Main Article Content
The objective of this study is to obtain a model that maximize production of γ-linolenic acid (GLA) by the moss Physcomitrella patens, employing response surface methodology (RSM). The RSM with a three-variables (sucrose, potassium nitrate and glutamate concentrations) and five-level (-2, -1, 0, 1 and 2) central composite design (CCD) including 20 experimental runs was employed to optimize the medium composition. Results showed that the experimental data could be appropriately fitted into a second-order polynomial model with a coefficient of determination (R2) more than 0.95 for GLA production. Analysis of variance (ANOVA) revealed that the model was highly significant (p<0.0001) and the effects of the sucrose (20-100 g/L) and glutamate (0.5-2.5 g/L) concentrations on GLA production were significant (p<0.05). The GLA production with the optimized culture medium (sucrose concentration of 62.92 g/L, potassium nitrate of 0.80 g/L and glutamate concentration of 1.42 g/L) by RSM increased 4.61 folds when compared with the standard BCD medium. This experimental value (16.37 mg GLA/L) fit well with the predicted values (16.20 mg GLA/L).
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.ลิขสิทธ์ ของมหาวิทยาลัยเทคโนโลยีราชมงคลพระนคร
 K. Youn, J. Lee, E.-Y. Yun, C.-T. Ho, M.V. Karwe, W.-S. Jeong and M. Jun, “Biological evaluation and in silico docking study of γ-linolenic acid as a potential BACE1 inhibitor,” Journal of Functional Foods, vol. 10, pp. 187-191, Sep. 2014.
 K. Youn, S. Lee and M. Jun, “Gamma-linolenic acid ameliorates Aβ-induced neuroinflammation through NF-κB and MAPK signalling pathways,” Journal of Functional Foods, vol. 42, pp. 30-37, Mar. 2018.
 W.H. Choi, “Evaluation of anti-tubercular activity of linolenic acid and conjugated-linoleic acid as effective inhibitors against Mycobacterium tuberculosis,” Asian Pacific Journal of Tropical Medicine, vol. 9, no. 2, pp. 125-129, Feb. 2016.
 G.J. Macfarlane, A. El-Metwally, V. De Silva, E. Ernst, G.L. Dowds and R.J. Moots, “Evidence for the efficacy of complementary and alternative medicines in the management of rheumatoid arthritis: a systematic review,” Rheumatology, vol. 50, pp. 1672-1683, Sep. 2011.
 A. Kawamura, K. Ooyama, K. Kojima, H. Kachi, T. Abe, K. Amano and T. Aoyama, “Dietary supplementation of gamma-linolenic acid improves skin parameters in subjects with dry skin and mild atopic dermatitis,” Journal of Oleo Science, vol. 60, no. 12, pp. 597-607, 2011.
 D. Simon, P.A. Eng, S. Borelli, R. Kägi, C. Zimmermann, C. Zahner, J. Drewe, L. Hess, G. Ferrari, S. Lautenschlager, B. Wüthrich and P. Schmid-Grendelmeier, “Gamma-linolenic acid levels correlate with clinical efficacy of evening primrose oil in patients with atopic dermatitis,” Advances in Therapy, vol. 31, pp. 180-188, Jan. 2014.
 M. Chas, C. Goupille, F. Arbion, P. Bougnoux, M. Pinault, M.L. Jourdan, S. Chevalier and L. Ouldamer, “Low eicosapentaenoic acid and gamma-linolenic acid levels in breast adipose tissue are associated with inflammatory breast cancer,” The Breast, vol. 45, pp. 113-117, Jun. 2019.
 K. Kavanagh, D.M. Flynn, K.A. Jenkins, M.D. Wilson and F.H. Chilton, “Stearidonic and γ-linolenic acids in echium oil improves glucose disposal in insulin resistant monkeys,” Prostaglandins, Leukotrienes and Essential Fatty Acids, vol. 89, no. 1, pp. 39-45, Jul. 2013.
 A. Freije, J. Alkhuzai and A.A. Al-Laith, “Fatty acid composition of three medicinal plants from Bahrain: New potential sources of γ-linolenic acid and dihomo-γ-linolenic,” Industrial Crops and Products, vol. 43, pp. 218- 224, May 2013.
 J.L. Guil-Guerrero, F. Gómez-Mercado, R.P. Ramos-Bueno, M.J. González-Fernández, M. Urrestarazu and M.A. Rincón-Cervera, “Sardinian Boraginaceae are new potential sources of gamma-linolenic acid,” Food Chemistry, vol. 218, pp. 435-439, Mar. 2017.
 J.L. Guil-Guerreroa, F. Gómez-Mercadob, R.P. Ramos-Buenoa, M.J. González-Fernándeza, M. Urrestarazuc, S. Jiménez-Beckerc and G. de Bélair, “Fatty acid profiles and sn-2 fatty acid distribution of γ-linolenic acid-rich Borago species,” Journal of Food Composition and Analysis, Anal, vol. 66, pp. 74-80, Mar. 2018.
 S. Piskernik, R. Vidrih, L. Demšar, D. Koron, M. Rogelj and T.P. Žontar, “Fatty acid profiles of seeds from different Ribes species,” LWT, vol. 98, pp. 424-427, Dec. 2018.
 J.L. Guil-Guerrero, M.A. Rincon-Cervera, F. Gomez-Mercado, R.P. Ramos-Bueno and E. Venegas-Venegas, “New seed oils of Boraginaceae rich in stearidonic and gamma-linolenic acids from the Maghreb region,” Journal of Food Composition and Analysis, vol. 31, no. 1, pp. 20-23, Aug. 2013.
 S. Lyashenko, M.J. González-Fernández, F. Gómez-Mercado, S. Yunusova, O. Denisenko and J.L. Guil-Guerrero, “Ribes taxa: A promising source of γ-linolenic acid-rich functional oils,” Food Chemistry, vol. 301, pp. 125309, Dec. 2019.
 S.U. Ahmed, K.K. Reddy, S.L. Swathy, S.K. Singh, S. Kanjilal, R.B.N. Prasad and A. Pandey, “Enrichment of γ-linolenic acid in the lipid extracted from Mucor zychae MTCC 5420,” Food Research International, vol. 42, no. 4, pp. 449-453, May. 2009.
 M.G. Sajilata, R.S. Singhal and M.Y. Kamat, “Fractionation of lipids and purification of γ-linolenic acid (GLA) from Spirulina platensis,” Food Chemistry, vol. 109, no. 3, pp. 580-586, Aug. 2008.
 M.L. Sun, C. Madzak, H.H. Liu, P. Song, L.J. Ren, H. Huang and X.J. Ji, “Engineering Yarrowia lipolytica for efficient γ-linolenic acid production,” Biochemical Engineering Journal, vol. 117, pp. 172-180, Jan. 2017.
 Y. Zhang, X. Luan, H. Zhang, V. Garre, Y. Song and C. Ratledge, “Improved γ-linolenic acid production in Mucor circinelloides by homologous overexpressing of delta-12 and delta-6 desaturases,” Microbial Cell Factories, vol. 16, pp. 113, Jun. 2017.
 S. Jeennor, J. Anantayanon, S. Panchanawaporn, S., Khoomrung, C. Chutrakul and K. Laoteng, “Reengineering lipid biosynthetic pathways of Aspergillus oryzae for enhanced production of γ-linolenic acid and dihomo-γ-linolenic acid,” Gene, vol. 706, pp. 106-114, Jul. 2019.
 S. Kaewsuwan, N. Bunyapraphatsara, D.J. Cove, R.S. Quatrano and P. Chodok, “High level production of adrenic acid in Physcomitrella patens using the algae Pavlova sp. ∆5-elongase gene,” Bioresource Technology, vol. 101, no. 11, pp. 4081-4088, Jun. 2010.
 P. Chodok, A. Kanjana-Opas and S. Kaewsuwan, “The plackett–burman design for evaluating the production of polyunsaturated fatty acids by Physcomitrella patens,” Journal of the American Oil Chemists' Society, vol. 87, pp. 521-529, Jan. 2010.
 P. Chodok, “Optimization of culture media for enhanced production of dihomo-γ-linolenic acid and eicosatetraenoic acid in Physcomitrella patens using multi-response surface methodology,” Songklanakarin Journal of Plant Science, vol. 5, no. 4, pp. 48-61, Oct.-Dec. 2018.
 O. Tehlivets, K. Scheuringer and S.D. Kohlwein, “Fatty acid synthesis and elongation in yeast,” Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, vol. 1771, no. 3, pp. 255-270, Mar. 2007.
 M.-J. Jin, H. Huang, A.-H. Xiao, Z. Gao, X. Liu, and C. Peng, “Enhancing arachidonic acid production by Mortierella alpina ME-1 using improved mycelium aging technology,” Bioprocess and Biosystems Engineering, vol. 32, pp. 117-122, May 2008.
 J.P. Wynn, A.A. Hamid and C. Ratledge, “The role of malic enzyme in the regulation of lipid accumulation in filamentous fungi,” Microbiology, vol. 145, no. 8, pp. 1911-1917, Aug. 1999.