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Lookup NU author(s): Jinzheng Wang, Dr Kheng-Lim GohORCiD
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© 2026 Elsevier Ltd. All rights are reserved, including those for text and data mining, AI training, and similar technologies. The fatty acid chain length of lipids plays a pivotal role in determining their nutritional value and industrial applicability. However, the synthesis of functional lipids with defined fatty acid compositions is often limited by the insufficient chain-length specificity of conventional lipases. In this study, a chain-length-selective lipase targeting palmitic (C16) acyl chains was engineered using an integrated computational–experimental strategy, and its performance was evaluated at a two-phase microfluidic interface. The engineered variants exhibited a 0.79 to 28.48-fold enhancement in hydrolytic activity toward p-nitrophenyl palmitate (p-NP C16:0), along with improved thermal stability (t1/2 of mutant I291Y increased from 0.40 to 2.04 h). Notably, the dominant mutant I291Y showed markedly enhanced selectivity, with hydrolytic activity toward p-NP C16:0 being 41.65-fold that toward p-nitrophenyl oleate (p-NP C18:1). Consistently, the Vmax of I291Y for p-NP C16:0 was 284.19% higher than that for p-NP C18:1. Molecular docking and molecular dynamics simulations lead to a mechanistic hypothesis in which the substitution of isoleucine 291 with tyrosine is suggested to strengthen the affinity of the substrate binding tunnel for C16 acyl chains while generating steric hindrance against oleic (C18) acyl chains, thereby potentially conferring C16 selectivity. This structural modification also enabled I291Y to selectively hydrolyze C16 acyl chains in glyceryl tripalmitate rather than the C18 acyl chains in glycerol trioleate at the microfluidic two-phase interface. This work demonstrates a computationally guided engineering approach for developing a lipase with enhanced C16 preference, which may facilitate targeted lipid modification and the tailored production of nutritional lipids.
Author(s): Wang J-Z, Tan L, Wu C-K, Wang X-Y, Goh KL, Herman RA, You S, Sheng S, Wu F-A, Wang J
Publication type: Article
Publication status: Published
Journal: Bioresource Technology
Year: 2026
Volume: 461
Print publication date: 01/12/2026
Online publication date: 18/07/2026
Acceptance date: 16/07/2026
ISSN (print): 0960-8524
ISSN (electronic): 1873-2976
Publisher: Elsevier Ltd
URL: https://doi.org/10.1016/j.biortech.2026.135431
DOI: 10.1016/j.biortech.2026.135431
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