Presentation Information
[2BRBP-17]FROM FLASK TO INDUSTRY: BOOSTING NANOCELLULOSE PRODUCTION BY Komagataeibacter sp. THROUGH MOLASSES BASED OPTIMIZATION AND SCALE-UP
○Diveyan Mohan1,2, Esther Qian Tung Ong1, Siti Nur Hazwani Oslan4, Mohammad Rizal Kapri2, Siti Nurbaya Oslan1,2,3 (1. Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia. (Malaysia), 2. Enzyme and Microbial Technology (EMTech) Research Centre, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia (Malaysia), 3. Enzyme Technology and X-Ray Crystallography Laboratory (VacBio 5), Laboratory of Vaccine and Biomolecules, Institute of Biosciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia (Malaysia), 4. Food Security Research Laboratory, Faculty of Food Science and Nutrition, Universiti Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah, Malaysia (Malaysia))
Keywords:
Bacterial nanocellulose (BNC),Komagataeibacter sp.,Scale-up fermentation,Sugarcane molasses,Bioprocess optimization
[Introduction & Purpose]
Bacterial nanocellulose (BNC) is a versatile biopolymer with high purity, mechanical strength and biocompatibility, making it attractive for various industrial applications. However, its scalability remains a critical bottleneck due to high costs and low yields, highlighting the need for robust and scalable bioprocess strategies. Therefore, this study aims to enhance BNC production by a high BNC-producing bacterium, Komagataeibacter sp., through sequential optimization of cultivation conditions and evaluation of scale-up performance in a 2 L bioreactor system using sugarcane molasses as a low-cost carbon source.
[Methodology & Results]
BNC production was first assessed at the shake flask level (500 mL) in Hestrin-Schramm (HS) medium by optimizing key physical parameters, including pH, temperature, agitation, aeration and surface area, as well as different carbon sources. Maximum BNC production was achieved under the following conditions: pH 6, 30°C, 50 rpm, 60% aeration rate, and 63.6 cm2 surface area. Among the carbon sources tested, heat-treated molasses yielded the highest BNC yield (>2.4 g/L), followed by crude molasses and acid-heat-treated molasses, while glucose showed the lowest performance. Using these optimized conditions, production was further evaluated in both shake flask and bioreactor systems. At the shake flask level, molasses supported higher BNC productivity (~0.08-0.09 g/L/day) and water-holding capacity (~240-250 g water/g dry BNC) compared to glucose. However, upon scale-up, no BNC formation was observed using molasses. This was likely due to excess nutrient availability and enhanced oxygen transfer, which redirected cellular metabolism toward biomass accumulation and by-product (gluconic acid) formation rather than cellulose synthesis. In contrast, bioreactor cultivation using glucose resulted in the highest BNC yield (~1.1-1.2 g/L), exceeding shake flask performance under similar conditions. Morphological analysis using scanning electron microscopy (SEM) revealed that BNC from molasses in shake flasks exhibited a more porous and hydrated structure, whereas bioreactor-produced BNC was denser with thicker fibres, reflecting process-dependent differences in morphology.
[Conclusion & Future Recommendations]
In conclusion, this study enhanced BNC yield and properties through sequential optimization of cultivation parameters and carbon source. While molasses is a promising low-cost substrate at shake flask scale, its performance was not directly transferable to bioreactor conditions, highlighting a key scale up limitation. Future work will focus on optimizing substrate composition and bioreactor parameters, particularly dissolved oxygen and agitation control, to enable efficient and scalable BNC production.
Bacterial nanocellulose (BNC) is a versatile biopolymer with high purity, mechanical strength and biocompatibility, making it attractive for various industrial applications. However, its scalability remains a critical bottleneck due to high costs and low yields, highlighting the need for robust and scalable bioprocess strategies. Therefore, this study aims to enhance BNC production by a high BNC-producing bacterium, Komagataeibacter sp., through sequential optimization of cultivation conditions and evaluation of scale-up performance in a 2 L bioreactor system using sugarcane molasses as a low-cost carbon source.
[Methodology & Results]
BNC production was first assessed at the shake flask level (500 mL) in Hestrin-Schramm (HS) medium by optimizing key physical parameters, including pH, temperature, agitation, aeration and surface area, as well as different carbon sources. Maximum BNC production was achieved under the following conditions: pH 6, 30°C, 50 rpm, 60% aeration rate, and 63.6 cm2 surface area. Among the carbon sources tested, heat-treated molasses yielded the highest BNC yield (>2.4 g/L), followed by crude molasses and acid-heat-treated molasses, while glucose showed the lowest performance. Using these optimized conditions, production was further evaluated in both shake flask and bioreactor systems. At the shake flask level, molasses supported higher BNC productivity (~0.08-0.09 g/L/day) and water-holding capacity (~240-250 g water/g dry BNC) compared to glucose. However, upon scale-up, no BNC formation was observed using molasses. This was likely due to excess nutrient availability and enhanced oxygen transfer, which redirected cellular metabolism toward biomass accumulation and by-product (gluconic acid) formation rather than cellulose synthesis. In contrast, bioreactor cultivation using glucose resulted in the highest BNC yield (~1.1-1.2 g/L), exceeding shake flask performance under similar conditions. Morphological analysis using scanning electron microscopy (SEM) revealed that BNC from molasses in shake flasks exhibited a more porous and hydrated structure, whereas bioreactor-produced BNC was denser with thicker fibres, reflecting process-dependent differences in morphology.
[Conclusion & Future Recommendations]
In conclusion, this study enhanced BNC yield and properties through sequential optimization of cultivation parameters and carbon source. While molasses is a promising low-cost substrate at shake flask scale, its performance was not directly transferable to bioreactor conditions, highlighting a key scale up limitation. Future work will focus on optimizing substrate composition and bioreactor parameters, particularly dissolved oxygen and agitation control, to enable efficient and scalable BNC production.
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