Enhanced production of insect raw-starch-digesting alpha-amylase accompanied by high erythritol synthesis in recombinant Yarrowia lipolytica fed-batch cultures at high-cell-densities |
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| Institution: | 1. College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, No. 30 South Puzhu Road, Nanjing 211816, People’s Republic of China;2. GMPA, AgroParisTech, INRA, Université Paris-Saclay, 78850, Thiverval-Grignon, France;3. School of Pharmaceutical Sciences, Nanjing Tech University, No. 30 South Puzhu Road, Nanjing 211816, People’s Republic of China;4. State Key Laboratory of Materials-Oriented Chemical Engineering, Nanjing Tech University, No.5 Xinmofan Road, Nanjing 210009, People’s Republic of China;5. Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), No.5 Xinmofan Road, Nanjing 210009, People’s Republic of China;1. Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand;2. Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand;3. Biofuels by Biocatalysts Research Unit, Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand;4. Innovation Institute, PTT Public Co., Ltd., Ayutthaya, 13170, Thailand;5. LISBP, Université de Toulouse, CNRS, INRA, INSA, Toulouse, 31077, France;1. Bioresource Center Russian National Collection of Industrial Microorganisms (BRC VKPM), State Research Institute of Genetics and Selection of Industrial Microorganisms National Research Centre “Kurchatov Institute” (NRC “Kurchatov Institute” - GosNIIgenetika), 1-st Dorozhny pr., 1, Moscow, 117545, Russian Federation;2. Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari “Aldo Moro”, Campus Universitario, Via Orabona 4, 70125, Bari, Italy;3. Department of Bioengineering, Imperial College London, London, SW7 2AZ, UK;4. CNR Institute of Biomembranes, Bioenergetics and Molecular Biotechnologies (IBIOM), Campus Universitario, Via Orabona 4, 70125, Bari, Italy |
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| Abstract: | Recently we reported on raw-starch-digesting ability of alpha-amylase from an insect Sitophilus oryzae (SoAMY) expressed in recombinant Yarrowia lipolytica cells, and demonstrated its usefulness in simultaneous saccharification and fermentation processes with industrial yeasts. In this study we applied fed-batch cultures of Y. lipolytica 4.29 strain reaching high-cell-densities (up to 70 gDCW/L]), to enhance SoAMY production. SoAMY activity in the medium reached the peak value of 22,979.23 ± 184 AU/L], at volumetric productivity of 121.58 ± 1.75 AU/L/h], and yield of 71.83 ± 3.08 AU/gglycerol], constituting roughly 160-fold improvement, compared to the best previous result. The cultivations were accompanied by high production of erythritol (83.58 g/L]), at the marginal production of mannitol (5.46 g/L]). Elementary analyses of media constituents, the enzyme and the yeast biomass gave better insight into carbon and nitrogen fluxes distribution. Due to application of genetic engineering and bioprocess engineering strategies, the insect-derived enzyme can be produced at the quantities competitive to microbial catalysts. |
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| Keywords: | Amylase Fed-batch cultures Heterologous over-expression High-cell-density cultures |
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