• Journal of Microbiology and Biotechnology
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• 제33권7호
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• pp.895-908
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• 2023

Disposal of waste containing heavy metals into the environment is a major threat to human health and can result in toxic or chronic poisoning in aquatic life. In the current study, metal-resistant Raoultella ornithinolytica was isolated from metal-contaminated samples collected from the Tanjaro River, located southwest of Sulaymaniyah, Iraq. R. ornithinolytica was identified by partial amplification of 16S rRNA. The uptake potency of heavy metals was assessed using inductively coupled plasma-optical emission spectroscopy (ICP-OES) and indicated that R. ornithinolytica removed 67, 89, 63.4, 55.6, 56.5, 65, and 61.9% of Cd, Pb, Cr, Ni, Zn, Co, and Fe, respectively. These removal rates were influenced by temperature, pH, and contact time; at 35℃ and pH 5 with a change in the incubation time, the reduction rate improved from 89 to 95% for Pb, from 36.4 to 45% for Cu, and from 55.6 to 64% for Ni. Gene analysis indicated that R. ornithinolytica contained pbrT, chrB, nccA, iroN, and czcA genes, but the pcoD gene was absent. Energy-dispersive X-ray spectroscopy (EDS) images showed evidence of metal ion binding on the cell wall surface with different rates of binding. Transmission electron microscopy (TEM) detected different mechanisms for metal particle localization; cell surface adsorption was the main mechanism for Pb, Zn, and Co uptake, while Cd, Ni, and Fe were accumulated inside the cell. The current study describes, for the first time, the isolation of R. ornithinolytica from metal-contaminated water, which can be used as an eco-friendly biological expedient for the remediation and detoxification of metals from contaminated environments.

• The Plant Pathology Journal
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• 제34권3호
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• pp.250-255
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• 2018

Pine wilt disease, caused by the nematode Bursaphelenchus xylophilus, is one of the most devastating conifer diseases decimating several species of pine trees on a global scale. Here, we report the draft genome of Raoultella ornithinolytica MG, which is isolated from mountain-cultivated ginseng plant as an bacterial endophyte and shows nematicidal activity against B. xylophilus. Our analysis of R. ornithinolytica MG genome showed that it possesses many genes encoding potential nematicidal factors in addition to some secondary metabolite biosynthetic gene clusters that may contribute to the observed nematicidal activity of the strain. Furthermore, the genome was lacking key components of avermectin gene cluster, suggesting that nematicidal activity of the bacterium is not likely due to the famous anthelmintic agent of wide-spread use, avermectin. This genomic information of R. ornithinolytica will provide basis for identification and engineering of genes and their products toward control of pine wilt disease.

• Journal of Microbiology and Biotechnology
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• 제28권12호
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• pp.1999-2008
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• 2018

The compound 2,5-furandicarboxylic acid (FDCA), an important bio-based monomer for the production of various polymers, can be obtained from 5-hydroxymethylfurfural (HMF). However, efficient production of FDCA from HMF via biocatalysis has not been well studied. In this study, we report the identification of key genes that are involved in FDCA synthesis and then the engineering of Raoultella ornithinolytica BF60 for biocatalytic oxidation of HMF to FDCA using its resting cells. Specifically, previously unknown candidate genes, adhP3 and alkR, which were responsible for the reduction of HMF to the undesired product 2,5-bis(hydroxymethyl)furan (HMF alcohol), were identified by transcriptomic analysis. Combinatorial deletion of these two genes resulted in 85.7% reduction in HMF alcohol formation and 23.7% improvement in FDCA production (242.0 mM). Subsequently, an aldehyde dehydrogenase, AldH, which was responsible for the oxidation of the intermediate 5-formyl-2-furoic acid (FFA) to FDCA, was identified and characterized. Finally, FDCA production was further improved by overexpressing AldH, resulting in a 96.2% yield of 264.7 mM FDCA. Importantly, the identification of these key genes not only contributes to our understanding of the FDCA synthesis pathway in R. ornithinolytica BF60 but also allows for improved FDCA production efficiency. Moreover, this work is likely to provide a valuable reference for producing other furanic chemicals.

• Journal of Dairy Science and Biotechnology
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• 제35권1호
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• pp.25-31
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• 2017

본 연구는 닭의 분뇨로부터 악취저감 방안으로, 쌀뜨물에 L. sakei의 배양조건과 저장 중 안정성을 검토하였다. 그리고 닭 분뇨로부터 미생물을 분리 동정하고, 이들 미생물에 대한 L. sakei의 항균능력을 검토하였다. 또한 L. sakei 함유 쌀뜨물 발효액이 닭 분뇨의 악취를 저감시키는지에 대하여 연구를 수행하였다. 연구결과, 쌀뜨물은 L. sakeiv의 생장에 좋은 영양소로 확인되었으며, 저장 기간 중 생존력도 강한 것으로 증명되었다. 또한 L. sakei는 닭의 분뇨로부터 분리된 악취유발 미생물의 생장 또한 억제하는 것으로 나타났다. 이러한 항균작용 또는 정균작용으로 사회적으로 문제가 되는 축산분뇨에 대하여, L. sakei를 이용해 배양된 쌀뜨물은 악취 유발을 저감할 수 있는 것으로 사료된다. 더욱이 쌀뜨물은 가치가 인정되지 않아 그대로 하수도로 버리게 되어 환경오염을 유발원으로 인식되었는데, 유산균 발효에 의한 악취 제거, 기능성 사료의 이용 등 산업적으로 많은 응용이 적용될 것으로 기대된다.

• Journal of Microbiology and Biotechnology
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• 제31권8호
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• pp.1154-1162
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• 2021

The transcriptional capacities of target genes are strongly influenced by promoters, whereas few studies have focused on the development of robust, high-performance and cross-species promoters for wide application in different bacteria. In this work, four novel promoters (P_k.rtufB, P_k.r1, P_k.r2, and P_k.r3) were predicted from Ketogulonicigenium robustum and their inconsistency in the -10 and -35 region nucleotide sequences indicated they were different promoters. Their activities were evaluated by using green fluorescent protein (gfp) as a reporter in different species of bacteria, including K. vulgare SPU B805, Pseudomonas putida KT2440, Paracoccus denitrificans PD1222, Bacillus licheniformis and Raoultella ornithinolytica, due to their importance in metabolic engineering. Our results showed that the four promoters had different activities, with P_k.r1 showing the strongest activity in almost all of the experimental bacteria. By comparison with the commonly used promoters of E. coli (tufB, lac, lacUV5), K. vulgare (Psdh, Psndh) and P. putida KT2440 (JE111411), the four promoters showed significant differences due to only 12.62% nucleotide similarities, and relatively higher ability in regulating target gene expression. Further validation experiments confirmed their ability in initiating the target minCD cassette because of the shape changes under the promoter regulation. The overexpression of sorbose dehydrogenase and cytochrome c551 by P_k.r1 and P_k.r2 resulted in a 22.75% enhancement of 2-KGA yield, indicating their potential for practical application in metabolic engineering. This study demonstrates an example of applying bioinformatics to find new biological components for gene operation and provides four novel promoters with broad suitability, which enriches the usable range of promoters to realize accurate regulation in different genetic backgrounds.