Ching T. Hou

A novel compound, 7,10-dihydroxy-8(E)-octadecenoic acid from oleic acid by bioconversion

Sixty-two cultures from the Agricultural Research Service (ARS) Culture Collection and 10 cultures isolated from soil and water samples in Illinois were screened for their ability to convert agricultural oils to value-added industrial chemicals. A new compound, 7,10-dihydroxy-8(E)-octadecenoic acid (DOD), was produced from oleic acid at a yield of greater than 60% by bacterial strain PR3 which was isolated from a water sample in Morton, IL. To our knowledge, DOD has not been previously reported. The optimum time, pH and temperature for the production of DOD were 2 days, 7.0, and 30°C, respectively. The production of DOD is unique in that it involves hydroxylation at two positions and rearrangement of the double bond of the substrate molecule.

Microbial conversion of oleic acid to 10-hydroxystearic acid

SummaryResting cell suspensions of seven Nocardia species catalyzed the production of 10-hydroxystearic acid from oleic acid. Nocardia cholesterolicum NRRL 5767 gave a good yield with optimum conditions at pH 6.5 and 40°C. Yields exceeding 90% can be obtained within 6 h with 0.1 g cells (dry weight) and 178 mg oleic acid in 10 ml of 0.05 M sodium phosphate buffer (pH 6.5). In addition, minor amounts of 10-ketostearic acid were formed as a by-product. The reaction proceeded via hydration of the double bond as shown by labeling experiments with deuterium oxide and 18O-labeled water. The system was specific for fatty acids with cis unsaturation at the 9 position.

Growth inhibition of plant pathogenic fungi by hydroxy fatty acids

Hydroxy fatty acids are plant self-defense substances (Masui et al, Phytochemistry1989). Three types of hydroxy fatty acids: 10-hydroxystearic acid (HSA), 7S,10S-dihydroxy-8(E)-octadecenoic acid (DOD), and 12,13,17-trihydroxy-9(Z)-octadecenoic acid (THOA) were tested against the following plant pathogenic fungi: Erysiphe graminis f sp tritici (common disease name, wheat powdery mildew); Puccinia recondita (wheat leaf rust); Pseudocercosporella herpotrichoides (wheat foot rot); Septoria nodorum (wheat glume blotch); Pyricularia grisea (rice blast); Rhizoctonia solani (rice sheath blight); Phytophthora infestans (potato late blight); and Botrytis cinerea (cucumber botrytis). At a concentration of 200 ppm, both HSA and DOD showed no fungal disease control activity. However, THOA at the same concentration showed weak activity and provided disease control (percent) of the following plant pathogenic fungi: Erysiphe graminis 77%; Puccinia recondita 86%; Phytophthora infestans 56%; and Botrytis cinerea 63%. The position of the hydroxy groups on the fatty acids seems to play an important role in activity against specific fungi. Journal of Industrial Microbiology & Biotechnology (2000) 24, 275–276.

Production of a new compound, 7,10-dihydroxy-8-(E)-octadecenoic acid from oleic acid byPseudomonas sp. PR3

SummarySixty-two cultures from the ARS Culture Collection and 10 cultures isolated from soil and water samples collected in Illinois were screened for their ability to convert agricultural oils to value-added industrial chemicals. A new compound, 7,10-dihydroxy-8-(E)-octadecenoic acid (DOD) was produced from oleic acid by a new strain,Pseudomonas sp. PR3 isolated from a water sample in Morton, IL. Strain PR3 is a motile, small rod-shaped, Gram-negative bacterium. It has multiple polar flagellae and is oxidase-positive. Strain PR3 grows aerobically and cannot grow anaerobically. The strain produces white, smooth colonies on agar plate and no water-soluble pigment. The yield of the product was greater than 60%. The optimum time, pH and temperature for the production of DOD were: 2 days, 7.0, and 30°C, respectively. Glycerol and dextrose support the growth of strain PR3, but the cells grown from the former failed to catalyse the conversion of oleic acid to DOD. The production of DOD is unique in that it involves a hydroxylation at two positions and a rearrangement of the double bond of the substrate molecule.

10(S)-Hydroxy-8(E)-octadecenoic acid, an intermediate in the conversion of oleic acid to 7,10-dihydroxy-8(E)-octadecenoic acid

The new microbial isolate Pseudomonas aeruginosa (PR3) has been reported to produce from oleic acid a new compound, 7,10-dihydroxy-8(E)-octadecenoic acid (DOD), with 10-hydroxy-8-octadecenoic acid (HOD) being a probable intermediate. The production of DOD involves the introduction of two hydroxyl groups at carbon numbers 7 and 10, and a rearrangement of the double bond from carbons 9–10 to 8–9. It has been shown that the 8–9 unsaturation of HOD was possibly in the cis configuration. Now we report that the rearranged double bond of HOD is trans rather than cis, as determined by spectral data. Also, it was found that the 10-hydroxyl was in the S-configuration as determined by gas chromatographic separation of R- and S-isomers after preparation of the (−)-menthoxycarbonyl derivative of the hydroxyl group followed by oxidative cleavage of the double bond and methyl esterification. This latter result coincides with our recent finding that the main final product, DOD, is in the 7(S),10(S)-dihydroxy configuration. In addition, a minor isomer of HOD (about 3%) with the 10(R)-hydroxyl configuration was also detected. From the data obtained herein, we concluded that 10(S)-hydroxy-8(E)-octadecenoic acid is the probable intermediate in the bioconversion of oleic acid to 7(S),10(S)-dihydroxy-8(E)-octadecenoic acid by PR3.

Production of a Novel Compound, 7,10,12-Trihydroxy-8(E)-Octadecenoic Acid from Ricinoleic Acid by Pseudomonas aeruginosa PR3

The production and its potential use of a novel trihydroxy unsaturated fatty acid, 7,10,12-trihydroxy-8(E)-octadecenoic acid (TOD), were investigated. TOD was formed by Pseudomonas aeruginosa PR3 (NRRL B-18602) in a culture supplied with exogenous ricinoleic acid. The yield of TOD production was always higher in a rich culture medium than in minimal screening medium. Extending the conversion time from 48 to 72 h prior to lipid extraction led to a 65% reduction in yield, indicating that TOD was further metabolized by strain PR3 and that control of reaction time is important to achieving a maximum yield. The optimum culture density, reaction time, pH, temperature, and substrate concentration for the production of TOD were: 20–24 h culture growth, 48 h, 7.0, 25°C, and 1% (vol/vol), respectively. Under optimum conditions, the yield of TOD production was greater than 45%. TOD was found to be an antifungal agent most active against the fungus that causes blast disease in rice plants, the most important fungal disease affecting rice production worldwide.

Screening of lipase activities with cultures from the agricultural research service culture collection

A simple, sensitive agar plate method was used to screen lipase activity from 1229 selected cultures, including 508 bacteria, 479 yeasts, 230 actinomycetes and 12 fungi, covering many genera and species. About 25% of the cultures tested are lipase-positive. These lipase-positive strains were further classified into three categories according to their enzyme activity: good, moderate and weak lipase producers for those having orange fluorescent halo zones greater than 10 mm, 7.5 mm or 5 mm diameters, respectively. The good lipase producers have the potential to be developed for industrial enzymes. The positional, fatty acid or enantiospecificity of each individual lipase requires further investigation. The data presented here are important for the discovery of new lipases.

Handbook of industrial biocatalysis.

Enzymes for the Transgenic Production of Long-Chain Polyunsaturated Fatty Acid-Enriched Oils, Y.-S. Huang, S.L. Pereira, and A.E. Leonard Screening for Unique Microbial Reactions Useful for Industrial Applications, J. Ogawa and S. Shimizu Biocatalyses in Microaquaeous Organic Media, T. Yamane Biocatalysts for Epoxidation and Hydroxylation of Fatty Acids and Fatty Alcohols, Steffen C. Maurer and Rolf D. Schmid Lipase-Catalyzed Kinetic Resolution for the Fractionation of Fatty Acids and Other Lipids, K.D. Mukherjee Protein Engineering of Recombinant Candida rugosa Lipases, G.-C. Lee, C.-J Shieh, and J.-F. Shaw Production of Value-Added Industrial Products from Vegetable Oils: Oxygenated Fatty Acids, C.T. Hou and M. Hosokawa Application of Lipases to Industrial-Scale Purification of Oil-and Fat-Related Compounds, Y. Shimada, T. Nagao, and Y. Watanabe Application of Lipases in Modification of Food Lipids, S. Sellappan and C.C. Akoh Lipase-Catalyzed Condensation in an Organic Solvent, S. Adachi Enzymatic Production of Diacylglycerol and Its Beneficial Physiological Functions, N. Yamada, N. Matsuo, T. Watanabe, and T. Yanagita The Use of Nonimmobilized Lipase for Industrial Esterification of Food Oils, S. Negishi Preparation of Polyunsaturated Phospholipids and Their Functional Properties, M. Hosokawa and K. Takahashi Production of Biosurfactants from Fermentation of Fats, Oils, and their Co-Products, Daniel K.Y. Solaiman, R.D. Ashby, and T.A. Foglia Fatty Acid-Modifying Enzymes: Implications for Industrial Applications, T. Aki, S. Kawamoto, S. Shigeta, and K. Ono Low-Calorie Fat Substitutes: Synthesis and Analysis, K-T. Lee, T.A. Foglia, and J.-H. Lee Microbial Polyunsaturated Fatty Acid Production, T. Nakahara Structured Triacylglycerols Comprising Omega-3 Polyunsaturated Fatty Acids, G.G. Haraldsson Biopolyesters Derived from the Fermentation of Renewable Resources, R.D. Ashby, Daniel K.Y. Solaiman, and T.A. Foglia Carbohydrate Active-Enzymes for the Production of Oligosaccharides, H. Taniguchi Microbial Hemicellulolytic Carbohydrate Esterases, P.Biely and G.L. Cote Application of Cyclodextrin Glucanotransferase to the Synthesis of Useful Oligosaccharides and Glycosides, H. Nakano and S. Kitahata Converting Herbaceous Energy Crops to Bioethanol: A Review with Emphasis on Pretreatment Processes, B.S. Dien, L.B. Iten, and C.D. Skory Enzymes as Biocatalysts for Conversion of Lignocellulosic Biomass to Fermentable Sugars, B. Saha Bioelectrocatalysis: Electroactive Microbial and Enzyme Technologies for Detection and Synthesis of Chemicals, Fuels, and Drugs, J.G. Zeikus Biocatalysis: Synthesis of Chiral Intermediates for Pharmaceuticals, R.N. Patel Nutrition Delivery System: A Novel Concept of Nutrient Fortification, T.P. Rao, N. Sakaguchi, and L.R. Juneja Renewable Resources for Production of Aromatic Chemicals, S. Sariaslani, T. Van Dyk, L. Huang, A. Gatenby, and A. Ben-Bassat Extremophiles from the Origin of Life to Biotechnological Applications, Chiara Schiraldi and Mario De Rosa Index

A novel compound, 12,13,17-trihydroxy-9(Z)-Octadecenoic acid, from linoleic acid by a new microbial isolateClavibacter sp. ALA2

A novel compound, 12,13,17-trihydroxy-9(Z)-octadecenoic acid (THOA), was produced from linoleic acid by microbial transformation at 25% yield. The newly isolated microbial strain that catalyzed this transformation was identified asClavibacter sp. ALA2. The product was purified by high-pressure liquid chromatography, and its structure was determined by1H and13C nuclear magnetic resonance, Fourier transform infrared, and mass spectroscopy. Maximum production of THOA was reached after 85 h of reaction. THOA was not further metabolized by strain ALA2. This is the first report on 12,13,17-trihydroxy unsaturated fatty acid and its production by microbial transformation.

Propylene oxide production from propylene by immobilized whole cells of Methylosinus sp. CRL 31 in a gas-solid bioreactor

SummaryMethanotrophic bacteria have been shown to oxidize gaseous alkenes to the corresponding epoxides utilizing an NADH2-dependent methane monooxygenase. A cell paste of methane-grown methylotrophs was coated on porous glass beads. The production of propylene oxide from propylene was performed in a gas-solid bioreactor to ensure continuous production and removal of product epoxide from the microenvironment of the biocatalyst. The amount of propylene oxide produced before cofactor regeneration was between 120–145 μmoles/20 mg cells in about 10 h depending on the microbial strains used. The conversion rate for propylene was 2.7%. Regeneration of cofactor NADH2 was performed in the bioreactor with the vapor of a cosubstrate, methanol.