Production of Polyunsaturated Fatty Acids by Polyketide Synthases in Both Prokaryotes and Eukaryotes

Metz, James G.; Roessler, Paul G.; Facciotti, Daniel; Levering, Charlene; Dittrich, Franziska; Lassner, Michael; Valentine, Ray; Lardizabal, Kathryn; Domergue, Frédéric; Yamada, Akiko; Yazawa, Kazunaga; Knauf, Vic; Browse, John

doi:10.1126/science.1059593

Cited by 632 publications

(547 citation statements)

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“…In contrast to the traditional desaturase-based PUFA biosynthesis pathway, M. marina MP-1 uses a distinct polyketide synthase (PKS)-type pathway that remains poorly characterized (Jenke-Kodama et al 2005;Metz et al 2001;Napier 2002). PKS-type pathways are important for a variety of complex natural products, such as cholesterol-lowering lovastatins (Campbell and Vederas), among others (Busch and Hertweck 2009;Calderone 2008;Shen 2003).…”

Section: Introductionmentioning

confidence: 99%

Growth condition optimization for docosahexaenoic acid (DHA) production by Moritella marina MP-1

Kautharapu

Rathmacher

Jarboe

2012

Appl Microbiol Biotechnol

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The marine organism Moritella marina MP-1 produces the polyunsaturated fatty acid docosahexaenoic acid (DHA). While the basic metabolic pathway for DHA production in this organism has been identified, the impact of growth conditions on DHA production is largely unknown. This study examines the effect of supplemental carbon, nitrogen and salts, growth temperature and media composition and pH on DHA and biomass production and the fatty acid profile. The addition of supplemental nitrogen significantly increased the overall DHA titer via an increase in biomass production. Supplemental glucose or glycerol increased biomass production, but decreased the amount of DHA per biomass, resulting in no net change in the DHA titer. Acidification of the baseline media pH to 6.0 increased DHA per biomass. Changes in growth temperature or provision of supplemental sodium or magnesium chloride did not increase DHA titer. This organism was also shown to grow on defined minimal media. For both media types, glycerol enabled more DHA production per biomass than glucose. Combination of these growth findings into marine broth supplemented with glycerol, yeast extract, and tryptone at pH 6.0 resulted in a final titer of 82 Â± 5 mg/L, a nearly eightfold increase relative to the titer of 11 Â± 1 mg/L seen in the unsupplemented marine broth. The relative distribution of other fatty acids was relatively robust to growth condition, but the presence of glycerol resulted in a significant increase in myristic acid (C14:0) and decrease in palmitic acid (C16:0). In summary, DHA production by M. marina MP-1 can be increased more than fivefold by changing the growth media. Metabolic engineering of this organism to increase the amount of DHA produced per biomass could result in additional increases in titer. ABSTRACTThe marine organism Moritella marina MP-1 produces the polyunsaturated fatty acid docosahexaenoic acid (DHA). While the basic metabolic pathway for DHA production in this organism has been identified, the impact of growth conditions on DHA production is largely unknown. This study examines the effect of supplemental carbon, nitrogen and salts, growth temperature and media composition and pH on DHA and biomass production and the fatty acid profile. The addition of supplemental nitrogen significantly increased the overall DHA titer via an increase in biomass production. Supplemental glucose or glycerol increased biomass production, but decreased the amount of DHA per biomass, resulting in no net change in the DHA titer.Acidification of the baseline media pH to 6.0 increased DHA per biomass. Changes in growth temperature or provision of supplemental sodium or magnesium chloride did not increase DHA titer. This organism was also shown to grow on defined minimal media. For both media types, glycerol enabled more DHA production per biomass than glucose. Combination of these growth findings into marine broth supplemented with glycerol, yeast extract and tryptone at pH 6.0 resulted in a final titer of 82+5 mg/L, a nearly 8-fold increase...

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Section: Introductionmentioning

confidence: 99%

Growth condition optimization for docosahexaenoic acid (DHA) production by Moritella marina MP-1

Kautharapu

Rathmacher

Jarboe

2012

Appl Microbiol Biotechnol

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“…The same compound (C31:9) was detected in marine and nonmarine, and psychrophilc and mesophilic bacteria, which contained EPA and/or DHA (Table 4). Based on our present results, previous studies on the bacterial distribution of PUFAs and pfa genes [3], and data on the predicted biosynthetic pathway of PUFAs in the polyketide synthesis mode [37,39,40], we will discuss the biosynthetic routes of C31:9.…”

Section: Discussionmentioning

confidence: 60%

“…6 and 7). With regard to the decarboxylation activity to synthesize C31:9, such activity would be present only in bacteria carrying pfa genes, because there was no C31:9 detected in two PUFA-producing marine eukaryotes, which are expected to have eukaryote type pfa genes ( [37] and Table 4). Acetonitrile was used as the ionization reagent in CI/MS.…”

Section: Discussionmentioning

confidence: 99%

“…To investigate effects of temperature on growth and production of C31:9, strain osh08 was cultivated at 0, 4,15,20,25,30,37,40, and 43 °C.…”

Section: Strains and Culture Conditions Of Bacteria And Eukaryotesmentioning

confidence: 99%

“…These eukaryotic microorganisms are expected to have pfa-like genes that are responsible for the biosynthesis of PUFAs because such genes have been cloned from Schizochytrium sp. ( [37] and see accession numbers AF378327, AF378328, and AF378329). Although E. coli DH5 does not inherently produce EPA or DHA, the cells transformed with the pfa genes responsible for the biosynthesis of EPA and DHA produced EPA and DHA, respectively, but not C31:9 (Table 4 and see [3]).…”

Section: Distribution Of C31:9 In Various Microorganisms and Its Possmentioning

confidence: 99%

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Possible Biosynthetic Pathways for all cis‐3,6,9,12,15,19,22,25,28‐Hentriacontanonaene in Bacteria

Sugihara

Hori

Nakanowatari

et al. 2009

Lipids

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A very long chain polyunsaturated hydrocarbon, hentriacontanonaene (C31:9), was detected in an eicosapentaenoic acid (EPA)-producing marine bacterium, which was isolated from the mid-latitude seashore of Hokkaido, Japan, and was tentatively identified as mesophilic Shewanella sp. strain osh08 from 16S rRNA gene sequencing. The geometry and position of the double bonds in this compound were determined physicochemically to be all cis at positions 3, 6, 9, 12, 15, 19, 22, 25, and 28. Although C31:9 was detected in all of the seven EPA-or/and docosahexaenoic acid-producing bacteria tested, an EPA-deficient mutant (strain IK-18) of one of these bacteria had no C31:9. Strain IK-18 had defects in the pfaD gene, one of the five pfa genes responsible for the biosynthesis of EPA.Although Escherichia coli DH5 does not produce EPA or DHA inherently, cells transformed with the pfa genes responsible for the biosynthesis of EPA and DHA produced EPA and DHA, respectively, but not C31:9. These results suggest that the Pfa protein complex is involved in the biosynthesis of C31:9 and that pfa genes must not be the only genes responsible for the formation of C31:9. In this report, we determined for the first time the molecular structure of the C31:9 and discuss the possible biosynthetic pathways of this compound. Key words

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Oils from Microorganisms

Wynn¹,

Ratledge

2005

Bailey's Industrial Oil and Fat Products

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All microorganisms, bacteria, yeasts, fungi, and algae produce a wide variety of lipids. Some species, the oleaginous ones, produce large amounts, sometimes up to 70% of the cell mass, of triacylglycerol oils, which, depending on the organism, are similar in fatty acyl constituents to most of the conventional plant seed oils. These are known as single‐cell oils (SCO). They may, if the economics are right, be considered as alternative sources to these commodities. However, because of the high cost of fermentation technology to cultivate microorganisms on a large scale, only the very highest valued microbial oils can be considered economically viable. This chapter gives a brief background to microbial technology and gives examples of how SCOs have been produced as sources of gamma‐linolenic acid, being alternatives to evening primrose and borage oils, and for the production of cocoa butter equivalent oils for possible use in chocolate confectionery. Current commercial productions of arachidonic acid (20:4,n‐4) using the filamentous fungus, Mortierella alpina , and of docosahexaenoic acid (22:6,n‐3) by the marine dinoflagellate, Crypthecodinium cohnii , and by the chytrid, Schizochytrium spp. are described in which oils are produced that are then incorporated into infant baby formulas and are taken for the prevention of coronary heart disease in adults. The prospect of using microorganisms to produce other polyunsaturated fatty acids, such as eicosapentaenoic acid, for possible medical applications, concludes this chapter.

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Production of Polyunsaturated Fatty Acids by Polyketide Synthases in Both Prokaryotes and Eukaryotes

Cited by 632 publications

References 25 publications

Growth condition optimization for docosahexaenoic acid (DHA) production by Moritella marina MP-1

Growth condition optimization for docosahexaenoic acid (DHA) production by Moritella marina MP-1

Possible Biosynthetic Pathways for all cis‐3,6,9,12,15,19,22,25,28‐Hentriacontanonaene in Bacteria

Oils from Microorganisms

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