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Journal : Natural Sciences Engineering and Technology Journal

Targeted Metabolic Engineering of Saccharomyces cerevisiae for High-Efficiency Valorization of Lignocellulosic Biomass into Superior-Quality Bioplastics Diana, Nur; Ahmad, Zaki; Fajic, Selma
Natural Sciences Engineering and Technology Journal Vol. 5 No. 2 (2025): Natural Sciences Engineering and Technology Journal
Publisher : HM Publisher

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37275/nasetjournal.v5i2.70

Abstract

The global transition towards a sustainable circular bioeconomy urgently requires innovative platforms for converting renewable waste streams into value-added products. Lignocellulosic biomass, particularly agricultural residue like rice straw, stands as a vast, underutilized carbon source. This study details the systematic metabolic engineering of Saccharomyces cerevisiae for the high-efficiency production of poly(3-hydroxybutyrate) (PHB), a biodegradable bioplastic, from rice straw hydrolysate. A multi-faceted synthetic biology approach was implemented in S. cerevisiae CEN.PK2-1C. A robust xylose co-utilization pathway was integrated using codon-optimized genes from Scheffersomyces stipitis. The PHB biosynthesis pathway from Cupriavidus necator was introduced using a cassette of strong, constitutive yeast promoters (pTDH3, pTEF1, pPGK1). To maximize carbon flux towards PHB, key competing pathways were eliminated via CRISPR-Cas9-mediated gene knockouts of the primary alcohol dehydrogenase (ADH1) and glycerol-3-phosphate dehydrogenase (GPD1) genes. The performance of the final engineered strain was evaluated in high-cell-density fed-batch fermentation using detoxified rice straw hydrolysate sourced from Palembang, Indonesia. The final engineered strain, YL-PHB-05 (Δadh1 Δgpd1), demonstrated superior performance. In fed-batch bioreactor cultivation, it achieved a final cell dry weight of 33.8 ± 1.5 g/L and a PHB titer of 15.2 ± 0.7 g/L, with an intracellular PHB accumulation of 45.0 ± 1.2% of cell dry weight. This corresponds to a high yield of 0.28 g PHB per gram of consumed sugars. Crucially, the produced PHB exhibited a superior weight-average molecular weight (Mw) of 1.2 x 10⁶ Da with a polydispersity index of 2.1. In conclusion, this work successfully demonstrates a robust strategy for engineering S. cerevisiae into an efficient cell factory for producing high-quality bioplastics from a globally relevant agricultural waste stream. The high titers, yields, and superior polymer properties achieved present a significant advancement towards establishing an economically viable and sustainable process for bioplastic production within a circular bioeconomy.
Co-Authors Ababil, Ressy Yulia Abdullah Syakur Novianto Ahmad, Zaki Ainun Jariyah Ainun Muthoharoh, Ainun Alfarobi, Abdul Qodir Jailani Anggriani, Lailla Ardhelia, Devi Arista Fauzi Kartika Sari Ashari, Ahmad Aziz Santoso Asriani Asriani, Asriani Audita Nuvriasari Barumbun, Mardyanto Bastomi, Mohamad DEWI DIAH FAKHRIYYAH Dewi, Anastasya Vivian Risna Dwiyani Sudaryanti Dyah Arini Rudiningtyas Eko Handayanto Eko Purnomo Erfan Efendi F, Dewi Diah Fadhilah, Gita Maulani Fadhilla, Yuka Fajar Ari Nugroho Fajic, Selma Hamida, Rahma Nur Hattaradzani, Shafiki Helmi Kurniawan Helmilia Putri, Nur Hermawan, Ismi Istihanah Hidayat, Amelia Putri Eka Nur Hidayati, Irma Hofifah, Nur Imam Syafi'i Irawan, Pristanto Ria Iskandaria, Weyni Jabar, Aryuni Salpiana jamaluddin hos Juliati, Ratih Junaidi Junaidi Kasanah, Nengnis Uswatul Khoiroh, Annis Duwi Rokhmatul Kinaza, Tsibat Vozi Kirana, Denny Candra Komariyah, Rizkiyatul Lestari, Ida Ayu M. Cholid Mawardi Mafaza, Arini Magviroh, Oktavia Maimatul Maslichah, Maslichah Masruroh, Hanik Maulida, Rizki Sofa Maulidiyah, Nailin Nikmatul MF. Arrozi Adhikara Miswanto Miswanto Mubarok Syam, Abdi Ningrum, Wulan Agustin NOVA Nurhayati Nurhayati Nurjana, Nurjanah Prabowo, Hernanda Bagus Prameswari, Gita Anggun Purbawati, Cindy Puspaharisti, Larasati Putri, Berliana Imandari Putri, Diah Utami Qorry Fatona, Arifa R. Shauki, Elvia Rahayu, Fika Putri Rahman, Maulidah Rahmawati, Rahmawati Rasendriya, Renata Ucca Regina Riskawati Riskawati, Riskawati Rizka Mayang Saputri Ginting, Rizka Mayang Saputri Sabirin, Rizky Sal Sabila, Asa Aisyi Septiani, Linda Sudarmiatin Supri Supri Syafi’i, Imam Thoyibah, Habibatut Tonang, Aliya Putri Umi Nandiroh Utami, Putrilia Dwi Shintya Adi Yekti, Muhammad Yoga Pratita Yulianto, Wahyudi