Construction of Efficient Recombinant Strain Through Genome Shuffling in Marine Endophytic Fusarium sp. ALAA-20 for Impr
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RESEARCH ARTICLE-BIOLOGICAL SCIENCES
Construction of Efficient Recombinant Strain Through Genome Shuffling in Marine Endophytic Fusarium sp. ALAA-20 for Improvement Lovastatin Production Using Agro-Industrial Wastes Alaa Ahmed Mohamed El-Bondkly1 · Mervat Morsy Abbas Ahmed El-Gendy2 · Ahmed M. A. El-Bondkly3 Received: 7 March 2020 / Accepted: 29 August 2020 © King Fahd University of Petroleum & Minerals 2020
Abstract During searching for marine endophytic fungi that have the potential to effectively produce lovastatin using agro-industrial residues, the endophytic Fusarium sp. ALAA-20 was isolated from marine sponge Aplysina fistularis with lovastatin production of 4.51 mg/gds in the solid-state fermentation of rice straw. Lovastatin yield was improved by enhancing approaches including mutation induction and three consecutive cycles of genome shuffling of the producing strain ALAA-20 to obtain the hyperactive recombinant strain FR3/1 with a higher yield of lovastatin (52.1 mg/gds), which is 11.55- and 3.1-fold of the parent strain ALAA-20 and the hyperactive mutant NE21 (16.8 mg/gds), respectively. Moreover, optimization of the solidstate fermentation of oil cakes by the recombinant FR3/1 significantly increased the lovastatin production by 64.19-, 17.23and 5.56-fold over the selected wild strain (ALAA-20), mutant (NE21) and fusant (FR3/1) prior to the optimization process. Lovastatin obtained from the recombinant FR3/1 strain showed pleiotropic effects such as antibacterial activity against drug-resistant Staphylococcus, Pseudomonas and Klebsiella species and antifungal activity against Candida, Aspergillus, Fusarium and Trichophyton species. New recombinant strain FR3/1 lovastatin showed potent antitumor activity against liver (HepG-2), colon (HCT-116), breast (MCF-7) and lung (A-549) cancer cell lines with IC50 equal to 8.0, 7.2, 4.8 and 9.1 µM, respectively. Keywords Lovastatin · Genome shuffling · SSF · Optimization · Antibacteria · Antifungal · Antitumor
1 Introduction Statins compounds such as lovastatin are fungal fermentation products and have been used as a lipid-lowering medication through a significant degree of competitive inhibition for HMG-CoA reductase enzyme in the biosynthesis pathway of cholesterol [1]. In addition to their role in lowering blood lipid levels, they are also characterized by anti-inflammation, anticancer, neuroprotective, antibacterial and antifungal properties [2, 3]. Filamentous fungi are well known for their abilities to produce an enormous amount
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Ahmed M. A. El-Bondkly [email protected]; [email protected]
1
Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, Cairo 11562, Egypt
2
Chemistry of Natural and Microbial Products Department, National Research Centre, Dokki, Giza 12622, Egypt
3
Genetics and Cytology Department, National Research Centre, Dokki, Giza 12622, Egypt
of secondary metabolites from low-value bulk chemicals to high-value drugs such as immunosuppressed, cholesterollowering and anti-osteoporosis drugs [4–6]. Marine sponge is kno
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