Biodesulfurization using thermophilic bacteria and identification of bds genes

嗜热菌生物脱硫及bds基因鉴定

• 批准号: 13650859
• 负责人: KIRIMURA Kohtaro
• 金额: $ 2.3万
• 依托单位: Waseda University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13650859/
• 关键词: Biotechnology; Bioengineering; Bioprocess; Microbial function; Biodesulfurization; Thermophilic bacteria; Dibenzothiophene; Flavin reductase; 環境微生物; Mycobacterium; 有機硫黄化合物; biodesulfurization; Bacillus subtilis; bioconversion; dibenzothiophe

Recalcitrant organosulfur compounds such as dibenzothiophene (DBT) derivatives in light gas oil (LGO) cannot be removed by hydrodesulfurization using metallic catalysts. The thermophilic bacteria, Bacillus subtilis WU-S2B and Mycobacterium phlei WU-F1, were isolated for its ability to grow on DBT as the sole source of sulfur at 50℃. In addition to DBT, WU-S2B and WU-F1 also could desulfurize alkylated DBTs such as 4, 6-dimethyl DBT through a sulfur-specific degradation pathway with the selective cleavage of carbon-sulfur bonds. Moreover, when resting cells of WU-F1 were incubated at 45℃ with two types of hydrodesulfurized LGOs in the reaction mixtures containing 50%(v/v)oils, the biodesulfurization reduced the sulfur contents from 120 to 50 ppm S (F-LGO) and from 34 to 15 ppm S (X-LGO), respectively. Gas chromatography analysis with an atomic emission detector revealed that the peaks of alkylated DBTs including 4-methyl DBT, 4, 6-dimethyl DBT, and 3, 4, 6-trimethyl DBT significantly decreased after the biodesulfurization. The DBT-desulfurization genes, bdsABC, were cloned from these two bacteria, and the flavin reductase genes, frb and frm, were also cloned from WU-S2B and Wu-F1, respectively. The recombinant WU-F1 carrying one more set of bdsABC and frm was constructed, and the desulfurizing activity of the recombinant strain was 2. 2-fold higher than that of the wild type strain WU-F1.

顽固的有机硫化合物，例如轻质气油（LGO）中的二苯唑噻吩（DBT）衍生物，无法使用金属催化剂去除氢化硫化。嗜热细菌，枯草芽孢杆菌WU-S2B和分枝杆菌Wu-F1分离出来，其在DBT上生长的能力是50℃的唯一硫磺来源。除DBT外，WU-S2B和WU-F1还可以通过硫磺特异性的降解途径，诸如4、6-二甲基DBT等烷基化的DBT，并具有选择性切割碳硫键。此外，将Wu-F1的静止细胞与含有50％（v/v）油的反应混合物中的两种类型的氢硫化LGO孵育时，生物硫化可将硫含量从120 ppm S（F-LGO）和34 ppm s（x-ppm s（x-lgo）降低至50 ppm s（f-lgo）。使用原子发射检测器进行气相色谱分析表明，烷基化DBT的峰在内，包括4-甲基DBT，4、6-二甲基DBT和3、4、6-三甲基DBT在生物硫化后显着降低。从这两种细菌克隆了DBT-二硫化基因BDSABC，而黄素还原基因FRB和FRM也分别从WU-S2B和WU-F1克隆。构建了携带一组BDSABC和FRM的重组WU-F1，重组菌株的脱硫活性比野生型菌株WU-F1高2倍。

项目成果

Kohtaro Kirimura, et al.: "Thermophilic biodesulfurization of hydrodesulfurized light gas oils by Mycobacterium Phlei WU-F1"FEMS Microbiol.Lett.. 221. 137-142 (2003)

Kohtaro Kirimura 等：“通过 Mycobacter Phlei WU-F1 对加氢脱硫轻质瓦斯油进行嗜热生物脱硫”FEMS Microbiol.Lett.. 221. 137-142 (2003)

Kohtaro Kirimura et al.: "Identification and functional analysis of the genes encoding dibenzothiophene-desulfurizing enzymes from thermophilic bacteria"Appl.Microbiol.Biotechnol.. (In press). (2004)

Kohtaro Kirimura 等人：“来自嗜热细菌的编码二苯并噻吩脱硫酶的基因的鉴定和功能分析”Appl.Microbiol.Biotechnol..（正在出版）。

Kohtaro Kirimura, et al.: "Thermophilic biodesulfurization of dibenzothiophene and its derivatives by Mycobacterium phlei WU-F1"FEMS Microbiol.Lett.. 204. 129-133 (2001)

Kohtaro Kirimura 等：“草分枝杆菌 WU-F1 对二苯并噻吩及其衍生物的嗜热生物脱硫”FEMS Microbiol.Lett.. 204. 129-133 (2001)

Kohtaro Kirimura, et al.: "Thermophilic biosedulfurization of naphthothiophene and 2-ethylnaphthothiophene by a dibenzothiophene-desulfurizing bacterium, Mycobacterium phlei WU-F1"Appl.Microbiol.Biotechnol.. 58. 237-240 (2002)

Kohtaro Kirimura 等人：“二苯并噻吩脱硫细菌，草分枝杆菌 WU-F1 对萘并噻吩和 2-乙基萘并噻吩的高温生物硫化”Appl.Microbiol.Biotechnol.. 58. 237-240 (2002)

T.Furuya, K.Kirimura, K.Kino, S.Usami: "Thermophilic Biodesulfurization of Naphthothiophene and 2-Ethylnaphthothiophene by -Mycobacterium phlei WU-F1"Appl. Microbiol. Biotechnol.. 58(2). 237-240 (2002)

T.Furuya、K.Kirimura、K.Kino、S.Usami：“草分枝杆菌 WU-F1 对萘并噻吩和 2-乙基萘并噻吩的高温生物脱硫”应用。