between the double bond and the methyl terminus. Monohydroxy products were used as starting material in subsequent allylic hydroxylation reactions to increase the yield of dihydroxy product, although this reaction is slow. Coinciding with the known mechanism, cis double bonds of starting materials isomerized nearly quantitatively to trans double bonds in the products while trans double bonds did not isomerize. Resonance differences of the olefinic carbons in 13C NMR of the unsaturated monohydroxy compounds show on which side of the double bond the hydroxy group is located. The magnitude of these differences depends on the nature of the group at C-1 and the distance of the double bond from C-1. Corresponding saturated hydroxy fatty acids were synthesized with the hydrazine–air system. 13C NMR of the saturated compounds showed that the dihydroxy products were erythro/threo diastereoisomers. With this assignment, 1H NMR of the unsaturated allylic dihydroxy compounds may be used to distinguish these diastereoisomers. The olefinic protons of the erthryo dihydroxy diastereoisomer resonate downfield from those in the threo form. The threo diastereoisomers are formed in higher yields than their erythro counterparts. Compounds with allylic keto group (enones) analogous to the monohydroxy products arose as side products. The 13C NMR spectra of these enones are discussed.

Identification of NRRL strain B-18602 (PR3) as Pseudomonas aeruginosa and effect of phenazine 1-carboxylic acid formation on 7,10-dihydroxy-8(E)-octadecenoic acid accumulation

A new compound, 7,10-dihydroxy-8(E)-octadecenoic acid (DOD), produced from oleic acid by a new bacterial isolate PR3, was discovered in 1991. We have now identified isolate PR3 as a strain of Pseudomonas aeruginosa by DNA reassociation studies. Strain PR3 also produced a crystalline yellowish compound the structure of which, as determined by GC/MS and NMR, is phenazine 1-carboxylic acid (PCA). In cultures of PR3, high PCA production was associated with low DOD accumulation.

Oleic acid transformations by selected strains of Sphingobacterium thalpophilum and Bacillus cereus from composted manure.

In a survey of 186 randomly selected microbial strains isolated from composted manure, 63 transformed oleic acid into three types of products: hydroxy fatty acid, fatty amide, and less polar oleyl lipid. Selection of oleic acid-transforming microorganisms was enhanced in nutrient agar supplemented with 0.1% (vol/vol) oleic acid at pH 7.2. Most of the 63 diverse isolates elicited inconsistent and poorly reproduced transformations. However, strains 142b (NRRL B-14797) transformed oleic acid to 10-hydroxystearic acid consistently, and strain 229b (NRRL B-14812) produced an octadecenamide. Taxonomic studies indicated that NRRL strain B-14797, possessing 1,3-dihydroxy-2-amino-15-methylhexadecane and sphinganine bases, was closely related to Sphingobacterium thalpophilum, and NRRL B-14812 was identified as Bacillus cereus.

Oleyl oleate and homologous wax esters synthesized coordinately from oleic acid by Acinetobacter and coryneform strains

Abstract. Newly isolated Acinetobacter (NRRL B-14920, B-14921, B-14923) and coryneform (NRRL B-14922) strains accumulated oleyl oleate and homologous liquid wax esters (C30:2–C36:2) in culture broths. Diunsaturated oleyl oleate preponderated in 75 mg liquid wax esters (280 mg lipid extract) recovered from 100-ml cultures of Acinetobacter B-14920 supplemented with 810 mg oleic acid–oleyl alcohol. With soybean oil instead of oleic acid, wax esters (260 mg) were increased to approximately 50% of the lipid extract. Production of wax esters by cultures supplemented with combined fatty (C8–C18) alcohols and acids suggests a coordinated synthesis whereby the exogenous alcohol remains unaltered, and the fatty acid is partially oxidized with removal of C2 units before esterification. Consequently, C8–C18 primary alcohols control chain lengths of the wax esters. Exogenous fatty acids are presumed to enter an intracellular oxidation pool from which is produced a homologous series of liquid wax esters.

2-Hydroxyhexadecanoic and 8,9,13-trihydroxydocosanoic acid accumulation by yeasts treated with fumonisin B1.

Fumonisin B1 is a sphingolipid-like compound that enhances the accumulation of yeast sphingolipids and 2-hydroxy fatty acids. These lipids occur both as freely extractable and cell bound components in yeast fermentations. Both free and bound 2-hydroxy fatty acids produced byPichia sydowiorum NRRL Y-7130 were increased when fumonisin B1 (50 mg/L) was added to the usual growth medium containing yeast extract/malt extract/peptone/glucose. Fumonisin-treated cultures contained 38 mg/L more 2-hydroxyhexadecanoic and 15 mg/L more 2-hydroxyoctadecanoic acids than did untreated cultures. By contrast, fumonisin inhibited the accumulation of free 8,9,13-trihydroxydocosanoic acid inRhodotorula sp. YB-2501 cultures, leading to 240 mg/L lower trihydroxy acid production than by untreated cultures.