石油作為現代工業中廣泛應用的原料之一，其可能造成的生態汙染也不容小覷。相較傳統物化方法，尋找直接使用碳氫化合物作為本身碳源的微生物，用於分解油汙的生態除汙法，愈發受到青睞。本研究旨在尋找石油污染土壤中，具有相關降解能力的微生物，並透過全基因體定序分析其功能基因。我們從台南市永康區一塊油汙污染土壤，分離出兩株細菌。經由PacBio平臺進行全基因體定序與功能註解，使用Flye演算法重組成基因體讀序，依據Ribosomal Multilocus Sequence Typing (rMLST)鑒定出兩種菌種：Alcaligenes ammonioxydans (Aa)及Paenibacillus lactis (Pl)。定序結果顯示，Aa株擁有豐富的芳香烴與烷烴降解基因，包括1,2-phenylacetyl-CoA epoxidase、phenol hydroxylase、醇/醛脫氫酶與β-氧化相關酶。Pl株則擁有cytochrome P450氧化酶等，與芳香烴及側鏈分解相關的酵素。雖然Pl株相較Aa株降解能力較弱，對後者卻有著潛在的補強功能。在代謝功能方面，兩者具有高度互補性，Aa偏向于環狀芳香烴降解，Pl則可加強對其他類烷烴的氧化能力。系統發育與基因序列比對結果指出，Aa株中的降解基因相較於近緣物種，具有演化新穎性，而Pl株的P450基因則與同屬菌種相似，展現穩定降解特性。整體而言，Aa株與Pl株的聯合應用，有潛力構建高效的石油烴污染生物修復微生物體系，未來可望進一步應用於油汙地生物復育實驗。

Crude oil is an essential industrial resource, but its leakage poses significant threats to ecosystems. Compared to traditional physical and chemical remediation methods, bioremediation using microorganisms capable of directly metabolizing hydrocarbons offers a more sustainable and efficient approach. This study focuses on the isolation and genomic characterization of such bacteria from petroleum-contaminated soil in Tainan, Taiwan. Whole-genome sequencing was conducted using the PacBio platform, and the genomes were assembled with the Flye algorithm, followed by functional gene annotation. Ribosomal multilocus sequence typing (rMLST) identified two bacterial strains: Alcaligenes ammonioxydans (Aa) and Paenibacillus lactis (Pl). A broad range of hydrocarbon-degrading genes were identified, including those encoding monooxygenases, dehydrogenases, and cytochrome P450 enzymes. The genome of Aa revealed a diverse repertoire of genes responsible for the degradation of both alkanes and aromatic hydrocarbons—such as 1,2-phenylacetyl-CoA epoxidase, phenol hydroxylase, and alcohol/aldehyde dehydrogenases—as well as enzymes involved in β-oxidation pathways. These features indicate a strong capability for aromatic hydrocarbon degradation. In contrast, Pl, though less potent overall, contains cytochrome P450 monooxygenases that support the oxidative breakdown of hydrocarbon side chains, aiding in linear alkane degradation. Functionally, the two strains exhibit strong metabolic complementarity: Aa specializes in the ring-cleaving degradation of aromatic compounds, while Pl enhances aliphatic hydrocarbon oxidation. Comparative genomic analysis revealed that Aa possesses evolutionarily novel degradation genes, while Pl’s P450 genes are conserved among related species, indicating functional stability. Collectively, these findings highlight the potential application of the Aa–Pl microbial consortium in the bioremediation of petroleum-contaminated environments.

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