Department of Biotechnology and Biochemistry, University of Zimbabwe, Harare, Zimbabwe.
ScientificWorldJournal. 2024 May 9;2024:3350591. doi: 10.1155/2024/3350591. eCollection 2024.
The challenge in polystyrene disposal has caused researchers to look for urgent innovative and ecofriendly solutions for plastic degradation. Some insects have been reported to use polystyrene as their sole carbon source, and this has been linked to the presence of microbes in their guts that aid in plastic digestion. Thus, this study focuses on the molecular detection and phylogenetic analysis of the alkane-1-monooxygenase (alkB) gene in strains isolated from the gut of . The alkB gene encodes for alkane-1-monooxygenase, an enzyme involved in the oxidation of inactivated alkanes. This gene can be used as a marker to assess bacteria's ability to biodegrade polystyrene. Three bacterial strains were isolated from the guts of mealworms and were confirmed using polymerase chain reaction (PCR) of the 16S ribosomal RNA gene. The primers used in the amplification of the 16S ribosomal RNA region were designed using NCBI, a bioinformatics tool. To detect the presence of the alkB gene in the isolated bacterial strains, a set of primers used in the amplification of this gene was manually designed from the conserved regions of the alkB nucleotide sequences of eleven bacterial species from GenBank. TCOFFE online tool was used to align the alkB sequences of the bacteria, while Jalview and ConSurf were used to view the alignment. The amplified alkB gene was then sequenced using the Sanger sequencing technique, blasted on NCBI to look for similar sequences, and a phylogenetic tree was constructed. Based on the 16S ribosomal RNA gene sequences, the isolated bacterial strains were confirmed to be NBRC 102593, JCM 1665, and ATCC 13182. The alkB gene sequence identical to fourteen alkB gene sequences derived from whole genome was detected in for the first time to the best of our knowledge. The novel nucleotide sequence was published in the NCBI database under accession number OP959069. This gene sequence was found to be for the enzyme alkane-1-monooxygenase and may be one of the enzymes responsible for polystyrene degradation by the putative ATCC 13182 in .
聚苯乙烯处理面临的挑战促使研究人员寻找紧急的创新和环保解决方案来降解塑料。据报道，一些昆虫将聚苯乙烯作为其唯一的碳源，这与它们肠道中存在有助于塑料消化的微生物有关。因此，本研究专注于从 肠道中分离出的 菌株中烷烃 1-单加氧酶 (alkB) 基因的分子检测和系统发育分析。alkB 基因编码烷烃 1-单加氧酶，该酶参与失活烷烃的氧化。该基因可用作评估细菌降解聚苯乙烯能力的标志物。从 黄粉虫的肠道中分离出 3 株细菌，并通过聚合酶链反应 (PCR) 对 16S 核糖体 RNA 基因进行了确认。用于扩增 16S 核糖体 RNA 区域的引物是使用生物信息学工具 NCBI 设计的。为了检测分离出的细菌菌株中 alkB 基因的存在，使用来自 GenBank 的 11 种细菌的 alkB 核苷酸序列保守区手动设计了一组用于扩增该基因的引物。使用 TCOFFE 在线工具对齐细菌的 alkB 序列，然后使用 Jalview 和 ConSurf 查看对齐结果。然后使用 Sanger 测序技术对扩增的 alkB 基因进行测序，在 NCBI 上进行 Blast 以寻找相似序列，并构建系统发育树。根据 16S 核糖体 RNA 基因序列，分离出的细菌菌株被确认为 NBRC 102593、JCM 1665 和 ATCC 13182。据我们所知，首次在 中检测到与 14 条源自 全基因组的 alkB 基因序列完全相同的 alkB 基因序列。新的核苷酸序列已在 NCBI 数据库中以 OP959069 号发布。该基因序列被发现是烷烃 1-单加氧酶的酶，并可能是 假定的 ATCC 13182 降解聚苯乙烯的酶之一。
