Arachidonic acid production by the oleaginous fungus Mortierella alpina 1S-4 : A review

The filamentous fungus Mortierella alpina 1S-4 is capable of accumulating a large amount of triacylglycerol containing C20 polyunsaturated fatty acids (PUFAs). Indeed, triacylglycerol production by M. alpina 1S-4 can reach 20 g/L of culture broth, and the critical cellular signaling and structural PUFA arachidonic acid (ARA) comprises 30%–70% of the total fatty acid. The demonstrated health benefits of functional PUFAs have in turn encouraged the search for rich sources of these compounds, including fungal strains showing enhanced production of specific PUFAs. Screening for mutants and targeted gene manipulation of M. alpina 1S-4 have elucidated the functions of various enzymes involved in PUFA biosynthesis and established lines with improved PUFA productivity. In some cases, these strains have been used for indistrial-scale production of PUFAs, including ARA. In this review, we described practical ARA production through mutant breeding, functional analyses of genes encoding enzymes involved in PUFA biosynthesis, and recent advances in the production of specific PUFAs through molecular breeding of M. alpina 1S-4

Characterization of galactose-dependent promoters from an oleaginous fungus Mortierella alpina 1S-4.

An inducible promoter is a useful tool for the controlled expression of a given gene. In this report, we describe galactose-dependent promoters for potential use in an oleaginous fungus Mortierella alpina. We cloned the putative promoter regions of two genes encoding galactose metabolic enzymes, GAL1 and GAL10, from the genome of M. alpina 1S-4. The β-glucuronidase (GUS) reporter gene assay in M. alpina 1S-4 revealed that regulation of these promoters was dependent on the presence of galactose in the medium both with and without other sugars. With the GAL10 promoter, an approximately 50-fold increase of GUS activity was demonstrated by addition of galactose into the culture media at any cultivation phase. The 5' deletion analysis of the GAL10 promoter revealed that a promoter region of over 2, 000 bp length was required for its high-level activity and sufficient inducible response. Significantly, this is the first report of inducible promoters of zygomycetes. The GAL10 promoter will be a valuable tool for gene manipulation in M. alpina 1S-4

Selection and characterization of promoters based on genomic approach for the molecular breeding of oleaginous fungus Mortierella alpina 1S-4.

To express a foreign gene effectively, a good expression system is required. In this study, we investigated various promoters as useful tools for gene manipulation in oleaginous fungus Mortierella alpina 1S-4. We selected and cloned the promoter regions of 28 genes in M. alpina 1S-4 on the basis of expression sequence tag abundance data. The activity of each promoter was evaluated using the β-glucuronidase (GUS) reporter gene. Eight of these promoters were shown to enhance GUS expression more efficiently than a histone promoter, which is conventionally used for the gene manipulation in M. alpina. Especially, the predicted protein 3 and the predicted protein 6 promoters demonstrated approximately fivefold higher activity than the histone promoter. The activity of some promoters changed along with the cultivation phase of M. alpina 1S-4. Seven promoters with constitutive or time-dependent, high-level expression activity were selected, and deletion analysis was carried out to determine the promoter regions required to retain activity. This is the first report of comprehensive promoter analysis based on a genomic approach for M. alpina. The promoters described here will be useful tools for gene manipulation in this strain

MECHANICAL BEHAVIOR OF ANTI-SYMMETRICALLY LAMINATED COMPOSITE BLADES

ABSTRACT Anti-symmetrically laminated composites have coupling effects between tensile stress and twisting deformation, and are very attractive as fan blade materials of aircraft engines. Blades fabricated by anti-symmetrically laminated composites can automatically adjust the stagger angle to better aerodynamic conditions with change of axial force or rotational speed owing to the coupling effects. Thus, the anti-symmetrically laminated composite blades are expected to improve aerodynamic efficiency and the stability of aircraft engines. In this paper, the mechanical behavior of antisymmetrically laminated composite blades is evaluated by spin tests and finite element analyses

Big-Volume SliceGAN for Improving a Synthetic 3D Microstructure Image of Additive-Manufactured TYPE 316L Steel

A modified SliceGAN architecture was proposed to generate a high-quality synthetic three-dimensional (3D) microstructure image of TYPE 316L material manufactured through additive methods. The quality of the resulting 3D image was evaluated using an auto-correlation function, and it was discovered that maintaining a high resolution while doubling the training image size was crucial in creating a more realistic synthetic 3D image. To meet this requirement, modified 3D image generator and critic architecture was developed within the SliceGAN framework.Sugiura K., Ogawa T., Adachi Y., et al. Big-Volume SliceGAN for Improving a Synthetic 3D Microstructure Image of Additive-Manufactured TYPE 316L Steel. Journal of Imaging 9, 90 (2023); https://doi.org/10.3390/jimaging9050090

Manipulation of charge carrier flow in Bi₄NbO₈Cl nanoplate photocatalyst with metal loading

Separation of photoexcited charge carriers in semiconductors is important for efficient solar energy conversion and yet the control strategies and underlying mechanisms are not fully established. Although layered compounds have been widely studied as photocatalysts, spatial separation between oxidation and reduction reaction sites is a challenging issue due to the parallel flow of photoexcited carriers along the layers. Here we demonstrate orthogonal carrier flow in layered Bi₄NbO₈Cl by depositing a Rh cocatalyst at the edges of nanoplates, resulting in spatial charge separation and significant enhancement of the photocatalytic activity. Combined experimental and theoretical studies revealed that lighter photogenerated electrons, due to a greater in-plane dispersion of the conduction band (vs. valence band), can travel along the plane and are readily trapped by the cocatalyst, whereas the remaining holes hop perpendicular to the plane because of the anisotropic crystal geometry. Our results propose manipulating carrier flow via cocatalyst deposition to achieve desirable carrier dynamics for photocatalytic reactions in layered compounds

Medium-Scale Traveling Ionospheric Disturbances and Plasma Bubbles Observed by an All-Sky Airglow Imager at Yonaguni, Japan

We report on night time air glow imaging observations of the low latitude ionosphere by means of a 630-m all-sky imager in stalled in March 2006 at Yonaguni, Japan (24.5°N, 123.0°E; 14.6°N geomagnetic), about 100 km east of Taiwan. The imager detected medium-scale traveling ionospheric disturbances (MSTIDs) for about 7 hours on the night of 26 May 2006. A dense GPS net work in Japan also ob served the same MSTID event on this night. The imager and GEONET data indicate that most of the MSTIDs prop a gated south west ward from the north of Japan to the south of Yonaguni and Taiwan over 4000 km, with a southern limit of 19°N (geomagnetic latitude 9°N) or lower. On the night of 10 November 2006, the imager observed two weak emission bands that were embedded on the F-region anomaly crest to the south of Yonaguni. The simultaneous electron density profiles from the FORMOSAT-3/COS MIC mission demonstrate that the weak emission bands are due to density depletions in equatorial plasma bubbles. These case studies suggest that the Yonaguni imager in collaboration with other instruments is very suit able for the study of ionospheric disturbances in and around the northern F-region anomaly crest

Metabolic engineering of oleaginous fungus Mortierella alpina for high production of oleic and linoleic acids

The aim of this work was to study the molecular breeding of oleaginous filamentous Mortierella alpina for high production of linoleic (LA) or oleic acid (OA). Heterologous expression of the Δ12-desaturase (DS) gene derived from Coprinopsis cinerea in the Δ6DS activity-defective mutant of M. alpina increased the LA production rate as to total fatty acid to 5 times that in the wild strain. By suppressing the endogenous Δ6I gene expression by RNAi in the Δ12DS activity-defective mutant of M. alpina, the OA accumulation rate as to total fatty acid reached 68.0%. The production of LA and OA in these transformants reached 1.44 and 2.76 g/L, respectively, on the 5th day. The Δ6I transcriptional levels of the RNAi-treated strains were suppressed to 1/10th that in the parent strain. The amount of Δ6II RNA in the Δ6I RNAi-treated strain increased to 8 times that in the wild strain

Gastric Metastasis from a Primary Renal Leiomyosarcoma

Primary leiomyosarcoma of the kidney is rare. Here we report a case of metastasis of this tumor to the stomach. A 73-year-old man visited our hospital suffering from general weakness and intermittent tarry stools. He had undergone right nephrectomy for renal leiomyosarcoma 2 years previously. There had been no local recurrence or distant metastasis in the 2-year follow-up period. Endoscopy revealed two submucosal tumors in the stomach. These tumors were diagnosed histologically as leiomyosarcoma and distal gastrectomy was performed. Subsequent histochemical staining confirmed the diagnosis of gastric metastasis from renal leiomyosarcoma. The patient died due to metastases to the liver and bone 9 months after the operation. To the best of our knowledge, this is the first report of gastric metastasis from primary renal leiomyosarcoma

Production of ricinoleic acid-containing monoestolide triacylglycerides in an oleaginous diatom, Chaetoceros gracilis.

実用珪藻ツノケイソウによるリシノール酸の生産に成功. 京都大学プレスリリース. 2016-11-14.Ricinoleic acid (RA), a hydroxyl fatty acid, is suitable for medical and industrial uses and is produced in high-oil-accumulating organisms such as castor bean and the ergot fungus Claviceps. We report here the efficient production of RA in a transgenic diatom Chaetoceros gracilis expressing the fatty acid hydroxylase gene (CpFAH) from Claviceps purpurea. In transgenic C. gracilis, RA content increased at low temperatures, reaching 2.2 pg/cell when cultured for 7 d at 15 °C, without affecting cell growth, and was enhanced (3.3 pg/cell) by the co-expression of a palmitic acid-specific elongase gene. Most of the accumulated RA was linked with monoestolide triacylglycerol (ME TAG), in which one RA molecule was esterified to the α position of the glycerol backbone and was further esterified at its hydroxy group with a fatty acid or second RA moiety, or 1-OH TAG, in which RA was esterified to the glycerol backbone. Overall, 80% of RA was accumulated as ME TAGs. Furthermore, exogenous RA-methyl ester suppressed the growth of wild-type diatoms in a dose-dependent manner and was rapidly converted to ME TAG. These results suggest that C. gracilis masks the hydroxyl group and accumulates RA as the less-toxic ME TAG