Revealing the Native Multi-Enzyme Synergy of Acinetobacter calcoaceticus SDH15 for Ultra-Long-Chain Alkane Biodegradation

Xianrui Liu; Donghuan Su; Shaojie Wang; Zhenxia Du; Haijia Su

doi:10.1016/j.eng.2026.02.035

Engineering ›› :202602035 DOI: 10.1016/j.eng.2026.02.035

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Revealing the Native Multi-Enzyme Synergy of Acinetobacter calcoaceticus SDH15 for Ultra-Long-Chain Alkane Biodegradation

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Abstract

Polyethylene (PE) biodegradation is critical, yet microbial degradation of its ultra-long-chain alkane (ULCA, > C22) components remains limited and poorly understood beyond C40. This study identiﬁed Acinetobacter calcoaceticus ( A. calcoaceticus ) SDH15, a novel strain capable of degrading solid n -alkanes up to C56, thereby signiﬁcantly extending the known biological range of alkane catabolism. Integrated genomic and transcriptomic analyses revealed a four-module enzymatic system driving this process. The ﬁrst module, chemotaxis and adsorption, employs an upregulated PilH-Chp chemosensory system and type IV ﬁmbriae (T4P), with ﬁmF/G expression increased up to 332-fold, for substrate sensing and attachment. The second module, transmembrane transport, involves upregulated genes encoding outer membrane proteins ( fadL , ompW , and tonB ) and inner membrane transporters ( mfs / emrAB-tolC ; up to 18.9-fold) that facilitate alkane uptake. The third module, oxidative degradation, is initiated by the AlkB-type alkane monooxygenase AlkMa, which is encoded by the dramatically upregulated alkMa gene (327.7-fold for C22) and is essential for ULCAs oxidation, followed by alcohol and aldehyde dehydroge- nases (ALDHs). The fourth module, energy metabolism, couples b -oxidation with an adaptive glyoxylate cycle to conserve carbon. Furthermore, electrospray ionization mass spectrometry enabled direct detec- tion of C56 fatty acid intermediates, providing molecular evidence of the pathway’s broad substrate capacity. Functional validation through alkMa gene deletion conﬁrmed its indispensable role in the initial oxidation of C22–C30 alkanes. This study presents a comprehensive model of native multi-enzyme syn- ergy underlying ULCAs biodegradation, offering a framework for engineering microbial systems for PE remediation.

Keywords

Long-chain alkane / Polyethylene / Multi-enzyme synergy / Genomics / Transcriptomics

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Xianrui Liu, Donghuan Su, Shaojie Wang, Zhenxia Du, Haijia Su. Revealing the Native Multi-Enzyme Synergy of Acinetobacter calcoaceticus SDH15 for Ultra-Long-Chain Alkane Biodegradation. Engineering 202602035 DOI:10.1016/j.eng.2026.02.035

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